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利用中间锦鸡儿干旱转录组数据自主开发、筛选具有多态性的SSR标记;在此基础上,选取了内蒙古地区10个居群的中间锦鸡儿种质资源为试验材料,对其遗传多样性展开了系统研究。研究如下:中间锦鸡儿干旱转录组共搜索到404个SSR标记,分布于349条Unigenes序列上,出现频率为14.78%。其中二核苷酸重复是主要的类型,占总SSRs的64.90%;其次是三核苷酸重复,占总SSRs的33.70%;二核苷酸重复基元中以AG/CT和GA/TC为优势重复基元,占总SSRs的29.45%,三核苷酸重复基元以AAG/CTT为主,占总SSRs的5.69%。成功设计了76对引物并筛选出条带清晰、多态性好、重复性好的7对引物进行遗传多样性分析。共检测到29个等位基因(Na),有效等位基因数为1.101~2.007,Nei’s多样性指数为0.254;Shannon多样性指数(I)为0.193~0.709,平均每位点I值为0.482;种群间遗传距离(GD)的范围是0.002 7~0.068 8,遗传一致度(GI)的范围是0.933 5~0.997 3,采用Nei’s遗传关系用UPGMA进行聚类,10个居群分为2个亚群,包头市百灵庙镇和乌兰察布市凉城县聚为一个亚群,其余种群聚为另一个亚群。中间锦鸡儿种群间平均近交系数(Fis)大于0且基因流为3.726,遗传距离与地理距离相关系数R~2=0.023。分析表明供试材料的遗传多样性较低,可能是中间锦鸡儿居群受到自然和人为干预较多,遗传交流大所导致的。
Based on the data of drought-resistant transcriptome of Caragana intermedia, the SSR markers with polymorphism were independently developed. Based on these data, 10 Caragana intermedia populations in Inner Mongolia were selected as experimental materials and their genetic diversity System research. The results were as follows: A total of 404 SSR markers were found in the middle Caragana arid transcriptome, distributed in 349 Unigenes sequences, with a frequency of 14.78%. Among them, dinucleotide repeats were the main type, accounting for 64.90% of the total SSRs; followed by trinucleotide repeats, accounting for 33.70% of the total SSRs; dinucleotide repeat motifs with AG / CT and GA / TC The dominant repeat motifs accounted for 29.45% of the total SSRs. The trinucleotide repeat motifs were mainly AAG / CTT, accounting for 5.69% of the total SSRs. 76 pairs of primers were successfully designed and 7 pairs of primers with clear bands, good polymorphism and good repeatability were selected for genetic diversity analysis. A total of 29 alleles (Na) were detected. The number of effective alleles was 1.101 ~ 2.007, Nei’s diversity index was 0.254, Shannon’s diversity index (I) was 0.193 ~ 0.709, and the average I value per locus was 0.482. The genetic distance (GD) ranged from 0.002 7 to 0.068 8 and the genetic range (GI) ranged from 0.933 5 to 0.997 3. Nei’s genetic relationship was clustered by UPGMA, and the 10 populations were divided into 2 sub-populations Herds, Bailingmiao Town in Baotou and Liangcheng County in Wulanchabu were grouped into one subgroup and the rest were clustered into another subgroup. The average inbred coefficient (Fis) of Caragana intermedia populations was greater than 0 and the gene flow was 3.726. The correlation coefficient between genetic distance and geographic distance was R ~ 2 = 0.023. The analysis showed that the genetic diversity of the tested materials was low, which may be caused by the natural and man-made intervening Caragana intermedia populations.