National Institute of Plant Genome Research, Aruna Asaf Ali Marg, Post Box No, 10531, New Delhi 110067, India.
BMC Genomics. 2011 Feb 17;12:117. doi: 10.1186/1471-2164-12-117.
Chickpea (Cicer arietinum L.) is an economically important cool season grain legume crop that is valued for its nutritive seeds having high protein content. However, several biotic and abiotic stresses and the low genetic variability in the chickpea genome have continuously hindered the chickpea molecular breeding programs. STMS (Sequence Tagged Microsatellite Sites) markers which are preferred for the construction of saturated linkage maps in several crop species, have also emerged as the most efficient and reliable source for detecting allelic diversity in chickpea. However, the number of STMS markers reported in chickpea is still limited and moreover exhibit low rates of both inter and intraspecific polymorphism, thereby limiting the positions of the SSR markers especially on the intraspecific linkage maps of chickpea. Hence, this study was undertaken with the aim of developing additional STMS markers and utilizing them for advancing the genetic linkage map of chickpea which would have applications in QTL identification, MAS and for de novo assembly of high throughput whole genome sequence data.
A microsatellite enriched library of chickpea (enriched for (GT/CA)n and (GA/CT)n repeats) was constructed from which 387 putative microsatellite containing clones were identified. From these, 254 STMS primers were designed of which 181 were developed as functional markers. An intraspecific mapping population of chickpea, [ICCV-2 (single podded) × JG-62 (double podded)] and comprising of 126 RILs, was genotyped for mapping. Of the 522 chickpea STMS markers (including the double-podding trait, screened for parental polymorphism, 226 (43.3%) were polymorphic in the parents and were used to genotype the RILs. At a LOD score of 3.5, eight linkage groups defining the position of 138 markers were obtained that spanned 630.9 cM with an average marker density of 4.57 cM. Further, based on the common loci present between the current map and the previously published chickpea intraspecific map, integration of maps was performed which revealed improvement of marker density and saturation of the region in the vicinity of sfl (double-podding) gene thereby bringing about an advancement of the current map.
An arsenal of 181 new chickpea STMS markers was reported. The developed intraspecific linkage map defined map positions of 138 markers which included 101 new locations.Map integration with a previously published map was carried out which revealed an advanced map with improved density. This study is a major contribution towards providing advanced genomic resources which will facilitate chickpea geneticists and molecular breeders in developing superior genotypes with improved traits.
鹰嘴豆(Cicer arietinum L.)是一种经济上重要的冷季粮食豆科作物,因其高蛋白含量的营养种子而受到重视。然而,几种生物和非生物胁迫以及鹰嘴豆基因组中低遗传变异性不断阻碍了鹰嘴豆的分子育种计划。STMS(序列标记微卫星位点)标记已成为几种作物饱和连锁图谱构建的首选标记,也是检测鹰嘴豆等位基因多样性的最有效和可靠来源。然而,在鹰嘴豆中报道的 STMS 标记数量仍然有限,并且表现出较低的种内和种间多态性,从而限制了 SSR 标记在鹰嘴豆种内连锁图谱上的位置。因此,本研究旨在开发额外的 STMS 标记,并利用它们推进鹰嘴豆遗传连锁图谱的构建,这将有助于 QTL 鉴定、MAS 和高通量全基因组序列数据的从头组装。
从鹰嘴豆微卫星富集文库(富含(GT/CA)n 和(GA/CT)n 重复序列)中鉴定出 387 个含有微卫星的假定克隆,从中设计了 254 个 STMS 引物,其中 181 个被开发为功能标记。利用一个鹰嘴豆种内图谱群体 [ICCV-2(单荚)×JG-62(双荚)]和 126 个 RILs 进行了图谱定位。在 LOD 得分 3.5 的情况下,获得了 8 个连锁群,定义了 138 个标记的位置,跨越 630.9cM,平均标记密度为 4.57cM。此外,根据当前图谱和先前发表的鹰嘴豆种内图谱之间存在的共同位点,进行了图谱整合,提高了标记密度,并使 sfl(双荚)基因附近区域的饱和度得到了提高,从而使当前图谱得到了改进。
报告了 181 个新的鹰嘴豆 STMS 标记。所开发的种内连锁图谱定义了 138 个标记的图谱位置,其中包括 101 个新位置。与先前发表的图谱进行了图谱整合,得到了一个具有更高密度的改进图谱。本研究为提供先进的基因组资源做出了重大贡献,这将有助于鹰嘴豆遗传学家和分子育种家开发具有改良性状的优良基因型。