广西师范大学学报（自然科学版） ›› 2021, Vol. 39 ›› Issue (1): 128-135.doi: 10.16088/j.issn.1001-6600.2020021502

• 研究论文 • 上一篇    下一篇

基于单拷贝核基因PAL研究贵州金花茶遗传多样性

李爽1,2,3, 刘上丽1,2,3, 裴思玉1,2,3, 唐绍清1,2,3*   

  1. 1.珍稀濒危动植物生态和环境保护教育部重点实验室(广西师范大学), 广西 桂林 541006;
    2.广西师范大学生命科学学院, 广西 桂林 541006;
    3.广西珍稀濒危动物生态重点实验室(广西师范大学), 广西 桂林 541006
  • 收稿日期:2020-02-15 修回日期:2020-03-16 出版日期:2021-01-25 发布日期:2021-03-27
  • 通讯作者: 唐绍清(1965—), 男(瑶族), 广西恭城人, 广西师范大学教授, 博士。E-mail: shaoqing@mailbox.gxnu.edu.cn
  • 基金资助:
    国家自然科学基金(31760088); 生态环境部生物多样性调查、观测和评估项目(837208)

A Study on Genetic Diversity of Camellia huana by Single-copy Nuclear Gene PAL

LI Shuang1,2,3, LIU Shangli1,2,3, PEI Siyu1,2,3, TANG Shaoqing1,2,3*   

  1. 1. Key Laboratory of Ecology of Rare and Endangered Species and Environmental Protection (Guangxi Normal University), Ministry of Education, Guilin Guangxi 541006, China;
    2. College of Life Science, Guangxi Normal University, Guilin Guangxi 541006, China;
    3. Guangxi Key Laboratory of Rare and Endangered Animal Ecology (Guangxi Normal University), Guilin Guangxi 541006, China
  • Received:2020-02-15 Revised:2020-03-16 Online:2021-01-25 Published:2021-03-27

PDF (PC)

353

摘要： 贵州金花茶是一濒危物种,分布在广西北部与贵州南部交界的石灰岩山的常绿阔叶林内。本文对贵州金花茶12个居群共120个个体的单拷贝核基因苯丙氨酸解氨酶(phenylalanine ammonia-lyase, PAL)进行扩增与测序,利用相关软件计算相应的遗传多样性和遗传分化系数。结果表明:1)序列总长为660 bp,包括7个多态位点,共检测到8个单倍型,总的核苷酸多样性指数Pi和单倍型多样性指数Hd分别为0.001 31和0.631。2)AMOVA分析结果显示91.47%的遗传变异存在于居群内,8.53%的遗传变异存在于居群间,遗传分化系数FST值为0.085 31。3)Mantel test结果显示,遗传距离和地理距离之间没有显著的相关性。鉴于贵州金花茶具有低水平的遗传多样性和中等水平的居群间遗传分化,建议在加强其自然居群保护的同时,从多个栽培居群取样进行迁地保护,以最大程度地保护其遗传多样性。

关键词: 贵州金花茶, 单拷贝核基因, 遗传多样性, 遗传分化

Abstract: Camellia huana is an endangered species which distributed in the evergreen,broad-leaved forests in the limestone hills bordering the northern Guangxi and southern Guizhou province. A single-copy nuclear gene PAL (phenylalanine ammonia-lyase,PAL) was employed to amplify and sequence 120 individuals in 12 populations of C. huana plant,and relevant softwares were used to calculate the corresponding genetic diversity and genetic differentiation coefficients. The results were as follows. The aligned nDNA (PAL) with a consensus length of 660 bp contained seven polymorphic sites. A total of eight haplotypes were identified,and genetic diversity indices of total nucleotide (Pi) and haplotype (Hd) diversity were 0.001 31 and 0.631,respectively. Analysis of molecular variance (AMOVA) further revealed that the majority of genetic variation (91.47%) was examined among populations,while only small amount (8.53%) was partioned within populations. The value of FST was 0.085 31. No significant correlation was detected between genetic and geographic distance which was supported by the result of Mantel test. Low level of genetic diversity and moderate level of genetic differential were detected for C. huana. It is suggested that while strengthening the protection of its natural populations,sampling from multiple cultivated populations for ex-situ conservation to maximize the protection of its genetic diversity.

Key words: Camellia huana, single-copy nuclear gene, genetic diversity, genetic differentiation

中图分类号: 

  • Q94
[1] MING T L,BARTHOLOMEW B.Theaceae[M] // WU Z Y, HONG D Y. Flora of China. Beijing: Science Press,2007: 370.
[2] 贺栋业,李晓宇,王丽丽,等. 金花茶化学成分及药理作用研究进展[J]. 中国实验方剂学杂志,2016,22(3): 231-234.
[3] 安明态. 贵州自然分布山茶属植物的资源现状[J]. 贵州林业科技,2005(2): 26-29.
[4] 谢代祖,牙正高,韩俊严,等. 天峨金花茶分布现状及保护策略研究[J]. 绿色科技,2014(4): 89-91.
[5] 覃海宁,杨永,董仕勇,等. 中国高等植物受威胁物种名录[J]. 生物多样性,2017,25(7): 696-744.
[6] ZHANG X, SU H L, YANG J, et al. Population genetic structure,migration,and polyploidy origin of a medicinal species Gynostemma pentaphyllum (Cucurbitaceae)[J]. Ecology and Evolution,2019,9(19): 11145-11170. DOI:10.1002/ece3.5618.
[7] RIVERS M C,BRUMMITT N A,LUGHADHA E N,et al. Genetic variation in Delonix s.l. (Leguminosae) in Madagascar revealed by AFLPs: fragmentation,conservation status and taxonomy[J]. Conservation Genetics,2011,12(5): 1333-1344. DOI:10.1007/s10592-011-0234-9.
[8] ANDIEGO K,DANGASUK O,ODEE D,et al. Genetic diversity of endangered sandalwood (Osyris lanceolata) populations in Kenya using ISSR molecular markers[J]. East African Agricultural and Forestry Journal,2019,83(2): 80-93. DOI:10.1080/00128325.2019.1605964.
[9] VARDARELI N,DOAROLU T,DOAÇ E,et al. Genetic characterization of tertiary relict endemic Phoenix theophrasti populations in Turkey and phylogenetic relations of the species with other palm species revealed by SSR markers[J]. Plant Systematics and Evolution,2019,305(6): 415-429. DOI:10.1007/s00606-019-01580-8.
[10] 张永德,曾兰,宾石玉,等. 尼罗罗非鱼选育家系的遗传多样性研究[J]. 广西师范大学学报(自然科学版),2014,32(3): 94-101.
[11] GAO Y D, GAO X F, HARRIS A. Species boundaries and parapatric speciation in the complex of alpine shrubs,Rosa sericea (Rosaceae),based on population genetics and ecological tolerances[J]. Frontiers in Plant Science,2019,10: 321. DOI:10.3389/fpls.2019.00321.
[12] HUANG J,ZENG S Y, ZHAO J H, et al. Genetic variation and phylogenetic relationships among Rehmannia (Scrophulariaceae) species as revealed by a novel set of single-copy nuclear gene markers[J]. Biochemical Systematics and Ecology,2016,66: 43-49. DOI:10.1016/j.bse.2016.03.011.
[13] XIAO S Y, JI Y H, LIU J,et al. Genetic characterization of the entire range of Cycas panzhihuaensis (Cycadaceae)[J]. Plant Diversity,2020,42(1): 7-18. DOI:10.1016/j.pld.2019.10.001.
[14] DOYLE J J,DOYLE J L. Isolation of plant DNA from fresh tissue[J]. Focus,1990,12: 13-15.
[15] MATSUMOTO S,TAKEUCHI A,HAYATSU M,et al. Molecular cloning of phenylalanine ammonia-lyase cDNA and classification of varieties and cultivars of tea plants ( Camellia sinensis ) using the tea PAL cDNA probe[J]. Theoretical and Applied Genetics,1994,89(6): 671-675. DOI:10.1007/BF00223703.
[16] WEI S J, LU Y B, YE Q Q, et al. Population genetic structure and phylogeography of Camellia flavida (Theaceae) based on chloroplast and nuclear DNA sequences[J]. Frontiers in Plant Science,2017,8: 718. DOI:10.3389/fpls.2017.00718.
[17] 张文驹,闵天禄. 山茶属古茶组植物的细胞学研究[J]. 云南植物研究,1995, 17(1): 48-54.
[18] CLARK A G. Inference of haplotypes from PCR-amplified samples of diploid populations[J]. Molecular Biology and Evolution,1990,7(2): 111-122.
[19] EDGAR R C. MUSCLE: multiple sequence alignment with high accuracy and high throughput[J]. Nucleic Acids Research,2004,32(5): 1792-1797. DOI:10.1093/nar/gkh340.
[20] TAMURA K,DUDLEY J,NEI M,et al. MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0[J]. Molecular Biology and Evolution,2007,24(8): 1596-1599. DOI:10.1093/molbev/msm092.
[21] LIBRADO P,ROZAS J. DnaSP v5: a software for comprehensive analysis of DNA polymorphism data[J]. Bioinformatics,2009,25(11): 1451-1452. DOI:10.1093/bioinformatics/btp187.
[22] TAJIMA F. Statistical method for testing the neutral mutation hypothesis by DNA polymorphism[J]. Genetics,1989,123(3): 585-595.
[23] FU Y X, LI W H. Statistical tests of neutrality of mutations[J]. Genetics,1993,133(3): 693-709.
[24] FU Y X. Statistical tests of neutrality of mutations against population growth,hitchhiking and background selection[J]. Genetics,1997,147(2): 915-925.
[25] PONS O,PETIT R J. Measuring and testing genetic differentiation with ordered versus unordered alleles[J]. Genetics,1996,144(3): 1237-1245.
[26] EXCOFFIER L,LAVAL G,SCHNEIDER S. Arlequin (version 3.0): an integrated software package for population genetics data analysis[J]. Evolutionary Bioinformatics,2005,1: 47-50. DOI:10.1177/117693430500100003.
[27] WRIGHT S. Evolution in mendelian populations[J]. Genetics,1931,16(2): 97-159.
[28] PEAKALL R, SMOUSE P E. GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research:an update[J]. Bioinformatics,2012,28(19): 2537-2539.
[29] BANDELT H J,FORSTER P,RÖHL A. Median-joining networks for inferring intraspecific phylogenies[J]. Molecular Biology and Evolution,1999,16(1): 37-48. DOI:10.1093/oxfordjournals.molbev.a026036.
[30] CHUNG M Y,LÓPEZ-PUJOL J,SON S,et al. Patterns of genetic diversity in rare and common orchids focusing on the korean peninsula: implications for conservation[J]. The Botanical Review,2018,84(1): 1-25. DOI:10.1007/s12229-017-9190-5.
[31] GUSTAFSSON S,SJÖGREN-GULVE P. Genetic diversity in the rare orchid Gymnadenia odoratissima and a comparison with the more common congener,G. conopsea[J]. Conservation Genetics,2002,3(3): 225-234. DOI:10.1023/A:1019969014333.
[32] LEVY E,BYRNE M,COATES D J,et al. Contrasting influences of geographic range and distribution of populations on patterns of genetic diversity in two sympatric pilbara acacias[J]. PLoS ONE,2016,11(10): e0163995. DOI:10.1371/journal.pone.0163995.
[33] LIU Y,YANG S X, JI P Z, et al. Phylogeography of Camellia taliensis (Theaceae) inferred from chloroplast and nuclear DNA: insights into evolutionary history and conservation[J]. BMC Evolutionary Biology,2012,12(1): 92. DOI:10.1186/1471-2148-12-92.
[34] 陈海玲. 薄叶金花茶及其近缘种遗传多样性和遗传结构研究[D]. 桂林:广西师范大学,2018.
[35] 路雪林. 金花茶的谱系地理学研究[D]. 桂林:广西师范大学,2018.
[36] 陈莉萍. 平果金花茶的保护遗传学研究[D]. 桂林:广西师范大学,2019.
[37] NYBOM H,BARTISH I V. Effects of life history traits and sampling strategies on genetic diversity estimates obtained with RAPD markers in plants[J]. Perspectives in Plant Ecology,Evolution and Systematics,2000,3(2): 93-114. DOI:10.1078/1433-8319-00006.
[38] YANG H S, LI X P, LIU D J, et al. Genetic diversity and population structure of the endangered medicinal plant Phellodendron amurense in China revealed by SSR markers[J]. Biochemical Systematics and Ecology,2016,66: 286-292. DOI:10.1016/j.bse.2016.04.018.
[1] 张永德, 曾兰, 宾石玉, 杜雪松, 杨慧赞, 陈忠, 余艳玲, 林勇. 尼罗罗非鱼选育家系的遗传多样性研究[J]. 广西师范大学学报（自然科学版）, 2014, 32(3): 94-101.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] 杨丽婷, 刘学聪, 范鹏来, 周岐海. 中国非人灵长类声音通讯研究进展[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 1 -9 .
[2] 宾石玉, 廖芳, 杜雪松, 许艺兰, 王鑫, 武霞, 林勇. 罗非鱼耐寒性能研究进展[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 10 -16 .
[3] 刘静, 边迅. 直翅目昆虫线粒体基因组的特征及应用[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 17 -28 .
[4] 李兴康, 钟恩主, 崔春艳, 周佳, 李小平, 管振华. 西黑冠长臂猿滇西亚种鸣叫行为监测[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 29 -37 .
[5] 和鑫明, 夏万才, 巴桑, 龙晓斌, 赖建东, 杨婵, 王凡, 黎大勇. 滇金丝猴主雄应对配偶雌性数量的理毛策略[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 38 -44 .
[6] 付文, 任宝平, 林建忠, 栾科, 王朋程, 王宾, 黎大勇, 周岐海. 济源太行山猕猴种群数量和保护现状[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 45 -52 .
[7] 郑景金, 梁霁鹏, 张克处, 黄爱面, 陆倩, 李友邦, 黄中豪. 基于木本植物优势度的白头叶猴食物选择研究[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 53 -64 .
[8] 杨婵, 万雅琼, 黄小富, 袁旭东, 周洪艳, 方浩存, 黎大勇, 李佳琦. 基于红外相机技术的小麂(Muntiacus reevesi)活动节律[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 65 -70 .
[9] 李友邦, 农娟丽, 杨婉琳, 赵佳佳, 朱琪琪. 弄岗同域分布赤腹松鼠和红颊长吻松鼠活动节律研究[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 71 -78 .
[10] 程瑞, 何明先, 钟春英, 罗树毅, 武正军. 野生与人工繁育鳄蜥游泳能力比较[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 79 -86 .
版权所有 © 广西师范大学学报（自然科学版）编辑部
地址：广西桂林市三里店育才路15号　邮编：541004
电话：0773-5857325　E-mail: gxsdzkb@mailbox.gxnu.edu.cn
本系统由北京玛格泰克科技发展有限公司设计开发