天麻基因组微卫星特征分析与分子标记开发

该研究基于第二代测序技术建立了天麻的基因文库,筛选微卫星序列,并对微卫星位点的类型、丰度、长度、偏好性等进行了分析与比较;并为60条重复次数高的微卫星序列设计了引物,运用4个种群80个样本进行了PCR扩增和聚丙烯酰胺凝胶电泳检测。结果表明:(1)天麻基因组测序得到61 048条基因序列,检测出微卫星位点12 107个,其中二核苷酸重复最多、长度变异大。(2)设计的60对微卫星引物中的20对能扩增出清晰条带且有多态性,每个位点的复等位基因数(N_a)在4~14之间,平均为8.40;多态性信息含量(PIC)平均为0.77。该研究开发的天麻微卫星分子标记为开展天麻遗传学研究及种质资源鉴定等工作奠定了基础。

Genomic Microsatellite Characteristic Analysis and Molecular Marker Development for Gastrodia elata Bl.

In this study, the genomic library of G. elata was set up by means of the second generation of DNA sequencing, and the microsatellite loci in genomic sequences was screened. The types, abundance, length and preference of microsatellite locus were analyzed and compared. Primer pairs for 60 microsatellite loci with high repetition were designed, PCR and polyacrylamide gels electrophoresis were carried out on 80 individuals sampled from 4 populations in order to test the polymorphism of the markers. (1) 61 048 sequences of G. elate were gotten in the genomic sequencing, 12 107 microsatellite loci were detected out, occupying 8.16% of total sequences. (2) Among these SSR loci, the repetition of dinucleotide was the highest. 20 loci among those 60 were detected amplifiable, stable, and polymorphism after PCR and electrophoresis; the number of polymorphic alleles of each locus (N_a) ranged from 4 to 14 with the average of 8.40, while the average of polymorphism information content (PIC) was 0.77. The microsatellite markers developed for G. elate in this study can make good basement for its genetic study, germplasm identification.