利用软件MISA对水生植物蓄菜转录组测序所获得的79536条EST序列进行分析,共检测出12319个EST—SSR位点。在蓄菜的EST—SSR中,二核苷酸和三核苷酸重复单元是主导类型,分别占总SSR的57.31%和30.87%,其中AG/CT、AAG/C1_r分别是二、三重复单元类型的优势重复基元,分别占总SSR的29.76%和8.66%。随机挑选了130对EST-SSR引物对蓄菜两个居群进行遗传多样性检测,结果发现:78对引物能扩增出清晰可分辨的条带,其中37对能成功检测出多态性,引物多态率为47.44%。这些多态性引物共检测出114个等位基因,每个位点2—6个,平均3.08个。观测杂合度(心)及预期杂合度(He)分别在0.229~1.000和0.351-0.756之间,多态信息含量PIC值在0.286~O.698之间,平均达0.495。以上研究结果表明,通过萏菜转录组测序产生的EsT数据来开发SSR标记是一种简单而高效的途径,这些新的SSR分子标记为研究苔菜的居群遗传多样性及其遗传结构提供了工具。
Using the program MISA, a total of 12319 EST-SSRs were sought out from 79536 ESTs of aquatic plant Nymphoides peltata. Dinucleotide and trinucleotide SSRs were the two main motif types, accounting for 57.31% and 30.87%, respectively. The most abundant di- and tri- motifs were AG/CT (29.76%) and AAG/CTT (8.66%), respectively. We randomly selected 130 primer pairs to evaluate their application and polymorphism across two populations of N. peltata. Seventy-eight SSR markers were amplified successfully, yielding clear and discernible bands, among which 37 were found to be polymorphic and the polymorphic rate was 47.44%. Using the polymorphic SSR markers, a total of 114 alleles were detected. The number of alleles per locus ranged from 2 to 6 (mean 3.08). Observed heterozygosity ( Ho) and expected value ( He) per locus varied from 0. 229 to 1. 000 and from 0. 351 to 0. 756, respectively. Polymorphism information content (PIC) ranged from 0. 286 to 0.698, with an average of 0. 495. The results indicated that the development of SSR molecular markers from the EST dataset .qenerated by next .qeneration sequencin.cl in the aouatic plant N.peltata is valuable and effective. These newly generated SSR makers will provide new tools for studying genetic diversity and population genetic structure of N. peltata.