Polymorphism in spring and winter rapeseed varieties (<i>Brassica napus</i> L.) identified by ssr markers
Keywords:varietal diversity, rapeseed, microsatellite markers, cluster analysis
Purpose. To assess the genetic diversity of rapeseed varieties using SSR markers in order to create breeding material and use that material in complex in vitro selection for drought and salt tolerance.
Methods. PCR analysis, cluster analysis.
Results. The results of analysis of rapeseed varieties polymorphism based on molecular-genetic markers is presented.As a result of the analysis of rapeseed varieties, 41 alleles were detected using the studied markers, that is, an average of 10.3 alleles per marker. The number of polymorphic loci identified by four microsatellite markers (Ra3-H09, Na12-A02, FITO-063 and Na10-B07) was 24. The polymorphism level of the studied varieties was 51% on average and varied between 33% (identified by FITO-063) and 87% (identified by Na12-A02). According to the frequency distribution of the obtained alleles, the highest frequency by SSR marker Ra3-H09 had a 117 bp allele identified in three varieties: ‘Senator Liuks’, ‘Danhal’ and ‘Chornyi Veleten’. It was found that the unique alleles identified by Ra3-H09 were the alleles at a frequency of 0.06 and size of 135 bp (variety ‘Aliot’) and 156 bp (variety ‘Kliff’). FITO-063 marker identified the smallest number of alleles (5) at a frequency distribution ranging from 0.11 to 0.33. The unique alleles identified by FITO-063 marker were the ones at a frequency of 0.1 and size of 258 bp (variety ‘Geros’) and 273 bp (variety ‘Chornyi Veleten’). The maximum number of alleles was obtained using Na12-A02 marker. The distribution showed the highest frequency (0.11) for the 158 bp and 192 bp alleles. Using Na10-B07 marker, three alleles were identified at a frequency of 0.04. These 144, 156 and 194-bp alleles were found in varieties ‘Kliff’, ‘Geros’ and ‘Nelson’. Cluster analysis revealed four variety clusters: ‘Senator Liuks’ and ‘Danhal’, ‘NK Technik’ and ‘NK Petrol’, ‘Geros’ and ‘Aliot’, ‘Kliff’ and ‘Nelson’. ‘Chornyi Veleten’ variety did not enter any cluster. The most distant varieties are ‘Kliff’ and ‘Nelson’ with a genetic distance value of 3.32. Foreign varieties ‘NK Technik’ and ‘NEC Petrol’ with the value of genetic distances between them equal 1.41 appeared to be the most similar by the four studied SSR markers. Other varieties differed by at least one marker.
Conclusions. Consequently, using the set of four microsatellite markers provides an assessment of rapeseed varietal diversity that can be used in complex in vitro selection for drought and salt tolerance.
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Copyright (c) 2018 О. Л. Кляченко, Л. М. Присяжнюк, Н. В. Шофолова, О. В. Піскова
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