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  • RAPD  (64)
  • gene expression  (49)
  • Springer  (113)
  • American Chemical Society (ACS)
  • American Institute of Physics (AIP)
  • Frontiers Media
  • 2015-2019
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  • 2000-2004  (113)
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  • 1
    ISSN: 1572-9702
    Schlagwort(e): Panonychus citri ; spider mite ; microsatellite ; RAPD ; PCR ; DNA polymorphism
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Genetic markers were searched using PCR with 40 kinds of decanucleotide primers to investigate DNA polymorphism in Panonychuscitri. A region consisting of a variable number of CT tandem repeats (microsatellite) was found in a fragment amplified with the OPB10 primer. The microsatellite differed in size by ca. 100 bp among several P. citri populations screened and was derived from at least seven alleles. This region was characteristic of P. mori and P. osmanthi, but was lacking in P. ulmi. The flanking regions were highly conserved among these species.
    Materialart: Digitale Medien
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  • 2
    ISSN: 1572-9737
    Schlagwort(e): allozyme ; microgeographic divergence ; microsatellite ; natural selection ; RAPD ; Triticum dicoccoides ; wild emmer wheat
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The levels of genetic diversity were compared by means of 35 allozyme, 60 RAPD, and 25 microsatellite (SSR) markers for 75–175 individuals of tetraploid wild emmer wheat (Triticum dicoccoides) collected in 1993 from a microgeographic microsite, Ammiad, north of the Sea of Galilee, Israel. This microsite included four major habitats, which showed highly significant differentiation in ecological factors, in particular with respect to rock cover, proximity and height, and surface soil moisture in the early growing season of T. dicoccoides. Higher within-subpopulation genetic diversity was found in the primarily non-coding DNA regions (RAPD and SSR) rather than in the protein-coding (allozymes) regions. However, much larger gene differentiation (G ST) among the subpopulations was observed in the protein-coding allozymes than in the RAPDs and SSRs. Larger genetic distance was found at SSR loci, followed by allozyme and RAPD loci. The subpopulations in drier habitats tend to have higher allozyme, RAPD and SSR diversities (He), the relatively wet Karst subpopulation showed only about half He of the other relatively drier habitats. The subpopulations with larger difference of soil moisture between habitats tend to show larger genetic distances at allozyme, RAPD and SSR loci. These results suggest that climatic selection through aridity stress may be an important factor acting on both structural protein-coding and presumably partly regulatory non-coding DNA regions, resulting in microscale adaptive patterns, although hitchhiking and random drift may also intervene. These results have profound implications for genetic conservation both in situ and ex situ.
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  • 3
    ISSN: 1572-9737
    Schlagwort(e): Australia ; conservation strategy ; cpDNA ; Euphorbiaceae ; Fontainea ; nrDNA ; RAPD
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Four new eastern Australian Fontainea species have beenrecently described and all have a limited distribution. F.oraria is the rarest, being restricted to 10 adult individualswithin a single site in regrowth littoral rainforest. In order todevelop adequate management strategies, this study was aimed atsurveying the genetic variability remaining within the species by usingRAPD analysis. To assist with the correct interpretation of the results,a matching study was conducted on four populations of the closelyrelated F. australis. Similar amounts of within-populationgenetic diversity were recorded for both species. The RAPD-based studysuggested that adult plants are contributing unevenly to successivegenerations. RAPD analysis also recognised a close evolutionaryrelationship between F. oraria and F. australis.Sequencing of cpDNA (trnL-F) and nrDNA (ITS2) regions,confirmed recent divergence and possibly some historical reticulationbetween these two species and two other members of the genus. Ofparticular interest was the recognition that one of the F.australis populations (Limpinwood) represented a novel genotypiccombination in need of conservation attention. The implications of theRAPD and sequencing results are discussed in reference to theirinfluence upon the development of adequate conservation strategies forall important conservation units.
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  • 4
    Digitale Medien
    Digitale Medien
    Springer
    Sexual plant reproduction 12 (2000), S. 353-359 
    ISSN: 1432-2145
    Schlagwort(e): Key words Rosa sect Caninae ; Heterogamy ; Apomixis ; RAPD ; Pollen viability
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  All members of Rosa section Caninae, dogroses are polyploid and characterized by their unbalanced meiosis, which in most cases leads to a pronounced morphological influence from the maternal parent. In a previous investigation on a pair of reciprocal crosses between two species in this section, Rosa dumalis and R. rubiginosa (2n=35), nine offspring plants (approximately 10%) did not receive any of the 21 RAPD markers present in the respective pollen parent. This was interpreted as a possible occurrence of apomixis. These nine plants have now been subjected to a further study with additional markers. Thirteen new RAPD markers showed the same result as in the previous investigation: none of the nine plants inherited any of the pollen donor markers. The reproducibility of the RAPD markers was checked by mixing DNA samples to obtain a series of artificial hybrids between the two parent plants. Twelve RAPD markers gave the expected result, whereas one marker appeared only 50% of the time. In addition, pollen viability, mean number of seeds per hip, mean seed weight, and mean weight of fruit flesh per hip have been studied on the four progeny groups: R. dumalis×R. rubiginosa plants which received pollen donor markers (PM plants), R. dumalis×R. rubiginosa plants which did not receive any pollen donor markers (NPM plants), R. rubiginosa×R. dumalis PM plants and R. rubiginosa×R. dumalis NPM plants. A canonical discriminant analysis based on these four reproductive characters separated the four progeny groups. There were significant differences between the two PM groups in all investigated characters, and also between the PM and the NPM groups in pollen viability. The result from the RAPD markers together with the differences in pollen viability between the PM and NPM progeny groups is taken as an indication that apomixis occurs within the Caninae section.
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  • 5
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 100 (2000), S. 1155-1166 
    ISSN: 1432-2242
    Schlagwort(e): Key words Citrus ; RAPD ; SCAR ; cpDNA ; Phylogeny ; Origin
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  Citrus phylogeny was investigated using RAPD, SCAR and cpDNA markers. The genotypes analyzed included 36 accessions belonging to Citrus together with 1 accession from each of the related genera Poncirus, Fortunella, Microcitrus and Eremocitrus. Phylogenetic analysis with 262 RAPDs and 14 SCARs indicated that Fortunella is phylogenetically close to Citrus while the other three related genera are distant from Citrus and from each other. Within Citrus, the separation into two subgenera, Citrus and Papeda, designated by Swingle, was clearly observed except for C. celebica and C. indica. Almost all the accessions belonging to subgenus Citrus fell into three clusters, each including 1 genotype that was considered to be a true species. Different phylogenetic relationships were revealed with cpDNA data. Citrus genotypes were separated into subgenera Archicitrus and Metacitrus, as proposed by Tanaka, while the division of subgenera Citrus and Papeda disappeared. C. medica and C. indica were quite distant from other citrus as well from related genera. C. ichangensis appeared to be the ancestor of the mandarin cluster, including C. tachibana. Lemon and Palestine sweet lime were clustered into the Pummelo cluster led by C. latipes. C. aurantifolia was located in the Micrantha cluster. Furthermore, genetic origin was studied on 17 cultivated citrus genotypes by the same molecular markers, and a hybrid origin was hypothesized for all the tested genotypes. The assumptions are discussed with respect to previous studies; similar results were obtained for the origin of orange and grapefruit. Hybrids of citron and sour orange were assumed for lemon, Palestine sweet lime, bergamot and Volkamer lemon, while a citron × mandarin hybrid was assumed for Rangpur lime and Rough lemon. For Mexican lime our molecular data indicated C. micrantha to be the female parent and C. medica as the male one.
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  • 6
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 100 (2000), S. 1209-1216 
    ISSN: 1432-2242
    Schlagwort(e): Key words Digitalis spp. ; AMOVA ; Genetic relationships ; RAPD ; Scrophulariaceae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  RAPD markers were used to study inter-specific variation among six species of the genus Digitalis: D. obscura, D. lanata, D. grandiflora, D. purpurea, D. thapsi and D. dubia, and the hybrid D. excelsior (D. purpurea×D. grandiflora). A total of 91 highly reproducible bands amplified with four arbitrarily chosen decamer primers were obtained. Homology of the co-emigrating RAPD markers was tested by blot hybridisation and sequencing of selected bands. The application of a range of statistical approaches for RAPD data analysis, including distance and parsimony methods, family clustering and the analysis of molecular variance (AMOVA), indicated that these molecular markers were taxonomically informative in Digitalis. The species relationships revealed were fully consistent with those previously obtained using morphological affinities. The hybrid D. excelsior seems to have stronger affinity to the section Digitalis than to Grandiflorae. This is the first known report of the application of RAPD markers for the study of genetic relationships among species of the genus Digitalis.
    Materialart: Digitale Medien
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  • 7
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 101 (2000), S. 70-79 
    ISSN: 1432-2242
    Schlagwort(e): Key words Poa annua L. ; Genetic diversity ; RAPD ; Turfgrass weeds ; Selection pressure ; Analysis of molecular variance ; AMOVA ; POPGENE
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  The genetic diversity of Poa annua L.populations collected from western Oregon grass-seed fields was surveyed using 18 randomly amplified polymorphic DNA (RAPD) markers. Markers from 1357 individual plants from 47 populations collected at three sampling dates (fall, winter, and spring) for 16 sites were used to measure genetic diversity within and among populations. Site histories varied from low to high herbicide selection pressure, and some sites were subdivided by 3 years of differing post-harvest residue management. Gene diversity statistics, simple frequency of haplotype occurrence, and analysis of molecular variance (AMOVA) revealed the presence of significant variability in P. annua among sites, among collection dates within sites, and within collection dates. Nei gene-diversity statistics and population-differentiation parameters indicated that P. annua populations were highly diverse. Mean Nei gene diversity (h) for all 47 populations was 0.241 and total diversity (HT) was 0.245. A greater proportion of this diversity, however, was within (HS=0.209) rather than among (GST=0.146) populations. When populations were grouped by season of collection, within-group diversity was HS=0.241, while among-group diversity was GST=0.017. When populations were grouped by site, within-group diversity was HS=0.224, while among-group diversity was GST=0.087. The diversity among populations within season for fall, winter, and spring collections was GST=0.121, 0.142, and 0.133, respectively. Populations collected from fields with histories of high herbicide selection pressure showed low differentiation among collection dates, with GST as low as 0.016, whereas those collected from fields with low herbicide selection pressure showed greater differentiation among collection dates, with GST as high as 0.125. At high selection-pressure sites, populations were also lower in gene diversity (as low as h=0.155), while at low selection-pressure sites there was higher gene diversity (as high as h=0.286). The site to site variability was greater for the high selection-pressure sites (GST=0.107 or 69% of the total among-population variance), while the season of germination variability was greater at sites of low herbicide-selection pressure (GST=0.067, or 70% of the total among-population variance). High initial diversity coupled with a long-term re-supply of genotypes from the seed bank must have been factors in maintaining the genetic diversity of this weed despite the intensive use of herbicides. Knowledge of the genetic diversity of Willamette Valley P. annua should help in formulating more effective strategies for managing this weed.
    Materialart: Digitale Medien
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  • 8
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 101 (2000), S. 90-94 
    ISSN: 1432-2242
    Schlagwort(e): Key words Molecular map ; AFLP ; RAPD ; Optimisation algorithm
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  A computer algorithm is presented which allows selection of a subset of multiplex markers based on the minimisation of an optimality criterion for a genetic linkage map. It could be applied for choosing a subset of primers (e.g. RAPD, IMA or AFLP), each of which provides several unevenly spaced genetic markers. The goal is to achieve a saturated map of evenly spaced markers, using as few primers as possible to minimise cost and labour. Minimising the average map distance between markers is trivial, but simply leads to selection of those primers which provide the greatest number of markers. However, minimising the standard deviation of interval length ensures that weight is given both to the number of markers and to the evenness of their distribution on the linkage map. This criterion was found empirically to give a result fairly close to the optimum. A stepwise-like selection procedure is therefore implemented, which stops when the optimality criterion does not decrease any more. An example is given of a molecular map of perennial ryegrass with 463 markers obtained from 17 AFLP primers. It is demonstrated that this can be safely reduced to a 175 marker map with only 6 primers. Genetic diversity studies may also benefit from using such a subset of less-redundant markers in genetic distance estimation.
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  • 9
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 101 (2000), S. 292-300 
    ISSN: 1432-2242
    Schlagwort(e): Key words AFLP ; DNA markers ; Early germination ; ISSR ; ISTR ; RAPD
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  Different DNA marker types were used to construct linkage maps in coconut (Cocos nucifera L.; 2n = 32) for the two parents of the cross Malayan Yellow Dwarf (MYD) × Laguna Tall (LAGT). A total of 382 markers was sufficient to generate 16 linkage groups for each parent. The total genome length corresponded to 2226 cM for the LAGT map and 1266 cM for the MYD map with 4–32 markers per linkage group. Common markers allowed the association of 9 linkage groups for the two parents MYD and LAGT. QTL analysis for the trait early germination identified six loci. These QTLs correlate with early flowering and yield, representing characters which are important in coconut breeding. The co-segregation of markers with these QTLs provides the first opportunity for marker-assisted selection in coconut breeding programmes.
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  • 10
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 100 (2000), S. 614-620 
    ISSN: 1432-2242
    Schlagwort(e): Key words Aigeiros ; Leuce ; Marssonina brunnea ; Poplar ; RAPD ; Tacamahaca
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  A broad collection was made for 42 isolates of Marssonina brunnea affecting poplar trees from three different sections (Leuce, Aigeiros and Tacamahaca) within the same Populus genus in China. Genetic diversity among these isolates was analyzed for morphological traits, cultural features, pathogenicity, hyphal anastomosis and randomly amplified polymorphic DNA markers (RAPDs). No significant difference was found in conidial morphological features, such as size, shape and septum location. Yet, considerable differences occur in other characteristics, which leads to the classification of the 42 isolates into two distinct groups, M. brunnea f.sp. monogermtubi and M. brunnea f.sp. multigermtubi. Isolates of M. brunnea f.sp. monogermtubi, derived from section Leuce, germinate only one germ tube, grow fast, produce dark-reddish conidiosorus clusters on the PDA medium, and are highly pathogenic to Populus tomentosa of section Leuce. By contrast, isolates of M. brunnea f.sp. multigermtubi, derived from sections Aigeiros and Tacamahaca, germinate 1–5 germ tubes, grow slowly, produce yellow-greenish conidiosorus clusters on PDA medium, and are pathogenic to Populus ×euramericana cv I-45 and Populus canadensis of section Aigeiros. DNA amplification using 11 RAPD primers generate 78 polymorphic bands among isolates. Cluster analyses based on RAPD markers broadly support such a classification by phenotypes, but provide a new insight into the possible origins of M. brunnea. It is proposed that the pathogen co-evolves with the poplars of section Leuce and has been subsequently distributed to the poplars of sections Aigeiros and Tacamahaca. An isolate from Populus adenopoda of section Leuce is placed in the third group, which is most likely a transmission type from M. brunnea f.sp. monogermtubi to M. brunnea f.sp. multigermtubi.
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  • 11
    ISSN: 1432-2242
    Schlagwort(e): Key words SCAR ; RAPD ; Bulked segregant analysis ; Marker-assisted selection ; Orobanche cumana ; Helianthus annuus
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  A consensus molecular linkage map of 61.9 cM containing the Or5 gene, which confers resistance to race E of broomrape orobanche cumana, five SCAR markers (three dominant, two codominant) and one RAPD marker were identified based on segregation data scored from two F2 populations of susceptible×resistant sunflower line crosses. Bulked segregant analysis was carried out to generate the five SCAR markers, while the single RAPD marker in the group was identified from 61 segregating RAPD markers that were directly screened on one of the two F2 populations. The five SCAR markers, RTS05, RTS28, RTS40, RTS29 and RTS41, were significantly (LOD≥4.0) linked to the Or5 gene and mapped separately at 5.6, 13.6, 14.1, 21.4 and 39.4 cM from the Or5 locus on one side, while the RAPD marker, UBC120_660, was found at 22.5 cM (LOD=1.4) on the opposite side. These markers should facilitate the efficient transfer of the resistance gene among sunflower breeding lines. As the first report on molecular markers linked to a broomrape resistance gene, the present work provides a starting point to study other genes and to examine the hypothesis of the clustering of broomrape resistance genes in sunflower.
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  • 12
    ISSN: 1432-2242
    Schlagwort(e): Key words TGMS ; RAPD ; AFLP ; Microsatellites ; STS ; Marker-assisted selection ; Bulked seg analysis
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  The present study of genetic analysis is an attempt to precisely characterize diverse temperature-sensitive genic male-sterile (TGMS) lines so as to explore the possibilities of utilizing the most promising in large-scale hybrid seed production. Genetical studies revealed that the TGMS segregants derived from crosses involving TGMS lines ID24 and SA2 expressed differential fertility levels at low-temperature conditions. A majority of these progenies expressed transgressive segregation towards either sterility of fertility, causing instability of sterility and low reversibilty of fertility which may be due to large numbers of single-locus QTLs and their epistatic interactions. We identified two putative genes imparting temperature-sensitive male sterility after observing crosses involving diverse TGMS sources. To identify suitable molecular markers closely linked to the trait we used RAPD, AFLP and microsatellites which generated polymorphism through bulked segregant analysis. AFLP analysis using a smaller genome kit resulted in enormous polymorphism, out of which the combination EAA/MCAG amplified a 330-bp fragment, which closely segregated with the gene at a distance of 5.3 cM. This fragment was eluted for cloning and from the sequence a STS primer (TS200) was developed which produced a dominant polymorphism specific to TGMS. The microsatellite RM257, located earlier on chromosome 9, was linked with the TGMS trait in SA2 at a distance of 6.2 cM. RM257 produced a codominant polymorphism with 145-bp (sterile) and 132-bp (fertile) products. Both individually and collectively, the markers TS200 and RM257 located on either side of the TGMS locus are very useful for marker-assisted selection.
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  • 13
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 100 (2000), S. 965-970 
    ISSN: 1432-2242
    Schlagwort(e): Key words Russian wheat aphid ; Near-isogenic lines ; Restriction digests ; RAPD ; SCAR
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstracts  Through random amplified polymorphic DNA (RAPD) analysis we identified a putative marker linked to the Dn5 resistance gene. This marker was converted to a more reliable sequence-characterised-amplified regions (SCAR) marker. The initial SCAR marker amplified the correct amplification product but failed to discern between the susceptible and resistant individuals. Hence, it was utilised to sequence the internal fragment. All nested primers designed from the internal sequences were also unable to produce any polymorphism between the susceptible and resistant cultivars. Restriction digests were then performed on these fragments, and the restriction enzyme EcoRI was able to discern between the susceptible and resistant F2 individuals of the Dn5 population. This granted one marker amplified with the internal SCAR primer set OPF141083 the ability to differentiate between parental individuals carrying the Dn5 genes. This marker was tested in a segregating F2 population carrying the Dn5 resistance gene and proved able to differentiate between the segregating individuals. This marker may prove useful in marker assisted selection (MAS), although performing restriction digests may hamper the throughput of a high number of samples.
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  • 14
    ISSN: 1432-2242
    Schlagwort(e): Key words Pinus pinaster ; AFLP ; RAPD ; Protein ; Linkage map ; QTL
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  TheAFLP (amplified fragment length polymorphism) technique was adapted to carry out genetic analysis in maritime pine, a species characterized by a large genome size (24 pg/C). A genetic linkage map was constructed for one F1 individual based on 239 AFLP and 127 RAPD (randomly amplified polymorphic DNA) markers. Markers were scored on megagametophytes (1n) from 200 germinated F2 seedlings. Polymorphism rate, labour time and cost of both AFLP and RAPD techniques were compared. The AFLP technique was found to be twice as fast and three-times less costly per marker than the RAPD technique. Thirteen linkage groups were identified with a LOD score ≥6 covering 1873 cM, which provided 93.4% of genome coverage. Proteins were extracted from needles (2n) of the F2 progeny and revealed by 2-DE (two-dimensional electrophoresis). Thirty one segregating proteins were mapped using a QTL detection strategy based on the quantification of protein accumulation. Two framework maps of the same F1 individual are now available. The first map (Plomion et al. 1996) uses RAPD markers and the second map, presented in this study, uses mostly AFLP markers. Although the total genetic length of both maps was almost identical, differences among homologous groups were observed.
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  • 15
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 100 (2000), S. 63-70 
    ISSN: 1432-2242
    Schlagwort(e): Key words Elaeis guineensis ; RAPD ; Pseudo-testcross ; Genetic linkage map ; bulked segregant analysis ; Shell thickness
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  Shell thickness is an important trait in oil palm breeding programs and is the basis for the classification of the varieties of oil palm into the types dura, tenera and pisifera. This trait seems to be controlled by a single locus, with two alleles (sh + and sh −) showing codominant expression. Two single-tree linkage maps were constructed for a maternal tenera (sh + sh −) palm and for a paternal pisifera (sh − sh −) palm using the pseudo-testcross mapping strategy in combination with RAPD markers through the analysis of an F1 tenera×pisifera progeny. A total of 308 arbitrary primers were screened in a sample of eight F1 plants and 121 markers were detected in a testcross configuration. An average of 1.66 polymorphic marker per selected primer were identified in this cross. At LOD 5.0 (with some few exceptions) and θ=0.25 the maternal tenera map included a total of 48 markers distributed in 12 linkage groups or pairs of markers (449.3 cM) while the paternal pisifera map included 42 markers distributed in 15 linkage groups or pairs of markers (399.7 cM). We used RAPD and bulked segregant analysis (BSA) to identify markers more tightly linked to the sh + locus. A total of 174 new primers not previously used in the linkage analysis were screened using bulks of DNA extracted from plants selected for the contrasting shell-thickness phenotypes. Two RAPD markers (R11–1282 and T19–1046) were identified to be linked on both sides of the sh + locus on linkage group 4. The estimated map distances from sh + to R11–1282 and to T19–1046 were 17.5 cM and 23.9 cM, respectively. The results demonstrate the usefulness of RAPD markers and the pseudo-testcross mapping strategy for developing genetic linkage information, and constitute an important step towards early marker-assisted selection for shell thickness in oil palm.
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  • 16
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 100 (2000), S. 249-255 
    ISSN: 1432-2242
    Schlagwort(e): Key words QTL ; Earliness ; CAP ; RAPD ; Lycopersicon esculentum
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  Using RAPD marker analysis, two quantitative trait loci (QTLs) associated with earliness due to reduced fruit-ripening time (days from anthesis to ripening = DTR) were identified and mapped in an F2 population derived from a cross between Lycopersicon esculentum’E6203’ (normal ripening) and Lycopersicon esculentum’Early Cherry’ (early ripening). One QTL, on chromosome 5, was associated with a reduction in both ripening time (5 days) and fruit weight (29.3%) and explained 15.8 and 13% of the total phenotypic variation for DTR and fruit weight, respectively. The other QTL, on chromosome 12, was primarily associated with a reduction only in ripening time (7 days) and explained 12.3% of the total phenotypic variation for DTR. The gene action at this QTL was found to be partially dominant (d/a=0.41). Together, these two QTLs explained 25.1% of the total phenotypic variation for DTR. Additionally, two QTLs associated with fruit weight were identified in the same F2 population and mapped to chromosomes 4 and 6, respectively. Together, these two QTLs explained 30.9% of the total phenotypc variation for fruit weight. For all QTLs, the ’Early Cherry’ alleles caused reductions in both ripening time and fruit weight. The polymorphic band for the most significant RAPD marker (OPAB-06), linked to the reduced ripening time QTL on chromosome 12, was converted to a cleaved amplified polymorphism (CAP) assay for marker-aided selection and further introgression of early ripening time (DTR) into cultivated tomato.
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  • 17
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 100 (2000), S. 299-307 
    ISSN: 1432-2242
    Schlagwort(e): Keywords Larix ; Linkage map ; RAPD ; AFLP ; ISSR ; Genetic mapping
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  Genetic linkage maps have been increasingly developed for a wide variety of plants, using segregating populations such as F2s or backcrosses between inbred lines. These pedigrees are rarely available in outbred species like forest trees which have long generation times. Thus genetic mapping studies have to use peculiar pedigrees and markers in appropriate configurations. We constructed single-tree genetic linkage maps of European larch (Larix decidua Mill.) and Japanese larch [Larix kaempferi (Lamb.) Carr.] using segregation data from 112 progeny individuals of an hybrid family. A total of 266 markers (114 AFLP, 149 RAPD and 3 ISSR loci) showing a testcross configuration, i.e.heterozygous in one parent and null in the other parent, were grouped at LOD 4.0, θ=0.3. The maternal parent map (L. decidua)consisted of 117 markers partitioned within 17 linkage groups (1152 cM) and the paternal parent map (L. kaempferi) had 125 markers assembled into 21 linkage groups (1206 cM). The map distance covered by markers was determined by adding a 34.7-cM independence distance at the end of each group and unlinked marker. It reached 2537 cM and 2997 cM respectively for European larch and Japanese larch, and represented respectively a 79.6% and 80.8% coverage of the overall genome. A few 3:1 segregating markers were used to identify homologous linkage groups between the European larch and the Japanese larch genetic maps. The PCR-based molecular markers allowed the construction of genetic maps, thus ensuring a good coverage of the larch genome for further QTL detection and mapping studies.
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  • 18
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 101 (2000), S. 1145-1154 
    ISSN: 1432-2242
    Schlagwort(e): Keywords Altitude differentiation ; Environmental selection ; Phytolacca dodecandra ; RAPD
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  The extent of genetic differentiation among 17 Ethiopian populations (249 individuals) of Phytolacca dodecandra (Endod) sampled along altitudinal gradients that varied from 1600 to 3000 m was investigated using random amplified polymorphic DNA (RAPD). The populations were classified into three altitude groups: lowland (1600–2100 m), central-highland (2101–2500 m) and highland (2500–3000 m). Seventy polymorphic loci scored from 12 RAPD primers, singly or in combination with ecogeographical variables (altitude, longitude, latitude, temperature and rainfall), were used for principal component, discriminant, correlation, and stepwise multiple regression analyses. Principal component analysis (PCA) clearly differentiated lowland and the central-highland populations from those of the highlands independent of their geographical regions. Canonical discriminant analysis separated the lowland plants from those of the highlands with the central-highland plants being intermediate. Classificatory discriminant analysis corrected classification of 92.8% of the 249 plants into their respective three altitude groups. Multiple regression analysis identified a strong association between some RAPDs and altitude, temperature and rainfall, while the variation in most RAPDs was explained by combinations of the different ecogeographical variables. It is hypothesised that the different altitude groups may be (1) chemical and/or physiological ecotypes produced as a result of complex interactions of altitude with climatic and/or edaphic factors, or (2) different in ploidy levels. The significant correlations obtained between population means from some RAPDs and altitude and temperature as well as the strong association of some RAPDs with the ecogeographical variables in the multiple regression analysis suggest that part of the RAPD polymorphism could be adaptive, and responsive to environmental selection.
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  • 19
    ISSN: 1432-2242
    Schlagwort(e): Key words Gene diversity ; Isozyme ; Non-neutrality ; RAPD ; Rosaceae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  RAPD and isozyme analyses based on numerous markers have been used for the first time to investigate patterns of phenetic and genetic differentiation among and within nine wild populations of the genus Chaenomeles represented by the species C. japonica, C. speciosa, C. cathayensis and C. thibetica. Highly significant correlations were found between the two different marker systems for both phenetic distances and gene diversity estimates. In agreement with previous studies on cultivated Chaenomeles material, C. japonica was clearly differentiated from C. speciosa and C. cathayensis. The recently recognised species C. thibetica appeared to be rather closely related to C. cathayensis. Populations of C. japonica and C. speciosa were considerably more diverse than populations of C. cathayensis and C. thibetica. Correspondingly, most of the total variability could be attributed to the within-population differentiation in the case of C. japonica and C. speciosa, and to the between-population differentiation in the case of C. cathayensis. Differences in mating systems among the species can be suggested as a possible explanation of the results. A discordant pattern was found between RAPDs and isozymes in the analyses of population structure within C. japonica. This may be explained by a higher proportion of non-neutral markers for isozymes than for RAPDs. This finding also shows the importance of using multiple molecular marker systems in studies of population structure.
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  • 20
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 100 (2000), S. 506-511 
    ISSN: 1432-2242
    Schlagwort(e): Key words Rye ; DNA instability ; Hypervariable sequences ; Somaclonal variation ; RAPD
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  RAPD analysis was performed to assess DNA variation among rye plants regenerated from immature embryos and inflorescences. From the studied plants, 40% showed at least one variation, and the number of mutations per plant was quite high, ranging from 1 up to 12. On some occasions (2.9% of the scored bands) the modified band was observed in only one plant or in several but originated from the same callus (variable band). In other cases (5.25%) the same band varied in several plants obtained from different calli. We call these hypervariable bands and they could vary between plants belonging to different cultivars and/or with different origins, inflorescences or embryos. Thus, they must originate through independent mutational events. We assume that these bands represent hypervariable regions of the rye genome and so detect hot spots of DNA instability. Some of these bands proved to be unique sequences, others were present in a low copy number while the remaining ones were moderately or highly repetitive.
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  • 21
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 101 (2000), S. 1194-1201 
    ISSN: 1432-2242
    Schlagwort(e): Key words ’Folle blanche’ ; Hybridization ; RAPD ; SCAR ; Sequence-specific primer pair ; Vitis vinifera L
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  Among 34 grapevine cultivars (Vitis vinifera L.), eight putative genotype-specific RAPD markers, from ’Albariño’, ’Caíño blanco’, ’Chardonnay’, ’Folle blanche’, ’Grenache blanc’, ’Malvasía Sitges’, ’Torrontés’ and ’Treixadura’ respectively, were selected to transform into SCAR markers. Of these, seven markers were cloned and then five which showed a positive specific hybridization signal were sequenced. For these five markers, 30 sequence-specific primers ranging from 14 to 29 bases were designed to amplify genomic DNA from 64 grapevine cultivars under more-stringent PCR conditions. Only, two primer pairs, OpA111175p17R/ p17F and OpD10800p14R/p14F, still produced a specific SCAR marker, the ’Folle blanche’ ScA111175 and the ’Malvasía Sitges’ ScD10800 respectively. Moreover, the ScA111175 marker was amplified only in ’Folle blanche’ among the 64 cultivars tested with a large annealing temperature range using either two different Taq DNA polymerases or two separate thermocyclers. In addition, we discuss the initial polymorphism originated by the RAPD technique and suggest a new design of SCAR primers to obtain reliable cultivar-specific SCAR markers from single PCR-based bands for identification purposes.
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  • 22
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 101 (2000), S. 860-864 
    ISSN: 1432-2242
    Schlagwort(e): Key words Melon ; AFLP ; RFLP ; RAPD ; Genetic similarity
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  Three different types of molecular markers, RAPD, AFLP and RFLP were used to measure genetic diversity among six genotypes of Cucumis melo L. Each line represented a different melon genotype: Piel de Sapo, Ogen, PI161375, PI414723, Agrestis and C105. A number of polymorphic RAPD, AFLP and RFLP bands were scored on all materials and the genetic similarity measured. Clustering analysis performed with the three types of markers separated the genotypes into two main groups: (1) the sweet type, cultivated melons and (2) the exotic type, not cultivated melons. While the data obtained suggest that all three types of markers are equally informative, AFLPs showed the highest efficiency in detecting polymorphism.
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  • 23
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 101 (2000), S. 1056-1065 
    ISSN: 1432-2242
    Schlagwort(e): Key words Plantain ; Musa ; RAPD ; Phenotype ; Breeding
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  Genetic diversity amongst 76 plantain landraces has been studied using RAPD analysis at two levels of intensity and compared with groupings based on phenotypic indices and morphotype. There was a good correlation (R2=0.78) between estimates of genetic diversity based on 76 RAPD bands and 164 RAPD bands. However, there was a poor correlation between RAPD-based estimates of genetic diversity and a phenotypic index based on agronomic characters. There was also a poor correlation between RAPD analyses and morphotype group (based on bunch type and stature). These results suggest that the traditional designations of plantain landraces based on morphotype do not provide a true reflection of overall genetic divergence. Similarly, classification systems using phenotypic indices based on agronomic characters may not provide accurate taxonomic differentiation. The level of genetic divergence within morphogroups based on bunch type suggests that True Horn plantains are derived from False Horn plantains which in turn are derived from French plantains. Genetic divergence was found to be generally quite low within the plantain landrace genepool, which is consistent with the proposed evolution of this germplasm through somatic mutation of a relatively small number of introductions. However, putative synonyms/duplicates have been shown to be genetically distinct. In contrast, a group of 12 landraces have been identified that are highly distinct from one another (showing 20–35% dissimilarity). Fertile members of this group may be useful for generating genetically diverse 2x and 4x breeding populations that can be used in breeding secondary triploid hybrid plantain varieties.
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  • 24
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 101 (2000), S. 931-938 
    ISSN: 1432-2242
    Schlagwort(e): Key words Gossypium species ; RAPD ; Phylogeny ; Cluster ; Diversity
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  Total genomic DNA from 31 available Gossypium species, three subspecies and one interspecific hybrid, were analysed to evaluate genetic diversity by RAPD, using 45 random decamer primers. A total of 579 amplified bands were observed, with 12.9 bands per primer, of which 99.8% were polymorphic. OPJ-17 produced the maximum number of fragments while the minimum number of fragments was produced with primer OPA-08. Cluster analysis by the unweighted paired group method of arithmetic means (UPGMA) showed six main clusters. Cluster ’A’ consisted of two species and one subspecies of the A-genome, with a 0.78–0.92 Nei’s similarity range. Cluster B, composed of all available tetraploid species and one interspecific hybrid, showed the same sister cluster. Nei’s similarity ranged from 0.69 to 0.84. The B-genome formed the UPGMA sister cluster to the E-genome species. Cluster ’C’ consisted of five Gossypium species of which three belong to the B-genome, with Nei’s similarity values of 0.81 to 0.86. Although there was considerable disagreement at lower infra-generic ranks, particularly among the D- genome (diploid New World species) and C-genome (diploid Australian species) species. The sole F-genome species Gossypium longicalyx was resolved as a sister group to the D-genome species. Gossypium herbaceum and G. herbaceum Africanum showed the maximum Nei’s similarity (0.93). Minimum similarity (0.29) was observed between Gossypium trilobum and Gossypium nelsonii. The average similarity among all studied species was 50%. The analysis revealed that the interspecific genetic relationship of several species is related to their centre of origin. As expected, most of the species have a wide genetic base range. The results also revealed the genetic relationships of the species Gossypium hirsutum to standard cultivated Gossypium barbadense, G. herbaceum and Gossypium arboreum. These results correspond well with previous reported results. The level of variation detected in closely related genotypes by RAPD analysis indicates that it may be a more efficient marker than morphological marker, isozyme and RFLP technology for the construction of genetic linkage maps.
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  • 25
    ISSN: 1570-0267
    Schlagwort(e): cDNA ; PCR cDNA ; TaqMan Analysis ; gene expression ; Pearson's correlation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Comparative gene expression studies are often limited by low availability of tissue and poor quality of extractable mRNA. Collective PCR amplification of minute quantities of mRNA has great potential for overcoming these limitations. However, there remains significant concern about the effects of amplification on the absolute and relative abundance of individual mRNAs that could complicate subsequent gene expression studies. To address this problem, we systematically compared the relative abundance of many specific mRNAs from complex cDNA preparations (from tissue and cultured cells) both before and after amplification by PCR. Our results demonstrated that, as expected, the absolute abundance of different mRNAs in a cDNA library is altered in an unpredictable manner by PCR amplification. However, we found that the concentration ratios of specific mRNAs among different cDNA preparations were routinely well conserved after PCR amplification. Thus, for the purpose of comparative expression studies for specific mRNAs in two (or more) complex cDNAs, PCR-amplified cDNA is equally useful as unamplified cDNA. These results provide a rigorous experimental validation and offer a theoretical treatment to support the utility of PCR amplified cDNA for differential gene expression studies. We conclude that the inherent difficulties in performing differential screening studies such as gene chip and array analyses on limited amounts of biological materials can be overcome by a PCR amplification step without compromising data quality.
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  • 26
    ISSN: 1573-4919
    Schlagwort(e): myosin heavy chain ; gene expression ; hypertrophy ; dexamethasone ; promoter function
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract Cardiac hypertrophy has been observed in newborn infants treated with dexamethasone (DEX). This study was undertaken to examine whether DEX-induced hypertrophy in newborn rats is associated with redistribution of cardiac myosin heavy chain (MHC) isoforms and if so, the effects involve transcriptional regulation. Newborn rats were injected with either DEX (1 mg/kg/day; s.c.) or equivalent volume normal saline for 1, 3, 5, 7 or 9 days. Hypertrophy was quantified by heart dry/wet wt ratios, heart/body wt ratios, and total protein content of the myocardium. Changes in the expression of cardiac MHC mRNA were characterized by northern blot and slot blot analyses, using isoform specific probes for a- and β-MHC genes. DEX effect on α-MHC gene transcription was analyzed by transiently transfecting various α-MHC promoter/CAT reporter constructs into primary cultures of cardiac myocytes derived from one day old rat pups. DEX administration into newborn rats produced significant cardiac hypertrophy ranging from 23% at day 1 to 59% at 9 days. The hypertrophy was accompanied by immediate increase (83%) in steady state level of the α-MHC mRNA within one day and a maximum increase (148%) at 7 days of treatment. The steady state level of β-MHC mRNA declined by 25% at day 1 and a maximum decrease of 54% at day 7 of DEX treatment. The changes in MHC mRNA were also reflected in their protein levels as determined by V1 and V3 isozyme analysis. DEX treatment of primary cultures of cardiomyocytes following transfection with a-MHC promoter/CAT reporter constructs resulted in increased CAT expression in a dose dependent manner. The minimum α-MHC gene sequences responding to DEX treatment were located between the -200 to -74-bp region of the gene, resulting in 2-fold and 6-fold activation of CAT reporter after 0.05 and 0.1 mM doses of DEX, respectively. Our data indicate that DEX induced cardiac hypertrophy in newborn rats is accompanied by increased expression of α-MHC and decreased expression of β-MHC. The α-MHC effects are mediated in part through transcriptional mechanisms.
    Materialart: Digitale Medien
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  • 27
    Digitale Medien
    Digitale Medien
    Springer
    Molecular and cellular biochemistry 212 (2000), S. 5-9 
    ISSN: 1573-4919
    Schlagwort(e): transcriptional regulation ; gene expression ; coactivator ; repressor
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract The CREB-CREM transcription factors are the main gene regulatory effectors of the cAMP signaling pathway. The investigations of this family of transcription factors had a profound impact on the understanding of signaling-induced gene transcription. Here we discuss some key aspects of the underlying biology, review transcriptional activation by CREB proteins through transcription cofactors and present novel insights into the context- and position-specific function of CREB on complex genes.
    Materialart: Digitale Medien
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  • 28
    ISSN: 1573-4919
    Schlagwort(e): AP-1 ; cobalt chloride ; gene expression ; heme oxygenase ; oxidative stress ; sodium arsenite
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract Previously, chick heme oxygenase-1 (cHO-1) gene was cloned by us and two regions important for induction by sodium arsenite were identified. These two regions were found to contain consensus sequences of an AP-1 (-1580 to -1573) and a MRE/cMyc complex (-52 to -41). In the current study, the roles of these two elements in mediating the sodium arsenite or cobalt chloride dependent induction of cHO-1 were investigated further. DNA binding studies and site-directed mutagenesis studies indicated that both the AP-1 and MRE/cMyc elements are important for the sodium arsenite induction, while cobalt chloride induction involves only the AP-1 element. Electrophoretic mobility shift assays showed that nuclear proteins binding to the AP-1 element was increased by both sodium arsenite or cobalt chloride treatment, whereas the binding of proteins to the MRE/cMyc element showed a high basal expression in untreated cells and the binding activity was only slightly increased by sodium arsenite treatment. Site-directed mutagenesis studies showed that, to completely abolish sodium arsenite induction, both the AP-1 and MRE/cMyc elements must be mutated; mutation of either element alone resulted in only a partial effect. In contrast, a single mutation at AP-1 element was sufficient to reduce the cobalt chloride induction almost completely. The MRE/cMyc complex plays a major role in the basal level expression, and shares some similarities to the upstream stimulatory factor element (USF) identified in the promoter regions of mammalian HO-1 genes and other stress regulated genes. Because sodium arsenite is known to cause oxidative stress and because activation of AP-1 proteins has been shown to be a key step in the oxidative stress response pathway, we also explored the possibility that the induction of the cHO-1 gene by sodium arsenite is mediated through oxidative stress pathway(s) by activation of AP-1 proteins. We found that pretreatment with antioxidants (N-acetyl cysteine or quercetin) reduced the induction of the endogenous cHO-1 message or cHO-1 reporter construct activities induced by sodium arsenite or cobalt chloride. These antioxidants also reduced the protein binding activities to the AP-1 element in the electrophoretic mobility shift assays. In summary, induction of the cHO-1 gene by sodium arsenite or cobalt chloride is mediated by activation of the AP-1 element located at -1,573 to -1,580 of the 5′ UTR.
    Materialart: Digitale Medien
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  • 29
    Digitale Medien
    Digitale Medien
    Springer
    Molecular and cellular biochemistry 205 (2000), S. 1-11 
    ISSN: 1573-4919
    Schlagwort(e): kidney ; ischemia-reperfusion injury ; free radicals ; reactive oxygen species ; gene expression ; antioxidant enzymes
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract Reactive oxygen species (ROS; O2-, H2O2, and OH·), normal by-products of cellular metabolic processes, are kept in control by antioxidant enzymes, such as catalase, glutathione peroxidase (GPX) and superoxide dismutases (SODs). To understand the role of antioxidant enzymatic defenses against ROS injury following ischemia-reperfusion, we examined the effect on kidney exposed to varying periods (30, 60 or 90 min) of ischemia followed by different periods of reperfusion. The enzymatic activities and protein levels of catalase, GPX, CuZnSOD and MnSOD were relatively unaffected at 30 min of ischemia followed by 0, 2 or 24 h reperfusion. However, 60 or 90 min of ischemia followed by 0, 2 or 24 h of reperfusion resulted in a decrease in activities and protein levels which paralleled the duration of ischemic injury. MnSOD activity tended to recover towards normal during reperfusion. Examination of the mRNA levels of these antioxidant enzymes demonstrated a severe decrease in mRNA levels of catalase and GPX at a time point of minimal ischemic injury (30 min of ischemia followed by reperfusion) suggesting that loss of mRNA of catalase and GPX may be the first markers of alterations in cellular redox in ischemia-reperfusion injury. Greater loss of mRNA for catalase, GPX and CuZnSOD were observed following longer periods (60 or 90 min) of ischemia. The mRNA for MnSOD was upregulated at all time points of ischemia-reperfusion injury. Actually, the greater decrease in mRNAs for catalase, GPX and CuZnSOD in the acute phase (within 24 h) subsequently showed a further decrease in these enzyme activities in the subacute phase (72 or 120 h after ischemia). These enzyme activities in the 30 min ischemia group, but not in the 90 min group, already showed tendencies for normalization at 120 h after ischemia. To understand the molecular basis of the loss of mRNA of these antioxidant enzymes during ischemia-reperfusion injury, we examined the rate of transcription by nuclear run-on assays. The similar rates of transcription in control and kidney exposed to ischemia-reperfusion indicates that the loss of mRNA for catalase, GPX and CuZnSOD are possibly due to the increased rate of turnover of their mRNAs. These studies suggest that expression of antioxidant genes during ischemia-reperfusion are not coordinately expressed and the differential loss of antioxidant enzymes may be the contributing factor(s) towards the heterogeneous renal tissue damage as a result of ischemia-reperfusion induced oxidative stress.
    Materialart: Digitale Medien
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  • 30
    ISSN: 1573-4919
    Schlagwort(e): prostaglandin ; cyclooxygenase ; transcriptional regulation ; gene expression ; promotor activation ; transcription ; endothelial cells
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract Interleukin-1β (IL-1) is a potent inducer of cyclooxygenase-2 (COX-2) and prostaglandin biosynthesis in many types of cells, yet little is known about the molecular mechanisms regulating IL-1 mediated prostanoid biosynthesis in the endothelium of the microvasculature. Therefore, we examined the cis- and trans-acting factors regulating IL-1-induced COX-2 expression in the human microvascular endothelial cell line, HMEC-1. IL-1 enhanced steady state levels of COX-2 protein and mRNA synthesis by ≈ 2-fold which preceded a 2-fold increase in PGFα biosynthesis. Expression of a series of COX-2 promoter-luciferase constructs in IL-1 treated HMEC-1 cells revealed that the 'full length' (-1432/+59 bp) promoter was 10 times more active than the SV-40 promoter/enhancer and that it could be further activated by IL-1. Surprisingly however, all except for the shortest COX-2 promoter construct retained the ability to respond to IL-1 and luciferase activity driven by -191/+59 bp COX-2 promoter was as responsive to IL-1 as the full-length promoter. Moreover, site-directed promoter mutagenesis and electophoretic mobility shift assays (EMSA) indicate that the combinatorial actions of AP2, NF-IL6, and CRE elements are critical for both constitutive and IL-1-inducible COX-2 promoter activity. Understanding the mechanism(s) regulating COX-2 gene expression and prostaglandin biosynthesis in the microvasculature has important implications with regard to inflammation and angiogenesis in vivo.
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  • 31
    Digitale Medien
    Digitale Medien
    Springer
    Molecular and cellular biochemistry 203 (2000), S. 163-167 
    ISSN: 1573-4919
    Schlagwort(e): thymosin β-4 ; gene expression ; chloramphenicol acetyltransferase ; NIH3T3 cells ; interferon response
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract Expression of the gene coding for thymosin β-4 (Tβ-4), the major G-actin sequestering peptide in the cell, is regulated mainly at the level of transcription. In this study, we examined the nucleotide sequence of the 5′-flanking region (from - 2202 to - 881) of the mouse Tβ-4 gene, and demonstrated that the DNA fragment from -278 to +410 of this gene was capable of directing the expression of a chloramphenicol acetyltransferase reporter gene in NIH3T3 cells. However, expression of the reporter gene in cells cannot be induced by interferon-a treatment even though a rapid activation of endogenous Tβ-4 gene by this cytokine was observed. These results suggest that the projected interferon-stimulated response element (ISRE) might reside in other parts of the mouse Tβ-4 gene.
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  • 32
    Digitale Medien
    Digitale Medien
    Springer
    Molecular and cellular biochemistry 211 (2000), S. 103-110 
    ISSN: 1573-4919
    Schlagwort(e): thioacetamide ; glutathione-S-transferase ; rat liver ; transcription ; gene expression
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract The effect of thioacetamide (TA), an hepatotoxic and hepatocarcinogenic compound, on the expression and activity of the cytosolic enzyme glutathione-S-transferase (GST) was studied in rat liver. Four h following the administration of 14C-labeled thioacetamide (10 mg/Kg), several subunits of GST were found to be radioactively labeled. A single sublethal dose of TA (250 mg/Kg) decreased by three-fold the expression of classα GST at 24-48 h of treatment, but did not significantly affect the transcription of class μ GST. The activity of the enzyme toward 1-chloro-2,4-dinitrobenzene was mildly inhibited (66% of the control) by a 24 h TA treatment and gradually increased thereafter. It is proposed that the covalent binding of TA or its derivative to the GST subunits does not affect the activity of the enzyme. Nevertheless, GST activity inhibition is due to the deleterious effect of TA on GST transcription.
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  • 33
    Digitale Medien
    Digitale Medien
    Springer
    Molecular and cellular biochemistry 209 (2000), S. 125-129 
    ISSN: 1573-4919
    Schlagwort(e): apolipoprotein E ; apolipoprotein A-I ; gene expression ; transgenic mouse
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract The levels of plasma apolipoprotein (apo) E, an anti-atherogenic protein involved in mammalian cholesterol transport, were found to be 2-3 fold lower in mice over-expressing human apoA-I gene. ApoE is mainly associated with VLDL and HDL-size particles, but in mice the majority of the apoE is associated with the HDL particles. Over-expression of the human apoA-I in mice increases the levels of human apoA-I-rich HDL particles by displacing mouse apoA-I from HDL. This results in lowering of plasma levels of mouse apoA-I. Since plasma levels of apoE also decreased in the apoA-I transgenic mice, the mechanism of apoE lowering was investigated. Although plasma levels of apoE decreased by 2-3 fold, apoB levels remained unchanged. As expected, the plasma levels of human apoA-I were almost 5-fold higher in the apoAI-Tg mice compared to mouse apoA-I in WT mice. If the over-expression of human apoA-I caused displacement of apoE from the HDL, the levels of hepatic apoE mRNA should remain the same in WT and the apoAI-Tg mice. However, the measurements of apoE mRNA in the liver showed 3-fold decreases of apoE mRNA in apoAI-Tg mice as compared to WT mice, suggesting that the decreased apoE mRNA expression, but not the displacement of the apoE from HDL, resulted in the lowering of plasma apoE in apoAI-Tg mice. As expected, the levels of hepatic apoA-I mRNA (transgene) were 5-fold higher in the apoAI-Tg mice. ApoE synthesis measured in hepatocytes also showed lower synthesis of apoE in the apoAI-Tg mice. These studies suggest that the integration of human apoA-I transgene in mouse genome occurred at a site that affected apoE gene expression. Identification of this locus may provide further understanding of the apoE gene expression.
    Materialart: Digitale Medien
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  • 34
    Digitale Medien
    Digitale Medien
    Springer
    Molecular and cellular biochemistry 212 (2000), S. 29-34 
    ISSN: 1573-4919
    Schlagwort(e): cAMP ; transcription factor-decoy oligonucleotides ; CRE ; Ap-1 ; p53 ; tumor growth ; gene expression
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract Nucleic acid molecules with high affinities for a target transcription factor can be introduced into cells as decoy cis-elements to bind these factors and alter gene expression. This review discusses a synthetic single-stranded palindromic oligonucleotide, which self-hybridizes to form a duplex/hairpin and competes with cAMP response element (CRE) enhancers for binding transcription factors. This oligonucleotide inhibits CRE- and Ap-1-directed gene transcription and promotes growth inhibition in vitro and in vivo in a broad spectrum of cancer cells, without adversely affecting normal cell growth. Evidence presented here suggests that the CRE-decoy oligonucleotide can provide a powerful new means of combating cancers, viral diseases, and other pathological conditions by regulating the expression of cAMP-responsive genes.
    Materialart: Digitale Medien
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  • 35
    Digitale Medien
    Digitale Medien
    Springer
    Molecular and cellular biochemistry 212 (2000), S. 73-79 
    ISSN: 1573-4919
    Schlagwort(e): adrenergic receptors ; renin-angiotensin system (RAS) ; gene expression ; kidney
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract To investigate the molecular mechanism(s) of action of catecholamines on the expression of the angiotensinogen (ANG) gene in kidney proximal tubular cells, we used opossum kidney (OK) cells with a fusion gene containing the 5′-flanking regulatory sequence of the rat ANG gene fused with a human growth hormone (hGH) gene as a reporter, pOGH (rANG N-1498/+18), permanently integrated into their genomes. The level of expression of the ANG-GH fusion gene was quantified by the amount of immunoreactive-hGH (IR-hGH) secreted into the medium. The addition of norepinephrine (NE), isoproterenol (a β1/β2-adrenergic receptor (AR) agonist) and iodoclonidine (an α2-AR agonist) stimulated the expression of the ANG-GH fusion gene in a dose-dependent manner, whereas the addition of epinephrine and phenylephrine (α1-AR agonist) had no effect. The stimulatory effect of NE was blocked by the presence of propranolol (β-AR blocker), atenolol (β1-AR blocker), yohimbine (α2-AR blocker), Rp-cAMP (an inhibitor of cAMP-dependent protein kinase AI & AII) and staurosporine (an inhibitor of protein kinase C), but was not blocked by ICI 118, 551 (β2-AR blocker) and prazosin (α1-AR blocker). The addition of a combination of isoproterenol and iodoclonidine or a combination of 8-Bromo-cAMP (8-Br-cAMP) and phorbol 12-myristate (PMA) synergistically stimulated the expression of the ANG-GH fusion gene as compared to the addition of isoproterenol, iodoclonidine, 8-Br-cAMP or PMA alone. Furthermore, the addition of NE, 8-Br-cAMP or PMA stimulated the expression of pOGH (rANG N-806/-779/-53/+18), a fusion gene containing the putative cAMP responsive element (CRE, ANG N-806/-779) upstream of the ANG promoter (ANG N-53/+18) in OK cells, but had no effect on the expression of fusion genes containing the mutant of the CRE. Gel mobility shift assays revealed that the ANG-CRE binds with the DNA-binding domain (bZIP 254-327) of the cAMP-responsive binding protein (CREB). The binding of the labeled ANG-CRE to CREB (bZIP254-327) was displaced by unlabeled ANG-CRE and the CRE of the somatostatin gene but not by the mutants of the ANG-CRE. Finally, NE stimulated the phosphorylation of CREB in OK cells. These studies demonstrate that the molecular mechanism(s) of NE action on the expression of the ANG gene in OK cells may be mediated via both the PKA and PKC signalling pathways and via the phosphorylation of CREB. The phosphorylated CREB then interacts with the CRE in the 5′-flanking region of the ANG gene and subsequently stimulates the gene expression.
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  • 36
    Digitale Medien
    Digitale Medien
    Springer
    Molecular and cellular biochemistry 212 (2000), S. 135-142 
    ISSN: 1573-4919
    Schlagwort(e): gene expression ; catecholamines ; angiotensin II ; heart failure ; myosin ; hypertension ; eprosartan
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract Pressure overload of the heart is associated with a perturbed gene expression of the cardiomyocyte leading to an impaired pump function. The ensuing neuro-endocrine activation results in disordered influences of angiotensin II and catecholamines on gene expression. To assess whether angiotensin II type 1 receptor inhibition can also counteract a raised sympathetic nervous system activity, spontaneously hypertensive rats fed a hypercaloric diet were treated with eprosartan (daily 90 mg/kg body wt) and cardiovascular parameters were monitored with implanted radiotelemetry pressure transducers. Both, blood pressure and heart rate were increased (p 〈 0.05) by the hypercaloric diet. Although eprosartan reduced (p 〈 0.05) the raised systolic and diastolic blood pressure, the diet-induced rise in heart rate was blunted only partially. In addition to drugs interfering with the enhanced catecholamine influence, compounds should be considered that selectively affect cardiomyocyte gene expression via 'metabolic' signals.
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  • 37
    ISSN: 1573-4919
    Schlagwort(e): angiotensinogen ; fibronectin ; gene expression ; transcriptional regulation ; cardiomyocytes ; vascular smooth muscle cells
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract Angiotensinogen (AGT) is a unique substrate of the renin-angiotensin system and fibronectin (FN) is an important component of the extracellular matrix. These play critical roles in the pathophysiological changes including cardiovascular remodeling and hypertrophy in response to hypertension. This study was performed to examine the regulation of AGT and FN gene in cardiac myocytes (CMs) and vascular smooth muscle cells (VSMCs) in response to mechanical stretch. Mechanical stretch significantly increased the AGT mRNA expression in CMs, while these stimuli did not affect FN mRNA levels. On the other hand, Mechanical stretch upregulated FN mRNA levels in VSMCs, whereas no increase in AGT mRNA levels was observed in response to stretch stimuli. An angiotensin II type 1 (AT1) receptor antagonist (CV11974) significantly decreased these stretch-mediated increases in mRNA level and promoter activity of the AGT and FN gene, whereas angiotensin II type 2 (AT2) receptor antagonist (PD123319) did not affect the induction. These results indicate that mechanical stretch activates transcription of the AGT and FN gene mainly via AT1 receptor-pathway in CMs and VSMCs. Furthermore, mechanisms regulating AGT and FN gene seem to be different between CMs and VSMCs.
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  • 38
    Digitale Medien
    Digitale Medien
    Springer
    Molecular and cellular biochemistry 212 (2000), S. 211-217 
    ISSN: 1573-4919
    Schlagwort(e): angiotensin receptor ; medullary thick ascending limb ; sodium intake ; primary cell culture ; gene expression
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract Angiotensin II (Ang II) is an important regulator of the function of medullary thick ascending limb of loop of Henle (MTAL). Recent studies showed that changes in Ang II receptor expression occur and underlie changes in the function of proximal tubules during altered sodium intake. The present experiment was designed to determine (1) whether expression of the type 1 Ang II (AT1) receptor in the MTAL is regulated by altered sodium intake, and (2) the specific pathway(s) mediating sodium-induced AT1 expression in the MTAL. Wistar rats were fed a normal sodium (0.5%, NS), low sodium (0.07%, LS), or high sodium (4%, HS) diet for 2 weeks. Northern blot analysis and radioligand binding showed that in rats fed a normal sodium diet the rank of order for both AT1 mRNA expression and receptor density was outer medulla 〉 cortex 〉 inner medulla. Sodium restriction significantly increased both AT1 mRNA expression and receptor density in the outer medulla. In contrast, neither AT1 mRNA expression nor receptor density in the outer medulla was altered by sodium loading. Losartan treatment (3 mg/kg/per day by oral gavage for 2 weeks) prevented low sodium-induced upregulation of the AT1 receptor in the outer medulla, but it had no effect on AT1 expression in the outer medulla of rats fed a normal sodium diet. Highly purified suspensions of MTAL were isolated from rats fed a normal or low sodium diet. Low sodium intake significantly increased AT1 mRNA level by 184% and AT1 receptor density by 58% in MTALs. Primary cultures of MTAL cells were treated with PBS, Ang II (10-8 M), and Ang II + 17 octadecynoic (17 ODYA, 10 μM). Ang II caused about 2-fold increase in AT1 mRNA levels, and this increase was diminished by about 30% by the addition of 17 ODYA. We conclude that (1) sodium restriction but not sodium loading increases AT1 receptor expression in the MTAL, (2) low sodium-induced upregulation of the AT1 receptor in the MTAL is Ang II-dependent, and (3) Ang II-induced upregulation of the AT1 receptor in the MTAL is mediated, at least in part, by cytochrome P450 pathways.
    Materialart: Digitale Medien
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  • 39
    ISSN: 1573-4919
    Schlagwort(e): renin angiotensin system ; sarcoplasmic reticulum ; Ca2+-handling ; gene expression ; ischemia-reperfusion ; cardioprotection
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract The aim of this study was to explore the possible participation of cardiac renin-angiotensin system (RAS) in the ischemia-reperfusion induced changes in heart function as well as Ca2+-handling activities and gene expression of cardiac sarcoplasmic reticulum (SR) proteins. The isolated rat hearts, treated for 10 min without and with 30 μM captopril or 100 μM losartan, were subjected to 30 min ischemia followed by reperfusion for 60 min and processed for the measurement of SR function and gene expression. Attenuated recovery of the left ventricular developed pressure (LVDP) upon reperfusion of the ischemic heart was accompanied by a marked reduction in SR Ca2+-pump ATPase, Ca2+-uptake and Ca2+-release activities. Northern blot analysis revealed that mRNA levels for SR Ca2+-handling proteins such as Ca2+-pump ATPase (SERCA2a), ryanodine receptor, calsequestrin and phospholamban were decreased in the ischemia-reperfused heart as compared with the non-ischemic control. Treatment with captopril improved the recovery of LVDP as well as SR Ca2+-pump ATPase and Ca2+-uptake activities in the postischemic hearts but had no effect on changes in Ca2+-release activity due to ischemic-reperfusion. Losartan neither affected the changes in contractile function nor modified alterations in SR Ca2+-handling activities. The ischemia-reperfusion induced decrease in mRNA levels for SR Ca2+-handling proteins were not affected by treatment with captopril or losartan. The results suggest that the improvement of cardiac function in the ischemic-reperfused heart by captopril is associated with the preservation of SR Ca2+-pump activities; however, it is unlikely that this action of captopril is mediated through the modification of cardiac RAS. Furthermore, cardiac RAS does not appear to contribute towards the ischemia-reperfusion induced changes in gene expression for SR Ca2+-handling proteins.
    Materialart: Digitale Medien
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  • 40
    ISSN: 1573-4919
    Schlagwort(e): pressure overload ; gene expression ; subcellular remodeling ; sarcoplasmic reticulum Ca2+-handling ; anti-hypertensive therapy
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract The effects of propranolol and verapamil on contractile dysfunction, subcellular remodeling and changes in gene expression in cardiac hypertrophy due to pressure overload were examined. Rats were subjected to banding of the abdominal aorta and then treated with either propranolol (10 mg/kg daily), verapamil (5 mg/kg daily) or vehicle for 8 weeks after the surgery. Depression of the left ventricular function in the hypertrophied heart was associated with decreases in myofibrillar and myosin CA2+ ATPase activities as well as Ca2+-pump and Ca2+-release activities of the sarcoplasmic reticulum (SR). The level of a-myosin heavy chain (α-MHC) mRNA was decreased while that of β-MHC mRNA was increased in the pressure-overloaded heart. The level of SR Ca2+-pump ATPase (SERCA2) mRNA and protein content for SERCA2 were decreased in the pressure overloaded heart. Treatment of the hypertrophied animals with propranolol or verapamil resulted in preservation of the left ventricular function and prevention of the subcellular alterations. Shift in the α- and β-MHC mRNA levels and changes in the expression in SERCA2 mRNA level and protein content were also attenuated by these treatments. The results suggest that blockade of β-adrenoceptors or voltage-dependent calcium channels normalizes the cardiac gene expression, prevents subcellular remodeling and thus attenuates heart dysfunction in rats with cardiac hypertrophy. Furthermore, both cardiac β-adrenoceptors and L-type Ca2+-channels may be involved in the genesis of cardiac hypertrophy due to pressure overload.
    Materialart: Digitale Medien
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  • 41
    Digitale Medien
    Digitale Medien
    Springer
    Molecular and cellular biochemistry 213 (2000), S. 119-126 
    ISSN: 1573-4919
    Schlagwort(e): TIS11 ; an immediate early gene ; gene cloning ; gene expression ; gene organization ; promoter
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract The TIS11 gene is an immediate early gene that is induced rapidly and transiently by phorbol 12-myristate 13-acetate and various growth factors. To study transcriptional regulation of the gene, a genomic clone of rat TIS11 was isolated, and the organization of exon-intron structure and transcriptional initiation site were determined. The rat TIS11 gene consisted of 2 exons spanning approximately 2.5 kb. Several canonical sequences for binding of transcriptional factors were found in the 5′-flanking region. The 5.3 kb of the 5′-flanking region fused to a luciferase reporter gene showed promoter activity when introduced into rat pheochromocytoma PC12 cells. Analyses with serial 5′-deletion mutants suggested that the major positive regulatory region is located at the region of -241 to -76, and that the minimum promoter region is within the 76-bp upstream of the transcriptional initiation site. Gel mobility shift assays revealed that PC12 cell nuclear proteins specifically bind to the major positive regulatory region of the TIS11 gene. The identified nuclear protein components may act as the positive trans-acting factors in the basal expression of the TIS11 gene in PC12 cells.
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  • 42
    ISSN: 1432-2242
    Schlagwort(e): Key words Pinus thunbergii ; Pine needle gall midge ; RAPD ; Bulked segregant analysis ; Linkage
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  Linkage of RAPD markers to a single dominant gene for resistance to pine needle gall midge was investigated in Japanese black pine (Pinus thunbergii). Three primers that generated linked markers were found after 1160 primers were screened by bulked segregant analysis. The distances between the resistance gene, R, and the marker genes OPC06580, OPD01700, and OPAX192100 were 5.1 cM, 6.7 cM and 13.6 cM, respectively. OPC06580 was in coupling phase to R, whereas OPD01700 and OPAX192100 were in repulsion phase to R. A linkage map for a resistant tree was constructed using 96 macrogametophytes. In linkage analysis, 98 out of 127 polymorphic markers were assigned to 17 linkage groups and six linked pairs. The total length of this map was 1469.8 cM, with an average marker density of 15.6 cM. The genome length was estimated to be 2138.3 cM, and the derived linkage map covered 67.5% of the genome. Although the linked markers OPC06580, OPAX192100, and OPD01700, belonged to the same linkage group, no precise positions were found for OPC06580 or OPD01700.
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  • 43
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 101 (2000), S. 1250-1258 
    ISSN: 1432-2242
    Schlagwort(e): Key words Micronuclei ; Microprotoplasts ; Chromosome transfer ; RAPD ; Helianthus
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  Asymmetric somatic hybrid (ASH) plants were obtained by PEG-mediated mass fusion of microprotoplasts from perennial Helianthus species and hypocotyl protoplasts of Helianthus annuus. The formation of micronuclei in perennial sunflower cell cultures was induced, at early log phase, by addition of the herbicides amiprophos-methyl or oryzalin. Sub-diploid microprotoplasts were isolated by high-speed centrifugation and the smallest enriched by sequential filtration through nylon sieves of decreasing pore size. Fusion products were cultured and the regenerated plants phenotypically, genetically and cytologically characterized. DNA analysis using RAPD markers revealed that 28 out of 53 regenerated plants were asymmetric hybrids. Subsequent nuclear-DNA flow cytometric analysis showed that these plants had a higher DNA content than the receptor H. annuus, suggesting that they represented addition lines. Cytological investigation of the metaphase cells of 16 hybrids revealed an addition of 2–8 extra chromosomes in these plants. The phenotype of most ASH plants resembled H. annuus. These results indicate that micronuclear induction and asymmetric somatic hybridization represent a potent tool for partial genome transfer aimed at the specific transfer of economically important traits in breeding programs.
    Materialart: Digitale Medien
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  • 44
    ISSN: 1871-4528
    Schlagwort(e): somatic hybrids ; potato leafroll virus ; RAPD ; Solanum wild species ; pollen fertility
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Summary Somatic fusions between an accession of the diploid wild speciesSolanum verrucosum and a dihaploid S.tuberosum genotype were produced in order to incorporate resistance to potato leafroll virus (PLRV). In total 15 somatic hybrids out of 16 regenerants were obtained. Identification of hybrids was based on additive RAPD patterns, general morphological characteristics, chromosome numbers and chloroplast counts in stomata guard cells. A field trial was performed with the hybrids, their two parents and the control cultivar Kennebec to assess field performance and phenotypic variability. Yield parameters varied considerably among somatic hybrids. Some of the hybrids gave significantly higher yields, tuber numbers and tuber weights than both parents. Pollen fertility of hybrids ranged from 19 to 77%. Twelve hybrids were found to be resistant to PLRV.
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  • 45
    Digitale Medien
    Digitale Medien
    Springer
    Potato research 43 (2000), S. 383-393 
    ISSN: 1871-4528
    Schlagwort(e): Solanum tuberosum L. ; plant development ; antioxidant genes ; gene expression
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Summary The expression of antioxidant genes has been analyzed in a potato plant and during tuber dormancy. Manganese superoxide dismutase (MnSOD), cytosolic copper and zinc superoide dismutase (Cu/ZnSOD), catalase class II, cytosolic ascorbate peroxidase (APX) and glutathione peroxidase (GPX) are expressed at the RNA level in all the contexts analyzed. By contrast, the expression of the iron superoxide dismutase (FeSOD) and plastidic Cu/ZnSOD seems to be limited to green tissues, as shown by northern blots and native gels. A complex DAB-peroxidase isozyme pattern (using diaminobenzidine as substrate) has been observed in different developmental contexts analyzed, but hardly observed in tubers. During tuber dormancy, MnSOD and cytosolic Cu/ZnSOD activity was relatively constant in both Désirée and Bintje varieties while catalase activity decreases. Moreover, tuber dormancy breakage did not involve significant changes in the activity of these enzymes. On the basis of these results, the possible link between active oxygen species (AOS) metabolism and dormancy is discussed.
    Materialart: Digitale Medien
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  • 46
    ISSN: 1871-4528
    Schlagwort(e): gene expression ; cDNA-AFLP ; RNA-fingerprinting ; organogenesis ; tuberisation ; dormancy ; sprouting ; cluster analysis ; metabolic pathways
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Summary Potato tuber life-cycle is composed of many individual developmental stages including tuber formation, tuber development, dormancy and sprouting. We have used cDNA-AFLP fingerprinting to analyse gene expression in 24 individual stages of development, over the period from stolon formation through sprouting. In addition to these developmental stages, different tissues were analysed to assess tissue specificity and various controls were incorporated to determine process specificity. In total around 18000 transcript derived cDNA fragments (TDFs) were visualised from which circa 2600 were included in a statistical analysis allowing general conclusions about gene expression during development. More than 200 process specific TDFs were isolated and sequenced throughout the potato tuber life-cycle. The sequence similarities of these TDFs to known genes give an insight into the kinds of processes occurring during tuberisation, dormancy and sprouting.
    Materialart: Digitale Medien
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  • 47
    ISSN: 1611-4663
    Schlagwort(e): Lentinula edodes ; Heterozygous DNA marker ; RAPD ; de-dikaryotization ; Protoplast
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft , Maschinenbau
    Notizen: Abstract A suitable screening method for heterozygous DNA markers in shiitake,Lentinula edodes (Berk.) Pegler, is reported. Monokaryons were derived from a dikaryon by de-dikaryotization via protoplast formation. Compatibility of the monokaryons was determined by pairwise culture on agar plates. We selected the primers to amplify polymorphic fragments among the original strain (Hokken600∶H600) and two monokaryons (H600PP-39 and H600PP-67) showing compatibility. A total of 135 fragments were selected as specific random amplified polymorphic DNAs (RAPDs) resulting from 56 primers of the 147 primers tested. Furthermore, we tested whether the polymorphic fragments segregated into 2∶2 among four strains isolated from a basidium. Most of the polymorphic fragments (about 97.8%) showed 2∶2 segregation among the four strains. We concluded that the polymorphic fragments were heterozygous if they were detected in either of the monokaryons (H600PP-39 and H600PP-67) and segregated to 2∶2 among four meiotic strains (H600B-1,-2, -3, and -4). A total of 132 heterozygous DNA markers were therefore selected from a dikaryon of shiitake (Hokken600∶H600).
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  • 48
    Digitale Medien
    Digitale Medien
    Springer
    Mycoscience 41 (2000), S. 145-148 
    ISSN: 1618-2545
    Schlagwort(e): mycelial compatibility ; RAPD ; Sclerotium rolfsii
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Sclerotium rolfsii isolates from peanut fields in Ibaraki were classified into mycelial compatibility groups (MCGs) based on the barrage zone formation. A total of 132 isolates collected from four fields within a 120 m radius in 1994 comprised four MCGs; MCG A (71 isolates), B (34 isolates), C (26 isolates) and D (one isolate). Fields 1 and 2 were occupied exclusively by MCG A. MCG A also predominated in field 3. In field 4, MCGs A, B and C were dominant. Population structure in 3 additional fields was determined in 1997. All 11 isolates from Field 5, which was 400 m distant from field 1, belonged to MCG C. A total of 42 isolates from fields 6 and 7, 2.5 km distant from other fields and 100 m distant from each other, were all MCG A. These results suggested that the population structure ofS. rolfsii was simple. RAPD fingerprintings showed that most isolates of the same MCG were clonal.
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  • 49
    ISSN: 1573-4927
    Schlagwort(e): AFLP ; RAPD ; polymorphism ; Mystus nemurus.
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie
    Notizen: Abstract This work represents the first application of the amplified fragment length polymorphism (AFLP) technique and the random amplified polymorphic DNA (RAPD) technique in the study of genetic variation within and among five geographical populations of M. nemurus. Four AFLP primer combinations and nine RAPD primers detected a total of 158 and 42 polymorphic markers, respectively. The results of AFLP and RAPD analysis provide similar conclusions as far as the population clustering analysis is concerned. The Sarawak population, which is located on Borneo Island, clustered by itself and was thus isolated from the rest of the populations located in Peninsular Malaysia. Both marker systems revealed high genetic variability within the Universiti Putra Malaysia (UPM) and Sarawak populations. Three subgroups each from the Kedah, Perak, and Sarawak populations were detected by AFLP but not by RAPD. Unique AFLP fingerprints were also observed in some unusual genotypes sampled in Sarawak. This indicates that AFLP may be a more efficient marker system than RAPD for identifying genotypes within populations.
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  • 50
    Digitale Medien
    Digitale Medien
    Springer
    Genetica 110 (2000), S. 267-276 
    ISSN: 1573-6857
    Schlagwort(e): Clarias ; RAPD ; sex determination ; siluroid ; teleost
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We searched for sex-specific DNA sequences in the male and female genomes of African catfish, Clarias gariepinus (Burchell, 1822) by comparative random amplified polymorphic DNA (RAPD) assays performed on pooled DNA samples. Two sex-linked RAPD markers were identified from the male DNA pool and confirmed on individual samples, showing good agreement with phenotypic sex. Both markers were isolated, cloned and characterized. The first marker (CgaY1) was nearly 2.6 kb long, while the length of second one (CgaY2) was 458 bp. Southern blot analysis with a CgaY1 probe showed strong hybridizing fragments only in males and not in females under stringent conditions, indicating the presence of multiple copies of CgaY1 in the male genome. When tested by zoo blot on the genomes of two closely related species from the Clariidae family, CgaY1 hybridized to the DNA of Heterobranchus longifilis and generated a faint male-specific band at low stringency. CgaY2 produced similar hybridization pattern in both sexes of C. gariepinus, C. macrocephalus and H. longifilis. Specific primers were designed to the sequences and the markers were amplified in multiplex PCR reactions together with a control band common to all individuals. This allowed for rapid, molecular sexing of the species on the basis of a simple three band (male) versus one band (female) pattern. According to our knowledge these are the first sex-specific DNA markers isolated from a siluroid fish species.
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  • 51
    ISSN: 1573-6857
    Schlagwort(e): boll weevil ; dispersal ; origin ; RAPD ; South America
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract RAPD technique provides useful information on the geographic origin and dispersal of the boll weevil Anthonomus grandis in South America. Nine populations from Argentina, Brazil, Paraguay, Mexico and USA were analyzed. Weevils were captured on native plants (Misiones province, Argentina) and on cotton cultures, except the sample from the United States (USDA laboratory-reared colony). A sample of the ‘Peruvian square weevil’, A. vestitus, from Ecuador, was included in the analysis in order to compare interspecific variation. The four primers used in the analysis revealed 41 ‘anonymous loci’. The neighbor-joining tree based on Nei's distances and values of Nm (migrants per generation), indicate that genetic similarity between samples from Tecomán (Mexico) and Puerto Iguazú (Argentina), is higher than among remaining South American populations. This result supports an hypothesis of natural occurrence of the boll weevil in South America, prior to extensive cotton cultivation. Population outbreaks of the species would be associated with increase of agricultural lands.
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  • 52
    ISSN: 1573-8469
    Schlagwort(e): diagnostics ; RAPD ; root-knot nematodes
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract This study describes the development of species-specific pairs of PCR primers for the root-knot nematodes Meloidogyne chitwoodi, M. fallax and M. hapla that amplify species-specific RAPD fragments. After sequencing the fragments, longer primers were designed to complement the terminal sequences of the polymorphic DNA fragments. The resulting pairs of primers were used to generate the sequence-characterized amplified regions (SCARs). Using the developed pairs of SCAR primers, SCAR fragments of M. chitwoodi, M. fallax or M. hapla were easily amplified from DNA extracts from juveniles, egg masses, females of the particular nematode species investigated, either present alone, in a mixture with other nematode species or in infested plant material. A specially designed multiplex assay using three pairs of SCAR primers enabled the identification of multiple species in a mixture in a single PCR step. Single juveniles were easily identified by applying this multiplex assay followed by a subsequent multiplex PCR using three pairs of nested primers. The SCAR-PCR-based assays described have potential to be optimized for routine practical diagnostic tests. The usefulness of converting RAPD markers into SCAR markers is discussed.
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  • 53
    ISSN: 1615-6110
    Schlagwort(e): Acacia ; classification ; Leguminosae ; morphology ; phenetics ; RAPD ; software ; taxonomy ; UPGMA
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The phenetic analysis of non-nodulatingAcacia species by Harrier et al. (1997) was repeated to illustrate how different computer programs may generate alternative UPGMA trees for the very same data, even in the absence of data input order effects (ties). For example, all Harrier et al.'s UPGMA dendrograms produced by software from the Scottish Agricultural Statistics Service differed from those obtained by the packages NTSYS and MVSP87. Particularly, the positions ofA. albida, A. rovumae, andA. pentagona, as well as the relationships betweenDiacanthae andTriacanthae were affected by this phenomenon. Hence, whenever clustering techniques are used, care should be taken to consider possible software-dependent caveats and artefacts. Nevertheless, all programs provided clusterings that largely coincided with the subgeneric and sectional groupings proposed by Vassal (1972) although the positions of some species varied depending on whether morphological or molecular data were considered (e.g.A. albida andA. rovumae).
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  • 54
    Digitale Medien
    Digitale Medien
    Springer
    Plant systematics and evolution 225 (2000), S. 85-101 
    ISSN: 1615-6110
    Schlagwort(e): Elaeagnaceae ; Hippophae ; sea buckthorn ; Systematics ; taxonomy ; genetic variation ; RAPD
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Genetic diversity has been investigated by the application of molecular markers in, for the first time, all the taxa recognised in recent treatises of the genusHippophae. RAPD (random amplified polymorphic DNA) analyses were conducted with 9 decamer primers, which together yielded 219 polymorphic markers. We found 16 fixed RAPD markers, i.e. markers that either occurred in all plants of a population or were absent from all plants. Several of these markers were useful for analysis of interspecific relationships, whereas others can be considered as taxon-specific markers. Clustering of taxa and populations in our neighbour-joining based dendrogram was in good agreement with some recently suggested taxonomic treatises ofHippophae. Amount and distribution of genetic variability varied considerably between species. Partitioning of molecular variance withinH. rhamnoides supported earlier findings that a considerable part of the total variance resides among subspecies (59.6%) Within-population variability also differed considerably. Percentage polymorphic RAPD loci and Lynch and Milligan within-population gene diversity estimates showed relatively high values for some species close to the geographic centre of origin in Central Asia, e.g.H. tibetana and the putatively hybridogenousH. goniocarpa. Spatial autocorrelation analyses performed on 12 populations ofH. rhamnoides revealed positive autocorrelation of allele frequencies when geographic distances ranged from 0 to 700 km, and no or negative autocorrelation at higher distances. At distances between 700 and 1900 km, we observed deviations from the expected values with strongly negative autocorrelation of allele frequencies. A corresponding relationship between geographic and genetic distances could not be found when the analysis instead was based on one population from each of 8 species.
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  • 55
    ISSN: 1573-5028
    Schlagwort(e): gene expression ; germination ; NADPH-cytochrome P450 reductase ; Pseudotsuga menziesii
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract NADH-cytochrome P450 is a key enzyme that transfers electrons from NADPH to the cytochrome P450 family of enzymes. To begin to determine the regulation of CPR gene expression and enzyme activity in Douglas-fir a full-length cDNA was isolated from a seedling λZAP cDNA library and the ORF was used to develop a synthetic CPR-peptide-based antiserum. Northern blot analysis indicated CPR expression was regulated both developmentally prior to seed maturation and during germination, and differentially in the cotyledons, radicle and megagametophyte of seed and seedling tissues. The CPR-peptide antiserum detected a single CPR in seed and seedling microsomes analyzed by western blot of two-dimensional SDS-polyacrylamide gels. In microsomal extracts from whole seeds and seedlings, the amount of CPR protein remained constant while NADPH:cytochrome c reductase activity increased during stratification, germination and early seedling development. In contrast to cotyledons and megagametophyte, the level of CPR protein detected in radicles was higher than expected when compared to the amount of CPR transcript.
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  • 56
    ISSN: 1573-5060
    Schlagwort(e): Cheyenne ; polymorphism ; RAPD ; recombinant inbred chromosome line(RICL) RFLP ; STS ; SSR ; Triticum aestivum ; Wichita
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Previously chromosome 3A of wheat (Triticum aestivum L.) was reported to carry genes influencing yield, yield components, plant height, and anthesis date. The objective of current study was to survey various molecular marker systems for their ability to detect polymorphism between wheat cultivars Cheyenne(CNN) and Wichita (WI), particularly for chromosome3A. Seventy-seven `sequence tagged site' (STS), 10simple sequence repeat (SSR), 40 randomly amplified polymorphic DNA (RAPD) markers, and 52 restriction fragment length polymorphism (RFLP) probes for wheat homoeologous group 3 chromosomes, were investigated. Three (3.9%) STS-PCR primer sets amplified polymorphic fragments for the two cultivars, of which one was polymorphic for chromosome 3A. Sixty percent of SSR markers detected polymorphism between CNN and WI of which 50% were polymorphic for chromosome 3A. Twenty percent of RAPD markers detected polymorphism between CNN and WI in general, but none of these detected polymorphism for chromosome 3A. Of the fifty-two RFLP probes, 78.8% detected polymorphism between CNN and WI for group 3 chromosomes with one or more of seven restriction enzymes and 42% of the polymorphic fragements were for chromosome 3A. These high levels of RFLP and SSR polymorphisms between two related wheat cultivars could be used to map and tag genes influencing important agronomic traits. It may also be important to reconsider RFLP as the most suitable marker system at least for anchor maps of closely related wheat cultivars.
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  • 57
    Digitale Medien
    Digitale Medien
    Springer
    Euphytica 111 (2000), S. 127-135 
    ISSN: 1573-5060
    Schlagwort(e): barley varieties ; genetic variability ; Hordeumvulgare ssp. vulgare ; molecular characterization ; RAPD
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Thirteen varieties of Brazilian barley selected for malting qualities were analysed by RAPD (Random Amplified Polymorphic DNA). Amplification with 18 random primers generated 221 reproducible bands, of which 206 bands were polymorphic (93%):of this number, 137 fragments (62%) detected diversity among varieties and 56 bands (25.34%) allowed the distinction of varieties or groups of them. Variation was detected in all Brazilian varieties studied. Within-variety similarities estimated by Jaccard Similarity Coefficient ranged from 0.28 to 0.94, with averages ranging from 0.57 to 0.83, and an overall average of 0.72. Nevertheless, in the cluster analysis representatives of the same variety always fell into the same group and only later joined the other varieties. The average intervarietal similarities estimated by Jaccard Similarity Coefficient ranged from 0.45 to 0.62, with an overall average of 0.52. Many bands or combinations of bands which were responsible for the differentiation of all varieties were detected. Nevertheless, the majority of these bands cannot be considered as diagnostic markers because a great number of them did not occur in many representatives of the variety or had low intensity or even because they were not easily identified in the total pattern of bands.
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  • 58
    ISSN: 1573-5060
    Schlagwort(e): cabbage (Brassica oleracea var. capitata) ; ERPAR ; RAPD ; male sterility gene ; molecular marker
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Similar to SCAR, an extended random primer amplified region (ERPAR) marker is a PCR amplified genomic DNA fragment at a single genetically defined locus. However, ERPAR uses specific primer pairs derived from RAPD primers by adding bases sequentially to their 3′-ends. As an example, an ERPAR marker was derived from a RAPD marker (OT11900) linked to a dominant male sterility gene in cabbage (Brassica oleracea var. capitata). After two cycles of base adding and primer pair screening, a primer pair (5′-TTCCCCGCGACT-3′and 5′-TTCCCCGCGAGA-3′) amplified a single intense band with the same size as OT11900. The identity of the new marker and OT11900 was verified by segregation analysis. The new marker amplified by this extended primer pair was named as EPT11900. The development of ERPAR exploits the importance of 3′-end bases of primers in PCR ERPAR shares advantages of SCAR, but eliminates the need for cloning and sequencing. It is a fast and universal way of converting RAPD markers into stable markers.
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  • 59
    Digitale Medien
    Digitale Medien
    Springer
    Plant and soil 221 (2000), S. 33-38 
    ISSN: 1573-5036
    Schlagwort(e): ammonium ; birch ; gene expression ; nia promoter ; nitrate ; nitrate reductase
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract A 1535 bp promoter of the nitrate reductase gene (nia) from birch (Betula pendula) and a series of 5′ deletions were fused to the β-glucuronidase (GUS) gene and introduced into Nicotiana plumbaginifolia. In transgenic plants the NR promoter sequences directed strong GUS expression in the root epidermal hair cells, and in phloem cells of leaf and stem vascular tissue. The NR promoter confers also a significant stimulation of the GUS gene expression by nitrate. These findings might indicate that nitrate flow is one of the signals involved into tissue and cell specific expression of the NR promoter GUS fusions.
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  • 60
    Digitale Medien
    Digitale Medien
    Springer
    Plant and soil 226 (2000), S. 219-225 
    ISSN: 1573-5036
    Schlagwort(e): arbuscular mycorrhizas ; gene expression ; Glomus mosseae ; nutrient transport processes ; plasma membrane H+-ATPases
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract The activity of H+-ATPases of plant and fungi generates an electrochemical gradient of H+ across the cell plasma membrane that drives a number of secondary transport systems, including those responsible for the translocation of cations, anions, amino acids and sugars. During the last years, several studies have been aimed at elucidating the role of plasma membrane H+-ATPases in the nutrient exchange processes taking place between the plant and the fungus in arbuscular mycorrhizal (AM) symbiosis. This paper reviews present knowledge about plasma membrane H+-ATPases and experimental evidence supporting the involvement of H+-ATPases of both organisms in the bidirectional transport of nutrients between partners. Molecular strategies that will provide further information on the function and regulation of plasma membrane H+-ATPases in AM symbiosis are presented and discussed.
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  • 61
    ISSN: 1573-5060
    Schlagwort(e): Camellia sinensis ; genetic diversity ; RAPD
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract The diversity of 27 superior tea (Camellia sinensis var. sinensis) accessions from Korea, Japan and Taiwan was examined with RAPD-PCR (Random Amplified Polymorphic DNA Polymerase Chain Reaction) markers. Out of the 50 primers screened, 17 primers generated 58 polymorphic and reproducible bands. A minimum of 3 primers was sufficient to distinguish all the 27 accessions studied. The Shannon's index used to partition diversity into inter- and intra-group, revealed that 71 percent of variability resided within groups and 29 percent between groups. Diversity was greatest within the Korean group followed by Taiwan and Japan. The relatively high diversity observed in Korea might reflect the larger genetic base of its plantations while the low diversity in Japan could be explained by the long and intensive tea selection programme in this country. A dendrogram based on the UPGMA-link method using Jaccard's distances and multivariate Factorial correspondence analysis clustered the tea accessions into two main groups, regrouping the Taiwan cultivars on the one side and the Korean and Japanese accessions on the other side. This suggests that the Taiwan tea studied here may have a different origin from that of Korea and Japan.
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  • 62
    Digitale Medien
    Digitale Medien
    Springer
    Euphytica 116 (2000), S. 281-285 
    ISSN: 1573-5060
    Schlagwort(e): Citrus ; early flowering ; electrofusion ; RAPD ; somatic hybrids
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Protoplasts from cell suspension cultures of ‘Bonnaza’ navel orange (Citrus sinensis (L.) Osbeck) were electrically fused with mesophyll protoplasts isolated from seedless ‘Red Blush’ grapefruit (Citrusparadisi). After 6 months of culture, a total of 20 plants were regenerated. Root tip chromosome counting revealed that 4 of them were tetraploids (2n = 4x = 36)and the rest were diploids (2n = 2x = 18) morphologically resembling the mesophyll parent. After 6 months of transplantation into the greenhouse, 4 of the diploidmesophyll regenerants unexpectedly flowered, but this phenomenon disappeared in the next year. This is the first report of precocious flowering in citrus via protoplast fusion. RAPD analysis further confirmed that the tetraploid regenerants were somatic hybrids while the diploid regenerants were mesophyll parent type. This somatic hybrid will be utilized as a possible pollen parent for improving the seedy pummelo cultivars in China by producing triploid seedless pummelo hybrid. The mechanism of early flowering was also discussed.
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  • 63
    Digitale Medien
    Digitale Medien
    Springer
    Euphytica 111 (2000), S. 121-125 
    ISSN: 1573-5060
    Schlagwort(e): DNA fingerprinting ; Mentha ; phylogenetic tree ; RAPD
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract A set of 60 random primers was used to analyse 11accessions from six taxa of Mentha developed byCIMAP. These accessions were maintained in the nationalgene bank for medicinal and aromatic plants at CIMAP.A total of 630 bands could be detected as amplifiedproducts upon PCR amplification, out of which 589 werepolymorphic (93.5%). Further analysis of these RAPDprofiles for band similarity indices clearlydifferentiated five of the Mentha arvensis L.accessions from the rest. Among two accessions of Mentha spicata L. CIMAP/C33 could bedistinguished from CIMAP/C32. Mentha × gracilis Sole cv. cardiaca showed a muchhigher similarity with Mentha spicata L. as wellas Mentha arvensis L. which amongst themselvesshowed rather a greater distance indicating that Mentha × gracilis Sole cv. cardiaca might have evolved as a natural hybridbetween M arvensis L. and M. spicataL. In terms of uniqueness of amplified bands fordeveloping RAPD markers, it was observed that at taxalevel 298 bands were unique to one of the six taxa,singly amounting to 47.3% of total amplifiedfragments. Primers MAP 10 and 17 produced polymorphismonly in case of M. spicata L. and Menthaspicata L. cv. viridis while MAP 08 producedpolymorphic bands in all 4 other species than thesetwo. Similarly unique patterns were observeddifferentiating all six species and could be used asRAPD markers for differentiating Mentha species.
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  • 64
    ISSN: 1573-5060
    Schlagwort(e): AFLP ; isozyme ; ISSR ; molecular marker ; RAPD ; rice
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract We have examined the effectiveness of similar numbers of markers from four molecular marker systems (AFLP, isozymes, ISSR and RAPD) for revealing genetic diversity and discriminating between infraspecific groups of Oryza sativa germplasm. Each marker system classifies the germplasm into three major groups (most effectively with isozymes and AFLPs), but with differences (primarily with ISSR) between the precise classifications generated. However, at the highest levels of genetic similarity there was only partial agreement as to relationships between individual accessions when different markers were used. When variance was partitioned among and within the three subspecific groups, although the differences were not significant, greater variation was found among than within groups using AFLP and isozymes, with the reverse for RAPD and ISSR. Measurement of polymorphism using average heterozygosity and effective number of alleles gave similar results for each marker system. These results are discussed in relation to various genetic resources conservation activities, and the advisability of extrapolating to other sets of germplasm particularly of other crop species.
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  • 65
    ISSN: 1573-5060
    Schlagwort(e): AP-PCR ; cultivar identification ; Oleaeuropaea ; olive tree ; RAPD
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Málaga is a province of Spain where olive-trees are cultivated in a large range of environments, climates and soils. We have developed a reliable and reproducible method to detect RAPD and AP-PCR polymorphisms, using DNA from olive-tree (Olea europaea L.) leaves. Starting from their natural orchards, fifty-six olive-tree cultivars throughout Málaga province, including oil and table olive cultivars, were screened and grouped into 22 varieties. A total of 62 informative polymorphic loci that provide 601 conspicuous bands were enough to differentiate the varieties. Clustering analyses managing 3 different pairwise distances, as well as phylogenetic analyses, led to the same result: olive-trees in Málaga can be divided into three main groups. Group I (90% of certainty) contains wild type and two introduced varieties, group II (83% of certainty) covers some native olive-trees, and group III (58% of certainty) is an heterogeneous cluster that includes varieties originating and cultivated in a number of Andalusian locations. Geographic location seems to be the first responsible of this classification, and morphological traits are needed to justify the group III subclustering. These results are consistent with the hypothesis of autochthonic origin of most olive-tree cultivars, and have been used to support a Label of Origin for the olive oil produced by the varieties included in group II.
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  • 66
    ISSN: 1573-5060
    Schlagwort(e): genetic markers ; marker-assisted selection ; multiple location analysis ; RAPD
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Marker assisted selection (MAS) may improve the efficiency of breeding downy mildew resistant cucumber cultivars. A study was conducted to identify random amplified polymorphic DNA (RAPD) markers linked to the downy mildew resistance gene (dm) which would be suitable for MAS. A total of 145 F3 families from two populations (55 from the WI 1983G × Straight 8 population and 90 from the Zudm1 × Straight 8 population) were evaluated over five locations in North America and Europe. Resistant and susceptible F3 families were identified and mean family resistance ratings were used to type individual F2 plants. No evidence for race differences in the pathogen (Psuedoperonospora cubensis (Berk. & Curt.) Rostow) between North America and Europe was found. Phenotypic correlations between locations ranged from 0.3 to 0.7. Of the 135 polymorphic RAPD markers identified from 960 primers, five were linked to dm - G14800, X151100, AS5800, BC5191100, and BC5261000. In the WI 1983G × Straight 8 population, G14800 was linked to dm at 16.5 cm, AS5800 at 32.8 cm, BC5191100 at 9.9 cm, and BC5261000 at 19.2 cm. In the Zudm1 ×Straight 8 population, G14800 was linked at 20.9 cm, X151100at 14.8 cm, AS5800 at 24.8 cm, and BC526_1000 at 32.9 cm. MarkersG14800 and BC5191100 were linked in repulsion to the dm allele, and X151100, AS5800, and BC5261000 were linked in coupling phase. These genetic markers may be exploited to develop an efficient MAS strategy for breeding resistant cucumber cultivars.
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  • 67
    ISSN: 1573-5060
    Schlagwort(e): genetic distance ; germplasm management ; multi-dimensional scaling ; plant germplasm ; RAPD ; SSR
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers were used to characterize genetic relationships among 46 accessions in two C. melo L. subsp. melo (Cantalupensis, Inodorus) and subsp.agrestis (Conomon, and Flexuosus) groups. Genetic distance (GD) estimates were made among and between accessions in four melon market classes [Galia, Ogen, Charentais, and Shipper (European and U.S. types)] of Cantalupensis, one market class of Inodorus (Cassaba and Honey Dew), one accession of Conomon, and one accession of Flexuosus by employing three GD estimators; simple matching coefficient, Jaccard's coefficient, and Nei's distance-D. Differences detected among 135 RAPD bands and 54 SSR bands (products of 17 SSR primers) were used to calculate GD. Band polymorphisms observed with 21 RAPD primers and 7 SSR primers were important (p =0.01) in the detection of genetic differences. Estimators of GD were highly correlated (p 0.0001; rs = 0.64 to0.99) when comparisons were made between estimation methods within a particular marker system. Lower correlations (rs = 0.17 to 0.40) were detected (P 〉 0.001) between marker systems using any one estimator. The GD of the Conomon and Flexuosus accessions was significantly different (p〉 0.001)from the mean GD of all the market classes examined. The mean GD (Jaccard's coefficient) among accessions of Ogen, Galia, Cassaba, Charentais, European shipper, and U.S. shipper groups was 0.11 ± 0.04, 0.33± 0.09, 0.21 ± 0.04, 0.26 ± 0.10, 0.17± 0.05 and 0.22 ± 0.08, respectively. Market classes were distinct (p 〉 0.001), such that GDs between Galia and other accessions were the largest(mean GD 0.34 to 0.35), and GDs between Ogen and other accessions were the smallest (mean GD 0.29 to 0.30). Contrasts between the U.S. shipper cultivar Top Mark and accessions within any market class was relatively large (mean GD = 0.42 ± 0.06). Empirical estimations of variances associated with each marker type in the accessions examined indicated that, per band, lower coefficients of variation can be attained in the estimation of GD when using RAPDs compared to SSRs. Nevertheless, the genetic relationships identified using these markers were generally similar. The disparity between the analyses of the two markers made may be related to the amount of genome coverage which is characteristic of a particular marker system and/or its efficiency in sampling variation in a population. Results of RAPD marker analysis suggest that 80 marker bands were adequate for assessing the genetic variation present in the accessions examined.
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  • 68
    ISSN: 1573-5060
    Schlagwort(e): Arachis pintoi ; Arachis repens ; peanut ; variation ; phylogeny ; RAPD
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Wild Arachis germplasm includes potential forage species, such as the rhizomatous Arachis glabrata and the stoloniferous A. pinto and A. repens. Commercial cultivars of A. pintoi have already been released in Australia and in several Latin American countries, and most of these cultivars were derived from a single accession of A. pintoi (GK 12787). Arachis repens is less productive as a forage plant than is A. pintoi. However, it can be crossed with A. pintoi, and thus has good potential as germplasm for the improvement of A. pintoi. Arachis repens is also used as an ornamental plant and ground cover. Many new accessions of these two stoloniferous species are now available, and they harbor significant genetic variability beyond that available in the few older accessions, previously available. Therefore, these new accessions need to be conserved, documented and considered in terms of their potential for crop improvement and direct commercial use. Sixty-four accessions of this new germplasm were analyzed using RAPD analysis. Most of the accessions of A. repens grouped together into a clearly distinct group. In general, the accessions from the distinct valleys of the Jequitinhonha, São Francisco and Paranã rivers did not group together, suggesting there is not a tight relation between dispersion by rivers and the geographic distribution of genetic variation in these species.
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  • 69
    Digitale Medien
    Digitale Medien
    Springer
    Plant growth regulation 32 (2000), S. 27-39 
    ISSN: 1573-5087
    Schlagwort(e): ethylene ; gene expression ; jasmonic acid ; reactive oxygen species ; salicylic acid ; ultraviolet-B radiation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Depletion of the stratospheric ozone layer is leadingto an increase in ultraviolet-B (UV-B: 280–320 nm)radiation reaching the earth's surface. This hasraised interest in the possible consequence ofincreased UV-B levels on plant growth and developmentand the mechanisms underlying these responses. Although the effects of UV-B are now wellcharacterised at the physiological level, little isknown about the cellular and molecular mechanismsinvolved. Recent studies have shown that UV-B affectsa number of important physiological processes, such asphotosynthesis, through effects on gene expression. In addition, induction of a number of defensemechanisms, such as production of UV-B screeningpigments, increase in antioxidant enzymes andinduction of pathogenesis-related proteins, are alsomediated at the level of gene expression. The signaltransduction pathways by which UV-B regulates geneexpression are at present poorly understood. Thestudies carried out to date have, however, indicateda pivotal role for reactive oxygen species as keysecond messengers acting up-stream of a number ofpathways involving the plant hormones salicylic acid,jasmonic acid and ethylene. The transduction pathwaysidentified to date and the role of intermediates inregulating tolerance to UV-B damage are discussed inthis review.
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  • 70
    Digitale Medien
    Digitale Medien
    Springer
    Genetic resources and crop evolution 47 (2000), S. 515-526 
    ISSN: 1573-5109
    Schlagwort(e): Capsicum ; core collection ; genetic diversity ; RAPD ; Theobroma cacao
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract The development of a core collection, one which represents the genetic diversity of a crop with minimal redundancy and increases utility of the collection as a whole, is especially important as the funding for germplasm collections decreases. With limited resources, it is difficult to manage large germplasm collections and disperse genetically diverse germplasm to plant breeders. An algorithm was developed to assist in selection of core collections based on estimates of genetic distance. The criteria for selection of the maximum genetically diverse set were based on rankings of genetic distance between an accession with respect to all other accessions. Depending on the size core which a user wished, a zone around each selected accession was determined and no other accession within these limits was selected. The premise for the algorithm was that the genetic variability represented in the core must be representative of the distribution of genetic distances within the population of interest. In the present study, the algorithm was used with RAPD-marker-based estimates of genetic distance for 270 Theobroma cacao L. accessions and 134 Capsicum accessions that chose a set representing 18.5% of the population and representing the breadth of RAPD-based variation.
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  • 71
    Digitale Medien
    Digitale Medien
    Springer
    Genetic resources and crop evolution 47 (2000), S. 603-610 
    ISSN: 1573-5109
    Schlagwort(e): azuki bean ; domestication ; RAPD ; wild ancestor
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract RAPD (Random Amplified Polymorphic DNA) variation was assessed in 42 accessions of azuki bean (Vigna angularis) including wild, weedy and cultivated races and in three accessions of two related species used as outgroups. A much lower level of genetic variation was observed in cultivated and weedy azuki beans compared to wild azuki bean. Wild azuki bean (V. angularis var. nipponensis) has relatively high genetic variation in subtropical highlands of Asia compared to the Far East. Although cultivated azuki bean has low RAPD variation, accessions from subtropical highlands and Southeast Asia showed different RAPD features compared to those of the Far East. It is hypothesized that the cultivated azuki bean has been derived from wild azuki bean in the Far East; the high variation in wild azuki bean has been created through its natural dissemination; and the relatively low variation in cultivated azuki bean has come about through human dissemination after genetic bottleneck reduced by domestication. In addition, high genetic diversity in wild azuki bean in subtropical highlands of Asia is regarded as an important genetic resource in azuki improvement.
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  • 72
    ISSN: 1573-5109
    Schlagwort(e): cluster analysis ; genetic variation ; germplasm ; RAPD ; Sorghum bicolor
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract The extent and patterns of distribution of genetic variation among 80 sorghum (Sorghum bicolor (L.) Moench) germplasm accessions from Ethiopia and Eritrea were investigated using RAPD with 20 oligonucleotide primers. The primers generated a total of 147 polymorphic bands across the 80 accessions with a mean of 7.35 bands per primer. Estimation of the extent of variation by the Shannon-Weaver diversity index revealed an intermediate level of overall variation (H = 53), although the levels varied among regions of origin of the accessions. Partitioning of the total variation revealed considerable variation (77%) within the region of origin of the accessions and the remainder (23%) among regions of origin. Similarly, a large portion (94%) of the total variation was found within the adaptation zones compared to among the adaptation zones (6%). The results suggest a weak differentiation of the sorghum material both on regional and agro-ecological bases, which could be ascribed to the high rate of outcrossing in cultivated sorghum and its free natural hybridization with its wild and weedy relatives, as well as to seed movement by humans. The average genetic dissimilarity was found to be 36% among the 80 accessions and 13% among the 15 regions of origin. Cluster analysis failed to group accessions of the same region or the same adaptation zone, which further confirmed the weak differentiation of the material studied. The clustering pattern of the regions of origin was broadly concordant with previous clustering patterns obtained using morphological characters, in which regions with broad agro-climatic conditions were grouped together.
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  • 73
    Digitale Medien
    Digitale Medien
    Springer
    Plant growth regulation 31 (2000), S. 35-42 
    ISSN: 1573-5087
    Schlagwort(e): β-1,4-endoglucanase ; ethylene ; fruit ; gene expression ; polygalacturonase
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Fruitlet abscission during fruit development is due to the activation ofpre-differentiated abscission zones (AZs) located between twig andpedicel, and/or pedicel and pericarp. Major advances on biochemicaland molecular aspects are related to β-1,4-endoglucanase (EG) andpolygalacturonase (PG), two cell hydrolases involved in the cell walldisassemblement responsible for fruit shedding. AZ activation isaccompanied by an increase in activity and transcript accumulation ofone or both enzymes. Expression of PG genes specifically related toabscission has been found in tomato flower AZ. In peach, an EG genehighly expressed in leaf and fruitlet AZs has been isolated. AZactivation is preceded by an induction of ethylene biosynthesis,paralleled by a stimulation of ACO activity and transcript accumulation.Ethylene, besides a dramatic stimulation of PG and EG, up or downregulates several other abscission related genes. The specificexpression of genes encoding for ethylene receptors in the AZ wouldsupport the hypothesis that fruitlet AZ specificity may depend on theability of this region to sense ethylene.
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  • 74
    Digitale Medien
    Digitale Medien
    Springer
    Genetic resources and crop evolution 47 (2000), S. 115-121 
    ISSN: 1573-5109
    Schlagwort(e): diversity ; genebank ; germplasm ; potato ; RAPD ; Solanum sucrense
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Genetic characterization of germplasm is important for setting objective guidelines for conservation. One common problem found in genebanks is determining the value of populations with insufficient or unreliable data regarding their geographic origin. In this study, a genetic analysis based on RAPD markers was conducted to characterize a `mystery' population of Solanum sucrense, a polysomic tetraploid potato (2n=4x=48), for which adequate documentation was lacking. The comparative analysis of genetic similarities between this mystery population and each one of 30 other S. sucrense populations in the genebank revealed that all populations within this species, including the mystery population, are significantly different from being duplicates, and are therefore worthy of separate conservation. RAPD markers also distinguished the mystery population from closely related tetraploid species S. oplocense, S. gourlayi and S. tuberosum ssp. andigena, suggesting that it is also not a duplicate of a population of these species. If RAPDs can clearly differentiate populations within highly heterogeneous tetraploids like S. sucrense, they should be generally useful for determining germplasm organization within potato species.
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  • 75
    ISSN: 1573-5109
    Schlagwort(e): core collection ; cultivar identification ; Dioscorea cayenensis/Dioscorea rotundata complex ; Guinea yam ; identification of duplicates ; RAPD
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract DNA from twenty-three late maturing cultivars of Guinea yams (D. cayenensis/D. rotundata complex) from the Benin Republic that could not be separated using isozyme markers, were examined using randomly amplified polymorphic DNA (RAPD) markers with decamer primers of arbitrary sequence. All the twelve primers tested were informative and yielded 63 amplified DNA bands from which 47 (75%) were polymorphic. Although no single primer produced polymorphic bands in all cultivars, the great majority of the cultivars were separated with the combinations of polymorphic bands generated by various primers. Putative duplicates and cultivar misclassifications were identified. Many morphologically distinct cultivars were close. The dwarf cultivar Tam-Sam considered as derived from Tabane, appeared more distant from the latter than was believed. RAPD analysis was found as a practical tool for the identification of duplicates toward establishment of an accurate core collection of Guinea yams in Benin Republic and in the other countries of the African yam belt.
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  • 76
    ISSN: 1573-5109
    Schlagwort(e): Fragaria chiloensis ; Fragaria virginiana ; genetic resources ; morphology ; RAPD
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Variation for 24 morphological traits measured in a greenhouse environment and 36 randomly amplified polymorphic DNA (RAPD) markers was assessed among 318 wild octoploid strawberry (Fragariaspp.) genotypes from diverse habitats across the northern USA. RAPD marker frequencies and certain leaf and flower morphology traits (petiole color, leaf mass/area ratio, leaflet length and width, flower and receptacle diameter, petal width, flowers/inflorescence) were significantly different between the F. chiloensis-platypetala and F. virginiana-glauca species complexes. The proportion of variation accounted for by provenance effects was lower for the RAPD markers than for most morphological traits, especially in the F. virginiana-glauca species complex. Morphological traits of potential adaptive importance group the collection into provenances within each species-complex, and reflect the significant habitat and geographic differences across the region from which the germplasm was collected. Variation among populations within provenances was low for the molecular and most morphological traits, with a much larger amount of variability among plants within populations. Most of the variation for the presumably more selectively-neutral RAPD data was among plants within populations and populations within provenances rather than among the provenances that were recognized based on morphological traits, especially in the F. virginiana-glauca complex. Patterns of diversity for morphological traits must be considered, along with more selectively-neutral molecular characters such as RAPDs, to formulate effective sampling strategies and to properly estimate the quantity and apportionment of diversity within this germplasm.
    Materialart: Digitale Medien
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  • 77
    Digitale Medien
    Digitale Medien
    Springer
    Genetic resources and crop evolution 47 (2000), S. 257-265 
    ISSN: 1573-5109
    Schlagwort(e): AFLP ; genetic similarity ; molecular markers ; pears ; Pyrus ; RAPD
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Twenty-five Pyrus communis L. cultivars including eight traditional Portuguese pears, and four commercial Pyrus pyrifolia (Burm.) Nak. (Japanese pear or `nashi') cultivars were analysed by RAPD and AFLP techniques focusing on their molecular discrimination and the assessment of their genetic relatedness. Twenty-five primers generated 324 RAPD markers, among which 271 (84%) were polymorphic. The AFLP technique, using seven primer combinations, revealed a similar level of molecular polymorphisms (87%), representing 418 polymorphic bands among a total of 478 scored in autoradiographs. The high reproducibility of RAPD and AFLP techniques was confirmed comparing DNA samples from different extractions and different digestions of DNA from the same plant. Three genetic similarity matrices and respective dendrograms were elaborated on using RAPD, AFLP or joint RAPD and AFLP data. Both molecular marker techniques proved their reliability to assess genetic relationships among pear cultivars. P. pyrifolia cultivars exhibit a closer genetic relatedness, clustering apart from P. communis cultivars. Within P. communis, `William's', as well as `Doyenne du Comice', cluster close to their hybrids. Most of the Portuguese cultivars tend to cluster together, indicating to constitute a relatively independent genetic pool, which can be of interest in pear breeding programs.
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  • 78
    Digitale Medien
    Digitale Medien
    Springer
    Hydrobiologia 421 (2000), S. 157-164 
    ISSN: 1573-5117
    Schlagwort(e): Bosmina ; RAPD ; genetic variation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We used RAPDs (Random Amplified Polymorphic DNA) to test genetic divergence between two populations of Bosmina spp. in Lake Östersjön, Sweden. Previous taxonomic studies on European species within the genus Bosmina have been based on morphological characters alone. RAPD markers distinguished the two populations and supported the specific status of B. coregoni and B. longispina based on morphological characters. Furthermore, juveniles with a long antennule and a mucro were classified as B. coregoni. RAPDs also revealed genetic differences among the tested individuals, suggesting several clones within each species.
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  • 79
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis thaliana ; β-glucuronidase (GUS) reporter gene ; gene expression ; isoprenoids ; mevalonate kinase ; transgenic plants
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Mevalonate kinase (MVK), the enzyme that catalyzes the phosphorylation of mevalonate to produce mevalonate 5-phosphate, is considered as a potential regulatory enzyme of the isoprenoid biosynthetic pathway. The Arabidopsis thaliana MVK gene corresponding to the MVK cDNA previously isolated has been cloned and characterized. RNAse protection analysis indicated that the expression of the MVK gene generates three mRNA populations with 5′ ends mapping 203, 254 and 355 nt upstream of the MVK ATG start codon. Northern blot analysis showed that the MVK mRNA accumulates preferentially in roots and inflorescences. Histochemical analysis, with transgenic A. thaliana plants containing a translational fusion of a 1.8 kb fragment of the 5′ region of the MVK gene to the β-glucuronidase (GUS) reporter gene, indicated that the MVK 5′-flanking region directs widespread expression of the GUS gene throughout development, although the highest levels of GUS activity are detected in roots (meristematic region) and flowers (sepals, petals, anthers, style and stigmatic papillae). The expression pattern of the MVK gene suggests that the role of the encoded MVK is the production of a general pool of mevalonate-5-phosphate for the synthesis of different classes of isoprenoids involved in both basic and specialized plant cell functions. Functional promoter deletion analysis in transfected A. thaliana protoplasts indicated that regulatory elements between positions −295 and −194 of the MVK 5′-flanking region are crucial for high-level MVK gene expression.
    Materialart: Digitale Medien
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  • 80
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 42 (2000), S. 387-396 
    ISSN: 1573-5028
    Schlagwort(e): gene expression ; hypersensitive responses ; plant defense responses ; salicylic acid ; tobacco mosaic virus ; WRKY DNA-binding proteins
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A pathogen- and salicylic acid (SA)-induced DNA-binding activity has been recently identified in tobacco that is related to a previously identified class of WRKY DNA-binding proteins. To identify members of the WRKY gene family associated with this DNA-binding activity, we have attempted to isolate those WRKY genes that are induced by pathogen infection. Using a domain-specific differential display procedure, we have isolated two tobacco WRKY genes, tWRKY3 and tWRKY4, that are rapidly induced in resistant tobacco plants after infection by tobacco mosaic virus (TMV). Both tWRK3 and tWRKY4 encode proteins with a single WRKY domain that contain the conserved WRKYGQK sequence. Unlike other isolated WRKY proteins that contain the Cys2His2 zinc motif, tWRKY3 and tWRKY4 appear to contain the Cys2HisCys zinc motif. Nonetheless, both tWRKY3 and tWRKY4 are capable of binding DNA molecules with the W-box (TTGAC) element recognized by other WRKY proteins. Expression of the tWRKY3 and tWRKY4 genes could be rapidly induced not only by TMV infection but also by SA or its biologically active analogues that are capable of inducing pathogenesis-related genes and enhanced resistance. Interestingly, induction of both genes by TMV infection was still observed in resistant tobacco plants expressing the bacterial salicylate hydroxylase gene (nahG), although the levels of induction appeared to be reduced. Identification of pathogen- and SA-induced genes encoding WRKY DNA-binding proteins should facilitate future studies on the regulation and functions of this novel group of DNA-binding proteins.
    Materialart: Digitale Medien
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  • 81
    ISSN: 1573-5028
    Schlagwort(e): gene expression ; glutamine synthetase ; legume-Rhizobium symbiosis ; nitrogen assimilation ; root nodules
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract In this paper we have studied the localisation of expression of the two functional cytosolic glutamine synthetase (GS) genes, MtGSa and MtGSb, in root nodules of the model legume Medicago truncatula. We have used a combination of different techniques, including immunocytochemistry, in situ hybridisation and promoter β-glucuronidase (GUS) fusions in transgenic plants, to provide the means of correlating gene expression with protein localisation. These studies revealed that transcriptional regulation (mRNA synthesis) plays an important part in controlling GS protein levels in nodules of M. truncatula. The major locations of cytosolic GS mRNA and protein are the central tissue, the parenchyma and the pericycle of the vascular bundles. These findings indicate that in nodules, GS might be involved in other physiological processes in addition to the primary assimilation of ammonia released by the bacterial nitrogenase. The two genes show different but overlapping patterns of expression with MtGSa being the major gene expressed in the infected cells of the nodule. Promoter fragments of 2.6 kb and 3.1 kb of MtGSa and MtGSb, respectively, have been sequenced and primer extension revealed that the MtGSb promoter is expressed in nodules from an additional start site that is not used in roots. Generally these fragments in the homologous transgenic system were sufficient to drive GUS expression in almost all the tissues and cell types where GS proteins and transcripts are located except that the MtGSa promoter fragment did not express GUS highly in the nodule infected cells. These results indicate that the cis-acting regulatory elements responsible for infected-cell expression are missing from the MtGSa promoter fragment.
    Materialart: Digitale Medien
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  • 82
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 42 (2000), S. 857-869 
    ISSN: 1573-5028
    Schlagwort(e): epidermis ; gene expression ; glycine-rich protein ; lipid transfer protein ; proline-rich protein ; stomata
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Guard cells are specialized and metabolically active cells which arise during the differentiation of the epidermis. Using Nicotiana glauca epidermal peels as a source of purified guard cells, we have constructed a cDNA library from guard cell RNA. In order to isolate genes that are predominantly expressed in guard cells, we performed a differential screen of this library, comparing the hybridization of a radiolabeled cDNA probe synthesized from guard cell RNA to that from a mesophyll cell cDNA probe. Sixteen clones were isolated based on their greater level of hybridization with the guard cell probe. Of these, eight had high homology to lipid transfer protein (LTP), two were similar to glycine-rich protein (GRP), and one displayed high homology to proline-rich proteins from Arabidopsis thaliana (AtPRP2, AtPRP4) and from potato guard cells (GPP). Northern analysis confirmed that one or more NgLTP genes, NgGRP1, and NgGPP1 are all differentially expressed, with highest levels in guard cells, and low or undetectable levels in mesophyll cells and in roots. In addition, all are induced to some degree in drought-stressed guard cells. NgLTP and NgGRP1 expression was localized by in situ hybridization to the guard cells and pavement cells in the epidermis. NgGRP1 expression was also detected in cells of the vasculature. Genomic Southern analysis indicated that LTP is encoded by a family of highly similar genes in N. glauca. This work has identified members of a subset of epidermis- and guard cell-predominant genes, whose protein products are likely to contribute to the unique properties acquired by guard cells and pavement cells during differentiation.
    Materialart: Digitale Medien
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  • 83
    ISSN: 1573-5028
    Schlagwort(e): chitinase function ; flower-predominant ; gene expression ; molecular cloning ; monocotyledon ; promoter
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A flower-predominant cDNA for a gene, termed OsChia1;175, was isolated from a cDNA library of rice pistils. Northern blot and RT-PCR analyses revealed that the OsChia1;175 gene is highly expressed in floral organs (pistils, stamens and lodicules at the heading stage) but not or at an extremely low level in vegetative organs. OsChia1;175 encodes a protein that consists of 340 amino acid residues, and the putative mature protein shows 52% to 63% amino acid identity to class I chitinases of rice or other plants. The phylogenetic tree shows that the OsChia1;175 protein is a new type of plant class I chitinase in rice. The expression of OsChia1;175 in vegetative organs is not induced by several chemicals, UV, and wounding. The soluble putative mature OsChia1;175 protein expressed in Escherichia coli exhibited chitinase activity in the assay with colloidal chitin as a substrate. Genomic Southern analysis revealed that the OsChia1;175 gene was organized as a low-copy gene family. The rice genomic library was screened and a genome clone corresponding to OsChia1;175 was isolated. The transcription start sites of the OsChia1;175 gene were mapped by primer extension analysis. The 1.2 kb putative promoter region of the OsChia1;175 gene was fused to the GUS (β-glucuronidase) gene, and this chimeric gene was introduced to rice by Agrobacterium-mediated transformation. The flower-predominant gene expression was identified also in the transgenic rice plants. The high promoter activity was detected in the stigmas, styles, stamens and lodicules in transgenic plants. The possible functions of OsChia1;175 are discussed.
    Materialart: Digitale Medien
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  • 84
    ISSN: 1573-5028
    Schlagwort(e): anaerobiosis ; electrophoretic mobility shift assays ; gene expression ; glyceraldehyde-3-phosphate dehydrogenase ; Nicotiana tabacum ; transgenic plants
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The promoter of the maize glyceraldehyde-3-phosphate dehydrogenase 4 gene (GapC4) confers strong, specific and ubiquitous anaerobic reporter gene expression in tobacco. To identify factors required for heterologous anaerobic gene expression, 19 progressive 5′ and 3′ promoter deletions were linked to a chimeric GapC4 TATA box-β-glucuronidase (GUS) reporter gene construct and transformed into tobacco. In all transgenic lines aerobic expression values were in the range obtained for negative controls while histochemical GUS assays reveal some weak expression in roots only. Anaerobic induction of about 100-fold to more than 1000-fold above unspecific background is mediated by a region of about 190 bp of the GapC4 promoter. Anaerobic reporter gene induction strongly decreases upon deletion of a 20 bp fragment from −286 to −266 relative to the transcription start point. This fragment harbours putative cis-acting sequences. Electrophoretic mobility shift assays with a 50 bp fragment harbouring these cis sequences reveal a high-mobility complex that is formed with nuclear extracts from aerobic and anaerobic leaf tissue while an additional low-mobility complex is anaerobiosis-specific. The formation of the high-mobility complex requires the sequence GTGGGCCCG. The 50 bp fragment alone confers weak and orientation-dependent anaerobic induction to a GapC4 TATA box-β-glucuronidase (GUS) reporter gene.
    Materialart: Digitale Medien
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  • 85
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 43 (2000), S. 659-675 
    ISSN: 1573-5028
    Schlagwort(e): A-type cyclins ; cell cycle ; gene expression ; regulation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Although the basic mechanisms which control the progression through the cell cycle appear to be conserved in all higher eukaryotes, the unique features of the plant developmental programme must be somehow reflected in a plant-specific regulation of the factors which control cell division. In the past few years, considerable progress has been achieved in identifying the major components of the cell cycle machinery in plants, especially the cyclin-dependent kinases (CDKs) and their regulatory subunits, the cyclins. The question of how these components direct expression of specific genes at specific stages of the cell cycle, and how they are themselves regulated, constitutes a challenge for the present and for the years to come. This review summarizes our current knowledge of a particular class of plant cyclins, the A-type cyclins, which can be further subdivided into three structural groups. The putative functions of these A-type cyclins are discussed in relation to the presence of remarkable motifs in their amino acid sequences, and to their specific transcriptional regulation, protein amount and subcellular localization.
    Materialart: Digitale Medien
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  • 86
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 44 (2000), S. 73-84 
    ISSN: 1573-5028
    Schlagwort(e): auxin ; Aux/IAA ; dgt ; gene expression ; Lycopersicon esculentum ; signal transduction
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The diageotropica (dgt) mutation has been proposed to affect either auxin perception or responsiveness in tomato plants. It has previously been demonstrated that the expression of one member of the Aux/IAA family of auxin-regulated genes is reduced in dgt plants. Here, we report the cloning of ten new members of the tomato Aux/IAA family by PCR amplification based on conserved protein domains. All of the gene family members except one (LeIAA7) are expressed in etiolated tomato seedlings, although they demonstrate tissue specificity (e.g. increased expression in hypocotyls vs. roots) within the seedling. The wild-type auxin-response characteristics of the expression of these tomato LeIAA genes are similar to those previously described for Aux/IAA family members in Arabidopsis. In dgt seedlings, auxin stimulation of gene expression was reduced in only a subset of LeIAA genes (LeIAA5, 8, 10, and 11), with the greatest reduction associated with those genes with the strongest wild-type response to auxin. The remaining LeIAA genes tested exhibited essentially the same induction levels in response to the hormone in both dgt and wild-type hypocotyls. These results confirm that dgt plants can perceive auxin and suggest that a specific step in early auxin signal transduction is disrupted by the dgt mutation.
    Materialart: Digitale Medien
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  • 87
    ISSN: 1573-5028
    Schlagwort(e): Arabidopsis thaliana ; calcium-binding protein ; caleosin ; EF-hand ; gene expression ; lipid bodies ; vesicles
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We have previously identified a rice gene encoding a 27 kDa protein with a single Ca2+-binding EF-hand and a putative membrane anchor. We report here similar genes termed caleosins, CLO, in other plants and fungi; they comprise a multigene family of at least five members in Arabidopsis (AtClo1–5). Northern hybridization demonstrated that AtClo2–4 mRNAs levels were low in various tissues, while AtClo1 mRNA levels were high in developing embryos and mature seeds. Analysis of transgenic Arabidopsis plants expressing the GUS reporter under control of the AtClo1 promoter showed strong levels of expression in developing embryos and also in root tip cells. Antibodies raised against AtCLO1 were used to detect caleosin in cellular fractions of Arabidopsis and rapeseed. This indicated that caleosins are a novel class of lipid body proteins, which may also be associated with an ER subdomain.
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  • 88
    Digitale Medien
    Digitale Medien
    Springer
    Journal of bioenergetics and biomembranes 32 (2000), S. 627-634 
    ISSN: 1573-6881
    Schlagwort(e): Mitochondrial biogenesis ; copy number ; gene expression ; mitochondrial transcription factor ; nuclear—mitochondrial communication ; stimulation ; endurance training
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Physik
    Notizen: Abstract Mitochondrial proliferation was studied in chronically stimulated rabbit skeletal muscle over a period of 50 days. After this time, subunits of COX had increased about fourfold. Corresponding mRNAs, encoded on mitochondrial DNA as well as on nuclear genes, were unchanged when related to total tissue RNA, however, they were elevated two- to fivefold when the massive increase of ribosomes per unit mass of muscle was taken into account. The same was true for the mRNA encoding mitochondrial transcription factor A. Surprisingly, tissue levels of mtTFA protein were reduced about twofold, together with mitochondrial DNA. In conclusion, mito chondria are able to maintain high rates of mitochondrial transcription even in the presence of reduced mtTFA protein and mtDNA levels. Therefore, stimulated mtTFA gene expression accompanies stimulated mitochondrial transcription, as in other models, but it is not sufficient for an increase of mtDNA copy number and other, yet unknown, factors have to be postulated.
    Materialart: Digitale Medien
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  • 89
    ISSN: 1573-6857
    Schlagwort(e): dendritic polymer ; reporter gene ; gene expression ; transfection
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Cyclic core dendritic polymer is a new type of synthetic polymers. The ability of generation 4 of the dendrimer with a core of 1,4,7,10-tetraazacyclododecane to function as an effective gene delivery vector was investigated. Results from fluorescence in situhybridization (FISH) show that the pCH 110 plasmid DNA was transferred into human small intestine cancer metastatic ascites (HICMA) cells induced by this kind of dendrimer as a vector. The transferred LacZ, GFP and luciferase genes were highly expressed in the transfected HICMA, COS-7 and 293 cells. These studies demonstrate that the dendrimer can transfect mammalian cells in vitrowhich offers an alternatively efficient method for mammalian gene transfer.
    Materialart: Digitale Medien
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  • 90
    Digitale Medien
    Digitale Medien
    Springer
    World journal of microbiology and biotechnology 16 (2000), S. 741-748 
    ISSN: 1573-0972
    Schlagwort(e): AFLP ; epidemiological typing ; Escherichia coli O157 ; molecular epidemiology ; PFGE ; RAPD ; sub-typing ; typing methods ; VTEC
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Werkstoffwissenschaften, Fertigungsverfahren, Fertigung
    Notizen: Abstract The epidemiological investigation of Escherichia coli O157 is complicated by the lack of heterogeneity between strains responsible for the majority of cases of infection. As a consequence it is difficult to reliably cluster together an outbreak strain and differentiate it from other sporadically occurring isolates. The methods available for the sub-typing of E. coli O157 vary in their speed, technical complexity, cost and ability to discriminate reliably between strains, with many of the recently developed methods targeting the genome to provide differentiation. No single typing method is individually superior, and ideally a combination of techniques should be employed depending on the level of discrimination required and time or resources available. The aim of this review is to consider the relative merits of the available typing methodologies with particular emphasis on those which may find application in a diagnostic laboratory.
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  • 91
    ISSN: 1572-9699
    Schlagwort(e): differentiation ; FtsZ ; gene expression ; septum ; SsgA ; transmission electron microscopy
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract This paper describes the effects of increased expression of the cell division genes ftsZ, ftsQ, and ssgA on the development of both solid- and liquid-grown mycelium of Streptomyces coelicolor and Streptomyces lividans. Over-expression of ftsZ in S. coelicolor M145 inhibited aerial mycelium formation and blocked sporulation. Such deficient sporulation was also observed for the ftsZ mutant. Over-expression of ftsZ also inhibited morphological differentiation in S. lividans 1326, although aerial mycelium formation was less reduced. Furthermore, antibiotic production was increased in both strains, and in particular the otherwise dormant actinorhodin biosynthesis cluster of S. lividans was activated in liquid- and solid-grown cultures. No significant alterations were observed when the gene dosage of ftsQ was increased. Analysis by transmission electron microscopy of an S. coelicolor strain over-expressing ssgA showed that septum formation had strongly increased in comparison to wild-type S. coelicolor, showing that SsgA clearly influences Streptomyces cell division. The morphology of the hyphae was affected such that irregular septa were produced with a significantly wider diameter, thereby forming spore-like compartments. This suggests that ssgA can induce a process similar to submerged sporulation in Streptomyces strains that otherwise fail to do so. A working model is proposed for the regulation of septum formation and of submerged sporulation.
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  • 92
    Digitale Medien
    Digitale Medien
    Springer
    Experimental and applied acarology 24 (2000), S. 751-774 
    ISSN: 1572-9702
    Schlagwort(e): AFLP ; allozymes ; DALP ; DNA sequencing ; genetic structure ; microsatellites ; mitochondrial DNA ; mites ; molecular systematics ; PCR ; phylogeny ; RAPD ; RFLP ; ribosomal DNA ; ticks
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The application of molecular markers to the study of ticks and mites has recently yielded new insights into their population structures and taxonomic relationships. Ticks have been studied at individual, population and species level. Mites are a more diverse group and those that have been studied to the same degree as the ticks include the Tetranychidae (spider mites), Phytoseiidae (predatory mites) and the Eriophyidae. Population variation has also been studied in the important bee parasitic mite Varroa jacobsoni Oudemans. The methods used to study these organisms have much in common. At the individual level these range from general approaches, such as AFLP, RAPD or DALP, to highly specific microsatellite analysis. Although these markers also work at the population and species level, additional analysis of specific nuclear or mitochondrial genes has been conducted either by RFLP or sequencing. Molecular applications have had particular success in facilitating the identification of taxonomically difficult species, understanding population structures and elucidating phylogenetic relationships.
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  • 93
    Digitale Medien
    Digitale Medien
    Springer
    Plant growth regulation 30 (2000), S. 193-200 
    ISSN: 1573-5087
    Schlagwort(e): ACC synthase ; ACC oxidase ; ethylene ; fruit ; gene expression ; regulation ; ripening
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Progress in ethylene regulating fruit ripening concerning itsperception and signal transduction and expression of ACC synthaseand ACC oxidase genes is reviewed. ACC synthase and ACC oxidasehave been characterized and their genes cloned from various fruittissues. Both ACC synthase and ACC oxidase are encoded bymultigene families, and their activities are associated withfruit ripening. In climacteric fruit, the transition toautocatalytic ethylene production appears to be due to a seriesof events in which ACC sythase and ACC oxidase genes have beenexpressed developmentally. Differential expression of ACCsynthase and ACC oxidase gene family members is probably involvedin such a transition that ultimately controls the onset of fruitripening.In comparison to ACC synthase and ACC oxidase, less is knownabout ethylene perception and signal transduction because of thedifficulties in isolating and purifying ethylene receptors orethylene-binding proteins using biochemical methods. However, theidentification of the Nr tomato ripening mutant as anethylene receptor, the applications of new potent anti-ethylenecompounds and the generation of transgenic fruits with reducedethylene production have provided evidence that ethylenereceptors regulate a defined set of genes which are expressedduring fruit ripening. The properties and functions of ethylenereceptors, such as ETR1, are being elucidated.Application of molecular genetics, in combination withbiochemical approaches, will enable us to better understand theindividual steps leading from ethylene perception and signaltransduction and expression of ACC synthase and ACC oxidase genefamily member to the physiological responses.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 94
    ISSN: 1573-5109
    Schlagwort(e): domestication ; genetic variation ; in situ conservation ; RAPD ; V. angularis complex
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract The present study, using RAPD analysis, was undertaken to characterize genetic variation in three forms of V. angularis, cultivated, wild and weedy forms, and their relationships. The materials used consisted of 171 individuals (plants) or cultivars from 23 populations including 5 wild populations, 6 weedy populations, 6 cultivated populations and 6 populations with plants having wild and weedy or intermediate morphology, denoted here as complex populations. The materials used were collected on Honshu Island, Japan and seeds collected directly from the field were germinated for DNA extraction. In addition, 6 landrace accessions of V. angularis from the genebank were also analyzed. Genetic variation was highest in the wild form (Hg= 0.132; GD = 0.388), followed by the weedy form (Hg= 0.124; GD = 0.341) and the least in the cultivated form (Hg= 0.079; GD = 0.274). Intra-population genetic variation was high in the weedy and in the wild populations. However, inter-population was greater than intra-population genetic variation for all groups of populations studied in the V. angularis complex. 93% of the total diversity in the present study was exhibited by plants from complex populations and specific RAPD bands were found in these populations. Our results provide evidence that complex populations would be a logical focus for efforts to conserve the V. angularis complex in situ. Our results suggest that weedy populations are usually an ecotype of the wild form adapted to a different habitat.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 95
    Digitale Medien
    Digitale Medien
    Springer
    Genetic resources and crop evolution 47 (2000), S. 135-140 
    ISSN: 1573-5109
    Schlagwort(e): Aegilops tauschii ; gene expression ; genetic inheritance ; Puccinia recondita f. sp. tritici ; rust resistance ; synthetic hexaploid wheats
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract A collection of 164 Aegilops tauschii accessions, obtained from Gatersleben, Germany, was screened for reaction to leaf rust under controlled greenhouse conditions. We have also evaluated a selection of synthetic hexaploid wheats, produced by hybridizing Ae. tauschii with tetraploid durum wheats, as well as the first and second generation of hybrids between some of these resistant synthetic hexaploid wheats and susceptible Triticum aestivum cultivars. Eighteen (11%) accessions of Ae. tauschii were resistant to leaf rust among which 1 was immune, 13 were highly resistant and 4 were moderately resistant. Six of the synthetic hexaploid wheats expressed a high level of leaf rust resistance while four exhibited either a reduced or complete susceptibility compared to their corresponding diploid parent. This suppression of resistance at the hexaploid level suggests the presence of suppressor genes in the A and/or B genomes of the T. turgidum parent. Inheritance of leaf rust resistance from the intercrosses with susceptible bread wheats revealed that resistance was dominant over susceptibility. Leaf rust resistance from the three synthetics (syn 101, syn 701 and syn 901) was effectively transmitted as a single dominant gene and one synthetic (syn 301) possessed two different dominant genes for resistance.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 96
    Digitale Medien
    Digitale Medien
    Springer
    Genetic resources and crop evolution 47 (2000), S. 191-196 
    ISSN: 1573-5109
    Schlagwort(e): Elymus ; genome ; Hystrix ; RAPD ; relationships ; specific marker ; variation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract To assess the generic delimitation and the interspecific relationships between Hystrix and Elymus, three Hystrix and 10 Elymus species were used for random amplified polymorphic DNA(RAPD) assay. Of the 54 primers tested, 26 (48%) produced polymorphic products. A total of 167 products amplified from 16 primers were selected for RAPD analysis, among which 156 (93.4%) amplified products were found to be polymorphic among the 13 species. The polymorphism produced by each primer ranged from 4 to 13, with an average of 9.8. Data were used to generate Jaccard's similarity coefficients and to construct a dendrogram using UPGMA in the NTSYS computer programs. It is concluded from this study that: (1) there were clear differences between Hystrix and Elymus, which possibly suggest that Hystrix is a valid genus; (2) great diversity existed among the species of Hystrix and Elymus; (3) the species similar to each other in morphological characters were grouped together; (4) the species from neighboring geographical regions were clustered; (5) the species with the same genomes and polyploidy level were clustered together; (6) RAPD results are comparable with those obtained from studies on morphology and cytology. It is a useful additional method for assessing the relationships among genera and species in Triticeae.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 97
    ISSN: 1573-5044
    Schlagwort(e): fall dormancy ; gene expression ; Medicago sativa L. ; protein ; starch ; sugar ; winter hardiness
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A major factor limiting persistence of alfalfa (Medicago sativa L.) in the northern US is poor winter hardiness. Our hypothesis is that suspension cell cultures derived from dormant, winter-hardy alfalfa cultivars would cold acclimate and survive sub-zero temperatures better than cell cultures derived from non-dormant, non-hardy cultivars. Our objectives were (1) to determine if genetic differences in winter hardiness between dormant and non-dormant alfalfa were retained by suspension cells derived from these contrasting cultivars; and (2) to determine the physiological and biochemical bases for differences in freezing tolerance of suspension cells. Cell suspensions derived from `5262' (fall dormant) and `5929' (fall non-dormant) were cold hardened at 2 °C for 14 days. Cells were frozen in a cooling bath and cell survival determined by measuring 2, 3, 5-triphenyltetrazolium chloride (TTC) reduction. Cold acclimation improved cell survival of both cultivars to −5 °C when compared to unacclimated cells. Only acclimated cells of 5262 survived temperatures of −10 °C to −25 °C. The freezing tolerance of cold-acclimated 5262 cells was associated with high sugar and starch concentrations, lower α-amylase activities and slightly lower cell protein levels when compared to 5929. No differences in polypeptide composition were evident when comparing acclimated and unacclimated cells of 5929, but polypeptide composition did change with acclimation of 5262 cells. As expected, expression of RootCAR1 in 5262 cells increased with cold acclimation, but high levels of RootCAR1 transcript were unexpectantly found in both cold acclimated and unacclimated 5929 cells. With the exception of the RootCAR1 expression, many of the physiological responses of these alfalfa cell lines to cold acclimation were similar to those that have been reported for field-grown plants.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 98
    Digitale Medien
    Digitale Medien
    Springer
    Euphytica 114 (2000), S. 87-91 
    ISSN: 1573-5060
    Schlagwort(e): Angelica ; Bupleurum ; intergenic transcribed sequences ; Peucedanum ; phylogeny ; RAPD ; Umbelliferae
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract The internal transcribed spacers ITS1 and ITS2 in the18S-5.8S-26S rDNA repeat units were amplified and cloned from Angelica gigas Nakai, Angelica acutiloba (Siebold & Zucc.) Kitagawa, A. dahurica Maxim, Angelica decursiva (Miq.) Franch. & Savat, Bupleurum falcatum L. and Peucedanum japonicum Thunb. Sequence analyses showed that ITS1 is approx. 215 bp, the 5.8S gene is 162 bp and the ITS2 approx. 221 bp in all six species. The sequences are deposited at the EMBL Nucleotide Database. By including these new sequences in the Apiaceae phylogenetic tree, a third branch consisting of P. japonicum, A. gigas, P. decursivum and A. decursiva is added to theAngelica clade. Peucedanum does not forma distinct branch. The sequence obtained from Angelica dahurica collected in S. Korea is identical to that reported for the same species originating from China. A Bupleurum clade of three species was added to the tree showing closer relationship to theDaucus Laserpitium clade than the Angelica clade. RAPD analysis of all six species showed that the 10-base primer OPC-17 only, out of the20 Kit-C primers from Operon gave polymorphic banding patterns.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 99
    ISSN: 1573-5060
    Schlagwort(e): Allium cepa var. ascalonicum ; Allium × wakegi ; PCR-RFLP ; RAPD ; shallot
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract RAPD and PCR-RFLP analyses were conducted to establish the phylogenetic relationships among collected accessions of shallot and Allium × wakegi, and to assess the origin of A. × wakegi. Twenty out of 100 primers were amplified with 112 scorable bands for cluster analysis. Two main cluster groups consisting of one group for shallot and another group for A. × wakegi were clearly separated. The sub-groups of clusters reflected the phenotype differentiation in shallots and regional specificity in some A. × wakegiaccessions. The present results were also in agreement with previous systematics by isozymes of which the highest genetic variation of A. × wakegi in Indonesia was found in West Java suggesting the possibility that this place might be one of the germplasm centers. From RFLP analysis of amplified matK gene of cpDNA it was demonstrated that A. × wakegi originated from shallot as a maternal plant × Welsh onion as a paternal plant as well as from reciprocal crosses.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 100
    ISSN: 1573-5060
    Schlagwort(e): Eragrostis curvula ; E. pilosa ; E. tef ; genetic diversity ; RAPD ; Tef
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Tef is one of the staple cereal crops in Ethiopia. To evaluate genetic diversity of tef and its relatives, 47 accessions of tef, three accessions of E. pilosa, and six accessions of E. curvulawere analyzed using random amplified polymorphic DNA (RAPD) markers. The level of polymorphism among the wild species was extremely high, while low polymorphism was detected among tef accessions. All cultivars and wild species under study could be distinguished with the help of different primers, thereby indicating the potential of RAPD in the genetic fingerprinting of tef. Accessions from E. curvula and E. pilosa can be differentiated by a single selected primer. In spite of low polymorphism within tef, accessions under study could be distinguished by a combination of selected primers. Cluster analysis indicated that tef is a very closely related species to E. pilosa with 45%similarity, supporting the hypothesis that tef originated from E. pilosa based on morphological data. Given that RAPD are relatively quick, simple to use, and are not subjected to environmental influences, they provide a valuable new approach for the genetic fingerprinting and study of genetic diversity in tef.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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