Sequence-tagged microsatellite site markers for chickpea (Cicer arietinum L.)
Sequence-tagged microsatellite site markers for chickpea (Cicer arietinum L.)
复制标题
DOI：
10.1139/gen-42-2-210
复制
发表时间：
1999-04-01
期刊：
影响因子：
3.1
通讯作者：
Kahl, G
中科院分区：
文献类型：
--
作者：
Hüttel, B;Winter, P;Kahl, G
用微卫星特异性寡核苷酸探针对鹰嘴豆（Cicer arietinum L.）的两个小插入基因组文库进行了筛选。在13000个镀菌落中共鉴定出121个阳性克隆。经（TAA）鉴定的无性系有39个(5),（GA）鉴定的有26个(8),（GT）鉴定的有18个(8)。27个由富含at的三核苷酸重复序列组成[(CAA)(5)、（CAT）(5)和（GAA）(5)]， 11个由富含gc的三核苷酸组成[(TCC)(5)、（CAC）(5)、tCAG(5)和（CGA）(5)]]。在选择测序的53个克隆中，有43个携带了一颗微卫星。侧翼引物对设计了28个位点，并用于一个小的测试集，其中包括1个网纹蒿和4个雀叶蒿。在琼脂糖或聚丙烯酰胺凝胶上分离PCR产物，发现22对引物具有预期大小的单条带。其中16个“黄花苜蓿序列标记微卫星位点”（CaSTMS）标记在种内水平上具有多态性，在4个被检测的材料中检测到2-4个等位基因。引物对CaSTMS10和CaSTMS15分别在63份不同地理位置的鸡蒿中发现25个和16个等位基因，基因多样性值分别为0.937和0.922。用一组重组自交系及其亲本证明了CaSTMS标记的孟德尔遗传。
Two small-insert genomic libraries of chickpea (Cicer arietinum L. were screened with a set of microsatellite-specific oligonucleotide probes. A total of 121 positive clones were identified among 13 000 plated colonies. Thirty-nine clones were recognized by (TAA)(5), 26 by (GA)(8), 18 by (GT)(8). 27 by a pool of AT-rich trinucleotide repeats [(CAA)(5), (CAT)(5), and (GAA)(5)], and 11 by a pool of GC-rich trinucleotides [(TCC)(5), (CAC)(5), tCAG)(5), and (CGA)(5)]. Of 53 clones selected for sequencing, 43 carried a microsatellite. Flanking primer pairs were designed for 28 loci, and used on a small test-set comprising one C. reticulatum and four C. arietinum accessions. Separation of the PCR products on agarose or polyacrylamide gels revealed single bands of the expected size with 22 of the primer pairs. Sixteen of these "Cicer arietinum sequence-tagged microsatellite site" (CaSTMS) markers were polymorphic at an intraspecific level, detecting 2-4 alleles within the four accessions examined. Primer pairs CaSTMS10 and CaSTMS15 revealed 25 and 16 alleles among 63 C. arietinum accessions from different geographic locations, reflecting gene diversity values of 0.937 and 0.922, respectively. Mendelian inheritance of CaSTMS markers was demonstrated using a set of recombinant inbred lines and their parents.