Winter P, Pfaff T, Udupa S M, Hüttel B, Sharma P C, Sahi S, Arreguin-Espinoza R, Weigand F, Muehlbauer F J, Kahl G
Plant Molecular Biology, Biocenter, University of Frankfurt, Germany.
Mol Gen Genet. 1999 Aug;262(1):90-101. doi: 10.1007/s004380051063.
A size-selected genomic library comprising 280,000 colonies and representing approximately 18% of the chickpea genome, was screened for (GA)n, (GAA)n and (TAA)n microsatellite-containing clones, of which 389 were sequenced. The majority (approximately 75%) contained perfect repeats; interrupted, interrupted compound and compound repeats were only present in 6%-9% of cases. (TAA)-microsatellites contained the longest repeats, with unit numbers from 9 to 131. For 218 loci primers could be designed and used for the detection of microsatellite length polymorphisms in six chickpea breeding cultivars, as well as in C. reticulatum and C. echinospermum, wild, intercrossable relatives of chickpea. A total of 174 primer pairs gave interpretable banding patterns, 137 (79%) of which revealed at least two alleles on native polyacrylamide gels. A total of 120 sequence-tagged microsatellite site (STMS) markers were genetically mapped in 90 recombinant inbred lines from an inter-species cross between C. reticulatum and the chickpea cultivar ICC 4958. Markers could be arranged in 11 linkage groups (at a LOD score of 4) covering 613 cM. Clustering as well as random distribution of loci was observed. Segregation of 46 markers (39%) deviated significantly (P > or = 0.05) from the expected 1:1 ratio. The majority of these loci (73%) were located in three distinct regions of the genome. The present STMS marker map represents the most advanced co-dominant DNA marker map of the chickpea genome.
构建了一个大小选择的基因组文库，包含280,000个菌落，约占鹰嘴豆基因组的18%，从中筛选含(GA)n、(GAA)n和(TAA)n微卫星的克隆，对其中389个进行了测序。大多数（约75%）包含完美重复序列；间断、间断复合和复合重复序列仅在6%-9%的情况下出现。(TAA) - 微卫星包含最长的重复序列，单元数从9到131。针对218个位点设计了引物，并用于检测六个鹰嘴豆栽培品种以及鹰嘴豆的野生可杂交近缘种网脉鹰嘴豆和刺籽鹰嘴豆中的微卫星长度多态性。总共174对引物产生了可解释的条带模式，其中137对（79%）在天然聚丙烯酰胺凝胶上显示至少两个等位基因。总共120个序列标签微卫星位点（STMS）标记被定位到来自网脉鹰嘴豆和鹰嘴豆品种ICC 4958种间杂交的90个重组自交系中。这些标记可以排列成11个连锁群（LOD值为4），覆盖613 cM。观察到位点的聚类以及随机分布。46个标记（39%）的分离显著偏离（P≥0.05）预期的1:1比例。这些位点中的大多数（73%）位于基因组的三个不同区域。目前的STMS标记图谱代表了鹰嘴豆基因组最先进的共显性DNA标记图谱。
