青藏高原植物手参的谱系地理学研究

生物技术通报 ›› 2016, Vol. 32 ›› Issue (12): 96-102.

青藏高原植物手参的谱系地理学研究

鲍武印¹,张阳²,林鹏程³,南蓬¹,黄艳燕¹，靳浩飞¹,钟扬¹

1. 复旦大学生命科学学院分子与进化生态学实验室，上海 200438；
2. 上海科技馆自然史研究中心，上海 200041；
3. 青海民族大学青海省青藏高原植物化学重点实验室，西宁 810007

收稿日期:2016-06-03 出版日期:2016-12-25 发布日期:2016-12-07
作者简介:鲍武印，男，硕士研究生，研究方向：系统与进化植物学，E-mail：13210700001@fudan.edu.cn；张阳和林鹏程为本文并列第一作者
基金资助:
青海省自然科学基金项目（2014-ZJ-923）

Phylogeography of Gymnadenia conopsea from the Qinghai-Tibet Plateau

BAO Wu-yin¹,ZHANG Yang²,LIN Peng-cheng³,NAN Peng¹,HUANG Yan-yan¹,JIN Hao-fei¹,ZHONG Yang¹

1. Molecular and Evolutionary Ecology Lab，School of Life Science，Fudan University，Shanghai 200438；
2. Natural History Research Center，Shanghai Science and Technology Museum，Shanghai 200041；
3. Key Laboratory for Tibet Plateau Phytochemistry of Qinghai Province，Qinghai University for Nationalities，Xining 810007

Received:2016-06-03 Published:2016-12-25 Online:2016-12-07

摘要/Abstract

摘要： 利用叶绿体基因（rbcl和psbA-trnH）对青藏高原地区的10个手参（Gymnadenia conopsea）种群进行谱系地理学研究，旨在揭示手参的遗传谱系分布格局及其演化过程。结果表明，共得到11个单倍型，分子方差分析显示65.93%的遗传变异来源于种群间，FST值为0.659（P<0.01），种群间存在显著的遗传分化；遗传分化系数NST（0.398）小于GST（0.626），说明青藏高原地区的手参没有明显的谱系地理结构；多峰的失配分析曲线和Tajima’s D中性检验（-1.455，P>0.1）表明手参种群近期没有经历扩张；系统发育分析和分歧时间估计表明，现代青藏高原上的手参为本地起源，手参单倍型在19.08 Mya的第三纪中新世时期开始分化，与青藏高原隆起关系密切，青藏高原上手参的遗传分布格局在第四纪冰期之前已基本形成。

关键词: 手参, 谱系地理学, 遗传多样性, 青藏高原

Abstract: Chloroplast DNA（rbcl and psbA-trnH）of Gymnadenia conopsea（G. conopsea）from the Qinghai-Tibet Plateau was used for phylogeography research，aiming at revealing the distribution pattern and evolutional process of genetic lineage of G. conopsea. Eleven haplotypes were identified in 199 samples of 10 G. conopsea populations. The analysis of molecular variance（AMOVA）showed 65.93% of the genetic variance was from inter-populations and FST value of 0.659（P<0.01）also indicated that there was significant genetic differentiation between different populations. The genetic differentiation coefficients NST（0.398）< GST（0.626）implied no significant phylogeographic structure for G. conopsea populations in the Qinghai-Tibet Plateau. Mismatch analysis of multi-peak and Tajima’s D neutral test（-1.455，P>0.1）demonstrated G. conopsea populations had not experienced abrupt expansion recently. Phylogenetic analysis and divergence time estimation showed that G. conopsea in the Qinghai-Tibet Plateau originated indigenously，haplotypes started to differentiate 19.08 million years ago（Mya）in Miocene，indicating that the genetic differentiation of G. conopsea was related to the uplift of the Qinghai-Tibetan Plateau，and the genetic distribution pattern of G. conopsea was mostly formed before the Quaternary glaciation.

Key words: Gymnadenia conopsea, phylogeography, genetic diversity, the Qinghai-Tibet Plateau

中图分类号:

10.13560/j.cnki.biotech.bull.1985.2016.12.016

鲍武印,张阳,林鹏程,南蓬,黄艳燕，靳浩飞,钟扬. 青藏高原植物手参的谱系地理学研究[J]. 生物技术通报, 2016, 32(12): 96-102.

BAO Wu-yin,ZHANG Yang,LIN Peng-cheng,NAN Peng,HUANG Yan-yan,JIN Hao-fei,ZHONG Yang. Phylogeography of Gymnadenia conopsea from the Qinghai-Tibet Plateau[J]. Biotechnology Bulletin, 2016, 32(12): 96-102.

参考文献

[1] Campbell VV, Rowe G, Beebee TJ, et al. Genetic differentiation amongst fragrant orchids（Gymnadenia conopsea sl）in the British Isles[J] . Botanical Journal of the Linnean Society, 2007, 155（3）：349-360.
[2] 杨永昌. 藏药志[M] . 西宁：青海人民出版社, 1991.
[3] 金洪, 王俊杰. 手掌参的组织培养研究[J] . 内蒙古农牧学院学报, 1995, 16（2）：74-77.
[4] Liu JQ, Sun YS, Ge XJ, et al.[4] Liu JQ, Sun YS, Ge XJ, et al. Phylogeographic studies of plants in China：advances in the past and directions in the future[J] . Journal of Systematics and Evolution, 2012, 50（4）：267-275.
[5] Wen J, Zhang JQ, Nie ZL, et al. Evolutionary diversifications of plants on the Qinghai-Tibetan Plateau[J] . Front Genet, 2014, 5（4）：1-16.
[6] Chen JM, Du ZY, Sun SS, et al. Chloroplast DNA phylogeography reveals repeated range expansion in a widespread aquatic herb Hippuris vulgaris in the Qinghai-Tibetan Plateau and adjacent areas[J] . PLoS One, 2013, 8（4）：60948-60948.
[7] Wei X, Wang R, Cao L, et al. Origin of Oryza sativa in China inferred by nucleotide polymorphisms of organelle DNA[J] . PLoS One, 2012, 7（11）：49546-49546.
[8] Zhao C, Wang CB, Ma XG, et al. Phylogeographic analysis of a temperate-deciduous forest restricted plant（Bupleurum longiradiatum Turcz.）reveals two refuge areas in China with subsequent refugial isolation promoting speciation[J] . Molecular Phylogenetics and Evolution, 2013, 68（3）：628-643.
[9] Gustafsson S, Lonn M. Genetic differentiation and habitat preference of flowering-time variants within Gymnadenia conopsea[J] . Heredity, 2003, 91（3）：284-292.
[10] Scacchi R, De Angelis G. Isoenzyme polymorphisms in Gymnaedenia conopsea and its inferences for systematics within this species[J] . Biochemical Systematics and Ecology, 1989, 17（1）：25-33.
[11] Gustafsson S, Thoren PA. Microsatellite loci in Gymnadenia conopsea, the fragrant orchid[J] . Molecular Ecology Notes, 2001, 1（1-2）：81-82.
[12] Stark C, Michalski SG, Babik W, et al. Strong genetic differentiation between Gymnadenia conopsea and G. densiflora despite morphological similarity[J] . Plant Systematics and Evolution, 2011, 293（1-4）：213-226.
[13] Little DP, Barrington DS. Major evolutionary events in the origin and diversification of the fern genus Polystichum（Dryopteridac-eae）[J] . American Journal of Botany, 2003, 90（3）：508-514.
[14] Taberlet P, Gielly L, Pautou G, et al. Universal primers for amplification of three non-coding regions of chloroplast DNA[J] . Plant Molecular Biology, 1991, 17（5）：1105-1109.
[15] Thompson JD, Gibson TJ, Plewniak F, et al. The CLUSTAL_X windows interface：flexible strategies for multiple sequence alignment aided by quality analysis tools[J] . Nucleic Acids Research, 1997, 25（24）：4876-4882.
[16] Tamura K, Stecher G, Peterson D, et al. MEGA6：molecular evolutionary genetics analysis version 6. 0[J] . Molecular Biology and Evolution, 2013, 30（12）：2725-2729.
[17] Nei M, Tajima F, Tateno Y. Accuracy of estimated phylogenetic trees from molecular data[J] . Journal of Molecular Evolution, 1983, 19（2）：153-170.
[18] Nei M. Molecular evolutionary genetics[M] . New York：Colum-bia University Press, 1987.
[19] Rozas J, Sanchez-Delbarrio JC, Messeguer X, et al. DnaSP, DNA polymorphism analyses by the coalescent and other methods[J] . Bioinformatics, 2003, 19（18）：2496-2497.
[20] Clement M, Posada D, Crandall KA. TCS：a computer program to estimate gene genealogies[J] . Molecular Ecology, 2000, 9（10）：1657-1659.
[21] Yang ZH. PAML 4：phylogenetic analysis by maximum likelihood[J] . Molecular Biology and Evolution, 2007, 24（8）：1586-1591.
[22] Excoffier L, Lischer HE. Arlequin suite ver 3. 5：a new series of programs to perform population genetics analyses under Linux and Windows[J] . Molecular Ecology Resources, 2010, 10（3）：564-567.
[23] Pons O, Petit R. Measwring and testing genetic differentiation with ordered versus unordered alleles[J] . Genetics, 1996, 144（3）：1237-1245.
[24] 陈伟烈. 西藏植被[M] . 北京：科学出版社, 1988.