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利用抗病基因类似物- EST标记评估小麦基因型的遗传多样性

Assessment of genetic diversity of wheat genotypes by resistance gene analog-EST markers.

作者信息

Karakas O, Gurel F, Uncuoglu A A

机构信息

The Scientific and Technological Research Council of Turkey, Marmara Research Center, Genetic Engineering and Biotechnology Institute, Gebze-Kocaeli, Turkey.

出版信息

Genet Mol Res. 2011 Jun 14;10(2):1098-110. doi: 10.4238/vol10-2gmr1065.

Abstract

Resistance gene analog-expressed sequence tag (RGA-EST)-based markers have been used for variety discrimination and studies of genetic diversity in wheat. Our aim is to increase the competitiveness of public wheat breeding programs through intensive use of modern selection technologies, mainly marker-assisted selection. The genetic diversity of 77 wheat nucleotide binding site (NBS)-containing RGA-ESTs was assessed. Resistant and susceptible bread wheat (Triticum aestivum) genotypes were used as sources of DNA for PCR amplifications. In our previous studies, the F₂ individuals derived from the combinations PI178383 x Harmankaya99, Izgi2001 x ES14, and Sonmez2001 x Aytin98 were evaluated for yellow rust resistance at both seedling and adult stages to identify DNA markers. We have now examined the genetic variability among the resistant and susceptible Turkish wheat cultivars for yellow rust disease and the mean genetic distance between the cultivars. The highest similarity was 0.500 between Harmankaya99 and Sonmez2001. The lowest similarity was 0.286 between Aytin98, PI178383 and Aytin98, ES14. A relatively high level (49.5%) of polymorphism was observed with 77 RGA-EST primers across the six wheat genotypes, despite the fact that all of them were local cultivars from geographically close locations. RGA-EST sequences were compared by BlastX algorithms for amino acid sequences to determine the polymorphic categories among the combinations. BlastX analyses of six RGA-ESTs that gave polymorphic patterns for all combinations were NBS-LRR class RGA, NB-ARC domain containing protein, NBS-type resistance protein RGC5, NBS-LRR-S/ TPK stem rust resistance protein, and putative MLA1 proteins, while 38 RGA-EST gave a monomorphic pattern.

摘要

基于抗病基因类似物表达序列标签(RGA-EST)的标记已被用于小麦品种鉴别和遗传多样性研究。我们的目标是通过密集使用现代选择技术,主要是标记辅助选择,来提高公共小麦育种项目的竞争力。对77个含小麦核苷酸结合位点(NBS)的RGA-EST的遗传多样性进行了评估。抗性和感病的面包小麦(普通小麦)基因型用作PCR扩增的DNA来源。在我们之前的研究中,对PI178383×Harmankaya99、Izgi2001×ES14和Sonmez2001×Aytin98组合产生的F₂个体在幼苗期和成年期进行了条锈病抗性评估,以鉴定DNA标记。我们现在研究了抗条锈病和感条锈病的土耳其小麦品种之间的遗传变异性以及品种间的平均遗传距离。Harmankaya99和Sonmez2001之间的相似性最高,为0.500。Aytin98、PI178383与Aytin98、ES14之间的相似性最低,为0.286。尽管所有六个小麦基因型均为来自地理位置相近的地方品种,但使用77个RGA-EST引物在这六个小麦基因型中观察到了相对较高水平(49.5%)的多态性。通过BlastX算法对RGA-EST序列的氨基酸序列进行比较,以确定各组合间的多态性类别。对所有组合均产生多态性模式的六个RGA-EST进行BlastX分析,结果表明它们属于NBS-LRR类RGA、含NB-ARC结构域的蛋白、NBS型抗病蛋白RGC5、NBS-LRR-S/T蛋白激酶杆锈病抗性蛋白以及假定的MLA1蛋白,而38个RGA-EST呈现单态模式。

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