目的 采用叶绿体基因(cp DNA)序列和ISSR分子标记技术对甘肃祁连山地区麻花秦艽进行序列变异和遗传分化分析。方法 利用试剂盒提取法提取麻花秦艽的基因组DNA,利用筛选出的trnS-trnG和rpl20-rps12引物进行扩增并测序,通过软件Chromas、ContigExpress对得到的序列进行校正、拼接,利用MegaX、DanSP5、GenALEx软件对拼接后的序列进行序列特征分析,最后利用PopART软件得到麻花秦艽的单倍型网状进化图。结果 18个麻花秦艽居群共计114个个体被成功扩增和测序;2个cp DNA片段经拼接后的长度为1 246 bp,共有676个多态性变异位点,459个单一突变位点、217个简约信息位点和107个插入-缺失位点;单倍型41个,其中特有单倍型34个;Tajima′s D检验在P>0.05水平上不显著,整体遵循中性进化模型。居群内变异百分比为89%,说明遗传变异主要存在于居群内;Fst=0.114(P<0.05),Nm=3.889说明居群间分化程度较低,居群间具有较强的基因交流。遗传距离与地理距离相关性分析r=-0.094(P<0.05),说明遗传距离与地理距离不具有明显的相关性。结论 甘肃祁连山地区麻花秦艽在物种水平上遗传多样性较高,具有丰富的单倍型类型,居群的遗传变异主要来自于遗传漂变。
Abstract
OBJECTIVE To analyze the sequence variation and genetic differentiation of Gentiana straminea Maxim. in Qilian mountain, Gansu province, using the chloroplast gene sequence and ISSR molecular marker technology in this experiment. METHODS Total DNA of G. straminea was extracted by kit extraction method, amplified and sequenced by two selected primers, the sequences were corrected and spliced by software Chromas and ContigExpress, the sequence characteristics of spliced sequences were analyzed by software MegaX, DanSP5, and GenALEx, and finally the haplotype network evolution diagram of G. straminea was drawn by software PopART. RESULTS A total of 114 individuals from 18 populations of G. straminea were successfully amplified and sequenced. The splice length of the two chloroplast DNA (cp DNA) fragments was 1 246 bp, and there were 676 polymorphic mutation sites, 459 single mutation sites, 217 condensed information sites, and 107 insert-deletion sites. The number of haplotypes was 41. Tajima′s D* test was not significant at P>0.05, which followed the neutral evolution model as a whole. The percentage of variation within the population was 89%, indicating that genetic variation mainly existed within the populations. The estimated pair-wise genetic differentiation (Fst) among the populations is 0.114 (P<0.05), and the gene flow was 3.889, respectively, indicating low genetic differentiation and a relatively high level of gene flow among populations. There was no obvious correlation between genetic differentiation among populations and geographical distance(r=-0.094, P<0.05). CONCLUSION The results show that G. straminea in the Qilian mountain area of Gansu province had high genetic diversity at the species level and rich haplotype types. The genetic variation of the population mainly come from genetic drift.
关键词
甘肃 /
麻花秦艽 /
叶绿体DNA /
遗传变异 /
遗传分化
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Key words
Gansu province /
Gentiana straminea Maxim /
cp DNA /
genetic variation /
genetic differentiation
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中图分类号：
R282
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基金
国家自然科学基金项目资助(81860673);甘肃省教育厅“双一流”科研重点项目资助(GSSYLXM-05);甘肃省高校中(藏)药化学与质量研究省级重点实验室开放基金项目资助(zzy-2019-04);中央本级重大增减支项目资助(2060302);甘肃省高等学校科学研究项目资助(2015A-098)
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