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  • 1
    ISSN: 1432-2242
    Schlagwort(e): Key words Early-methionine-labelled protein multigene family ; Heterologous PCR primers ; Grasses ; Sequence-tagged-sites ; Wheat-rye recombinants
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract  Two wheat consensus primer sets, directed to ”early-methionine-labelled” (Em) gene sequences, were tested for their ability to amplify beyond their original source. A range of widely diverse templates, including other Triticeae species and sample monocot and dicot species, was assayed. Primer set EMC5/EMC3, amplifying the entire coding region with its intron and part of the 3’ untranslated region, targets Triticeae and sorghum Em sequences. The other set, EMC5/EMCO31, directed to the coding region and its intron, amplifies templates from all the grass species. Both primer sets fail to amplify Em sequences from more distant monocots and the dicots. Using set EMC5/EMC3, we isolated and sequenced ten members of the rye Em gene family from five different rye sources. Significant DNA sequence variation between wheat and rye sequences in the non-coding regions was found, and this was used to develop seven sequence-specific primers. Twelve primer combinations were analysed, 7 of which were Em-R1-specific, amplifying a product in at least one of the tested rye or rye-carrying genotypes but not in wheat. Four sets exhibited clear amplification length polymorphisms which allowed discrimination between and within the rye sources. The primers also discriminated between wheat-rye recombinants with proximal 1RL rye chromatin and those carrying distal 1RL rye chromatin. These results show that wheat consensus primer sets can be used to isolate orthologous sequences, especially from species that are used for alien gene transfer in wheat. Subsequently, species-specific assays can be designed that are useful tools for this application.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    ISSN: 1573-5060
    Schlagwort(e): amplified fragment length polymorphism (AFLP) ; geneticdiversity ; geographical distribution ; Mexico ; Stylosanthes humilis
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Amplified fragment length polymorphism (AFLP) analysis was used to determine genetic relationships among 32 accessions of the species Stylosanthes humilis, representing 26 and 6 accessions of Mexican and South American origin, respectively. Seven AFLP primer pairs generated a total of 111 polymorphic bands with an average of 15.8 polymorphisms per combination and a polymorphic information content ranging from 0.89 to 0.79. Cluster analysis and principal coordinates analysis of the calculated similarity matrix clearly separated all the Mexican accessions, with one exception, from the accessions of South American origin. The observation of differentiation of these two main groups by AFLP analysis is highly congruent with previous studies based on morphological descriptors, cross-hybridization experiments, and chloroplast DNA sequence analysis, and may suggest that both groups evolved separately due to geographical isolation in the past, resulting in the existence of at least 2 principal gene pools. Within the Mexican collection, three groups were recognized, each corresponding to a geographical region with a low within-group and a high between-group genetic variability. Although most Mexican accessions formed one major group, one accession clustered with the South American gene pool. These findings show that Mexico may contain unique sources of S.humilis and therefore would merit attention for conservation and maintenance of S. humilis germplasm. Also, these results demonstrate that AFLP analysis is an efficient method for assessing genetic diversity among S. humilis accessions.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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