MOLECULAR CHARACTERIZATION OF COLD TOLERANT GERMPLASM OF PHASEOLUS BEANS WITH SEQUENCE RELATED AMPLIFIED POLYMORPHISM (SRAP) AND RETROTRANSPOSON-BASED INTERPRIMER BINDING SITES (iPBS) MARKERS

June 1 Pub Date : 2023-02-10 DOI:10.36899/japs.2023.3.0655
B. T. Certel, H. İkten, Y. Yilmaz, F. Kantar, V. Çiftçi, V. Gözen, A. Tepe
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Abstract

In this study genetic diversity of 55 Phaseolus sp. beans selected for cold tolerance from different regions of Türkiye was investigated by using sequence related amplified polymorphism (SRAP) and retrotransposon-based interprimer binding sites (iPBS)markers. Four commercially registered cultivars, one accession of Phaseolus coccineus and a tepary bean Phaseaolus acutifolius species were included for comparison. Genomic DNA was isolated from young fresh leaves and PCR reaction was carried out using 30 SRAP ( sequence related amplified polymorphism) and 12 iPBS (retrotransposon-based interprimer binding sites) primers. Similarity analyses were performed and dendrograms were produced according to the Unweighted Pair-Group Mean Arithmetic method (UPGMA). In PCR reactions, 331 total and 146 polymorphic bands were produced with 30 SRAP primer combinations. The number of polymorphic bands ranged between 1 and 12 with an average 4.86 polymorphic marker for each primer pair. Twelve iPBS primers produced 156 total bands and 72 of them were polymorphic. The highest polymorphism was obtained with SRAP primer combinations of Me8Em3 and Me7Em14 and iPBS primers 2270, 2394 and 2252. Characterization of germplasm with SRAP and iPBS primers was discussed in relation to cold tolerance, species, source, seed size, seed color and growth type. In conclusion, genetic variability of germplasm of 55 Phaseolus bean species, genotypes and cultivars selected for cold tolerance were effectively assessed by PCR based molecular techniques, SRAP and iPBS. High levels of polymorphism determined in the core collection may be used in breeding programs for the development of cold tolerant superior cultivars
利用序列相关扩增多态性(SRAP)和基于反转录转座子的引物间结合位点(iPBS)标记对菜豆耐寒种质进行分子鉴定
利用序列相关扩增多态性(SRAP)和基于反转录转座的引物间结合位点(iPBS)标记,研究了55份来自 rkiye基因不同区域的菜豆耐寒性的遗传多样性。选取4个商业登记品种、1个种属菜豆(Phaseolus coccineus)和1个赤豆菜豆(Phaseaolus acutifolius)进行比较。利用30条序列相关扩增多态性(SRAP)和12条基于反转录转座的引物间结合位点(iPBS)引物进行PCR反应。根据未加权对组平均算法(Unweighted Pair-Group Mean algorithm, UPGMA)进行相似性分析并生成树形图。在PCR反应中,30个SRAP引物组合共产生331个多态性条带和146个多态性条带。多态性条带数在1 ~ 12个之间,平均每对引物有4.86个多态性标记。12条iPBS引物共产生156条条带,其中72条为多态性条带。Me8Em3和Me7Em14的SRAP引物组合与iPBS引物2270、2394和2252的多态性最高。利用SRAP和iPBS引物从耐寒性、种类、来源、种子大小、种子颜色和生长类型等方面对种质进行了表征。综上所述,采用基于PCR的分子技术、SRAP和iPBS对55个菜豆品种、基因型和耐寒品种的遗传变异进行了有效评价。在核心种质中确定的高水平多态性可用于培育耐寒优良品种
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