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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Entomologia experimentalis et applicata 68 (1993), S. 219-229 
    ISSN: 1570-7458
    Keywords: Hymenoptera ; Aphidiidae ; Homoptera ; Aphididae ; Schizaphis graminum ; wheat ; tritrophic interactions ; learning ; host-habitat location
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of experience on the responsiveness of the aphidiid parasitoidLysiphlebus testaceipes (Cresson) (Hymenoptera: Aphidiidae) to host-associated cues was investigated using a wind-tunnel bioassay. Naive females were able to discriminate between uninfested wheat (Triticum aestivum L.) and wheat infested withSchizaphis gramimum (Rondani) (Homoptera: Aphididae), but oviposition experience significantly increased the parasitoid's propensity to respond to aphid-infested plants with upwind, targeted flight. The behavioural change associated with such experience was acquired rapidly (within five minutes) and persisted for at least 24 h. The parasitoid could be successfully conditioned to associate a novel odour with the presence of hosts, suggesting that the increase in response to aphid-infested plants which occurs as a result of experience is probably due to associative learning of olfactory cues from the plant-aphid complex.
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  • 2
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    Acta biotheoretica 39 (1991), S. 1-14 
    ISSN: 1572-8358
    Keywords: Hematological diseases ; first order partial differential equations ; stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To address the possibility that proliferative disorders may originate from interactions between multiple populations of proliferating and maturing cells, we formulate a model for this process as a set of coupled nonlinear first order partial differential equations. Using recent results for the asymptotic behaviour of the solutions to this model, we demonstrate that there exists a region of coupling coefficients, maturation rates, and proliferation rates that will guarantee the stable coexistence of coupled cellular populations. The analysis shows that increases in the coupling between populations may ultimately lead to a loss of stability. Furthermore, the analysis indicates that increases (decreases) in the maturation and/or proliferation rates above (below) critical levels will lead either to instability in the populations or the destruction of one population and the persistence of the other.
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  • 3
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    Aquatic sciences 52 (1990), S. 330-344 
    ISSN: 1420-9055
    Keywords: Flood ; phytoplankton succession ; reversion ; stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of the August 1987 River Reuss flood event on the phytoplankton biocoenosis in Lake Uri (Urnersee, part of Lake Lucerne, Switzerland) was investigated firstly by comparing biological, chemical and physical data sampled before the event with equivalent data sampled after the event; and secondly by comparing the phytoplankton succession in 1987 with that occurring in the “floodfree” year 1989. As a consequence of the flood, the physical and chemical environment of the phytoplankton was found to have undergone a change which resulted in an alteration in the composition of the phytoplankton community. The phytoplankton community existing previous to the flood event, which had been dominated byTabellaria fenestrata sensu Husted 1930 (K-strategist), was replaced by a biocoenosis characterized mainly by various species of flagellates, which represent a typical spring successional stage (r-strategists). After the externally-imposed perturbation, the return to stable physical and chemical conditions was followed by the re-establishment of the successional stage which had existed before the flood (termed “reversion” by Reynolds, 1980).
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  • 4
    ISSN: 1420-9071
    Keywords: Mitochondrial DNA ; RFLP ; Leptinotarsa decemlineata ; Colorado potato beetle ; population genetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary This study demonstrates variability in restriction enzyme cleavage sites of mitochondrial DNA (mtDNA) among four popalations of Colorado potato beetle (CPB). A suite of three enzymes (EcoRI,HpaI,PstI) was sufficient to discriminate among the populations tested. Individuals heteroplasmic for restriction enzyme patterns were found in some populations. Variability in CPB mtDNA should prove useful in efforts to trace the origin and dispersal of the species in North America.
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  • 5
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    Cellular and molecular life sciences 50 (1994), S. 571-575 
    ISSN: 1420-9071
    Keywords: Ancient DNA ; archaeobotany ; carbonized grain ; DNA sequences ; glutenin alleles ; seed proteins ; Triticum ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have used hybridization analysis to detect ancient DNA in wheat seeds collected from three archaeological sites in Europe and the Middle East. One of these samples, carbonizedT. spelta dated to the first millennium BC, has yielded PCR products after amplification with primers directed at the leader regions of the HMW (high molecular weight) glutenin alleles. Sequences obtained from these products suggest that the DNA present in the Danebury seeds is chemically damaged, as expected for ancient DNA, and also indicate that it should be possible to study the genetic variability of archaeological wheat by ancient DNA analysis. Finally, we describe a PCR-based system that enables tetraploid and hexaploid wheats to be distinguished.
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  • 6
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    Mycopathologia 111 (1990), S. 181-189 
    ISSN: 1573-0832
    Keywords: mycotoxin ; ochratoxin ; Penicillium ; storage ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Eleven-kilogram parcels of HY-320 wheat, a cultivar of the new Canada Prairie Spring class, were kept at 15 and 19% initial moisture contents (IMC) in simulated storage in a Manitoba farm granary for 60 weeks to determine biotic and abiotic changes and mycotoxin production. Ochratoxin A reached a maximum of 0.24 ppm by week 20 in the 19% IMC wheat, but was absent in the 15% IMC wheat; no other mycotoxins were detected. Temperature, moisture content, O2 and CO2 levels, fat acidity values, seed germination, microfloral incidence and abundance, and the presence of other mycotoxins were monitored. Principal component analysis of all variables showed that the first principal components accounted for 32–41% of the system variability, and contained the ochratoxin A variable. Ochratoxin A was produced in moist grain that had decreased seed germination andAltermaria activity, and high fungal activity byPenicillium andAspergillus versicolor. Compared to other stored cereals previously studied, HY-320 wheat would be ranked in a low-risk category for mycotoxin formation, based on the ochratoxin A levels observed.
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  • 7
    ISSN: 1573-0832
    Keywords: Penicillium griseofulvum ; patulin ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Sixty-four wheat samples from Spanish flour factories were screened for patulin and patulin-producing moulds. None of them was found to contain any patulin, whereas samples experimentally contaminated with this toxin proved it to be highly unstable. On the other hand, Penicillium griseofulvum was the only in vitro patulin-producing species found (19 samples). Mould growth in the samples was investigated by using yeast-sucrose medium (YES) and high-performance liquid chromatography (HPLC) to measure the amounts of toxin produced during 40 day's incubation at 20 and 28°C. The highest yield rate of patulin was obtained between the 20th and 30th day of incubation; such a rate, however, was very low throughout the vigorous growth phase, during the first 20 days of incubation. The more appropriate temperature for incubation and patulin production was 28 °C. We also investigated the influence of other incubation conditions in the yield and found stationary dark cultures to be more efficient that shaken or fermentation cultures in YES medium. The best patulin yield achieved was 11.9 mg in the culture broth and 6.3 mg in the mycelium from 100 ml of medium.
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  • 8
    ISSN: 1420-9098
    Keywords: Leptothorax acervorum ; mtDNA ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 27 nests ofLeptothorax acervorum were analysed for restriction fragment-length polymorphism (RFLP) in mitochondrial DNA (mtDNA), using four endonucleases. A substantial degree of variation was found between nests in the population (13 composite haplotypes). Intra-nest variation was detected in 15 % of the nests. The presence of occasional alien inseminated females indicates that polygyny in this species is caused by adoption of mated females. The occasional acceptance of alien females is difficult to explain, but interesting, since this behaviour could have given rise to inquilinism. Our results suggest that analysis of mtDNA RFLP is a method well suited for investigations of the population structure of ants.
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  • 9
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    Cellular and molecular life sciences 46 (1990), S. 1016-1017 
    ISSN: 1420-9071
    Keywords: In vitro absorption ; calcium ; wheat ; Bengal gram
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The in vitro absorption of calcium from the duodenum was significantly less in a group of rats fed on a wheat diet than in a group fed a wheat and Bengal gram (70∶30) diet.
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  • 10
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    Plant cell reports 12 (1993), S. 293-297 
    ISSN: 1432-203X
    Keywords: Lycopersicon esculentum ; Genetic marker ; Intraspecific genetic map ; DNA polymorphism ; Isozyme ; RFLP ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The existing molecular genetic maps of the tomato, Lycopersicon spp, are constructed based on isozyme and RFLP polymorphisms between tomato species. These maps are useful for certain applications but have few markers that exhibit sufficient polymorphisms for intraspecific analysis and manipulations within the cultivated tomato. The purpose of this study was to investigate the relative potential of RAPD technology, as compared to isozymes and RFLPs, to generate polymorphic DNA markers within cultivated tomatoes. Sixteen isozymes and 25 RFLP clones that were known to detect polymorphism between L. esculentum and L. pennellii, and 313 random oligonucleotide primers were examined. None of the isozymes and only four of the RFLP clones (i.e., 16%) revealed polymorphism between the cultivated varieties whereas up to 63% of the RAPD primers detected one or more polymorphic DNA fragments between these varieties. All RAPD primers detected polymorphism between L. esculentum and L. pennellii genotypes. These results clearly indicate that RAPD technology can generate sufficient genetic markers exploiting sequence differences within cultivated tomatoes to facilitate construction of intraspecific genetic maps.
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  • 11
    ISSN: 1432-203X
    Keywords: wheat ; rye ; embryogenesis ; growth ; tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The influence of the short arm of rye chromosome 1 (1RS) from Secale cereale var. Imperial on the growth and differentiation of callus cultures from wheat Triticum aestivum var. Chinese Spring immature embryos was analysed. This chromosome arm was found to stimulate both embryogenesis and the rate of growth of calli. Recombinant lines carrying segments of 1RS were used to delineate the regions of 1RS responsible for the tissue culture effects. The enhancement of embryogenesis and the stimulation of growth were shown to be associated with two distinct genetic regions of the chromosome arm; the former is located between the centromere and the Sec 1 locus, while the latter is situated in the immediate vicinity of the Sec 1 locus.
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  • 12
    ISSN: 1432-203X
    Keywords: Protoplast fusion ; RFLP ; Mitochondrial DNA ; Chloroplast DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Somatic hybrid plants were recovered following fusion of leaf mesophyll protoplasts isolated from tomato (Lycopersicon esculentum) cultivar UC82 with protoplasts isolated from suspension cultured cells of L. chilense, LA 1959. Iodoacetate was used to select against the growth of unfused tomato protoplasts. Two somatic hybrids were recovered in a population of 16 regenerants. No tomato regenerants were recovered; all of the non-hybrid regenerants were L. chilense. The L. chilense protoplast regenerants were tetraploid. The hybrid nature of the plants was verified using species-specific restriction fragment length polymorphisms for the nuclear, chloroplast and mitochondrial genomes. The somatic hybrids had inherited the chloroplast DNA of the tomato parent, and portions of the mitochondrial DNA of the L. chilense parent. The somatic hybrids formed flowers and developed seedless fruit.
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  • 13
    ISSN: 1432-2145
    Keywords: Gametophytic self incompatibilityself-compatibility ; Lycopersicon peruvianum Lycopersicon hirsutum ; S-associated proteins ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Self-compatibility was investigated separately in two species of tomato, Lycopersicon peruvianum and L. hirsutum. The codominant expression of self-compatibility (SC)/self incompatibility (SI) was established using intraspecific hybrids of SC and SI hybrids. In SC L. peruvianum, a major stylar protein of approximately 29 kDa cosegregates with self-compatibility in the progeny of SC/SI hybrids. The SC/SI hybrids are self-fertile, but only partially so, since the SI allele present in the hybrids is capable of eliminating certain genotypes in the resultant progeny. In L. hirsutum, the majority of hybrids between one accession of SI L. hirsutum f. hirsutum and one of SC L. hirsutum f. glabratum are self-fertile. Analysis of the progeny revealed that the SC and SI alleles are codominant in this species as well. A protein product for the SC allele is not obvious in style extracts of L. hirsutum f. glabratum. Segregating progeny from SC/SI hybrids of L. hirsutum were used to map the S locus against five RFLP markers on chromosome 1, and estimated map distances are given. In addition, evidence is presented that indicates that one of the DNA markers, CD15, is duplicated in L. hirsutum f. glabratum, and the duplication is not linked to the S locus.
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  • 14
    ISSN: 1432-203X
    Keywords: Rice (Oryza sativa) ; Genetic Marker ; Genetic Map ; Integrated Linkage Map ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Application of genetic linkage maps in plant genetics and breeding can be greatly facilitated by integrating the available classical and molecular genetic linkage maps. In rice, Oryza sativa L., the classical linkage map includes about 300 genes which correspond to various important morphological, physiological, biochemical and agronomic characteristics. The molecular maps consist of more than 500 DNA markers which cover most of the genome within relatively short intervals. Little effort has been made to integrate these two genetic maps. In this paper we report preliminary results of an ongoing research project aimed at the complete integration and alignment of the two linkage maps of rice. Six different F2 populations segregating for various phenotypic and RFLP markers were used and a total of 12 morphological and physiological markers (Table 1) were mapped onto our recently constructed molecular map. Six linkage groups (i.e., chr. 1, 3, 7, 9, 11 and 12) on our RFLP map were aligned with the corresponding linkage groups on the classical map, and the previous alignment for chromosome 6 was further confirmed by RFLP mapping of an additional physiological marker on this chromosome. Results from this study, combined with our previous results, indicate that, for most chromosomes in rice, the RFLP map encompasses the classical map. The usefulness of an integrated genetic linkage map for rice genetics and breeding is discussed.
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  • 15
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    Current genetics 26 (1994), S. 456-460 
    ISSN: 1432-0983
    Keywords: Parasitella parasitica ; Zygomycetes ; RAPD ; PCR ; RFLP ; Electrophoretic karyotype ; Molecular taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In addition to conventional methods for the identification of fungi, molecular techniques at the DNA level are increasingly being employed. In order to check the validity of such experimental approaches, we have analyzed the well-defined species Parasitella parasitica, which belongs to the family Mucoraceae (Mucorales, Zygometes). The seven strains of this species, which are available from international strain collections, were analyzed by several molecular methods: restriction fragment length polymorphism analysis (RFLP), the random primer-dependent polymerase chain reaction (RAPD-PCR), and electrophoretic karyotyping. Unexpectedly, these strains are highly diverse at the molecular level. By these techniques they can be divided consistently into two different groups. Nevertheless, all seven strains belong to a single species. They show no morphological differences and sexual spores (zygospores) were found in all possible combinations either within or between the two groups. Southern-blot analysis of genomic DNA of all P. parasitica strains with RAPD-PCR-derived labelled probes shows the existence of repetitive elements characteristic for only one group of P. parasitica. In addition, chromosome sizes, which were separated by rotating-field electrophoresis, were highly divergent, and ranged from 3 to 6.5 Mb in one group and between 2 and 4.5 Mb in the other. The RAPD-PCR patterns also discriminate both groups of P. parasitica. However, they are very similar if strains of a single group are compared. Therefore, we propose that the determination of fungal species by molecular techniques should be vetted at least by morphological and physiological parameters and, whenever possible, by mating experiments.
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  • 16
    ISSN: 1432-0983
    Keywords: Cytoplasmic male sterility ; Antisense RNA ; RFLP ; Cybrids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Asymmetric cell-fusion of the japonica cultivar ofOryza sativa (rice) with cytoplasmic-male-sterile (CMS) plants bearing cytoplasm derived from Chinsurah Boro II, resulted in two classes of cytoplasmic hybrids (cybrids), fertile and CMS. Southern-blot analysis of the mitochondrial DNA (mtDNA) indicates recombination events around a number of genes; however, the appearance of the CMS character is tightly correlated to reorganization around theatp6 gene, suggesting recombination downstream from theatp6 gene is involved in CMS. The nucleotide sequence downstream fromatp6 contains a pseudogene which was probably created by recombination of the mitochondrial genome. Sense and antisense transcripts of the downstream region ofatp6 were found in CMS-and restored CMS (fertile)-lines, but not in the normal (fertile) line. In the CMS line, several antisense transcripts of theatp6 gene were also found. However, in the restored line which contains a nuclear-encoded gene,Rf-1, the levels of these transcripts were lower than in the CMS line. These results suggest abnormal transcripts of theatp6 gene produced in the antisense direction may be involved in CMS, and that products of the nuclear-encoded restorer gene may reduce abnormal transcription in this region of the mitochondrial genome.
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  • 17
    ISSN: 1432-0983
    Keywords: DNA fingerprinting of Trichoderma ; Trichoderma reesei ; RFLP ; Strain classification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have analyzed nine different species of the filamentous fungus Trichoderma and three strains of T. reesei for the presence of hypervariable loci in their genomes by hybridization with simple repeat oligonucleotides [(CT)8, (GTG)5, and (GACA)4]. On the basis of the DNA-fingerprints obtained, the Trichoderma aggregate is re-classified into five groups: I (T. reesei, T. todica), II (T. polysporum, T. longibrachiatum, T. koningii, and T. pseudokoningii), III (T. virgatum), IV (T. saturnisporum) and V (T. harzianum). These results contradict the claim that T. reesei is a subspecies of T. longibrachiatum. Furthermore, hybridization with (CA)8 allowed a subdivision of group II, wherein T. pseudokoningii formed a subgroup, IIb, which is highly homologous with, but distinct from subgroup IIa. The results show that RFLP analysis may be used to re-classify the Trichoderma aggregate.
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  • 18
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    Journal of chemical ecology 16 (1990), S. 2203-2216 
    ISSN: 1573-1561
    Keywords: Yponomeuta cagnagellus ; caterpillars ; Lepidoptera ; Yponomeutidae ; trail following ; chemical marker ; trail pheromone ; stability ; pheromone secretory site
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Trail following in lepidopterous larvae is often attributed to chemical markers, but only a few clear-cut examples are found in the literature. In this paper evidence is presented for a chemical basis of the trail following behaviour ofYponomeuta cagnagellus. (Lepidoptera: Yponomeutidae) The marker is shown to be very persistent under laboratory conditions and is water soluble. Several possible secretory sites were investigated, and it is concluded that the marker is probably secreted together with the silk from the labial gland. Problems associated with the demonstration of trail markers in caterpillars are discussed.
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  • 19
    ISSN: 1573-1561
    Keywords: Allelochemicals ; no-tillage ; conventional-tillage ; soils ; wheat ; Triticum aestivum ; mass spectrometry ; Petri-dish bioassay ; fatty acids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Putative allelochemicals found in the soil of no-tillage and conventional-tillage wheat plots near Stillwater, Oklahoma, were obtained by a mild alkaline aqueous extraction procedure, bioassayed to determine their biological activity, purified, and analyzed with a capillary gas chromatography-mass spectrometry-data analysis system. The most significant inhibition was found in bioassays of extracts from soil collected immediately after harvest in June, July, and August. No-tillage soils produced significant inhibition during the rest of the year also. Mass spectrometry showed fatty acids as the most abundant compounds. However, when bioassayed authentic samples of the five free fatty acids showed no significant biological activity toward wheat.
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  • 20
    ISSN: 1573-1561
    Keywords: Cover crops ; wheat ; Triticum aestivum ; soybean ; Glycine max ; soil extracts ; germination bioassays ; phenolic acids ; hydroxamic acids ; allelopathy ; slope analysis ; ivy-leaved morning glory ; Ipomoea hederacea ; crimson clover ; Trifolium incarnalum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The primary objective of this research was to determine if soil extracts could be used directly in bioassays for the detection of allelopathic activity. Here we describe: (1) a way to estimate levels of allelopathic compounds in soil; (2) how pH, solute potential, and/or ion content of extracts may modify the action of allelopathic compounds on germination and radicle and hypocotyl length of crimson clover (Trifolium incarnatum L.) and ivyleaved morning glory (Ipomoea hederacea L. Jacquin.); and (3) how biological activity of soil extracts may be determined. A water-autoclave extraction procedure was chosen over the immediate-water and 5-hr EDTA extraction procedures, because the autoclave procedure was effective in extracting solution and reversibly bound ferulic acid as well as phenolic acids from wheat debris. The resulting soil extracts were used directly in germination bioassays. A mixture of phenolic acids similar to that obtained from wheat-no-till soils did not affect germination of clover or morning glory and radicle and hypocotyl length of morning glory. The mixture did, however, reduce radicle and hypocotyl length of clover. Individual phenolic acids also did not inhibit germination, but did reduce radicle and hypocotyl length of both species. 6-MBOA (6-methoxy-2,3-benzoxazolinone), a conversion product of 2-o-glucosyl-7-methoxy-1,4-benzoxazin-3-one, a hydroxamic acid in living wheat plants, inhibited germination and radicle and hypocotyl length of clover and morning glory. 6-MBOA, however, was not detected in wheat debris, stubble, or soil extracts. Total phenolic acids (FC) in extracts were determined with Folin and Ciocalteu's phenol reagent. Levels of FC in wheat-conventionaltill soil extracts were not related to germination or radicle and hypocotyl length of either species. Levels of FC in wheat-no-till soil extracts were also not related to germination of clover or morning glory, but were inversely related to radicle and hypocotyl length of clover and morning glory. FC values, solute potential, and acidity of wheat-no-till soil extracts appeared to be independent (additive) in action on clover radicle and hypocotyl length. Radicle and hypocotyl length of clover was inversely related to increasing FC and solute potential and directly related to decreasing acidity. Biological activity of extracts was determined best from slopes of radicle and hypocotyl length obtained from bioassays of extract dilutions. Thus, data derived from the water-autoclave extraction procedure, FC analysis, and slope analysis for extract activity in conjunction with data on extract pH and solute potential can be used to estimate allelopathic activity of wheat-no-till soils
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  • 21
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    Plant molecular biology 18 (1992), S. 423-427 
    ISSN: 1573-5028
    Keywords: Hordeum vulgare L. ; CM protein ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The primary structure of the insect α-amylase inhibitor CMa of barley seeds was deduced from a full-length cDNA clone pc43F6. Analysis of RNA from barley endosperm shows high levels 15 and 20 days after flowering. The cDNA predicts an amino acid sequence of 119 residues preceded by a signal peptide of 25 amino acids. Ala and Leu account for 55% of the signal peptide. CMa is 60–85% identical with α-amylase inhibitors of wheat, but shows less than 50% identity to trypsin inhibitors of barley and wheat. The 10 Cys residues are located in identical positions compared to the cereal inhibitor family with a Pro-X-Cys motif present in all.
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  • 22
    ISSN: 1573-5028
    Keywords: carboxy-terminal processing ; glycosylation ; Hordeum vulgare L. ; Prx locus ; RFLP ; signal peptide ; targeting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The major peroxidase of barley seed BP 1 was characterized. Previous studies showed a low carbohydrate content, low specific activity and tissue-specific expression, and suggested that this basic peroxidase could be particularly useful in the elucidation of the structure-function relationship and in the study of the biological roles of plant peroxidases (S.K. Rasmussen, K.G. Welinder and J. Hejgaard (1991) Plant Mol Biol 16: 317–327). A cDNA library was prepared from mRNA isolated from seeds 15 days after flowering. Full-length clones were obtained and showed 3′ end length variants, a G+C content of 69% in the translated region, a 90% G or C preference in the wobble position of the codons and a typical signal peptide sequence. N-terminal amino acid sequencing and sequence analysis of tryptic peptides verified 98% of the sequence of the mature BP 1 which contains 309 amino acid residues. BP 1 is the first characterized plant peroxidase which is not blocked by pyroglutamate. BP 1 polymorphism was observed. BP 1 is less than 50% identical to other plant peroxidases which, taken together with its developmentally dependent expression in the endosperm 15–20 days after flowering, suggests a unique biological role of this enzyme. The barley peroxidase is processed at the C-terminus and might be targeted to the vacuole. The single site of glycosylation is located near the C-terminus in the N-glycosylation sequon -Asn-Cys-Ser- in which Cys forms part of a disulphide bridge. The major glycan is a typical plant modified-type structure, Manα1-6(Xylβ1-2)Manβ1-4GlcNAcβ1-4(Fucα1-3)GlcNAc. The BP 1 gene was RFLP-mapped on barley chromosome 3, and we propose Prx5 as the name for this new peroxidase locus.
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  • 23
    ISSN: 1573-5028
    Keywords: gene duplication ; photosynthesis ; RFLP ; Southern blots
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A second locus (Lhb1B) encoding Photosystem II Type I chlorophyll a/b-binding (CAB) polypeptides was identified in Arabidopsis thaliana. This locus carries two genes in an inverted orientation. The predicted sequences of the polypeptides encoded by these two genes show substantial divergence in their amino termini relative to each other and to the proteins encoded by the three Lhb1 CAB genes previously characterized [10], but little divergence within the predicted primary structure of the mature protein. DNA probes derived from seven additional types of tomato CAB genes, encoding chlorophyll a/b-binding polypeptides of several antenna systems of the photosynthetic apparatus, were tested against A. thaliana. Each of these hybridized in Southern blots to unique DNA fragment(s), demonstrating the existence of each of these different types of CAB genes in the genome of A. thaliana. The number of genes encoding each CAB type in A. thaliana was estimated to be similar to that of tomato.
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  • 24
    ISSN: 1573-5028
    Keywords: bark storage protein ; gene clustering ; RFLP ; systemic response
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two wound-inducible cDNAs from poplar leaves show sequence identity to vegetative storage proteins (VSP) that accumulate seasonally in poplar bark tissues. We have compared the genomic organization, cDNA sequences and expression of the genes encoding the wound-inducible cDNAs (win4) with that of a bark VSP (called bark storage protein, or BSP). There appear to be several win4 genes in the poplar genome which segregate as a single locus and are therefore likely to be clustered. The same is true of the BSP genes. The win4 locus is linked (map distance of 5 cM) to the BSP locus, consistent with a common evolutionary origin of the genes. A near full-length win4 cDNA shows 75% sequence identity to BSP cDNAs. Both win4 and BSP are systemically wound-inducible; win4 transcripts accumulate in leaves and stems, whereas BSP transcripts accumulate almost exclusively in stems. A phloem transport-dependent signaling mechanism appears to be involved in systemic win4 expression after wounding. In contrast to BSP gene expression, win4 genes are not expressed in response to short day conditions. The data indicate win4 and BSP genes are differentially regulated, and their products may play important roles in the storage and reallocation of nitrogen in perennial plants.
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  • 25
    ISSN: 1573-5028
    Keywords: protein degradation ; ubiquitin conjugating enzymes ; DNA repair ; N-end recognition ; wheat ; Arabidopsis thaliana
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    Notes: Abstract Covalent attachment of ubiquitin to other cellular proteins has been implicated in a multitude of diverse physiological processes in eukaryotes including selective protein degradation. This attachment is carried out by a multi-enzyme pathway consisting of three classes of enzymes: ubiquitin-activating enzymes (E1s), ubiquitin-conjugating enzymes (E2s), and ubiquitin-protein ligases (E3s). E2s accept activated ubiquitin from E1 and conjugate it to target proteins with or without the participation of specific E3s. Previously, we have isolated wheat cDNAs encoding 16 and 23 kDa E2s, TaUBC1 and TaUBC4, respectively. TaUBC1 shows structural homology to the yeast RAD6 E2 that is essential for DNA repair whereas TaUBC4 is related to the yeast ScUBC8 E2, both of which effectively conjugate ubiquitin to histones in vitro but as yet are without a known in vivo function. Here, we report the isolation of genomic and cDNA homologues of these genes from Arabidopsis thaliana. In Arabidopsis, both of these E2s are encoded by three member gene families. Members of the AtUBC1 gene family, comprising AtUBC1, 2 and 3, encode 150–152 amino acid proteins that are 83–99% identical to each other and TaUBC1 and contain four introns that are conserved with respect to position. Members of the AtUBC4 gene family, comprising AtUBC4, 5 and 6, encode 187–191 amino acid proteins that are 73–88% identical to each other and TaUBC4 and contain five introns that are conserved with respect to position. In contrast, AtUBC1-3 gene products are only 31–36% identical to those derived from AtUBC4-6. mRNA for each family was detected in Arabidopsis roots, leaves, stems, and flowers indicating that members of each family are expressed in most if not all tissues.
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  • 26
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    Plant molecular biology 20 (1992), S. 849-856 
    ISSN: 1573-5028
    Keywords: GA regulation ; thiol-protease promoter ; wheat ; aleurone ; particle gun
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    Topics: Biology
    Notes: Abstract A wheat gene (A121) encoding a protein with sequence similarity to mammalian cathepsin B is regulated by gibberellic acid (GA) in aleurone layers of germinating grains. To analyse the mechanism of A121 regulation, its promoter was fused to the β-glucuronidase reporter gene (GUS) and introduced by micro-projectile bombardment into aleurone layers of oat. With 2.3 kb of promoter sequence, the GUS expression was enhanced by GA treatment. This effect was reversed by abscisic acid (ABA). This result showed for A121, like the α-amylase genes, that the regulation by GA and ABA was at the level of transcription. The GA responsiveness of the promoter was retained with as little as 276 bp of promoter sequence. Sequence comparison with a GA responsive promoter of an α-amylase gene identified the conserved element GCAACGGCAACGATGG which is required intact for full expression of both promoters. However, there was no identifiable similarity in the cathepsin-like promoter with the GA-responsive element of α-amylase promoters with the consensus sequence TAACAAA, suggesting that GA affects more than one mechanism of transcriptional control.
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  • 27
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    Plant molecular biology 20 (1992), S. 991-995 
    ISSN: 1573-5028
    Keywords: retrotransposon-like element ; sequence analysis ; wheat
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    Topics: Biology
    Notes: Abstract WIS-2-1A, a 8624 bp insertion in the Glu-1A-2 locus of chromosome 1A of wheat, consists of two 1755 bp long terminal repeats enclosing a 5114 bp internal region. No long open reading frames could be found, but inspection of the predicted amino acid sequence showed regions with homology to retrotransposon structures, including a methionine tRNA initiator binding site, a nucleotide binding domain, a protease, an integrase and a polymerase. DNA replication errors have resulted in frame-shifts in the protein coding region, suggesting that retrotransposition of WIS-2-1A, if it occurs, must be mediated by trans-acting factors.
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  • 28
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    Plant molecular biology 21 (1993), S. 99-108 
    ISSN: 1573-5028
    Keywords: fusion protein ; Oenothera ; plastome ; RFLP ; unidentified open reading frame
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    Topics: Biology
    Notes: Abstract The chloroplast DNA encodes genes for components of photosynthesis and the transcription-translation machinery; a number of unidentified open reading frames (ORFs) are also present. To determine whether a large ORF in the inverted repeat of chloroplast DNA of tobacco (ORF2280) encodes a chloroplast protein, a conserved region of the ORF was expressed in Escherichia coli. An antibody against the ORF protein was prepared using the purified fusion protein as an antigen. When incubated with proteins from the soluble fraction of tobacco, spinach and Oenothera chloroplasts, the antiserum detects relatively labile polypeptides, which have apparent molecular weights of 170 to 180 kDa. The ORF in tobacco and spinach is large enough to encode a protein of 240–250 kDa, thus it is possible that post-transcriptional or post-translational processing reduces the size of the expression product. Analysis of Oenothera chloroplasts representing four different plastome types revealed endonuclease restriction fragment length polymorphisms in chloroplast DNA indicative of insertion/deletion events in a region of the chloroplast DNA that shared significant sequence similarity with ORF2280. The ORF2280 antiserum was used to demonstrate that there are qualitative differences in the ORF proteins from different Oenothera plastome types.
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  • 29
    ISSN: 1573-5028
    Keywords: antinutritional factor ; pea lectin ; site-directed mutagenesis ; stability
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    Notes: Abstract Legume lectins are considered to be antinutritional factors (ANF) in the animal feeding industry. Inactivation of ANF is an important element in processing of food. In our study on the stability ofPisum sativum L. lectin (PSL), a conserved hydrophobic amino acid (Val103) in a surface loop was replaced with alanine. The mutant lectin, PSL V103A, showed a decrease in unfolding temperature (T m ) by some 10 °C in comparison with wild-type (wt) PSL, and the denaturation energy (ΔH) is only about 55% of that of wt PSL. Replacement of an adjacent amino acid (Phe104) with alanine did not result in a significant difference in stability in comparison with wt PSL. Both mutations did not change the sugarbinding properties of the lectin, as compared with wt PSL and with PSL from pea seeds, at ambient temperatures. The double mutant, PSL V103A/F104A, was produced inEscherichia coli, but could not be isolated in an active (i.e. sugar-binding) form. Interestingly, the mutation in PSL V103A reversibly affected sugar-binding at 37 °C, as judged from haemagglutination assays. These results open the possibility of production of lectins that are activein planta at ambient temperatures, but are inactive and possibly non-toxic at 37 °C in the intestines of mammals.
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  • 30
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    Plant molecular biology 22 (1993), S. 1173-1176 
    ISSN: 1573-5028
    Keywords: polymerase chain reaction ; LMW glutenin genes ; wheat ; quality
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    Topics: Biology
    Notes: Abstract Polymerase chain reaction (PCR) was used to amplify low-molecular-weight (LMW) glutenin sequences from genomic DNA extracted from a single germinating seed of several durum wheat genotypes. Electrophoretic analysis of PCR reactions showed the presence of amplified products characteristic of durum wheat cultivars with good and poor technological properties. This PCR-based approach is proposed as a very efficient and safe alternative to standard procedures for selecting durum wheat genotypes with good qualitative characteristics.
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  • 31
    ISSN: 1573-5028
    Keywords: retrotransposon ; Pisum ; segregation ; PCR ; RFLP ; mapping
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    Notes: Abstract A DNA sequence between two legumin genes in Pisum is a member of the copia-like class of retrotransposons and represents one member of a polymorphic and heterogeneous dispersed repeated sequence family in Pisum. This sequence can be exploited in genetic studies either by RFLP analysis where several markers can be scored together, or the segregation of individual elements can be followed after PCR amplification of specific members.
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  • 32
    ISSN: 1573-5028
    Keywords: tRNA-like sequences ; t-elements ; RNA processing ; mitochondria ; wheat
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    Notes: Abstract We have recently described the properties of a wheat mitochondrial extract that is able to process, accurately and efficiently, artificial transcripts containing wheat mitochondrial tRNA sequences, with the production of mature tRNAs (P.J. Hanic-Joyce and M.W. Gray, J. Biol. Chem., in press). Such processing involves 5′-endonucleolytic, 3′-endonucleolytic, and TRNA nucleotidyltransferase activities. Here we show that this system also acts on transcripts containing sequences corresponding to an unusual class of short repeats (‘t-elements’) in wheat mtDNA. These repeats are theoretically capable of assuming a tRNA-like secondary structure, although stable transcripts corresponding to them are not detectable in vivo. We find that t-element sequences are processed with the same specificity and with comparable efficiency as are authentic tRNA sequences. Because known t-elements are located close to and in the same transcriptional orientation as active genes (18S-5S, 26S, tRNAPro) in wheat mtDNA, our results raise the question of whether t-elements play a role in gene expression in wheat mitochondria.
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  • 33
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    Plant molecular biology 16 (1991), S. 335-337 
    ISSN: 1573-5028
    Keywords: Triticum ; wheat ; endosperm ; gliadin ; pseudogene ; duplication ; evolution
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  • 34
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    Plant molecular biology 16 (1991), S. 907-908 
    ISSN: 1573-5028
    Keywords: ubiquitin ; wheat ; heat shock protein
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  • 35
    ISSN: 1573-5028
    Keywords: α-amylase inhibitor ; expression inE. coli ; glycosylation versus activity ; insect α-amylase ; mutagenesis ; wheat
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    Notes: Abstract The wheat monomeric inhibitor WMAI-1 (syn. 0.28) produced inEscherichia coli using the pT7-7 expression ventor has the correct N-terminal sequence and the same electrophoretic mobility and specific activity towards the α-amylase from the insectTenebrio molitor as the native WMAI-1 isolated from wheat. This confirms that the native inhibitor is not glycosylated and contradicts claims that a putative glycosyl moiety was essential for inhibition. Thirteen mutants have been obtained at six different sites. Substitution of the highly conserved N-terminal S by the sequence ARIRAR increased the pre-incubation time required for maximum activity. A similar result was obtained by insertion of GPRLPW after position 4, while insertion of EPRAPW at the same position rendered the inhibitor inactive. The substitution D/EGPRL and insertions DGP or D, at position 58, produced complete inactivation. All other mutations had only minor effects on activity.
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  • 36
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    Plant molecular biology 17 (1991), S. 273-275 
    ISSN: 1573-5028
    Keywords: Tahsp17.3 ; low-molecular-weight HSP ; Triticum aestivum L. ; wheat
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  • 37
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    Plant molecular biology 21 (1993), S. 907-912 
    ISSN: 1573-5028
    Keywords: abscisic acid ; dehydration ; LEA ; water stress ; wheat
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    Topics: Biology
    Notes: Abstract A cDNA clone, pMA1949, detects two mRNA species in wheat seedling tissue that are late embryogenesis-abundant (LEA) and dehydration stress-inducible. Sequence analysis of the pMA1949 clone shows it to be a 991 bp partial cDNA encoding a polypeptide of 317 amino acids with homology to two group 3 LEA proteins, carrot (DC8) and a soybean protein encoded by pGmPM2 cDNA. Molecular analysis of the deduced protein reveals a 33 kDa acidic and extremely hydrophilic protein with potential amphiphilic α-helical regions. In addition, the protein contains eleven similar, contiguous repeats of 11 amino acids, which are separated by 118 amino acids from two additional and unique repeats of 36 residues each at the carboxyl end of the protein. Comparisons of sequences of reported group 3 LEA proteins revealed that there are two types, separable by sequence similarity of the 11 amino acid repeating motifs and by the presence or absence of a certain amino acid stretch at the carboxyl terminus. Based on resuls from these comparisons, we propose a second type of group 3 LEA proteins, called group 3 LEA (II).
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  • 38
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    Plant molecular biology 22 (1993), S. 547-552 
    ISSN: 1573-5028
    Keywords: acetyl-CoA carboxylase ; chloroplast ; haloxyfop ; herbicide ; wheat
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    Topics: Biology
    Notes: Abstract The acetyl-CoA carboxylase present in both wheat germ and total wheat leaf protein contains ca. 220 kDa subunits. It is the major biotin-dependent carboxylase present in wheat chloroplasts. Active acetyl-CoA carboxylase purified from wheat germ is a homodimer with an apparent molecular mass of ca. 500 kDa. The enzyme from wheat germ or from wheat chloroplasts is sensitive to the herbicide haloxyfop at micromolar levels. The incorporation of 14C-acetate into fatty acids in freshly cut wheat seedling leaves provides a convenient in vivo assay for both acetyl-CoA carboxylase and haloxyfop.
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  • 39
    ISSN: 1573-5028
    Keywords: amino acid sequence ; cDNA ; germination ; (1→3, 1→4)-β-glucanases ; wheat
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    Notes: Abstract A (1→3, 1→4)-β-glucan 4-glucanohydrolase [(1→3, 1→4)-β-glucanase, EC 3.2.1.73] was purified to homogeneity from extracts of germinated wheat grain. The enzyme, which was identified as an endohydrolase on the basis of oligosaccharide products released from a (1→3, 1→4)-β-glucan substrate, has an apparent pI of 8.2 and an apparent molecular mass of 30 kDa. Western blot analyses with specific monoclonal antibodies indicated that the enzyme is related to (1→3, 1→4)-β-glucanase isoenzyme EI from barley. The complete primary structure of the wheat (1→3, 1→4)-β-glucanase has been deduced from nucleotide sequence analysis of cDNAs isolated from a library prepared using poly(A)+ RNA from gibberellic acid-treated wheat aleurone layers. One cDNA, designated λLW2, is 1426 nucleotide pairs in length and encodes a 306 amino acid enzyme, together with a NH2-terminal signal peptide of 28 amino acid residues. The mature polypeptide encoded by this cDNA has a molecular mass of 32085 and a predicted pI of 8.1. The other cDNA, designated λLW1, carries a 109 nucleotide pair sequence at its 5′ end that is characteristic of plant introns and therefore appears to have been synthesized from an incompletely processed mRNA. Comparison of the coding and 3′-untranslated regions of the two cDNAs reveals 31 nucleotide substitutions, but none of these result in amino acid substitutions. Thus, the cDNAs encode enzymes with identical primary structures, but their corresponding mRNAs may have originated from homeologous chromosomes in the hexaploid wheat genome.
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  • 40
    ISSN: 1573-5028
    Keywords: wheat ; Triticum aestivum ; cDNA clone ; ADP-glucose pyrophosphorylase ; small-subunit ; starch
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    Notes: Abstract A full-length cDNA clone from hexaploid bread wheat, encoding the small subunit of ADP-glucose pyrophosphorylase, has been isolated from an endosperm cDNA library. The cDNA insert has an open reading frame which encodes a protein of 473 amino acids (52.1 kDa). The presence of a chloroplast/amyloplast transit peptide of 22 amino acids is proposed. The deduced amino acid sequence exhibits a high degree of homology with the small subunit ADP-glucose pyrophosphorylase proteins from rice (with 90% of identical amino acids) and potato (with 86% of identical amino acids) and contains conserved sequence elements which are thought to represent the substrate binding and allosteric activator sites. The genes are organised as single-copy loci on chromosomes 7A, 7B and 7D in the wheat genome and are highly expressed during grain development. Homologous transcripts are expressed in leaves and roots.
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  • 41
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    Plant molecular biology 26 (1994), S. 1065-1071 
    ISSN: 1573-5028
    Keywords: Brassica ; polyploid ; gene expression ; RT-PCR ; RFLP
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    Notes: Abstract One of the essential issues regarding evolution of polyploid species is how duplicate genes are expressed. Most studies on gene expression in polyploids have been based on isozyme analyses; RNA analysis has not been widely used partially due to difficulties in distinguishing homologous transcripts which usually have the same length and similar or almost identical sequences. In this study, a method combining RT-PCR with RFLP was used to analyze transcripts of homologous genes in natural and synthetic Brassica amphidiploids. Sequences coding for several known genes were selected and used to synthesize gene-specific primers. Total RNAs were used as templates for RT-PCR to amplify homologous transcripts in three diploid parental species, three cultivated amphidiploid species and six synthetic amphidiploids. For each gene, initial PCR products amplified in all species had identical length; however, homologous transcripts in the diploid and amphidiploid species could be distinguished after digesting the PCR products with restriction enzymes. Preliminary results based on three genes indicated that both transcripts from the diploid parents were expressed in the synthetic and natural amphidiploids. This study represents the first application of RT-PCR and RFLP analysis to investigate expression of homologous genes in higher plants. The technique is a sensitive, simple and efficient method for distinguishing homologous transcripts in a mixed RNA population and can be applied to many types of studies on expression of homologous genes.
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  • 42
    ISSN: 1573-5028
    Keywords: α-amylase tetrameric inhibitor ; cDNA cloning ; genetic mapping ; wheat
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    Notes: Abstract We have characterized three cDNA clones corresponding to proteins CM1, CM3 and CM16, which represent the three types of subunits of the wheat tetrameric inhibitor of insect α-amylases. The deduced amino acid sequences of the mature polypeptides are homologous to those of the dimeric and monomeric α-amylase inhibitors and of the trypsin inhibitors. The mature polypeptides are preceded by typical signal peptides. Southern blot analysis of appropriate aneuploids, using the cloned cDNAs as probes, has revealed the location of genes for subunits of the CM3 and of the CM16 type within a few kb of each other in chromosomes 4A, 4B and 4D, and those for the CM1 type of subunit in chromosomes 7A, 7B and 7D. Known subunits of the tetrameric inhibitor corresponding to genes from the B and D genomes have been previously characterized. No proteins of this class have been found to be encoded by the A genome in hexaploid wheat (genomes AA, BB, DD) or in diploid wheats (AA) and no anti α-amylase activity has been detected in the latter, so that the A-genome genes must be either silent (pseudogenes) or expressed at a much lower level.
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  • 43
    ISSN: 1573-5028
    Keywords: pathogen-induced ; peroxidase ; wheat
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    Notes: Abstract We report here the complete amino acid sequence of a pathogen-induced putative peroxidase from wheat (Triticum aestivum L.) as deduced from cDNA clones representing mRNA from leaves infected with the powdery mildew fungus Erysiphe graminis. The protein consists of 312 amino acids, of which the first 22 form a putative signal sequence, and has a calculated pI of 5.7. Sequence comparison revealed that the putative wheat peroxidase is most similar to the turnip (Brassica rapa) peroxidase, with which it shares 57% identical and 13% conserved amino acids.
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  • 44
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    Plant molecular biology 16 (1991), S. 831-839 
    ISSN: 1573-5028
    Keywords: RFLP ; Oryza ; rice ; genome evolution ; allotetraploid
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    Topics: Biology
    Notes: Abstract Restriction fragment length polymorphisms (RFLPs) were studied in fourteen accessions of CCDD genome allotetraploid wild rice species (Oryza latifolia, O. alta and O. grandiglumis). Fourteen nuclear RFLP markers previously mapped in AA genome-cultivated rice were used as probes. A phylogenetic tree, constructed by parsimony analysis based on RFLPs, grouped the accessions according to their geographic origin from Central or South America. Oryza alta, O. grandiglumis and one accession of O. latifolia grouped together as a subgroup, and our results suggested that the three taxa should be considered as populations of a single complex species. Duplicate loci, representing the two constituent genomes of the allotetraploid, were observed for most RFLP markers. By comparing RFLPs from the allotetraploids with those from a CC genome diploid wild species (O. officinalis), it was possible to detect RFLPs specific for both the CC and DD genomes of the allotetraploid. In inter-accession F2 populations, independent segregation of RFLP markers for CC and DD genomes was observed.
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  • 45
    ISSN: 1573-5028
    Keywords: peroxidase gene ; wheat
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    Notes: Abstract We have used a cDNA clone encoding a pathogen-induced putative wheat peroxidase to screen a genomic libary of wheat (Triticum aestivum L. cv. Cheyenne) and isolated one positive clone, lambda POX1. Sequence analysis revealed that this clone contains a gene encoding a putative peroxidase with a calculated pI of 8.1 which exhibits 58% and 83% sequence identity to the amino acid sequence of the turnip (Brassica rapa) peroxidase and a pathogen-induced putative wheat peroxidase, respectively. The two introns in the wheat gene are at the same positions as introns in the peroxidase genes of tomato and horseradish. Results of S1-mapping experiments suggest that this gene is neither pathogen-nor wound-induced in leaves but is constitutively expressed in roots.
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  • 46
    ISSN: 1573-5028
    Keywords: chromosome assignment ; dihaploid progenies ; heterozygosity ; potato ; rDNA ; RFLP
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    Notes: Abstract Restriction map and organization of rDNA was inferred from analysis of dihaploid progenies of two tetraploid genotypes of cultivated potato. Each tetraploid genotype was characterized by a specific distribution of different types of rDNA repetition units on their four homologous chromosomesII. The genotypes were heterozygous and differed by the kind of units carried by each chromosomeII. Models for the generation of the observed organization are discussed and supported by first cloning studies.
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  • 47
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    Plant molecular biology 16 (1991), S. 1073-1076 
    ISSN: 1573-5028
    Keywords: abscisic acid ; dehydration ; LEA ; water stress ; wheat
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    Topics: Biology
    Notes: Abstract A cDNA clone (pMA2005) of a Group 3 LEA (late embryogenesis abundant) protein has been sequenced from wheat. The wheat cDNA clone codes for a protein with ten tandem repeats of an 11 amino acid sequence and has homology to other Group 3 LEAs reported in barley, carrot, cotton and rape (L. Dure et al., Plant Mol Biol 12: 475–486, 1989). The deduced amino acid sequence indicates that the wheat protein has a molecular weight of 23 000 and is a basic, hydrophilic protein. Northern analysis with the cDNA clone shows that dehydration of wheat shoot tissue results in increased transcript levels that correlate with increases in endogenous ABA.
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  • 48
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    Plant molecular biology 26 (1994), S. 535-539 
    ISSN: 1573-5028
    Keywords: cytoplasmic male sterility ; coxI ; mitochondria ; membrane protein ; wheat
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    Topics: Biology
    Notes: Abstract Mitochondria derived from Triticum timopheevi have a chimeric gene, orf256, immediately upstream from coxI. Antibodies to a peptide corresponding to a part of the encoded amino acid sequence of orf256 detect a 7 kDa protein on western blots of mitochondrial proteins from cytoplasmic male-sterile (cms) wheat (T. aestivum nucleus, T. timopheevi mitochondria) but not in mitochondrial proteins from T. aestivum, T. timopheevi, or cms plants restored to fertility by introduction of nuclear genes for fertility restoration. The 7 kDa protein appears to serve as a marker for cms wheat. Its occurrence as an integral protein of the inner membrane may indicate a cms effect through an influence on mitochondrial membrane function.
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  • 49
    ISSN: 1573-5028
    Keywords: chromosome mapping ; inhibitors of trypsin/α-amylases ; wheat
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    Notes: Abstract Amino acid sequences for three members (CMx1, CMx2, and CMx3) of a new subfamily of trypsin/α-amylase inhibitors in wheat have been deduced from the nucleotide sequences of the corresponding cDNAs. A cDNA clone encoding CMx1 was selected from a wheat developing endosperm library using a probe that encoded barley trypsin inhibitor BTI-CMe at low stringency. Sequences corresponding to CMx2 and CMx3 were obtained from cDNA amplified by the polymerase chain reaction. The three CMx sequences contain a premature stop codon after 363 nt, as well as a second stop codon at the same position as in BTI-CMe (nt 439–441). Southern analysis of DNAs from diploid, tetraploid, and hexaploid wheats, as well as from aneuploid lines, indicate that there is a single CMx locus in each of the three genomes of hexaploid wheat, respectively associated with chromosomal arms 4AS, 4BS, and 4DL. These genes are expressed early during endosperm development and not expressed at detectable levels in other tissues. Evolutionary implications are discussed.
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  • 50
    ISSN: 1573-5028
    Keywords: cDNA ; cyanogenesis ; β-glucosidase ; RFLP ; Trifolium repens
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    Topics: Biology
    Notes: Abstract The Li locus in white clover controls the presence of cyanogenic β-glucosidase (linamarase) activity in leaf tissue, such that plants homozygous for the ‘null’ allele (li) have no linamarase activity in this tissue. The isolation of a cDNA clone from linamarase mRNA is described. The cDNA clone is used to further characterise alleles of the Li locus. Northern blot analysis shows that plants homozygous for the ‘null’ allele (li li) produce very reduced levels of mRNA which hybridises to the cDNA. Heterozygous plants (Li li), which have intermediate levels of enzyme activity, produce intermediate levels of mRNA. Southern blot analysis of Hind III digested genomic DNA shows that the white clover genome contains three genes with homology to the linamarase cDNA and that at least two of these genes segregate independently. Analysis of the cosegregation of linamarase activity and the presence of genomic restriction fragments identifies the genomic sequence specifying linamarase structure and indicates either a structural or cis acting control function of the Li locus.
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  • 51
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    Plant molecular biology 15 (1990), S. 793-795 
    ISSN: 1573-5028
    Keywords: mitochondrial gene ; ORF25 ; wheat
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    Topics: Biology
    Notes: Abstract The wheat mitochondrial orf25 nucleotide sequence of 576 pb has been determined. Its derived protein sequence shares 88% and 75% amino acid identity with those of maize and tobacco mitochondria, respectively. The wheat and tobacco orf25 sequences lack four inserts, of 6 bp to 36 bp, that are present in the maize homologue. The wheat orf25 gene is actively transcribed and is preceded by a regulatory sequence block very similar to those located upstream of the wheat coxII and atp6 genes. Our observations support the view that orf25 sequences encode a functional polypeptide in plant mitochondria.
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  • 52
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    Plant molecular biology 16 (1991), S. 663-670 
    ISSN: 1573-5028
    Keywords: gene expression ; proline-rich protein ; wheat
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    Topics: Biology
    Notes: Abstract A cDNA (WPRP1) encoding a wheat proline-rich protein has been isolated and sequenced. The amino acid composition shows 45% proline, with high levels of methionine, lysine and glutamic acid. The derived 378 residue amino acid sequence has a highly repetitive structure which is unlike those of other proline-rich proteins. The WPRP1 cDNA clone was used to determine the copy number and chromosomal location of the WPRP1 gene by restriction fragment length polymorphism analysis of wheat inbred lines. Although WPRP1 is encoded by a single-copy gene it is also a representative of a larger family of related sequences. RNA gel blot analysis showed that expression of WPRP1 is highest in rapidly growing tissue which together with its amino acid composition suggests a structural role for the encoded protein.
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  • 53
    ISSN: 1573-5028
    Keywords: wheat ; glutathione-S-transferase ; transposon-like sequence
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  • 54
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    Plant molecular biology 16 (1991), S. 1099-1101 
    ISSN: 1573-5028
    Keywords: wheat ; Triticum aestivum ; cDNA clone ; waxy protein
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  • 55
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    Plant molecular biology 17 (1991), S. 167-168 
    ISSN: 1573-5028
    Keywords: chloroplast ; monocot gene ; phosphoribulokinase ; wheat
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  • 56
    ISSN: 1573-5028
    Keywords: Tahsp26.6 ; chloroplast-localized HSP ; Triticum aestivum L. ; wheat
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  • 57
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    Plant molecular biology 22 (1993), S. 25-41 
    ISSN: 1573-5028
    Keywords: gliadin storage proteins ; wheat ; transcriptional regulation ; nuclear proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The promoter region (−524 to −46) of the wheat α/β-gliadin seed storage protein gene was analyzed for interactions with nuclear proteins from developing wheat seeds. Six complexes were detected within the first 165 bp upstream of the transcriptional start site. One of the proteins was a non-sequence specific AT-binding protein. The remaining five proteins bound in a sequence specific manner. One (CABP) mapped to a conserved CA-rich element at −134 to −112 while another (PalBP) mapped to an adjacent, palindromic sequence at −112 to −106. Three proteins (CTBPs 1–3) formed complexes at two, independent homologous sites. The activities of four of the binding proteins, CTBPs 1–3 and CABP, exhibited similar patterns of expression during seed development: they first appeared at early to mid stages, reached a maximum at mid stage and subsequently decreased, paralleling the pattern of gliadin mRNA accumulation. The non-specific AT-binding protein was detected at relatively high levels only at mid development. PalBP activity, on the other hand, first appeared at mid stage and was present at a constant level throughout later stages of development. The results suggest that the binding proteins may regulate gliadin expression in an antagonistic manner.
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  • 58
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    Plant molecular biology 22 (1993), S. 437-446 
    ISSN: 1573-5028
    Keywords: RFLP ; genome ; PFGE ; multicopy markers ; symbiosis
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    Topics: Biology
    Notes: Abstract Pulsed-field gel electrophoresis (PFGE) was used to study a cluster of molecular markers in the soybean genome. There were 550 kb per centimorgan (cM) in the cluster, which is close to the calculated average for the whole genome. The analysis was complicated by the presence of duplicated sequences, and some ambiguities arising from this were resolved by using second-dimension conventional electrophoresis to relate physical maps to the RFLP map of soybean. The results show that there is a high degree of conservation of ‘rare cutter’ sites between homoeologous regions. Finally, PFGE can confirm physical linkage of monomorphic copies of markers, which can aid in the study and comparison of homoeologous regions that are invisible to RFLP analysis.
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  • 59
    ISSN: 1573-5028
    Keywords: Calvin cycle genes ; DNA-binding proteins ; wheat
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    Notes: Abstract We have characterised a DNA-binding interaction common to the upstream sequences of the wheat fructose-1,6-bisphosphatase (FBPase) and sedoheptulose-1,7-bisphosphatase (SBPase) genes. The recognition site for this sequence-specific binding activity, designated wheat FBPase factor (WF-1), is located within 125 bp of the transcription start site of each gene. Within these regions there are no sequence motifs similar to those shown to be important for light-regulated expression in other species. The binding activity was not detected in wheat root nuclear extracts, or in pea leaf extracts. There was a higher level of binding activity in light-grown than in dark-grown wheat leaves. The level was also found to decline when light-grown plants were given an extended dark treatment, but could be reinduced by light. Utilising the gradient of developmental maturity which exists within the wheat leaf it was found that WF-1 activity increases during leaf development.
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  • 60
    ISSN: 1573-5028
    Keywords: TATA box ; TFIID ; transcription factor ; wheat
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    Notes: Abstract We isolated a complementary DNA (cDNA) encoding the TATA-binding factor ‘TFIID’ from a wheat seedling cDNA library. The wheat TFIID transcript of 1.2 kb poly(A)+ RNA was expressed at a low level early in germination, but gradually increased as the seedlings developed. In vitro binding experiments showed that the bacterially expressed wheat TFIID protein could specifically bind to the TATA boxes of the cauliflower mosaic virus (CaMV) 35S, wheat histone H3 and adenovirus major late genes with different affinity. A comparison with Arabidopsis TFIID showed the presence of a plant-specific region consisting of 13 amino acids at the divergent amino terminus and a conserved region (182 amino acids) at the carboxy terminus longer than that observed in yeasts (180 amino acids) and animals (181 amino acids).
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  • 61
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    Keywords: rRNA ; PCR ; ITS ; DNA sequence ; nucleotide ; Triticum speltoides ; wheat
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  • 62
    ISSN: 1573-5028
    Keywords: rRNA ; PCR ; ITS ; DNA sequence ; nucleotide ; Triticum aestivum ; wheat
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  • 63
    ISSN: 1573-5028
    Keywords: mitochondrial DNA ; repeated sequences ; ribosomal RNA ; t-elements ; Triticum aestivum ; wheat
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    Topics: Biology
    Notes: Abstract We report the sequence of a 7.2 kilobase pair DNA fragment containing a copy of the wheat mitochondrial gene (rrn26) that encodes the mitochondrial large-subunit ribosomal RNA (26S rRNA). The mature 26S rRNA was determined by direct RNA sequencing to be 3467 nucleotides long, and to share a 5′-terminal pentanucleotide (5′-AUCAU), thought to be important in post-transcriptional processing, with the wheat mitochondrial small-subunit (18S) rRNA. Two other prominent features of the sequence were noted. First, upstream of rrn26 are located two tandem copies of a 70 base pair element containing a putative mitochondrial promoter motif (TCGTATAAAAA). Second, downstream of rrn26 is a sequence element that, if transcribed, would produce and RNA with a secondary structure resembling that of tRNAs but differing sufficiently from the latter structure to preclude any transcript from functioning normally in translation. These upstream and downstream sequence elements may play a role in the expression of rrn26 in wheat mitochondria.
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  • 64
    ISSN: 1573-5028
    Keywords: apocytochrome b pseudogene ; pea cox1 ; plant mitochondria ; potato ; S10 ribosomal protein ; wheat
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    Notes: Abstract The S10 ribosomal protein gene (rps10), which has not been previously reported in any angiosperm mitochondrial genome, was identified by sequence analysis in the potato mitochondrial DNA. This gene is found downstream of a truncated non-functional apocytochrome b (cob) pseudogene, and is expressed as multiple transcripts ranging in size from 0.8 to 5.0 kb. Southern hybridization analysis indicates that rps10-homologous sequences are not present in the wheat mitochondrial genome. Sequence analysis of a single-copy region of the pea mitochondrial genome located upstream of cox1 [11] shows that a non-functional rps10 pseudogene is present in this species. These results suggest that the functional genes coding for wheat and pea mitochondrial RPS10 polypeptides have been translocated to the nucleus.
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  • 65
    ISSN: 1573-5028
    Keywords: chloroplast ; gene expression ; photosystem 2 ; transcription ; wheat
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    Notes: Abstract The time course of the accumulation of the transcripts from 13 psb genes encoding a major part of the proteins composing photosystem II during light-induced greening of dark-grown wheat seedlings was examined focusing on early stages of plastid development (0.5 h through 72 h). The 13 genes can be divided into three groups. (1) The psbA gene is transcribed as a single transcript of 1.3 kb in the dark-grown seedlings, but its level increases 5- to 7-fold in response to light due to selective increase in RNA stability as well as in transcription activity. (2) The psbE-F-L-J operon, psbM and psbN genes are transcribed as a single transcript of 1.1 kb, two transcripts of 0.5 and 0.7 kb and a single transcript of 0.3 kb, respectively, in the dark-grown seedlings. The levels of accumulation of every transcript remain unchanged or rather decrease during plastid development under illumination. (3) The psbK-I-D-C gene cluster and psbB-H operon exhibit fairly complicated northern hybridization patterns during the greening process. When a psbC or psbD gene probe was used for northern hybridization, five transcripts differing in length were detected in the etioplasts from 5-day old dark-grown seedlings. After 2 h illumination, two new transcripts of different length appeared. Light induction of new transcripts was also observed in the psbB-H operon.
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  • 66
    ISSN: 1573-5028
    Keywords: cDNA sequence ; cystine-rich proteins ; gene expression ; puroindolines ; tryptophan-rich domain ; wheat
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    Notes: Abstract From a mid-maturation seed cDNA library we have isolated cDNA clones encoding two Triticum aestivum puroindolines. Puroindoline-a and puroindoline-b, which are 55% similar, are basic, cystine-rich and tryptophan-rich proteins. Puroindolines are synthezised as preproproteins which include N- and C-terminal propeptides which could be involved in their vacuolar localization. The mature proteins have a molecular mass of 13 kDa and a calculated isoelectric point greater than 10. A notable feature of the primary structure of puroindolines is the presence of a tryptophan-rich domain which also contains basic residues. A similar tryptophan-rich domain was found within an oat seed protein and a mammalian antimicrobial peptide. The ten cysteine residues of puroindolines are organized in a cysteine skeleton which shows similarity to the cysteine skeleton of other wheat seed cystine-rich proteins. Northern blot analysis showed that puroindoline genes are specifically expressed in T. aestivum developing seeds. No puroindoline transcripts as well as no related genes were detected in Triticum durum. The identity of puroindolines to wheat starch-granule associated proteins is discussed as well as the potential role of puroindolines in the plant defence mechanism.
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  • 67
    ISSN: 1573-5028
    Keywords: alcohol dehydrogenase ; cDNA ; enzyme ; tepary ; stability
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    Topics: Biology
    Notes: Abstract The zinc-binding long-chain alcohol dehydrogenases from plants and animals exhibit a considerable level of amino acid sequence conservation. While the functional importance of many of the conserved residues is known, the role of others has not yet been determined. We have identified a naturally occurring Adh-1 allele in the legume Phaseolus acutifolius with several unusual characteristics. Individuals homozygous for this allele, Adh-1CN, possess a single isozyme starch gel electrophoretic pattern suggestive of a null allele, and exhibit ADH enzyme activity levels ca. 60% lower than the standard wild-type Adh-1F line. Interestingly, analysis of Adh-1CN homozygotes on an alternative gel system indicates that Adh-1CN does encode a polypeptide capable of forming functional homo- and heterodimers. However, the levels of ADH activity displayed by these isozymes are far lower than those observed for the corresponding wild type ADH-1F isozymes. Dialysis experiments indicate that isozymes containing the ADH-1CN polypeptide are inactivated by slightly acidic conditions, which may explain the apparent null phenotype on starch gels. Elevated temperatures cause a similar loss of enzyme activity. The deduced amino acid sequences of ADH-1CN and ADH-1F were obtained from their corresponding cDNA clones, and the only significant difference detected between the two is a single amino acid replacement substitution. Residue 144 is occupied by phenylalanine in the ADH-1F polypeptide, whereas serine occupies this position in the ADH-1CN polypeptide. The proximity of residue 144 to the catalytic zinc in the substrate-binding pocket, coupled with the fact that it is integral to a defined hydrophobic core of the ADH polypeptide, may explain the observed disruptive effect that the serine substitution has on both the activity and stability of the ADH-1CN polypeptide. It also provides an explanation for the maintenance of phenylalanine or the structurally similar tyrosine at this residue in Zn-binding long-chain ADHs.
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    Journal of mathematical biology 32 (1994), S. 395-426 
    ISSN: 1432-1416
    Keywords: Uniform persistence ; stability ; Lyapunov functional ; level-crossing
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    Topics: Biology , Mathematics
    Notes: Abstract Dynamical characteristics of an integrodifferential system modelling two species competition with hereditary effects are investigated; in particular we derive sufficient conditions for the persistence of the species, existence of an attracting periodic solution and ‘level-crossings’ of solutions about the periodic solution.
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    Journal of mathematical biology 32 (1994), S. 515-520 
    ISSN: 1432-1416
    Keywords: Gametophytic incompatibility ; model ; equilibrium ; stability
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    Topics: Biology , Mathematics
    Notes: Abstract The deterministic dynamics of the classical single-locus multiple-allele model of gametophytic incompatibility is analyzed with the intention to prove the conjecture that the symmetric state (uniform distribution of genotypes) is the only polymorphic equilibrium and that this equilibrium is globally asymptotically stable in the interior of the frequency simplex. It is shown that the minimum allelic frequency increases strictly over the generations as long as a uniform allelic distribution is not realized. Hence, the minimum allelic frequency is a Ljapunov function for the invariant set of genotypic frequencies characterized by a uniform allelic distribution. Within this set, the uniform genotypic distribution is approached in an exponential fashion, which proves the assertion. An evolutionary optimization rule associated with the global convergence to the symmetric state is implied by the fact that at this state the overall amount of pollen elimination resulting from incompatible crosses is minimized.
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    Theoretical and applied genetics 79 (1990), S. 577-582 
    ISSN: 1432-2242
    Keywords: Genetic marker ; RFLP ; Quantitative traits
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    Topics: Biology
    Notes: Summary Digestion of genomic DNA with the restriction endonuclease Avail disclosed a probable insertion deletion of approximately 200 base pairs (bp) near the prolactin gene. Two alleles were apparent as three distinct hybridization patterns. These alleles were statistically associated with quantitative trait loci among sons of one elite Holstein sire family. The favorable genotype was correlated with the presence of a 1.15-kb hybridization band inherited from the sire when genomic DNA was probed with a full-length cDNA for prolactin. Pedigree estimates of genetic merit among genotypes were similar, differing by only 19.3 kg for milk in ancestor merit. Comparisons of genetic estimates for quantitative yield traits in offspring of this heterozygous sire showed significant (P〈0.05) differences between homozygous genotypes for predicted difference milk (PDM), predicted difference dollars (PD$), cheese yield dollars, and protein dollars. The estimated differences between homozygous genotypes for USDA Transmitting Abilities of PDM, PD$, Cheese Yield $ and Protein $ were 282.93 kg, $74.35, $48.58 and $53.67, respectively. However, the estimated breeding values from progeny ranged over 900 kg in transmitting ability for milk. Frequency of the favorable marker allele was estimated to be 0.231 in the elite cow population used as dams of sons. These results demonstrate the potential of molecular biological techniques to discriminate between individuals within a family and to predict breeding values for selection schemes.
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    Theoretical and applied genetics 80 (1990), S. 673-679 
    ISSN: 1432-2242
    Keywords: Rice ; Tissue culture ; Somaclonal variation ; RFLP ; Methylation
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    Topics: Biology
    Notes: Summary Regenerants of rice were examined by RFLP analysis to determine the occurrence and extent of somaclonal variation. DNA polymorphisms were observed both among plants regenerated from different callus cultures as well as among sibling plants derived from a single callus. Regardless of the basal medium, a higher degree of genetic instability was found among plants regenerated from callus cultures maintained for longer incubation periods (67 days) than among those from shorter incubation periods (28 days). Detailed analysis showed that in several regenerants, there was a close correlation among those plants exhibiting DNA rearrangements and those with apparent methylation changes. Such alterations were observed with both structural and housekeeping genes.
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    Theoretical and applied genetics 80 (1990), S. 680-686 
    ISSN: 1432-2242
    Keywords: Retrotransposon ; Zea ; RFLP ; Bs1 ; Transposable element
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    Topics: Biology
    Notes: Summary Thirty-eight accessions from Zea and 20 accessions from related genera were probed for the presence of Bs1, a retrotransposon originally found in maize. All maize and teosinte plants tested show the presence of Bs1 in one to five densely hybridizing bands. The mean copy numbers of Bs1 elements among the maize and teosinte accessions were similar: 2.92 and 3.25, respectively, with no large differences between any subgroups. Most exotic maize samples exhibited two common bands of 7.8 kb and 4.7 kb. Section Zea teosintes (but not teosintes of section Luxuriantes) also show the presence of a common band of the same size as the smaller common band in maize. At reduced stringency, Tripsacum dactyloides exhibited a single hybridizing band at 6.9 kb. Results argue for the evolution of maize from a mexicana or parviglumis teosinte, and the evolution of the Bs1 element within the tribe Andropogoneae. Additionally, recombinant inbred lines were probed for the presence of Bs1, in order to map the chromosomal locations of Bs1 elements in four different maize lines. Two of the recombinant inbred parental lines had an element (Bs1-F) on chromosome 5, while the other two lines had an element (Bs1-S) on chromosome 8. Restriction site polymorphisms have apparently arisen in the vicinity of Bs1-S since its insertion. Segregation analysis of other lines was also performed; the data indicate that Bs1 has the distribution expected of a transposable element, different locations in different lines, and not that of a fixed gene locus. However, the common bands in the Zea mays lines and the recombinant inbred data imply that Bs1 is not highly mobile.
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    Theoretical and applied genetics 82 (1991), S. 393-398 
    ISSN: 1432-2242
    Keywords: Maize ; Helminthosporium turcicum race1 ; RFLP ; NILs ; Mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have identified tight linkage of an RFLP marker to theHt1 gene of maize that confers resistance to the fungal pathogenHelminthosporium turcicum race 1. This was accomplished by the use of four pairs of near isogenic lines (NILs; B73, A619, W153R, and CM105), each differing by the presence or the absence of the geneHt1. SinceHt1 maps to chromosome 2, 26 clones already mapped to this chromosome were labeled and probed against Southern blots of these NILs DNA digested with three restriction enzymes:EcoRI,BamHI, andHindIII. Six markers exhibited an RFLP for at least one pair of NILs. Presumptive linkage was further tested by analyzing the segregation of five of the six markers (one was monomorphic in the cross studied) and resistance toH. turcicum race 1 on 95 F2 individuals from the cross DF20 × LH146Ht. The results indicate a tight linkage between one of the DNA markers,UMC150B, and theHt1 gene.
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    Theoretical and applied genetics 82 (1991), S. 697-703 
    ISSN: 1432-2242
    Keywords: RFLP ; Bamboo ; Phyllostachys ; Chloroplast DNA ; Germplasm screening
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Bamboo species are difficult to identify because flowering material is seldom available and taxonomy is of necessity based on vegetative characters. To evaluate the utility of restriction fragment length polymorphism (RFLP) analysis in bamboo systematics and germplasm screening, a library of random genomic probes from a Phyllostachys nigra PstI library was constructed. Probes from the library were used to screen bamboo germplasm consisting mostly of temperate bamboos of the genus Phyllostachys. RFLP variation was abundant, and species-specific patterns were readily obtained. Chloroplast DNA showed little variation among the bamboo accessions analyzed.
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  • 75
    ISSN: 1432-2242
    Keywords: RFLP ; PCR ; Rice ; Inheritance ; Nonradioactive
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    Topics: Biology
    Notes: Summary Thirty mapped Indica rice genomic (RG) clones were partially sequenced from each end. From such sequence data, pairs of oligonucleotides were synthesized to act as primers for polymerase chain reaction (PCR) amplification of the corresponding loci in crude total DNA preparations. The PCR products from DNA of Indica varieties were of the sizes expected from the sizes of the corresponding RG clones. However, size polymorphisms were seen between PCR products from Indica and Japonica varieties, and among wildOryza species. Restriction fragment length polymorphism (RFLP) was observed between PCR products of Indica varieties simply by electrophoretic analysis of restricted products, without the need for Southern hybridization or radiolabelling. The RFLPs noted between varieties ARC6650 and Phalguna were inherited in recombinant inbred lines derived from a cross between them. The RFLPs were detectable in PCR products amplified from DNA extracted by a simple procedure from single seedlings or leaves, and revealed genetic heterogeneity in cultivated lines. An approach is described that is relevant to the acceleration of classical plant breeding through molecular techniques.
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    Theoretical and applied genetics 83 (1991), S. 89-96 
    ISSN: 1432-2242
    Keywords: RFLP ; Alfalfa ; Genetic diversity ; Phylogenetic tree ; Gene duplication
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Alfalfa (Medicago sativa L.) is a major forage crop throughout the world. Although alfalfa has many desirable traits, continued breeding is required to incorporate pest resistances and other traits. We conducted this study to determine the amount of restriction fragment length polymorphism (RFLP) variability present within and between diploid and tetraploid alfalfa populations, and whether or not this variability is sufficient for construction of an RFLP map. Diploid plants from M. sativa ssp. falcata, ssp. coerulea, and ssp. sativa and tetraploid spp. sativa cultivars ‘Apollo,’ ‘Florida 77,’ and ‘Spredor 2’ were included. A total of 19 cDNA clones was probed onto genomic Southern blots containing DNA digested by EcoRI, HindIII, or BamHI. Phylogenetic trees were produced, based on parsimony analysis of shared restriction fragments. Evidence for extensive gene duplication was found; most probes detected complex patterns of restriction fragments. Large amounts of variation are present within all diploid subspecies. M. sativa ssp. falcata plants formed clusters distinct from ssp. sativa or ssp. coerulea plants, which were not distinctly clustered. Some M. sativa ssp. falcata plants were more similar to the other groups than to other plants within ssp. falcata. Variation among tetraploid cultivars showed that Florida 77 and Apollo had more similarities than either showed with Spredor 2. All three cultivars showed large within-population variation, with Apollo being the most diverse and Spredor 2 the least. Based on these results, development of an RFLP map at the diploid level appears possible. Also, differentiation of cultivars, particularly ones of divergent origin, seems possible based on RFLP patterns.
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    Theoretical and applied genetics 83 (1992), S. 428-434 
    ISSN: 1432-2242
    Keywords: ‘Null’ forms ; RFLP ; Gliadins ; LMW glutenins ; Deletion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Wheat accessions lacking some of the ω- and γ-gliadin components encoded by the Gli-1 loci on the short arm of chromosome 1D in bread wheat and chromosome 1A in durum wheat were studied by two-dimensional polyacrylamide gel electrophoresis and restriction fragment analysis. Digested genomic DNAs of ‘normal’ and ‘null’ forms were probed with a cDNA clone related to ω-/γ-gliadins and with a genomic clone encoding an LMW subunit of glutenin. The hybridisation patterns with the ω-/γ-gliadin probe were similar to those of cvs ‘Chinese Spring’ and ‘Langdon’ used as standards for bread and durum wheats, respectively, but several restriction fragments located on the 1D chromosome of bread wheat and the 1A chromosome of durum wheat were absent in the ‘null’ forms. In addition, specific LMW glutenin fragments encoded by the same chromosomes were also absent in the ‘null’ forms, suggesting that simultaneous deletions of blocks of genes for both ω-/γ-gliadins and LMW glutenins had occurred. Comparisons of the protein and RFLP patterns enabled some proteins to be mapped to specific restriction fragments.
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    Theoretical and applied genetics 83 (1992), S. 895-902 
    ISSN: 1432-2242
    Keywords: Finger millet ; Eleusine ; rDNA ; RFLP ; Grasses
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Finger millet is an important cereal crop in the semi-arid regions of Africa and India. The crop belongs to the grass genus Eleusine, which includes nine annual and perennial species native to Africa except for the New World species E. tristachya. Ribosomal DNA (rDNA) variation in finger millet and related wild species was used to provide information on the origin of the genomes of this tetraploid crop and point out genetic relationships of the crop to other species in the genus. The restriction endonucleases used revealed a lack of variability in the rDNA spacer region in domesticated finger millet. All the rDNA variants of the crop were found in the proposed direct tetraploid ancestor, E. coracana subsp. africana. Wild and domesticated finger millet displayed the phenotypes found in diploid E. indica. Diploid Eleusine tristachya showed some similarity to the crop in some restriction sites. The remaining species were quite distinct in rDNA fragment patterns. The study supports the direct origin of finger millet from subspecies africana shows E. indica to be one of the genome donors of the crop, and demonstrates that none of the other species examined could have donated the second genome of the crop. The rDNA data raise the possibility that wild and domesticated finger millet could have originated as infraspecific polyploid hybrids from different varieties of E. indica.
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    Theoretical and applied genetics 84 (1992), S. 39-48 
    ISSN: 1432-2242
    Keywords: Genetic map ; Molecular markers ; RFLP ; Fruit breeding ; Citrus spp.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Genetic linkage analysis was performed using two segregating populations of citrus. One population arose from an intergeneric backcross of Citrus grandis (L.) Osb. cv ‘Thong Dee’ and Poncirus trifoliata (L.) Raf. cv ‘Pomeroy’, using the former as the recurrent (female) parent. The other population came from an interspecific backcross of C. reticulata Blanco cv ‘Clementine’ and C. x paradisi Macf. cv ‘Duncan’, using the former as the recurrent (male) parent. A total of 11 isozyme and 58 restriction fragment length polymorphisms were found to segregate in a monogenic fashion in one or both populations. Linkage analysis revealed that 62 of the loci examined mapped to 11 linkage groups, while 7 loci segregated independently from all other markers. Gene order was highly conserved between the maps generated from the two divergent segregating populations. Possible applications of the use of such maps in tree fruit breeding are discussed.
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    Theoretical and applied genetics 84 (1992), S. 113-117 
    ISSN: 1432-2242
    Keywords: Beet ; Somaclonal variation ; Isozyme ; RFLP ; Genetic stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Sugar, fodder and garden beet (Beta vulgaris L.) plants have been regenerated in culture from a range of expiant material. Of the regenerants 764 were subjected to isozyme analysis using eight enzyme-specific stains, and 60 were subjected to RFLP analysis using three cDNA probes. Both molecular techniques allowed the identification of somaclonal variant plants. Assessment of the numbers of variant isozymes and restriction fragments has allowed the calculation of the approximate percentage of variant alleles occurring in any one somaclonal regenerant, namely between 0.05% and 0.1%.
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  • 81
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    Theoretical and applied genetics 84 (1992), S. 286-290 
    ISSN: 1432-2242
    Keywords: Taxonomy ; Banana ; RFLP ; Musa ; Phylogeny
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Random genomic probes were used to detect RFLPs in 19 Musa species and subspecies. A total of 89 phylogenetically informative alleles were scored and analyzed cladistically and phenetically. Results were in general agreement with morphology-based phylogenetic analyses, with the following exceptions: our data unambiguously places M. boman in section Australimusa, and indicates M. beccarii is very closely related to M. acuminata. Additionally, no support was found for the separation of section Rhodochlamys from section Musa. A comparison of morphology-based and RFLP-based phylogenetic analyses is presented.
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  • 82
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    Theoretical and applied genetics 84 (1992), S. 845-850 
    ISSN: 1432-2242
    Keywords: Flax ; Rust ; RFLP ; Genetic segregation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Random cDNA sequences synthesized from poly A+ RNA extracted from germinated urediospores of the flax rust fungus, Melampsora lini, were used as probes to detect restriction fragment length polymorphisms (RFLPs) in three races of M. lini originating from cultivated flax, Linum usitatissimum, and one race originating from Australian native flax, L. marginale. Fourteen out of 22 probes tested detected RFLPs in the three races from cultivated flax while 19 of the probes detected polymorphisms between these three races and the race from L. marginale. The segregation of seven RFLPs was determined in a family of 19 F2 progeny derived from a cross between two of the rust races. With six of these the inheritance was consistent, in each case, with the segregation of alleles at a single locus. Inheritance of the seventh was unusual and an explanation involving two loci with null alleles at each was proposed. No linkage was detected between any of the RFLP loci and nine unlinked loci specifying avirulence.
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  • 83
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    Theoretical and applied genetics 85 (1992), S. 101-104 
    ISSN: 1432-2242
    Keywords: Marker assisted selection ; RFLP ; QTL ; Quantitative genetics ; Corn breeding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The objective of this paper is to present genetic theory demonstrating the conditions under which it should be possible to identify molecular marker-quantitative trait locus (QTL) associations in crosses of random-mating populations to inbreds. Using as an example the cross of a corn (Zea mays L.) population to an inbred, the expected disequilibrium for testcross and per se performance of F2, F3, BC1 (to the inbred) and recombinant inbred generations was derived for cases where a marker allele is linked to an unfavorable QTL allele in the inbred and where the marker allele is linked to a favorable QTL allele in the inbred. Disequilibrium in segregating generations was shown to be a function of disequilibrium in the parent population, the frequency of marker and QTL alleles in the parent population, and the recombination distance between the marker and the QTL. To maximize the opportunity to identify a favorable QTL the following procedures are suggested: (1) Select marker loci with alleles in the parent population which are not present in the inbred. (2) Select populations known to have favorable QTL alleles not present in the inbred. (3) Use as many marker loci as possible to enhance the probability of tight linkage between the marker and the QTL.
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  • 84
    ISSN: 1432-2242
    Keywords: Pea ; Pea seed-borne mosaic virus ; Disease resistance ; Genome mapping ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The location of sbm-1 on the Pisum sativum genetic map was determined by linkage analysis with eight syntenic molecular markers. Analysis of the progeny of two crosses confirmed that sbm-1 is on chromosome 6 and permitted a more detailed map of this chromosome to be constructed. The inclusion of Fed-1 and Prx-3 among the markers facilitated the comparison of our map with the classical genetic map of pea. The sbm-1 gene is most closely linked to RFLP marker GS185, being separated by a distance of about 8 cM. To determine the practical value of GS185 as a marker for sbm-1 in plant breeding programs, the GS185 hybridization pattern and virus-resistance phenotype were compared in of a collection of breeding lines and cultivars. Three GS185 hybridization patterns were discerned among the lines. A strong association was found between one of these patterns and resistance to PSbMV.
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  • 85
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    Theoretical and applied genetics 86 (1993), S. 181-188 
    ISSN: 1432-2242
    Keywords: Sugarcane ; Gramineae ; Tissue culture ; RFLP ; Molecular analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The genomic stability of tissue culture regenerants of sugarcane (Saccharum spp. hybrids, cvs ‘CP721210’, ‘CP68-1067’ and ‘B43-62’) was analyzed by DNA restriction fragment length polymorphism (RFLP). Plants regenerated from calli, cell suspensions, cryopreserved cell suspensions and protoplasts were used. Total DNA isolated from 19 different sources was digested with EcoRI, HindIII, BamHI, BamHI, EcoRI and PstI and probed with six known maize mitochondrial genes (coxI, coxII, atpA, atp6, atp9 and rrn18-rrn5), three random maize mitochondrial cosmid clones, two random maize chloroplast cosmid clones and a wheat Nor locus clone. Hybridization patterns indicated that the variation observed was minor and appeared only in the secondcycle regenerants. No differences were observed among the three cultivars and the regenerants from calli, suspension culture, cryopreserved suspension culture and protoplasts. Mitochondrial DNA (mtDNA) isolated from ‘CP72-1210’ plants and its embryogenic cell suspensions, and bulk samples from all ‘CP72-1210’ regenerants pooled together were digested with EcoRI, HindIII, PstI, BamHI and SalI and probed with three recombinationally active wheat mtDNA clones, K′, K3 and X2. No variation in the mtDNA restriction patterns was observed between the ‘CP72-1210’ plants and its regenerants. However, restriction pattern variation was observed only from EcoRI digestion, and hybridization patterns of K3, K′ and X2 revealed minor variations in the mtDNA of cell suspensions when compared with the DNA of the ‘CP72-1210’ plant. Except for a qualitative variation detected by the X2 probe and minor stoichiometric variations detected by the K3 probe, sugarcane DNAs were found to be stable after plant regeneration.
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  • 86
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    Theoretical and applied genetics 86 (1993), S. 301-307 
    ISSN: 1432-2242
    Keywords: RFLP ; Polyploid ; Nondisjunction ; Interspecific hybrid ; Cottonwood
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary While constructing a genetic linkage map of a hybrid poplar genome (Populus trichocarpa x P. deltoides), we identified several restriction fragment length polymorphismus (RFLPs) for which the parental trees are heterozygous. Although 8 of the 11 F1 hybrid offspring inherited, as expected, single RFLP alleles from each parent, 3 F1 trees in the mapping pedigree inherited both maternal alleles along with a single paternal allele at some loci. Aneuploidy or polyploidy in these 3 F1 trees due to partial or complete nondisj unction during female gametogenesis is the simplest explanation for this finding. Of the 3 f1 offspring with supernumerary RFLP alleles 2 have triploid nuclear DNA contents as measured by fluorescence flow cytometry; the 3rd F1 with supernumerary alleles has a sub-triploid nuclear DNA content and is probably aneuploid. Among the tri/aneuploid hybrids, leaf quantitative traits either are skewed toward those values characteristic of the P. trichocarpa female parent (adaxial stomate density, petiole length: blade length ratio; abaxial color) or show transgressive variation (epidermal cell size). Abaxial leaf color was used to screen a large population of P. trichocarpa x P. deltoides hybrids for further evidence of tri/aneuploidy. In each case where a “white” abaxial leaf surface was observed and the nuclear DNA content measured, the hybrid proved to be tri/aneuploid. All sexually mature female triploids examined were sterile, although the inflorescences completed their development in the absence of embryo formation. The (probably) aneuploid F1 hybrid is a fertile female. Of 15 female P. trichocarpa parents used in crosses to P. deltoides, 10 produced one or more tri/aneuploid hybrid offspring. In an intraspecific cross using a P. trichocarpa female that had produced triploid hybrids with five different P. deltoides males, no tri/aneuploid offpsring were found.
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  • 87
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    Theoretical and applied genetics 86 (1993), S. 329-332 
    ISSN: 1432-2242
    Keywords: RFLP ; Alfalfa ; Genetic map ; Segregation distortion ; Plant breeding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have developed a restriction fragment length polymorphism (RFLP) linkage map in diploid alfalfa (Medicago sativa L.) to be used as a tool in alfalfa improvement programs. An F2 mapping population of 86 individuals was produced from a cross between a plant of the W2xiso population (M. sativa ssp. sativa) and a plant from USDA PI440501 (M. sativa ssp. coerulea). The current map contains 108 cDNA markers covering 467.5 centimorgans. The short length of the map is probably due to low recombination in this cross. Marker order may be maintained in other populations even though the distance between clones may change. About 50% of the mapped loci showed segregation distortion, mostly toward excess heterozygotes. This is circumstantial evidence supporting the maximum heterozygote theory which states that relative vigor is dependent on maximizing the number of loci with multiple alleles. The application of the map to tetraploid populations is discussed.
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  • 88
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    Theoretical and applied genetics 86 (1993), S. 811-821 
    ISSN: 1432-2242
    Keywords: Brassica ; Interspecific hybridization ; Amphidiploid ; RFLP ; Evolution ; Nuclear ; cytoplasmic interaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In a previous study we proposed that cytoplasmic genomes have played an important role in the evolution of Brassica amphidiploid species. Based on this and other studies, we hypothesized that interactions between the maternal cytoplasmic genomes and the paternal nuclear genome may cause alterations in genome structure and/or gene expression of a newly synthesized amphidiploid, which may play an important role in the evolution of natural amphidiploid species. To test this hypothesis, a series of synthetic amphidiploids, including all three analogs of the natural amphidiploids B. napus, B. juncea, and B. Carinata and their reciprocal forms, were developed. These synthetic amphidiploids were characterized for morphological traits, chromosome number, and RFLPs revealed by chloroplast, mitochondrial, and nuclear DNA clones. The maternal transmission of chloroplast and mitochondrial genomes was observed in all of the F1 hybrids examined except one hybrid plant derived from the B. rapa x B. oleracea combination, which showed a biparental transmission of organelles. However, the paternal chloroplast and mitochondrial genomes were not observed in the F2 progeny. Nuclear genomes of synthetic amphidiploids had combined RFLP patterns of their parental species for all of the nuclear DNA clones examined. A variation in fertility was observed among self-pollinated progenies of single amphidiploids that had completely homozygous genome constitutions. Comparisons between natural and synthetic amphidiploids based on restriction fragment length polymorphism (RFLP) patterns indicated that natural amphidiploids are considerably more distant from the progenitor diploid species than the synthetic amphidiploids. The utility of these synthetic amphidiploids for investigating the evolution of amphidiploidy is discussed.
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  • 89
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    Theoretical and applied genetics 86 (1993), S. 833-836 
    ISSN: 1432-2242
    Keywords: T. aestivum ; RFLP ; Fingerprint
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    Topics: Biology
    Notes: Abstract Two probes, specific for HMW-glutenins and γ-gliadins have been used to identify 50 common wheat Italian cultivars, most of which are closely related, and four common wheat cultivars originating outside Italy. The probes revealed complex polymorphic patterns; three probe/enzyme combinations had the necessary sensitivity for the identification of all 54 cultivars. As already shown for potato and barley, the use of four-cutter restriction enzymes and polyacrylamide gels proved particularly useful for detecting polymorphism.
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  • 90
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    Theoretical and applied genetics 86 (1993), S. 383-388 
    ISSN: 1432-2242
    Keywords: Mitochondria ; Inheritance ; RFLP ; Larix ; Conifers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Restriction fragment length polymorphisms between Larix leptolepis and Larix decidua were identified in heterologous hybridization experiments, using wheat mitochondrial DNA probes specific for atp9, coxI, nad3/rps12, and orf25. Analysis of eight individuals of each reciprocal hybrid of these two species revealed that mitochondrial DNA was maternally inherited. Furthermore, sequences homologous to wheat orf25 were also identified in Larix gmelini, Larix siberica, Larix olgensis, and Larix laricina, as well as Ginkgo biloba, Picea mariana, Picea glauca and Pinus contorta.
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  • 91
    ISSN: 1432-2242
    Keywords: DNA ; RFLP ; Soybean ; Genetic map
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    Topics: Biology
    Notes: Abstract Genetic markers were mapped in segregating progeny from a cross between two soybean (Glycine max (L.) Merr.) cultivars: ‘Minsoy’ (PI 27.890) and ‘Noir 1’ (PI 290.136). A genetic linkage map was constructed (LOD ⩾ 3), consisting of 132 RFLP, isozyme, morphological, and biochemical markers. The map defined 1550cM of the soybean genome comprising 31 linkage groups. An additional 24 polymorphic markers remained unlinked. A family of RFLP markers, identified by a single probe (hybridizing to an interspersed repeated DNA sequence), extended the map, linking other markers and defining regions for which other markers were not available.
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  • 92
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    Theoretical and applied genetics 86 (1993), S. 573-578 
    ISSN: 1432-2242
    Keywords: COXI ; COXII ; Maternal inerhitance ; mtDNA ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have studied two mitochondrial DNA polymorphisms in 741 individuals from 16 allopatric populations ofPinus banksiana Lamb. andPinus contorta Dougl. Restriction fragments of both polymorphisms distinguished the two species qualitatively, except in aP. Banksiana population whose ancestors were involved in hybridization withP. contorta.COXI-associated restriction fragments were monomorphic within species, whileCOXII-associated restriction fragments were highly variable inP. contorta (Hes=0.68). Population differentiation was substantial inP. contorta (Fst=0.31 among subspecies; mean Fst=0.66 within subspecies) and consistent with predictions for maternally inherited markers. Plant mitochondrial markers appear to be useful for the investigation of seed migration routes, hybridization and introgression, breeding zone designation, and the development of germ plasm conservation sampling strategies.
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  • 93
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    Keywords: Hordeum vulgare ; Microdissection ; Microcloning ; Chromosome specific library ; RFLP
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    Notes: Abstract We have applied a refined microdissection procedure to create a plasmid library of the barley (Hordeum vulgare L.) chromosome arm 1HS. The technical improvements involved include synchronization of meristematic root tissue, a metaphase drop-spread technique, paraffin protection of the collection drop to avoid evaporation, and a motorized and programmable microscope stage. Thirteen readily-discernible telocentric chromosomes have been excised from metaphases of synchronized root-tip mitoses. After lysis in a collection drop (2 nl), the DNA was purified, restricted withRsaI, ligated into a vector containing universal sequencing primers, and amplified by the polymerase chain reaction. Finally, the amplified DNA was cloned into a standard plasmid vector. The size of the library was estimated to be approximately 44,000 recombinant plasmids, of which approximately 13% can be utilized for RFLP analysis. Tandem repetitive probes could be rapidly excluded from further analysis after colony hybridization with labelled total barley DNA. Analysis of 552 recombinant plasmids established that: (1) the insert sizes ranged between 70 and 1150 bp with a mean of 250 bp, (2) approximately 60% of the clones contained highly repetitive sequences, and (3) all single- or low-copy probes tested originate from chromosome 1HS. Four probes were genetically mapped, using an interspecificH. vulgare xH. spontaneum F2 population. One of these probes was found to be closely linked to theMla locus conferring mildew resistance.
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  • 94
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    Theoretical and applied genetics 86 (1993), S. 837-842 
    ISSN: 1432-2242
    Keywords: Androgenesis ; RFLP ; Maize ; Agronomic evaluation ; Gametoclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The homogeneity of anther culture-derived lines of maize has been evaluated by means of field observations and molecular markers. The homogeneity of the doubled haploid (DH) lines was shown by the absence of segregation for morphological oligogenic traits. The intravariance for polygenic traits for 42 DH and two conventionally derived lines was similar, which confirmed the homozygosity of the DH lines. More than 100 RFLP markers were tested on 189 DH lines derived from two crosses, DH5 x DH7 and A188 x DH7, and 60 single-seed descent (SSD, F6) lines derived from A188 x DH7. The overall rate of heterozygosity for all of the DH lines was approximatively 1% and pertained to 6 lines out of 189, while it was 8.5% for the SSD lines after four selfings. A precise description of the material used suggested that the events which led to this unexpected heterozygozity in DH lines were more likely to have occurred after rather than during the androgenetic process. Nine duplicated pairs of genotypes were found within the DH lines, indicating that a single microspore-derived structure can fragment to give two identical plantlets. Despite the extensive screening with more than 100 markers, only 2 lines showed unexpected banding profiles, and these were probably gametoclonal variants. The use of a direct regeneration system that avoids any callus phase might explain this low frequency of gametoclonal variation.
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  • 95
    ISSN: 1432-2242
    Keywords: DNA ; RFLP ; QTL ; Soybean ; Recombinant inbreds
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An experimental test is described for linkages between RFLP markers and quantitative trait loci (QTL). Two hundred and eighty-four F7-derived recombinant inbred lines (RIL) obtained from crossing the soybean cultivars (Glycine max L. Merr.) ‘Minsoy’ and ‘Noir 1’ were evaluated for maturity, plant height, lodging, and seed yield. RIL exhibiting an extreme phenotype for each trait (earliest and latest plants for maturity, etc.) were selected, and two bulked DNA samples were prepared for each trait. A Southern transfer of the digested bulked DNA was hybridized with restriction fragement length polymorphism (RFLP) probes, and linkages with QTL were established by quantitating the amount of radioactive probe that bound to fragments defining alternative parental RFLP alleles. When an RFLP marker was linked to a QTL, one parental allele predominated in the bulked DNA from a particular phenotype; the other allele was associated with the opposite phenotype. When linkage was absent, radioactivity was associated equally with both alleles for a given phenotype (or with both phenotypes for a given allele). These results confirmed RFLP-QTL associations previously discovered by interval mapping on a smaller segregating population from the same cross. New linkages to QTL were also verified.
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  • 96
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    Theoretical and applied genetics 87 (1993), S. 278-287 
    ISSN: 1432-2242
    Keywords: Maize ; RFLP ; Recombination ; Disturbed segregation ; DH/SSD lines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Doubled-haploid (DH) and single-seed-descent (SSD) lines in maize have been compared for quantitatively inherited traits and for RFLP markers. The comparisons of the distributions for agromorphological traits do not allow definite conclusions to be drawn on the similarity of the two reproductive systems. We have used more than 100 RFLP markers to provide a precise description of the parental allele frequency and the recombination fractions. A comparison of two DH populations shows that non-random meiotic reassortment is influenced by differences in the anther culture capacities of the two parental lines. For the DH lines derived from the cross DH5 x DH7, involving two responsive lines in anther culture, the distortion in segregation (P 〈 0.05) affected less than 20% of the genome with half of the deviations towards each parent. DH lines derived from the cross A188 x DH7, where A188 is a non-responsive line, showed more than twice this level of distortion and an excess of DH7 alleles was found for almost all of the skewed loci. The recombination fractions were homogeneous between the two DH populations for most of the genome. The genome sizes calculated with the DH and the SSD lines derived from the same cross, A188 x DH7, were also similar, which suggests that no selection against recombinant gametes occurs during anther culture. The observed recombination fraction after five meioses (SSD) is on average twice as large as after one meiosis (DH). No difference is observed for recombination fractions greater than 20%. Despite a precise description of the material at the molecular level, it has not been possible to make a definite conclusion as to whether or not the differences in some morphological characters are the consequences of differences in the segregation ratio and/or the recombination frequency. However, the agromorphological evaluation shows a narrow range in differences between the two types of lines and suggests that the use of DH lines is possible in breeding programmes.
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    Theoretical and applied genetics 88 (1994), S. 273-278 
    ISSN: 1432-2242
    Keywords: RFLP ; Genetic linkage mapping ; Pinus taeda L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A genetic linkage map for loblolly pine (Pinus taeda L.) was constructed using segregation data from a three-generation outbred pedigree consisting of four grandparents, two parents, and 95 F2 progeny. The map was based predominantly on restriction fragment length polymorphism (RFLP) loci detected by cDNA probes. Sixty-five cDNA and three genomic DNA probes revealed 90 RFLP loci. Six polymorphic isozyme loci were also scored. One-fourth (24%) of the cDNA probes detected more than 1 segregating locus, an indication that multigene families are common in pines. As many as six alleles were observed at a single segregating locus among grandparents and it was not unusual for the progeny to segregate for three or four alleles per locus. Multipoint linkage analysis placed 73 RFLP and 2 isozyme loci into 20 linkage groups; the remaining 17 RFLP and 4 isozyme loci were unlinked. The mapped RFLP probes provide a new set of codominant markers for genetic analyses in loblolly pine.
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  • 98
    ISSN: 1432-2242
    Keywords: Hevea brasiliensis ; RFLP ; Nuclear probes ; Isozyme ; Genetic diversity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Restriction fragment length polymorphism was assessed in wild and cultivated populations of Hevea brasiliensis using random probes from an Hevea nuclear library. One-hundred-and-sixty-four individuals were surveyed, and the results discussed in the light of previous work performed on isozyme variation. Both studies show that germplasm collections have led to an effective enrichment of the genetic resources available for Hevea breeding, and that cultivated clones have conserved a relatively high level of polymorphism, despite their narrow genetic base and their high level of inbreeding. An equivalent level of polymorphism is revealed by random nuclear probes and isozymes. However, the genetic structuring of the diversity appears more striking using RFLP markers. Wild accessions can be divided into three genetic groups according to their geographical origin. The present results are an essential guide to the incorporation of wild material in breeding schemes.
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    Theoretical and applied genetics 88 (1994), S. 383-394 
    ISSN: 1432-2242
    Keywords: Photosynthesis-related genes ; Copy numbers ; Chromosome assignments ; RFLP ; Origin of polyploid wheats
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Copy numbers of four photosynthesis-related genes, PhyA, Ppc, RbcS and Lhcb1 *1, in wheat genomes were estimated by slot-blot analysis, and these genes were assigned to the chromosome arms of common wheat by Southern hybridization of DNA from an aneuploid series of the cultivar Chinese Spring. The copy number of PhyA was estimated to be one locus per haploid genome, and this gene was assigned to chromosomes 4AL, 4BS and 4DS. The Ppc gene showed a low copy number of small multigenes, and was located on the short arm of homoeologous group 3 chromosomes and the long arm of chromosomes of homoeologous group 7. RbcS consisted of a multigene family, with approximately 100 copies in the common wheat genome, and was located on the short arm of group 2 chromosomes and the long arm of group 5 chromosomes. Lhcb1 *1 also consisted of a multigene family with about 50 copies in common wheat. Only a limited number of restriction fragments (approximately 15%) were used to determine the locations of members of this family on the long arm of group 1 chromosomes owing to the multiplicity of DNA bands. The variability of hybridized bands with the four genes was less in polyploids, but was more in the case of multigene families. RFLP analysis of polyploid wheats and their presumed ancestors was carried out with probes of the oat PhyA gene, the maize Ppc gene, the wheat RbcS gene and the wheat Lhcb1 *1 gene. The RFLP patterns of common wheat most closely resembled those of T. Dicoccum (Emmer wheat), T. urartu (A genome), Ae. speltoides (S genome) and Ae. squarrosa (D genome). Diversification of genes in the wheat complex appear to have occurred mainly at the diploid level. Based on RFLP patterns, B and S genomes were clustered into two major groups. The fragment numbers per genome were reduced in proportion to the increase of ploidy level for all four genes, suggesting that some mechanism(s) might operate to restrict, and so keep to a minimum, the gene numbers in the polyploid genomes. However, the RbcS genes, located on 2BS, were more conserved (double dosage), indicating that the above mechanism(s) does not operate equally on individual genes.
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    Theoretical and applied genetics 88 (1994), S. 472-478 
    ISSN: 1432-2242
    Keywords: RFLP ; PCR ; Chrysanthemum ; Genetic diversity ; Polyploidy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In order to study genetic variability at the DNA level in chrysanthemum (Dendranthema grandiflora Tzvelev) PstI and HindIII genomic libraries were constructed. Probes from both libraries were tested for the presence of restriction fragment length polymorphisms (RFLPs). Of the probes from the PstI library 91% appeared to hybridize to low-copy genes, while only 35% of those from the HindIII library appeared to do so. The PstI probes were used in further analyses as 79% of them showed RFLPs, whereas the HindIII low-copy number probes gave only 14% polymorphic patterns. Because of the hexaploid character of chrysanthemum, complex patterns generally consisting of 6–12 fragments were visible on a Southern blot after hybridization. To simplify the genetic analysis, locus-specific polymerase chain reaction (PCR) primers were developed that gave simple polymorphic patterns in a number of cases. The RFLP probes and primers developed will be used in future marker-assisted selection in this polyploid crop.
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