植物学报 ›› 2018, Vol. 53 ›› Issue (3): 313-321.doi: 10.11983/CBB17193

所属专题: 药用植物专辑

• 研究报告 • 上一篇    下一篇

锁阳ITS序列遗传多样性分析

任梦云1,2, 杜乐山1, 陈彦君1,2, 张盾2, 沈奇2, 关潇1,*(), 张银东2,*()   

  1. 1中国环境科学研究院, 北京 100012
    2海南大学热带农林学院, 海口 570228
  • 收稿日期:2017-10-18 接受日期:2018-04-30 出版日期:2018-05-01 发布日期:2018-09-11
  • 通讯作者: 关潇,张银东 E-mail:cynthia815@126.com;23300558@163.com

Analysis on Genetic Diversity of Cynomorium songaricum by ITS Sequence

Ren Mengyun1,2, Du Leshan1, Chen Yanjun1,2, Zhang Dun2, Shen Qi2, Guan Xiao1,*(), Zhang Yindong2,*()   

  1. 1Chinese Research Academy of Environmental Sciences, Beijing 100012, China
    2College of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, China
  • Received:2017-10-18 Accepted:2018-04-30 Online:2018-05-01 Published:2018-09-11
  • Contact: Guan Xiao,Zhang Yindong E-mail:cynthia815@126.com;23300558@163.com

摘要:

为阐明锁阳(Cynomorium songaricum)的遗传结构与遗传多样性, 以甘肃省河西走廊地区及青海省共18个居群的188个锁阳个体为研究对象, 利用现代分子生物学技术, 采用序列分析方法从核-质基因方面对遗传结构进行了分析。结果显示, 锁阳ITS序列总长度为687 bp, 含有7个变异位点, 定义9个单倍型, 整体单倍型多态性Hd=0.294 20, 核苷酸多样性π=0.000 49。在整个单倍型网络中介图中, 单倍型H1位于中心位置, 并在所有的居群中均有分布, 为核心的古老单倍型。分子方差分析结果显示, 锁阳种群变异主要来源于种群内。根据ITS序列得到的群体间遗传分化系数以及Mantel检验结果, 锁阳种群间的遗传距离与地理距离之间不存在相关性, 表明现存的锁阳居群是相对近期发生生境片段化的产物。中性检验结果表明, 锁阳拒绝中性进化, 群体历经扩张或者基因座位受到负选择作用, 其中性零假说不能被排除。该研究为锁阳的系统分类、资源鉴定以及保护措施的制定提供了分子证据。

关键词: 锁阳, ITS, 遗传多样性, 中性假说

Abstract:

To elucidate the genetic structure and genetic diversity of Cynomorium songaricum, modern molecular biology techniques at the DNA level were used to study the genetic structure of 188 C. songaricum individuals from 18 wild populations in the Hexi Corridor Region of Gansu and Qinghai. After alignment, all amplified sequence lengths were 687 bp. The 687 bp ITS sequence detected 7 mutation sites in 188 individuals, defining 9 haplotypes. Processing these 9 haplotype sequences led to a data matrix for calculating the haplotype diversity (Hd=0.294 20) and nucleotide diversity (π=0.000 49). In the haplotype network map, H1 is located in the center and distributed in all populations, and is the ancient and core haplotype. AMOVA revealed that the variation in C. songaricum mainly occurs in populations. According to the genetic differentiation coefficient and Mantel test of ITS sequences, we found no significant relation between genetic and geographic distances, so the current distribution of C. songaricum represents the fragmentation product in recent time. Detection of historical expansion of populations showed that the Tajima’s D test rejected a neutral mutation evolution, and the population expansion history or gene locus is under negative selection pressure, so the null hypothesis cannot be ruled out. Our study provides molecular evidence for the classification system, identification and protection measures of C. songaricum.

Key words: Cynomorium songaricum, ITS, genetic diversity, neutral hypothesis

表1

18个锁阳居群样地分布信息"

Sample No. Location Longitude E (°) Latitude N (°) Height (m) Number
R1 Zhangye, Gansu 102.522 36.822 1803 11
R2 Minqin, Gansu 100.772 39.223 1370 17
R3 Jinchang, Gansu 102.54 38.44 1466 7
R4 Sandan, Gansu 101.118 38.897 2020 10
R5 Gaotai, Gansu 99.086 39.787 1339 11
R6 Sunan, Gansu 99.176 39.604 1395 10
R7 Sunan, Gansu 99.474 39.445 1391 12
R8 Jiuquan, Gansu 98.55 40.3 1232 4
R9 Yumen, Gansu 97.196 40.516 1395 10
R10 Guazhou, Gansu 95.585 40.212 1343 11
R11 Dunhuang, Gansu 94.583 40.368 1033 11
R12 Subei, Gansu 96.619 39.397 2355 10
R13 Delingha, Qinghai 97.334 37.206 2780 11
R14 Geermu, Qinghai 92.839 36.701 2790 7
R15 Dulan, Qinghai 98.164 36.476 2972 12
R16 Wulan, Qinghai 98.622 36.458 2917 12
R17 Gonghe, Qinghai 100.183 36.446 2904 11
R18 Guide, Qinghai 101.639 36.219 2389 11

表2

锁阳核糖体ITS序列的单倍型多态性位点"

Haplotype Variable sites Abundance
63 86 197 274 421 605 623
H1 G G G T G C G 143
H2 A G G T G C T 3
H3 G A G T G C G 1
H4 G A G T G G G 2
H5 A G G T G C G 26
H6 G G A T G C G 5
H7 G G A G G C G 1
H8 G G G T G G G 4
H9 G G G T A C G 3

表3

锁阳18个居群的单倍型遗传多样性组成"

Population code Samples Haptotypes Hd
R1 11 H1, H2, H3, H4 0.25974
R2 17 H1, H5, H6 0.11586
R3 7 H1, H5 0.43956
R4 10 H1, H2, H5 0.42632
R5 11 H1, H6 0.24675
R6 10 H1, H7, H8 0.19474
R7 12 H1, H5 0.08333
R8 4 H1, H5, H9 0.60714
R9 10 H1, H8 0.10000
R10 11 H1, H5, H8 0.17749
R11 11 H1, H4, H6 0.17749
R12 10 H1 0
R13 11 H1 0
R14 7 H1, H6 0.14286
R15 12 H1, H9 0.08333
R16 12 H1, H9 0.08333
R17 11 H1, H8 0.09091
R18 11 H1 0
Total 188 0.29420

图1

基于锁阳ITS序列的严格一致性树步长=784, CI=0.949 0, RI=0.911 7, RCI=0.865 2。分支上的数字分别代表MP/ML/BI的支持率。"

图2

锁阳18个居群基于ITS序列的单倍型网络图圆圈的大小与单倍型的相对频率成正比; 不同颜色代表锁阳属不同种群。"

表4

锁阳种群遗传结构参数"

Sequence HS (SE) HT (SE) GST (SE) NST (SE)
ITS 0.179 (0.0390) 0.277 (0.0820) 0.353 (0.1914) 0.408 (0.2038)

表5

锁阳核糖体基因单倍型分子方差分析"

Source of variation df Sum of squares Variance components Percentage FST P
Among populations 17 28.833 0.07689 Va 44.57 0.44566 <0.001*
Within populations 358 34.239 0.09564 Vb 55.43
Total 375 63.072 0.17253

图3

基于ITS序列的锁阳种群失配分析柱形代表变异位点在群体扩张模型下的预期分布; 虚线代表变异位点的实际分布。"

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