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  • 1
    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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  • 2
    ISSN: 1432-1890
    Keywords: Key words PCR ; RFLP ; ITS sequence analysis ; Ectomycorrhizal fungi ; Cortinarius taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The ability of restriction fragment length polymorphism (RFLP) analysis of the rDNA internal transcribed spacer (ITS) region to discriminate 10 co-occurring Cortinarius and Dermocybe species at a southeastern Australian sclerophyll forest site was assessed. Using the basidiomycete-specific primers ITS1F and ITS4B, some taxa were separated on the basis of individual RFLP patterns derived using the restriction endonucleases Hae III or Hinf I. Combined data from both endonucleases were, however, required to separate all taxa [Dermocybe austro-veneta Clel. (Moser & Horak), C. rotundisporus Clel. & Cheel, C. archeri Berk., C. sinapicolor Clel., C. violaceus (L.: Fr.) S.F.Gray, C. radicatus Clel. and four morphologically-distinct, but unidentified Cortinarius spp.]. ITS sequence comparisons confirmed that D. austro-veneta belongs in Dermocybe, that C. rotundisporus is correctly placed in subgenus Phlegmacium, and suggest that Australian C. violaceus collections are not conspecific with northern hemisphere C. violaceus.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 14 (1988), S. 177-182 
    ISSN: 1432-0983
    Keywords: RFLP ; Tomato ; Plastid ; Mitochondria ; Protoplasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Restriction fragment length polymorphisms (RFLPs) were used to determine the transmission of organelle genomes in somatic hybrid plants of tomato and its wild relativeSolanum lycopersicoides. Biased frequencies of organelle combinations were observed in a population of 70 somatic hybrid plants each derived from a separate callus. The plastids in 68 of 70 hybrids examined were fromL. esculentum. One of the remaining hybrids, plant 240, hadS. lycopersicoides plastids and the other, plant 63, had a mixture of parental plastids. Forty-six of the same 70 plants were analyzed for mtDNA and all had that ofS. lycopersicoides including plant 240. One of these hybrids had novel mtDNA fragments which mayhave resulted from recombination or rearrangement. The biased transmission may have resulted from an initial unequal input of organelles, differential replication of organelles, or nucleo-organelle incompatibility.
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  • 4
    ISSN: 1432-1432
    Keywords: Escherichia coli ; RAPD ; RFLP ; Clonal theory ; Recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Analysis of the Escherichia coli population by multilocus enzyme electrophoresis (MLEE) has established its clonal organization, but there is increasing evidence that horizontal DNA transfer occurs in E. coli. We have assessed the genetic structure of the species E. coli and determined the extent to which recombination can affect the clonal structure of bacteria. A panel of 72 E. coli strains from the ECOR collection was characterized by random amplified polymorphic DNA (RAPD) and restriction-fragment-length polymorphism (RFLP) of the ribosomal RNA gene (rrn) regions. These strains have been characterized by MLEE and are assumed to reflect the range of genotypic variation in the species as a whole. Statistical analysis, including factorial analysis of correspondence (FAC) and hierarchical classifications, established that the data obtained with the three genetic markers are mutually corroborative, thus providing compelling evidence that horizontal transfer does not disrupt the clonal organization of the population. However, there is a gradient of correlation between the different classifications which ranges from the highly clonal structure of 132 group strains causing extraintestinal infections in humans to the less-stringent structure of B1 group strains that came mainly from nonprimate mammals. This group (B1) appears to be the framework from which the remaining non-A group strains have emerged. These results indicate that RAPD analysis is well suited to intraspecies characterization of E. coli. Lastly, treating the RAPD data by FAC allowed description of subgroup-specific DNA fragments which can be used, in a strategy comparable to positional cloning, to isolate virulence genes.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 100 (2000), S. 564-568 
    ISSN: 1432-2242
    Keywords: Key words Triticum aestivum ; Tritiam timopheevii ; Pm6 ; Introgression lines ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Pm6 in bread wheat (Triticum aestivum L.), which was transferred from Triticum. timopheevii L., is a gene conferring resistance to the powdery mildew disease caused by Erysiphe graminis f. sp. tritici. Six near-isogenic lines ( NILs ) of Pm6 in a cultivar ’Prins’ background were analyzed to map this gene using restriction fragment length polymorphism (RFLP). Each of the six NILs possessed a T. timopheevii-derived segment, varying in length, and associated with powdery mildew resistance. Lines IGV1–465 (FAO163b/ 7*Prins) and IGV1–467 (Idaed 59B/7*Prins) had the shortest introgressed segments, which were detected only by DNA probes BCD135 and PSR934, respectively. The polymorphic loci detected by both probes were mapped to the long arm of chromosome 2B. Lines IGV1–458 (CI13250/7*Prins) and IGV1–456 (CI12559/8*Prins) contained the longest T. timopheevii segments involving both arms of donor chromosome 2G across the centromere. All these introgressed segments had an overlapping region flanked by the loci xpsr934 and xbcd135 on 2BL. Thus, Pm6 was located in this region since the powdery mildew resistance in all the NILs resulted from the introgressed fragments. Using the F2 mapping population from a cross of IGV1–463 (PI170914/7*Prins)×Prins, Pm6 was shown to be closely linked to the loci xbcd135 and xbcd266 at a genetic distance of 1.6 cM and 4.8 cM, respectively. BCD135 was successfully used in detecting the presence of Pm6 in different genetic backgrounds.
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  • 6
    ISSN: 1572-9818
    Keywords: cpDNA ; DNA extraction ; fingerprinting ; forest trees ; M13 fingerprinting ; method ; PCR ; rDNA ; RFLP ; rhododendron ; plant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Four DNA extraction protocols were compared for ability to produce DNA from the leaves or needles of several species: oak, elm, pine, fir, poplar and maize (fresh materials) and rhododendron (silica dried or frozen material). With the exception of maize and poplar, the species are known to be difficult for DNA extraction. Two protocols represented classical procedures for lysis and purification, and the other two were a combination of classical lysis followed by anion exchange chromatography. The DNA obtained from all procedures was quantified and tested by PCR and Southern hybridisation.Test results indicated superiority of one of the four protocols; a combination of CTAB lysis followed by anion exchange chromatography which enabled DNA extraction from all seven species. A second protocol also produced DNA from leaves or needles of all species investigated and was well suited for PCR applications but not Southern hybridisations. The remaining protocols produced DNA from some but not all species tested. Abbreviations: CTAB, hexadecyltrimethylammonium bromide; EtOH, Ethanol; TBE, tris-borate-EDTA.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology reporter 16 (1998), S. 90-90 
    ISSN: 1572-9818
    Keywords: DNA extration ; DNA fingerprint ; half seed ; PCR ; RFLP ; target gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A single-seed DNA extraction method was developed for rapid identification of plant genotype. The method was applied to 12 plant species, including the oil seeds sesame and soybean. The results were comparable to those obtained for oil-less seeds such as rice. This method will be useful for genotypic selection which requires rapid screening of large populations. It can also be used to identify varietal purity of seed stocks by PCR and RFLP analysis. The method includes two major steps, (i) treatment by proteinase K in an SDS extraction buffer, and (ii) grinding of a single half seed in the buffer after incubation. About 1.5–2 µg of DNA per half seed (the endosperm part) of rice was obtained and more than 200 half seed samples could be handled by one person in a day. The DNA could be used for fingerprinting and detection of target genes in a transgenic plant by PCR. The amplified PCR products from the half seed DNA exhibited the same banding patterns as those from leaf DNA. Yield and quality of DNA extracted from half seeds of rice was also sufficient for RFLP analysis. The remnant half seeds containing the embryo can be maintained for later germination of selected genotypes.
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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
    ISSN: 1420-9071
    Keywords: ETS 1 proto-oncogene ; RFLP ; Saudi ; leukemia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We have cloned part of theETS 1 proto-oncogene and demonstrated the presence of two polymorphic Sst I restriction sites. A probe derived from one of our clones revealed the presence of 8.3 kb, 9.5 kb and/or 11.5 kb fragments on Southern blots of human DNA samples. The relative frequencies of these alleles appear to be significantly different between Saudi and Western populations, but there are no apparent differences in these frequencies between Saudi non-leukemic and leukemic individuals.
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  • 10
    ISSN: 1432-1211
    Keywords: Key words Interleukin (IL)-4 ; Promoter ; Polymorphism ; RFLP ; Linkage disequilibrium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
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  • 11
    ISSN: 1432-1211
    Keywords: Key words IL-5Rα ; Promoter ; Polymorphism ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
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  • 12
    Electronic Resource
    Electronic Resource
    Springer
    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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  • 13
    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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  • 14
    ISSN: 1432-2145
    Keywords: Key words Abies ; Egg cell ; Plastid inheritance ; RFLP ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The ultrastructure of egg cells in Abies alba was examined to elucidate the lack of maternal inheritance of plastids. Before fertilization, maternal plastids are absent in the perinuclar zone containing mainly mitochondria and smooth endoplasmic reticulum. During egg cell development the maternal plastids are transformed into large inclusions which are situated mostly towards the periphery of the egg cell, and finally disintegrate. As a consequence, they do not participate in zygote formation. RFLP analysis of cpDNA of parental trees and their F1 interspecific hybrids (A. alba×A. numidica, A. alba×A. nordmanniana, A. nordmanniana×A. Alba) using HindIII and BamHI showed a paternal mode of cpDNA inheritance. Paternal inheritance has also been found with PCR/RFLP analysis of cpDNA from parental trees and their hybrids (A. alba×A. pinsapo, A. pinsapo×A. alba, A. pinsapo×A. numidica) using ApaI and HaeIII digests, as well as in the crosses of A. cephalonica×A. nordmanniana, A. nordmanniana×A. cephalonica, A. cephalonica×A. numidica using TagI digests.
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  • 15
    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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  • 16
    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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  • 17
    Electronic Resource
    Electronic Resource
    Springer
    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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  • 18
    ISSN: 1432-0983
    Keywords: Genetic linkage mapping ; Segregation distortion ; RAPD ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The inheritance of DNA markers was investigated in 27 F2 progeny from a single F1 hybrid derived from a wide cross inUromyces appendiculatus. This cross was unusual because asexual spores were used to fertilize sexual fruiting structures. Sixty percent of the DNA markers failed to segregate according to simple Mendelian ratios. Segregation bias was evident, in that F2 progeny inherited on average 91 % of maternal bands and 52% of paternal bands, which deviates significantly from the expected value for each of 75% for dominant markers. Because of these distortions, linkage mapping was not possible with this population. Evaluation of two F1s from a second wide cross, reciprocals obtained by normal fertilization, also showed non-Mendelian inheritance of one of three co-dominant RFLPs and five of six isozyme markers, indicating that the method of crossing was probably not responsible for the abnormal segregation patterns in the first cross. Either genetic incompatibility, similar to that of an interspecific cross, or selection of particular genotypes could explain the genetic anomalies reported here.
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  • 19
    ISSN: 1432-0983
    Keywords: Cronartium flaccidum ; Pine blister rusts ; Ribosomal intergenic region polymorphism ; RFLP ; HPA analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mixed aeciospore isolates ofCronartium flaccidum andPeridermium pini were obtained from single-tree infections in Britain, Italy and Greece. The 5.8s ribosomal RNA gene and flanking intergenic transcribed spacer regions ITS 1 and ITS2 were found to be highly similar betweenC. flaccidum andP. pini. Within samples heterogeneity was detected at three nucleotide loci in the ITS1 and at four loci in the ITS2 suggesting that several fungal genotypes may occur at a single infection court. The heterogeneity was confirmed by heteroduplex polymorphism analysis of mixed aeciospore products. RFLP of the ribosomal intergenic spacer region 1 (IGSI) amplified from the same templates indicated limited sequence polymorphism in some copies of this repeated locus. Both the sexual and asexual forms ofC. flaccidum show evidence of sequence polymorphism in two independent, non-coding regions of the ribosomal gene array. Variation appears to be greater in the sexual formC. flaccidum, than in the monoaecious formP. pini.
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  • 20
    ISSN: 1432-0983
    Keywords: Key words  Cronartium flaccidum ; Pine blister rusts ; Ribosomal intergenic region polymorphism ; RFLP ; HPA analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract   Mixed aeciospore isolates of Cronartium flaccidum and Peridermium pini were obtained from single-tree infections in Britain, Italy and Greece. The 5.8s ribosomal RNA gene and flanking intergenic transcribed spacer regions ITS1 and ITS2 were found to be highly similar between C. flaccidum and P. pini. Within samples heterogeneity was detected at three nucleotide loci in the ITS1 and at four loci in the ITS2 suggesting that several fungal genotypes may occur at a single infection court. The heterogeneity was confirmed by heteroduplex polymorphism analysis of mixed aeciospore products. RFLP of the ribosomal intergenic spacer region 1 (IGS1) amplified from the same templates indicated limited sequence polymorphism in some copies of this repeated locus. Both the sexual and asexual forms of C. flaccidum show evidence of sequence polymorphism in two independent, non-coding regions of the ribosomal gene array. Variation appears to be greater in the sexual form C. flaccidum, than in the monoaecious form P. pini.
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  • 21
    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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  • 22
    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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  • 23
    ISSN: 1432-0983
    Keywords: Key words Genetic linkage mapping ; Segregation distortion ; RAPD ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The inheritance of DNA markers was investigated in 27 F2 progeny from a single F1 hybrid derived from a wide cross in Uromyces appendiculatus. This cross was unusual because asexual spores were used to fertilize sexual fruiting structures. Sixty percent of the DNA markers failed to segregate according to simple Mendelian ratios. Segregation bias was evident, in that F2 progeny inherited on average 91% of maternal bands and 52% of paternal bands, which deviates significantly from the expected value for each of 75% for dominant markers. Because of these distortions, linkage mapping was not possible with this population. Evaluation of two F1s from a second wide cross, reciprocals obtained by normal fertilization, also showed non-Mendelian inheritance of one of three co-dominant RFLPs and five of six isozyme markers, indicating that the method of crossing was probably not responsible for the abnormal segregation patterns in the first cross. Either genetic incompatibility, similar to that of an interspecific cross, or selection of particular genotypes could explain the genetic anomalies reported here.
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  • 24
    ISSN: 1432-072X
    Keywords: Key words Cyanobacteria ; Planktonic ; Total protein ; SDS-PAGE ; PCR ; RFLP ; 16S rRNA ; UPGMA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Planktonic, filamentous cyanobacterial strains from different genera, both toxic and nontoxic strains, were characterized by SDS-PAGE of whole-cell proteins and PCR/RFLP of the 16S rRNA gene. Total protein pattern analysis revealed the mutual relationships at the genus level. Restriction fragment length polymorphism (RFLP) of the 16S rRNA gene with reference strains proved to be a good method for the cyanobacterial taxonomy. The nonheterocystous strains outgrouped from the nitrogen-fixing ones. With both methods, Aphanizomenon clustered with Anabaena, and Nodularia with Nostoc. In the RFLP study of Anabaena, the neurotoxic strains were identical, but the hepatotoxic ones formed a heterogeneous group. Genetic distances found in the RFLP study were short, confirming that close genotypic relationships underlie considerable diversity among cyanobacterial genera.
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  • 25
    ISSN: 1432-072X
    Keywords: Key words Thermus oshimai ; Megaplasmids ; Pulsed-field gel electrophoresis ; RFLP ; Southern ; hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Megaplasmid DNA was detected in ten isolates belonging to the recently described thermophilic eubacterial species Thermus oshimai and isolated from hot springs in Portugal (eight isolates) and Iceland (two isolates). The estimated size of the large plasmids purified from T. oshimai SPS-18 from S. Pedro do Sul, Portugal, and from isolate JK-91 from Hveragerdhi-Hengill, Iceland, was 214 and 275 kb, respectively. No sequence homologous to isolate SPS-18 megaplasmid is present in chromosomal DNA as indicated by Southern hybridization analysis. Overall examination of the HindIII fragment profiles of megaplasmid DNAs purified from isolates from the same geographical area gave similar but not always identical restriction profiles on agarose gels. Restriction fragment length polymorphism (RFLP) was higher for megaplasmids present in isolates purified from the Portuguese and Icelandic isolates than for megaplasmids from the same hot spring. Megaplasmid RFLP correlated with previous results obtained on the polymorphism of macrorestriction patterns of whole genomic DNA and with the RFLP of co-resident small plasmid DNA that was found in one half of the isolates examined. The 16-kb HindIII–HindIII fragment from isolate SPS-18 megaplasmid showed DNA–DNA homology with restriction fragments of similar size generated by the large plasmids present in all the other isolates, even in those from hot springs of widely separated geographical areas. This suggests a high degree of sequence conservation in T. oshimai megaplasmids.
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  • 26
    ISSN: 1432-2242
    Keywords: Key words Chloroplast DNA ; Genetic diversity ; Ipomoeabatatas ; ISSR ; RFLP ; Sweetpotato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Genetic diversity and relationships of 40 accessions of Ipomoea, representing ten species of series Batatas, were examined using ISSR markers and restriction-site variation in four non-coding regions of chloroplast DNA. A total of 2071 ISSR fragments were generated with 15 primers in these accessions and, on average, 52 bands per accession were amplified. Most of the primers contained dinucleotide repeats. The ISSR fragments were highly polymorphic (62.2%) among the 40 accessions studied. Restriction analysis of chloroplast (cp) DNA revealed 47 informative restriction-site and length mutations. Phylogenetic analyses of ISSR and cpDNA datasets generally revealed similar relationships at the interspecific level, but the high polymorphism of ISSRs resulted in a better separation of intraspecific accessions. However, the combined ISSR and cpDNA dataset appeared to be appropriate in resolving both intra- and interspecific relationships. Of the species examined, I. trifida was found to be the most closely related to cultivated sweetpotato, the hexaploid I. batatas, while I. ramosissima and I. umbraticola were the most distantly related to I. batatas within the series. Ipomoea triloba, hitherto considered to be one of the ancestors of sweetpotato, was only distantly related to sweetpotato based on ISSR similarity index.
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  • 27
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    Theoretical and applied genetics 100 (2000), S. 899-905 
    ISSN: 1432-2242
    Keywords: Key words Petunia ; RFLP ; Genetic map ; Genome ; Segregation distortion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Due in large part to the data accumulated from years of classic genetic analysis, petunia (Petunia hybrida Vilm) has remained a useful model system, particularly for studies of gene regulation and genome structure. We have used three segregating populations of petunia, including those serving as the source of an earlier actin gene RFLP map, for RFLP mapping of several additional genes. Twenty-seven loci have been merged with 11 previously mapped morphological and biochemical markers. Our results contribute additional evidence to reports of a high degree of genome plasticity and segregation distortion in this species and suggest that petunia may be a useful plant system for detailed analysis of plant genome organization, activity and evolution.
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  • 28
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    Theoretical and applied genetics 100 (2000), S. 1018-1024 
    ISSN: 1432-2242
    Keywords: Key words cpDNA ; Cytoplasmic male sterility ; mtDNA ; Olea europaea ; Inheritance ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The olive tree is usually hermaphrodite but self-incompatible. In the Western Mediterranean some cultivars are totally male-sterile. Three different male-sterile phenotypes have been recognised. To infer the genetic basis of male sterility we studied its inheritance and cytoplasmic diversity in wild (oleaster) and cultivated Mediterranean olive. In the cross Olivière×Arbequina, the male-sterile trait was maternally inherited and affected all progenies. We also checked that both chloroplast and mitochondrial DNAs are maternally inherited. RFLP studies on chloroplast and mitochondrial DNAs revealed several cytotypes: two chlorotypes and four mitotypes in cultivars and oleaster (wild or feral Mediterranean olive). Furthermore, a total linkage desequilibrium between the CCK chlorotype and the MCK mitotype in cultivars and oleaster from different regions supports the fact that paternal leakage of organelles was not observed. The male sterility (ms 2) displayed by Olivière, plus six other cultivars and three oleaster was strictly associated with the CCK chlorotype and the MCK mitotype. These facts suggest that Olivière carries cytoplasmic male sterility. Male-fertile and male-sterile oleasters carrying this cytotype showed the presence of restorer alleles. This CMS might be due to a distant cross between olive taxa. The two other male-sterile phenotypes displayed by Lucques (ms 1) and Tanche (ms 3) were associated with the ME1 mitotype but we have not demonstrated CMS.
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  • 29
    ISSN: 1432-2242
    Keywords: Key words Triticum aestivum ; Aegilops speltoides ; Chromosome addition ; C-banding ; In situ hybridization ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Aegilops speltoides Tausch (2n = 2x = 14, SS) is considered as the closest living relative of the B and G genomes of polyploid wheats. A complete set of Triticum aestivum L. cv Chinese Spring-Ae. speltoides whole chromosomes and seven telosomic addition lines was established. A low pairing accession was selected for the isolation of the chromosome addition lines. Except for chromosomes 3S and 6S, which are presently only available as monosomic additions, all other lines were recovered as disomic or ditelosomic additions. The individual Ae. speltoides chromosomes isolated in the wheat background were assayed for their genetic effects on plant phenotype and cytologically characterized in terms of chromosome length, arm ratio, distribution of marker C-bands, and FISH sites using a Ae. speltoides-specific repetitive element, Gc1R-1, as a probe. The homoeology of the added Ae. speltoides chromosomes was established by using a standard set of RFLP probes. No chromosomal rearrangements relative to wheat were detected.
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  • 30
    ISSN: 1432-2242
    Keywords: Keywords Alfalfa ; Post-meiotic cytokinesis ; RFLP ; 4n pollen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The objective of this research was to map the jumbo-pollen trait in diploid alfalfa. Homozygous recessive (jpjp) plants are characterized by the complete failure of post-meiotic cytokinesis during microsporogenesis resulting in 100% 4n-pollen formation. Three F1 segregating populations were produced and analyzed for pollen-grain production and the segregation of RFLP markers. The first cross combination did not segregate for the jumbo-pollen trait, but showed a clear segregation for multinucleate (bi-, tri- and tetra-nucleate)-microspore formation. Cytological analysis showed that few plants produced 100% normal (uninucleate) microspores, whereas most of them produced multinucleate microspores at a variable frequency (0–75%). Plants with multinucleate microspores always showed a prevalence of binucleated microspores, even though some plants showed a background presence of tri- and tetra-nucleate microspores. QTL analysis based on ANOVA I and Stepwise Multiple Regression identified three QTLs with a highly significant effect on multinucleate-microspore formation. Two cross combinations, subsequently executed, showed Mendelian segregation for the jumbo-pollen trait and were effective in locating the jp gene on linkage group 6 close to the Vg1G1b RFLP locus. Interestingly, this RFLP locus was also linked to one QTL for multinucleate-microspore formation. Genetic models are discussed concerning the presence in linkage group 6 of a cluster of genes involved in multinucleate-microspore formation together with possible relationships between the jp gene and the Vg1G1b QTL.
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  • 31
    ISSN: 1432-2242
    Keywords: Key words Acacia ; RFLP ; Restriction enzymes ; Genome mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Random genomic probes were used to assess levels of restriction fragment length polymorphism (RFLP) in two 2-generation outbred pedigrees of Acacia mangium Willd. Probes were evaluated for their ability to detect polymorphic loci in each pedigree and to determine the relative efficiency of different restriction enzymes in revealing polymorphisms. Sixty two percent of the probes which detected single- or low-copy number sequences revealed polymorphisms with at least one restriction enyzme. HpaII was the most efficient in detecting polymorphism among first-generation individuals. The recognition sequence of HpaII contains a CpG dimer, suggesting that cytosines in the CpG sequence may be hotspots for mutation in plant genomes, as previously reported in bacterial and mammalian genomes. Mendelian inheritance of 230 loci was demonstrated based on single-locus segregation in second-generation individuals. Less than 5% of loci showed evidence of segregation distortion. The proportion of fully informative loci (15%) was lower than previously reported in eucalypts reflecting the lower level of genetic diversity in A. mangium. The RFLP probes are suitable for the construction of a high-density genetic linkage map in A. mangium. Cross-hybridisation of the A.mangium RFLPs to DNA from species representing the three subgenera of the genus Acacia indicates that these markers could be used in breeding programs of other diploid acacias, for comparative studies of genome organisation, and for phylogenetic studies.
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  • 32
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    Theoretical and applied genetics 100 (2000), S. 602-606 
    ISSN: 1432-2242
    Keywords: Key words Triticum ; Hexaploid ; Mapping ; RFLP ; RSL ; Spelt
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A dominant allele at the Q locus on chromosome 5A is believed to be the principal factor responsible for free-threshing, square-head spikes with a non-fragile rachis in bread wheat, Triticum aestivum ssp. aestivum. The spelt syndrome, resulting in pyramidal spikes with a brittle rachis and hulled grain in T. aestivum, is believed to be principally caused by the q allele. Chromosome 5A of European and Iranian spelt was substituted for 5A of bread wheat and the lines were characterized with molecular markers. The substitution of bread wheat chromosome 5A by 5A of European spelt resulted in weakly hulled, pyramidal spikes with a non-brittle rachis, whereas and the substitution of 5A by 5A of Iranian spelt did not alter spike morphology at all. It is concluded that the expression of the spelt syndrome depends, to a large extent, on the interactions of q with genes controlling glume tenacity and rachis fragility on other chromosomes. The genetic basis for the spelt syndrome and the apparent presence of the Q allele in Iranian spelt are discussed.
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  • 33
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    Theoretical and applied genetics 100 (2000), S. 934-941 
    ISSN: 1432-2242
    Keywords: Key words Zea mays L ; Maize ; Sugarcane mosaic virus ; SCMV ; Scm1 ; Scm2 ; AFLP ; RFLP ; SSR ; Pedigree relationship
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Sugarcane mosaic virus (SCMV) causes serious losses of grain and forage yield of maize (Zea mays L.) in Europe. Two dominant genes, Scm1 and Scm2, have been identified to confer resistance to SCMV. Scm1 is located on the short arm of chromosome 6 and Scm2 near the centromere region of chromosome 3. In the present study,resistant, partially resistant, and susceptible maize inbred lines, together with their ancestral lines, were evaluated with molecular markers to trace back the origin of Scm1 and Scm2. The banding patterns indicated that the Scm1 region, originally identified in resistant European line FAP1360A, was derived from its ancestral line FAP954A. The other two resistant European lines, D21 and D32, most likely carry the same Scm1 region, which originated from their common ancestral line A632. This Scm1 region was also present in three partially resistant lines, D09, FAP1396A and FAP693A, but not in the resistant U.S. inbred Pa405. Apart from FAP954A and A632, none of the remaining ancestral lines and none of the susceptible lines harbored the Scm1 region. The Scm2 region present in FAP1360A was obviously transmitted from its ancestral line Co125. However, the presence of the respective Scm2 region was not confirmed in the other three resistant lines (D21, D32 and Pa405), the remaining ancestral lines, and all partially resistant lines by using closely linked markers.
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  • 34
    ISSN: 1432-2242
    Keywords: Key words Map-based cloning ; RFLP ; YAC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  An ethylene-inducing xylanase (EIX) from Tricohoderma viride is a potent elicitor of ethylene biosynthesis, localized cell death and other defense responses in specific cultivars of tobacco (Nicotiana tabacum) and tomato (Lycopersicon esculentum). Wild species of tomato, such as Lycopersicon cheesmanii and Lycopersicon pennellii, do not respond to EIX treatment. The F1 progeny of a L. esculentum×L. cheesmanii and a L. esculentum×L. pennellii cross responded to EIX treatment with an increase in ethylene biosynthesis and the induction of localized cell death. The F2 progeny of the above mentioned crosses segregated 3:1 (responding:non-responding). We mapped the EIX-responding locus (Eix) to the short arm of chromosome 7 using a population of introgression lines (ILs), containing small RFLP-defined chromosome segments of L. pennellii introgressed into L. esculentum. RFLP analysis of 990 F2 plants that segregated for the introgressed segment mapped the Eix locus 0.1 cM and 0.9 cM from the flanking markers TG61 and TG131, respectively. Using the marker TG61 we isolated a yeast artificial chromosome (YAC) clone that carries 300-kb DNA segments derived from the Eix region. By mapping the ends of this YAC clone we show that it spans the Eix locus. Thus, positional cloning of the Eix locus appears feasible.
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  • 35
    ISSN: 1432-2242
    Keywords: Keywords Resistance ; Tomato powdery mildew ; Tomato ; Mapping ; Oidium lycopersicum ; RFLP ; Sequence characterised amplified region (SCAR)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Lycopersicon hirsutum G1.1560 is a wild accession of tomato that shows resistance to Oidium lycopersicum, a frequently occurring tomato powdery mildew. This resistance is largely controlled by an incompletely dominant gene Ol-1 near the Aps-1 locus in the vicinity of the resistance genes Mi and Cf-2/Cf-5. Using a new F2 population (n=150) segregating for resistance, we mapped the Ol-1 gene more accurately to a location between the RFLP markers TG153 and TG164. Furthermore, in saturating the Ol-1 region with more molecular markers using bulked segregant analysis, we were able to identify five RAPDs associated with the resistance. These RAPDs were then sequenced and converted into SCAR markers: SCAB01 and SCAF10 were L. hirsutum-specific; SCAE16, SCAG11 and SCAK16 were L. esculentum-specific. By linkage analysis a dense integrated map comprising RFLP and SCAR markers near Ol-1 was obtained. This will facilitate a map-based cloning approach for Ol-1 and marker-assisted selection for powdery mildew resistance in tomato breeding.
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  • 36
    ISSN: 1432-2242
    Keywords: Key words Genomic map ; Acacia mangium ; Recombination rate ; Microsatellites ; RFLP ; Legumes ; Polyads
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  An integrated genetic linkage map, comprised of 219 RFLP and 33 microsatellite loci in 13 linkage groups, was constructed using two outbred pedigrees of Acacia mangium Willd. The linkage groups ranged in size from 23 to 103 cM and the total map length was 966 cM. Individual maps were made for each pedigree and the ordering of loci was consistent with the integrated map. The use of two independent pedigrees allowed a comparison of recombination rates between linked loci in male and female meioses as well as between parents. Differences were confined to specific regions and were not uniform across the male and female genomes or between genotypes. The heterogeneity in recombination frequencies did not result in major differences in the ordering of loci between pedigrees; hence, the integrated map provides a sound basis for QTL detection, leading to marker-assisted selection in A. mangium. It also provides a reference map for comparative genome analysis in acacias. The co-dominant markers used for mapping provide a useful resource in population studies and for quality control in acacia breeding programs. Detection of a relatively high proportion of selfs in pods derived from flowers which were not emasculated (30%), compared with emasculated flowers (0.01%), indicates that emasculation is desirable for efficient delivery of control-crossed seed in acacia breeding programs.
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  • 37
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    Theoretical and applied genetics 101 (2000), S. 613-624 
    ISSN: 1432-2242
    Keywords: Keywords Melon (Cucumis melo L.) ; Fruit ripening ; Ethylene production rate ; Postharvest fruit decay ; Shelf-life ; ACC oxidase ; ACC synthase ; SSR ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Sixty three cultigens from eight market types of the melon (Cucumis melo L. subsp. melo) groups Cantaloupensis and Inodorus were evaluated for ethylene production rate, shelf-life (postharvest decay), and RFLP polymorphisms. The ethylene production rates of melon fruits at maturity and (after) postharvest decay were measured on individual genotypes. The ethylene production rates of individual genotypes ranged from undetectable to 103 nl/g per h. The mean ethylene production rates of the eight market types, ranked from highest to lowest, were Eastern U.S. type, Charentais, Western U.S. type, Long Shelf-Life cantaloupes (LSL), Galia, Ananas, Honeydew, and Casaba. Ethylene production and postharvest decay rating were positively significantly correlated (r 2=0.87, P=0.05). Orange-fleshed melon fruits produced significantly (P=0.05) more ethylene than did green- or white-fleshed types. Melon fruits with a netted rind had significantly (P=0.05 for orange-flesh fruits and 0.01 for green- or white-flesh fruits) higher ethylene production than did smooth-type fruits. Using probes made from cDNAs encoding ACC oxidase (MEL1) or ACC synthase (MEACS1) genes, RFLPs were detected melon cultigens of the eight marker types showing varying ethylene production rates and different flesh colors. Low ethylene production and green- and white-flesh color were associated (r 2=0.91; P=0.05) with the presence of a putative RFLP-MEL1 allele A 0 (15-kb), whereas high ethylene production and orange-flesh color were associated with allele B 0 (8.5-kb) in the homozygous condition, after probing MEL1 with EcoRV-digested genomic DNA. Also, after probing MEACS1 with NdeI-digested genomic DNA, RFLP polymorphism revealed five fragments denoted as A, B, C, D and E, with molecular sizes of 5.2-, 4.2-, 3.8-, 3.0- and 1.0-kb, respectively. A two-fragment pattern, AB, and a three-fragment pattern, ACE, the two predominant RFLP patterns, were also associated with low and high ethylene production, respectively. The ACE fragment pattern was also associated with orange-flesh melons. Scoring of both probes allowed for the unique classification of most melon market types consistent with ethylene production and the postharvest decay phenotypes. Therefore, these RFLPs might have utility in marker-assisted selection for the development of melons with enhanced postharvest keeping ability.
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  • 38
    ISSN: 1432-2242
    Keywords: Keywords Oryza sativa L. ; Gene mapping ; Magna porthe grisea ; Pyricularia grisea ; Disease resistance ; Complete resistance ; Resistance genes ; Pyramiding ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A framework linkage map was developed using 284 F10 recombinant inbred lines (RILs) from a ’Lemont’×’Teqing’ rice cultivar cross. Evaluation of a subset of 245 of these RILs with five races of the rice blast pathogen permitted RFLP mapping of three major resistance genes from Teqing and one major gene from Lemont. All mapped genes were found to confer resistance to at least two blast races, but none conferred resistance to all five races evaluated. RFLP mapping showed that the three resistance genes from Teqing, designated Pi-tq5, Pi-tq1 and Pi-tq6, were present on chromosomes 2, 6 and 12, respectively. The resistance gene from Lemont, Pi-lm2, was located on chromosome 11. Pi-tq1 is considered a new gene, based on its reaction to these five races and its unique map location, while the other three genes may be allelic with previously reported genes. Lines with different gene combinations were evaluated for disease reaction in field plots. Some gene combinations showed both direct effects and non-linear interaction. The fact that some of the lines without any of the four tagged genes exhibited useful levels of resistance in the field plots suggests the presence of additional genes or QTLs affecting the blast reaction segregating in this population.
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  • 39
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    Theoretical and applied genetics 100 (2000), S. 519-527 
    ISSN: 1432-2242
    Keywords: Key words Wheat ; Triticum aestivum ; Physical mapping ; Deletion lines ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Extended physical maps of chromosomes 6A, 6B and 6D of common wheat (Triticum aestivum L. em Thell., 2n=6x=42, AABBDD) were constructed with 107 DNA clones and 45 homoeologous group-6 deletion lines. Two-hundred and ten RFLP loci were mapped, including three orthologous loci with each of 34 clones, two orthologous loci with each of 31 clones, one locus with 40 clones, two paralogous loci with one clone, and four loci, including three orthologs and one paralog, with one clone. Fifty five, 74 and 81 loci were mapped in 6A, 6B and 6D, respectively. The linear orders of the mapped orthologous loci in 6A, 6B and 6D appear to be identical and 65 loci were placed on a group-6 consensus physical map. Comparison of the consensus physical map with eight linkage maps of homoeologous group-6 chromosomes from six Triticeaespecies disclosed that the linear orders of the loci on the maps are largely, if not entirely, conserved. The relative distributions of loci on the physical and linkage maps differ markedly, however. On most of the linkage maps, the loci are either distributed relatively evenly or clustered around the centromere. In contrast, approximately 90% of the loci on the three physical maps are located either in the distal one-half or the distal two-thirds of the six chromosome arms and most of the loci are clustered in two or three segments in each chromosome.
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  • 40
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    Theoretical and applied genetics 101 (2000), S. 860-864 
    ISSN: 1432-2242
    Keywords: Key words Melon ; AFLP ; RFLP ; RAPD ; Genetic similarity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Three different types of molecular markers, RAPD, AFLP and RFLP were used to measure genetic diversity among six genotypes of Cucumis melo L. Each line represented a different melon genotype: Piel de Sapo, Ogen, PI161375, PI414723, Agrestis and C105. A number of polymorphic RAPD, AFLP and RFLP bands were scored on all materials and the genetic similarity measured. Clustering analysis performed with the three types of markers separated the genotypes into two main groups: (1) the sweet type, cultivated melons and (2) the exotic type, not cultivated melons. While the data obtained suggest that all three types of markers are equally informative, AFLPs showed the highest efficiency in detecting polymorphism.
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  • 41
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    Theoretical and applied genetics 101 (2000), S. 948-955 
    ISSN: 1432-2242
    Keywords: Key words Theobroma cacao L. ; AFLP ; Microsatellites ; RFLP ; High-density genetic map
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The first linkage map established by Lanaud et al. (1995) was used as a starting point to produce a high-density molecular linkage map. A mapping population of 181 progenies resulting from a cross between two heterozygous genotypes, a Forastero and a Trinitario (hybrid between Forastero and Criollo), was used for the linkage analysis. A new DNA isolation protocol was established, which allows enough good quality DNA to construct a genetic map with PCR-based markers. The map comprises 424 markers with an average spacing between markers of 2.1 cM. The marker types used were five isozymes, six loci from known function genes, 65 genomic RFLPs, 104 cDNA RFLPs, three telomeric probes, 30 RAPDs, 191 AFLPs and 20 microsatellites. The use of new marker types, AFLP and microsatellites, did not disturb the original order of the RFLP loci used on the previous map. The genetic markers were distributed over ten linkage groups and cover 885.4 cM. The maximum distance observed between adjacent markers was 16.2 cM, and 9.4% of all loci showed skewed segregation.
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  • 42
    ISSN: 1432-2242
    Keywords: Key words Sugarcane ; Rust resistance gene ; Comparative mapping ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A major rust resistance gene has been identified in a self-progeny of the sugarcane cultivar R570. Until now, this gene was known to be linked to a marker revealed by the sugarcane probe CDSR29 but unassigned to any linkage group of the current genetic map. We used synteny relationships between sugarcane and three other grasses in an attempt to saturate the region around this rust resistance gene. Comparison of sugarcane, sorghum, maize and rice genetic maps led to the identification of homoeologous chromosome segments at the extremity of sorghum linkage group D, rice linkage group 2, maize linkage group 4 and in the centromeric region of maize linkage group 5. One hundred and eighty-four heterologous probes were selected and tested for cross-hybridization with sugarcane DNA; 106 produced a good hybridization signal and were hybridized on 88 individuals of the R570 selfed progeny. Two hundred and seventeen single-dose markers were added to the R570 genetic map, of which 66% mapped to linkage group VII, together with the rust resistance gene. This gene has now been mapped to the end of a co-segregating group consisting of 19 RFLP markers. None of the mapped loci were located closer to the gene than CDSR29. The gene thus appears to reside at the edge of a ’’synteny cluster’’ used to describe the different grass genomes.
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  • 43
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    Molecular biology reports 22 (1995), S. 33-35 
    ISSN: 1573-4978
    Keywords: fish ; repetitive DNA ; RFLP ; satellite DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A member of satellite repetitive DNA was isolated and sequenced from a saltwater fishSillago japonica (Percoidei). This sequence consists of several oligo-dA/dT tracts and two inverted repeats which resemble each other. Dot blot hybridization analysis using a satellite DNA clone pSJ2 among the species in the suborder Percoidei revealed that the pSJ2 sequence was amplified at least after the family Sillaginidae had been derived.
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  • 44
    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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  • 45
    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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  • 46
    ISSN: 1573-5028
    Keywords: bark storage protein ; gene clustering ; RFLP ; systemic response
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    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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  • 47
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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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  • 48
    ISSN: 1573-5028
    Keywords: chalcone synthase genes ; genomic linkage ; multigene family ; Petunia ; RFLP
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    Notes: Abstract Chalcone synthase (CHS) genes in Petunia hybrida comprise a multigene family containing at least 7 complete members in the strain Violet 30 (V30). Based on a high sequence homology in both coding and non-coding sequence, a number of CHS genes can be placed into two subfamilies. By restriction fragment length polymorphism (RFLP) analysis it was shown that both chromosomes II and V carry one of these subfamilies, in addition to the other CHS genes identified so far. Members of a subfamily were found to be closely linked genetically. Analysis of the Petunia species that contributed to the hybrid nature of P. hybrida (P. axillaris, P. parodii, P. inflata and P. violacea) shows that none of the CHS gene clusters is specific for either one of the parents and therefore did not arise as a consequence of the hybridization. The number of CHS genes within a subfamily varies considerably among these Petunia species. From this we infer that the CHS subfamilies arose from very recent gene duplications.
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  • 49
    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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  • 50
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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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    Topics: Biology
    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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  • 51
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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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  • 52
    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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  • 53
    ISSN: 1573-5028
    Keywords: cDNA ; cyanogenesis ; β-glucosidase ; RFLP ; Trifolium repens
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    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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  • 54
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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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  • 55
    ISSN: 1573-5028
    Keywords: chalcone synthase cDNA (chs cDNA) ; RFLP ; anthocyanins ; flavonoids ; synteny
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    Topics: Biology
    Notes: Abstract The chalcone synthase is a key enzyme that catalyses the first dedicated reaction of the flavonoid pathway in higher plants. The chs gene and its protein product in rice has been investigated. The presence of a chalcone synthase (CHS) protein in rice seedlings and its developmental stage-specific expression has been demonstrated by western analysis. The chalcone synthase of rice was found to be immunologically similar to that of maize. A rice cDNA clone, Os-chs cDNA, encoding chalcone synthase, isolated from a leaf cDNA library of an indica rice variety Purpleputtu has been mapped to the centromeric region of chromosome 11 of rice. It was mapped between RFLP markers RG2 and RG103. RG2 is the nearest RFLP marker located at a genetic distance of 3.3 cM. Some segments of chromosome 11 of rice including chs locus are conserved on chromosome 4 of maize. The markers, including chs locus on chromosome 11 of rice are located, though not in the same order, on chromosome 4 of maize. Genetic analysis of purple pigmentation in two rice lines, Abhaya and Shyamala, used in the present mapping studies, indicated the involvement of three genes, one of which has been identified as a dominant inhibitor of leaf pigmentation. The Os-chs cDNA shows extensive sequence homology, both for DNA and protein (deduced), to that of maize, barley and also to different monocots and dicots.
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  • 56
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    Theoretical and applied genetics 73 (1987), S. 364-370 
    ISSN: 1432-2242
    Keywords: Brassica ; Mitochondrial DNA (mtDNA) ; Chloroplast DNA (cpDNA) ; Cytoplasmic male sterility (cms) ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary An assay is described whereby Eco RI restriction fragment length polymorphisms of mitochondrial and chloroplast DNAs can definitively identify cytoplasms of interest in Brassica crop development. Restrictable mitochondrial and chloroplast DNA is extracted from as little as 2–3 g and 0.5 g leaf tissue, respectively, and the donor plants are able to continue to develop in a normal manner. An unknown cytoplasm can be identified in three days, which is a considerable saving in time and labor compared to the several years required by traditional methods. The assay is very inexpensive and should be established as a routine procedure in laboratories involved in sexual or somatic Brassica hybrid production.
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  • 57
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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 77 (1989), S. 839-843 
    ISSN: 1432-2242
    Keywords: Lens culinaris ; Lens nigricans ; RFLP ; Wildlentil ; Genetic distance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Thirty accessions of domesticated (Lens culinaris ssp. culinaris) and wild (L. culinaris ssp. orientalis, L. culinaris ssp. odemensis, L. nigricans ssp. ervoides and L. nigricans ssp. nigricans) lentil were evaluated for restriction fragment length polymorphisms (RFLPs) using ten relative low-copy-number probes selected from partial genomic and cDNA libraries of lentil. Nei's average gene diversity was used as a measure of genetic variability for restriction fragment lengths within subspecies and a dendrogram was constructed from genetic distance estimates between subspecies. The wild lentils L. culinaris ssp. orientalis and L. culinaris ssp. odemensis showed the greatest variability for restriction fragment lengths and were closely positioned to domesticated lentil in the dendrogram. Little variability for restriction fragment lengths was observed within accessions of L. nigricans ssp. ervoides and L. nigricans ssp. nigricans. This observation is consistent with a previously published proposal that nigricans may have been independently domesticated. Estimates of genetic variability based on RFLPs tended to be greater than estimates from isozymes.
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  • 59
    ISSN: 1432-2242
    Keywords: Sugar beet ; Beta nana ; Beta vulgaris ; RFLP
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    Topics: Biology
    Notes: Summary cDNA probes have been used to assess genetic variation in beet using hybridisation techniques that detect restriction fragment length polymorphism. Probes have been identified which differ in the levels of variation that they can detect (i) within closely related genetic material of sugar beet, and (ii) between sugar beet and a taxonomically distant Beta species.
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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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  • 63
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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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  • 64
    ISSN: 1432-2242
    Keywords: RFLP ; PCR ; Rice ; Inheritance ; Nonradioactive
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    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
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    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
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    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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    Theoretical and applied genetics 84 (1992), S. 286-290 
    ISSN: 1432-2242
    Keywords: Taxonomy ; Banana ; RFLP ; Musa ; Phylogeny
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    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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  • 71
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    Theoretical and applied genetics 84 (1992), S. 845-850 
    ISSN: 1432-2242
    Keywords: Flax ; Rust ; RFLP ; Genetic segregation
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    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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  • 72
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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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  • 73
    ISSN: 1432-2242
    Keywords: Pea ; Pea seed-borne mosaic virus ; Disease resistance ; Genome mapping ; RFLP
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    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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  • 74
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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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    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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    Theoretical and applied genetics 86 (1993), S. 329-332 
    ISSN: 1432-2242
    Keywords: RFLP ; Alfalfa ; Genetic map ; Segregation distortion ; Plant breeding
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    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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    Theoretical and applied genetics 86 (1993), S. 811-821 
    ISSN: 1432-2242
    Keywords: Brassica ; Interspecific hybridization ; Amphidiploid ; RFLP ; Evolution ; Nuclear ; cytoplasmic interaction
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    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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    Theoretical and applied genetics 86 (1993), S. 833-836 
    ISSN: 1432-2242
    Keywords: T. aestivum ; RFLP ; Fingerprint
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    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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    Theoretical and applied genetics 86 (1993), S. 383-388 
    ISSN: 1432-2242
    Keywords: Mitochondria ; Inheritance ; RFLP ; Larix ; Conifers
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    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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  • 80
    ISSN: 1432-2242
    Keywords: DNA ; RFLP ; Soybean ; Genetic map
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    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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  • 81
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    Theoretical and applied genetics 86 (1993), S. 573-578 
    ISSN: 1432-2242
    Keywords: COXI ; COXII ; Maternal inerhitance ; mtDNA ; RFLP
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    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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  • 82
    ISSN: 1432-2242
    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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    Theoretical and applied genetics 86 (1993), S. 837-842 
    ISSN: 1432-2242
    Keywords: Androgenesis ; RFLP ; Maize ; Agronomic evaluation ; Gametoclonal variation
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    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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  • 84
    ISSN: 1432-2242
    Keywords: DNA ; RFLP ; QTL ; Soybean ; Recombinant inbreds
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    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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    Theoretical and applied genetics 87 (1993), S. 278-287 
    ISSN: 1432-2242
    Keywords: Maize ; RFLP ; Recombination ; Disturbed segregation ; DH/SSD lines
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    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.
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    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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  • 87
    ISSN: 1432-2242
    Keywords: Hevea brasiliensis ; RFLP ; Nuclear probes ; Isozyme ; Genetic diversity
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    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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    Theoretical and applied genetics 89 (1994), S. 865-872 
    ISSN: 1432-2242
    Keywords: Comparative mapping ; RFLP ; Barley ; Triticum tauschii ; Genome evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Hordeum vulgare (barley) and Triticum tauschii are related, but sexually incompatible, species. This study was conducted to determine the extent of homology between the genomes of barley and T. tauschii using a common set of restriction fragment length polymorphism (RFLP) markers. Results showed that 〉95% of low-copy sequences are shared, but 42% of the conserved sequences showed copy-number differences. Sixty-three loci were mapped in T. tauschii using RFLP markers previously mapped in barley. A comparison of RFLP marker order showed that, in general, barley and T. tauschii have conserved linkage groups, with markers in the same linear orders. However, six of the seven linkage groups of T. tauschii contained markers which mapped to unrelated (i.e., non-homoeologous) barley chromosomes. Additionally, four of the T. tauschii linkage groups contained markers that were switched in order with respect to barley. All the chromosome segments differing between T. tauschii and barley contained markers that were detected by multi-copy probes. The results suggest that the observed differences between the T. tauschii and barley genomes were brought about by duplications or deletions of segments in one or both species. The implications of these findings for genetic mapping, breeding, and plant genome evolution are discussed.
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    Theoretical and applied genetics 88 (1994), S. 818-823 
    ISSN: 1432-2242
    Keywords: Diploid wheat ; Einkorn ; RFLP ; Taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To study the relationships between different species of the Einkorn group, 55 different accessions ofTriticum monococcum,T. boeoticum,T. urartu,T. sinskajae,T. thaoudar andT. aegilopoides were analyzed. Fifteen anonymous probes and four clones corresponding to storage protein genes were used for detecting restriction fragment length polymorphisms (RFLPs). The DNA was restricted with the restriction enzymesAluI,HaeIII,RsaI andTaqI. The 25 probe/enzyme combinations employed yielded a total of 488 polymorphic fragments. Statistical analyses were performed using Jaccard's coefficient of similarity and principal coordinate analysis. Different values of similarity within the three main taxa,monococcum,boeoticum andurartu, were obtained; the grouping at the species level was quite well reflected by the RFLP analysis done here. The coincidence between RFLP data and the subspecies classification of theT. monococcum group was only partial. OneT. urartu accession is clearly different from all of the other 54 accessions. The need for an RFLP based revision of the Einkorn taxonomy is evident.
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  • 92
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    Theoretical and applied genetics 88 (1994), S. 824-830 
    ISSN: 1432-2242
    Keywords: Pleurotus cornucopiae ; rDNA unit ; RFLP ; Ribosomal polymorphism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the ribosomal DNA unit ofPleurotus cornucopiae, the rDNA coding regions are in the order 5′, 5S-18S-5.8S-25S, 3′, with the 5′ location of the 5S gene differing from its 3′ location found in other basidiomycetes. The most discriminating probe used to study the rDNA polymorphism consisted of a fragment that included the 5S, 18S and part of the 5.8S and 25S genes flanking three intergenic sequences. A high degree of rDNA polymorphism was observed in the sevenP. cornucopiae dikaryons studied. For the first time within a basidiomycete species, the restrictions maps distinguished two types of rDNA units (I and II). In each rDNA type, length variations in the external intergenic sequence IGS 1 located between the 25S and 5S genes allowed characterization of two different rDNA units in type I and four rDNA units in type II. This suggested that theP. cornucopiae rDNA units were derived from two kinds of ancestors (type I and II) by insertion or deletion events (100–700 bp) in the IGS 1. In four dikaryotic strains, two rDNA units of the same type (I or II) differing only by the IGS 1 length, were found in a similar number of copies, and presented a meiotic segregation in homokaryotic progeny. In one progeny, some homokaryotic strains possessed two different rDNA units: one with a high copy number and another with a lower one, showing that two different rDNA units could coexist in a single nucleus.
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  • 93
    ISSN: 1432-2242
    Keywords: DNA ; RFLP ; RAPD ; Brassica ; Genetic relationships
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) markers are being used widely for evaluating genetic relationships of crop germplasm. Differences in the properties of these two markers could result in different estimates of genetic relationships among some accessions. Nuclear RFLP markers detected by genomic DNA and cDNA clones and RAPD markers were compared for evaluating genetic relationships among 18 accessions from six cultivated Brassica species and one accession from Raphanus sativus. Based on comparisons of genetic-similarity matrices and cophenetic values, RAPD markers were very similar to RFLP markers for estimating intraspecific genetic relationships; however, the two marker types gave different results for interspecific genetic relationships. The presence of amplified mitochondrial and chloroplast DNA fragments in the RAPD data set did not appear to account for differences in RAPD- and RFLP-based dendrograms. However, hybridization tests of RAPD fragments with similar molecular weights demonstrated that some fragments, scored as identical, were not homologous. In all these cases, the differences occurred at the interspecific level. Our results suggest that RAPD data may be less reliable than RFLP data when estimating genetic relationships of accessions from more than one species.
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  • 94
    ISSN: 1432-2242
    Keywords: Wheat ; Spelt ; RFLP ; Marker Genetic diversity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Fifty-two winter wheat (Triticum aestivum L.), nine spring wheat, and 20 spelt (Triticum spelta L.) lines representing part of the European breeding germplasm, were assayed for RFLPs (restriction fragment length polymorphisms) with 56 wheat DNA clones and two barley cDNA clones. Objectives of this study were to (1) determine the level of variation for RFLPs in the wheat and spelt breeding lines, (2) characterize the genetic diversity within the European winter wheat germplasm, and (3) evaluate the usefulness of RFLP markers for pedigree analysis and the grouping of wheat and spelt lines of various origins. Seventy-three of the 166 RFLP loci detected with 58 probes and one restriction enzyme were polymorphic for the 81 lines. The percentage of polymorphic loci was greatest for the B genome (58%) and smallest for the D genome (21%). Among the 81 lines, 271 different RFLP bands were detected. RFLP band frequencies of the winter wheat lines differed considerably (≥0.5) from those of the spring wheat lines at five loci, and from those of the spelt lines at 17 loci. Eight cultivars that had a major impact as progenitors on the development of improved winter wheat cultivars accounted for 93% of the observed RFLP bands in winter wheat. Genetic distance (GD) estimates between two lines ranged between 0.01 and 0.21. Mean GD estimates within winter wheat (0.083), within spring wheat (0.108) and within spelt (0.096) were smaller than between spring and winter wheat (0.114), and greatest between winter wheat and spelt (0.132) and spring wheat and spelt (0.148). Principal coordinate analysis performed on GD estimates revealed a clear separation of wheat and spelt germplasm. Novel spelt lines with various proportions of wheat germplasm were positioned between wheat and traditional spelt lines. The spring wheat lines formed a distinct group at the periphery of the distribution of the winter wheat lines. Subgroupings of the winter wheat lines according to the cluster analysis were in good agreement with their origin, and lines with common ancestors were grouped together.
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  • 95
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    Theoretical and applied genetics 90 (1995), S. 1063-1067 
    ISSN: 1432-2242
    Keywords: T. monococcum ssp. monococcum ; T. monococcum ssp. boeoticum ; T. urartu ; RFLP ; Diversity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The A genome of the Triticeae is carried by three diploid species and subspecies of the genus Triticum: T. monococcum ssp. monococcum, T. monococcum ssp. boeoticum, and T. urartu, the A-genome donor of bread wheat. These species carry many genes of agronomic interest, including disease resistances, and may also be used for the genetic mapping of the A genome. The aim of this study was to evaluate the variability present in a sample of 25 accessions representative of this group using RFLP markers. Twenty probes, consisting of genomic DNA or cDNA from wheat, were used in combination with four restriction enzymes. A high level of polymorphism was found, especially at the interspecific level. Selecting the most informative enzymes appeared to be of great importance in order to obtain a stable structure for the diversity observed with only 20 probes. The results are largely consistent with taxonomy and data relating to geographical origins. The probes were also tested on 14 wheat cutivars. A good correlation coefficient was found for their informative values on wheat cultivars and diploid lines. Whether the group of species studied here would be useful for genetic mapping remains to be determined. Nevertheless, RFLP markers will be useful to follow genes that can possibly be introgressed from these species into cultivated wheat.
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  • 96
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    Theoretical and applied genetics 90 (1995), S. 1198-1203 
    ISSN: 1432-2242
    Keywords: Diplospory ; RFLP ; Bulk-segregant analysis ; Genome similarity ; Intergeneric hybrids ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Polyploid plants in the genus Tripsacum, a wild relative of maize, reproduce through gametophytic apomixis of the diplosporous type, an asexual mode of reproduction through seed. Moving gene(s) responsible for the apomictic trait into crop plants would open new areas in plant breeding and agriculture. Efforts to transfer apomixis from Tripsacum into maize at CIMMYT resulted in numerou intergeneric F1 hybrids obtained from various Tripsacum species. A bulk-segregant analysis was carried out to identify molecular markers linked to diplospory in T. dactyloides. This was possible because of numerous genome similarities among related species in the Andropogoneae. On the basis of maize RFLP probes, three restriction fragments co-segregating with diplospory were identified in one maize-Tripsacum dactyloides F1 population that segregated 1∶1 for the mode of reproduction. The markers were also found to be linked in the maize RFLP map, on the distal end of the long arm of chromosome 6. These results support a simple inheritance of diplospory in Tripsacum. Manipulation of the mode of reproduction in maize-Tripsacum backcross generations, and implications for the transfer of apomixis into maize, are discussed.
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  • 97
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    Theoretical and applied genetics 89 (1994), S. 211-216 
    ISSN: 1432-2242
    Keywords: Hordeum vulgare ; RFLP ; Genetic map ; Recombination ; Crossover
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The pattern of recombination in barley with regard to (1) the distribution of crossover points among whole gametes, (2) the distribution of crossover points among individual chromosomes and (3) the distribution of crossover points within chromosomes has been analysed using data sets underlying two recently published restriction fragment length polymorphism (RFLP) linkage maps representing male and female meiosis, respectively. The data indicated that the process of recombination had been random with no interference. The two data sets gave similar results, indicating that male and female meiosis in barley do no differ significantly. The possibility of using RFLP data in studies of crossover distribution is stressed.
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  • 98
    ISSN: 1432-2242
    Keywords: Evolution ; Tandemly repeated DNA sequences ; Phylogenetic relationships ; RFLP ; Poaceae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The recombinant plasmid dpTa1 has an insert of relic wheat DNA that represents a family of tandemly organized DNA sequences with a monomeric length of approximately 340 bp. This insert was used to investigate the structural organization of this element in the genomes of 58 species within the tribe Triticeae and in 7 species representing other tribes of the Poaceae. The main characteristic of the genomic organization of dpTa1 is a classical ladder-type pattern which is typical for tandemly organized sequences. The dpTa1 sequence is present in all of the genomes of the Triticeae species examined and in 1 species from a closely related tribe (Bromus inermis, Bromeae). DNA from Hordelymus europaeus (Triticeae) did not hybridize under the standard conditions used in this study. Prolonged exposure was necessary to obtain a weak signal. Our data suggest that the dpTa1 family is quite old in evolutionary terms, probably more ancient than the tribe Triticeae. The dpTa1 sequence is more abundant in the D-genome of wheat than in other genomes in Triticeae. DNA from several species also have bands in addition to the tandem repeats. The dpTa1 sequence contains short direct and inverted subrepeats and is homologous to a tandemly repeated DNA sequence from Hordeum chilense.
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  • 99
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    Theoretical and applied genetics 89 (1994), S. 259-264 
    ISSN: 1432-2242
    Keywords: RFLP ; Bootstrap ; Sampling variance ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Knowledge of genetic relationships among genotypes is useful in a plant breeding program because it permits the organization of germplasm and provides for more efficient sampling. The genetic distance (GD) among genotypes can be estimated using random restriction fragment length polymorphisms (RFLPs) as molecular markers. Knowledge of the sampling variance associated with RFLP markers is needed to determine how many markers are required for a given level of precision in the estimate of GD. The sampling variance for GD among all pairs of 37 maize (Z. mays L.) inbred lines was estimated from 1202 RFLPs. The 1202 polymorphisms were generated from 251 enzyme-probe combinations (EPC). The sampling variance was used to determine how large a sample of RFLPs was required to provide a given level of precision. The coefficient of variation (CV) associated with GD has a nearly linear relationship between its expected standard deviation and mean. The magnitude of the decrease in the mean CV for GD with increasing numbers of bands was dependent upon the sampling unit; e.g., individual polymorphic bands vs EPC, and the degree of relatedness among the inbreds compared. The rate of reduction in mean CV with increasing sample size was the same regardless of the restriction enzyme used, BamHI, EcoRI or HindIII, when the bootstrap sampling units were individual polymorphic bands. In constrast, although the rate of reduction (slopes) was the same, the intercepts of the mean CVs were different when EPCs were used as the bootstrap sampling unit. This difference was due to the higher number of bands per EPC in BamHI (4.94) compared with EcoRI (4.83) and HindIII (4.63).
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  • 100
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    Theoretical and applied genetics 89 (1994), S. 265-270 
    ISSN: 1432-2242
    Keywords: Bamboo ; Phyllostachys ; RFLP ; Variation ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phylogenetic and taxonomic difficulties are common within the woody bamboos, due to their unique life cycle, which severely limits the availability of floral characters. To addresss some of these problems, 20 species of woody bamboos in the genus Phyllostachys were analyzed using nuclear restriction fragment length polymorphisms (RFLPs). The RFLP data were used to generate genetic distances between all pairs of taxa and to examine the degree of genetic variation within and among bamboo species. The genetic distances were also used to create dendrograms of accessions and species. These trees supported the current division of the genus into two sections and provided some information on the thorny taxonomic problems in this group. We show that RFLPs can be used for species identification and the delineation of species limits.
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