不同喀斯特小生境中小蓬竹的遗传多样性
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摘要
通过RAPD标记法对5类喀斯特小生境中小蓬竹的遗传结构及其多样性进行了分析。RAPD随机引物8个扩增共得87个清晰度高且重复性好的条带,分子量200～3 000 bp,含多态68条,物种PPL=78.16%,Nei’s基因多度H=0.291 3,Shannon多样性指数I=0.431 0。土面(TM)生长的小蓬竹遗传多样性水平最高(PPL=47.13%,H=0.195 3,I=0.283 4),石缝(SF)中的小蓬竹遗传多样性水平最低(PPL=39.08%,H=0.147 4,I=0.219 2)。POPGENE 32分析5类小生境中小蓬竹的遗传分化系数G_(st)=0.415 1,基因流N_m=0.704 5。5类小生境的小蓬竹间遗传相似系数(I)变化范围0.777 3～0.999 4,平均为0.865 6;遗传距离(D)变化范围0.000 6～0.251 9,平均为0.148 3。各类小生境的遗传多样性水平PPL均值为43.91%,5类小生境间的遗传分化G_(st)=0.405 4。研究结果说明,小生境间的差异增加了小蓬竹的遗传多样性。
By random amplified polymorphic DNA(RAPD) markers analysis,5 kinds of karst microhabitats of Ampelocalamus luodianensis was conducted to study genetic diversity and genetic structure.Through 8 RAPD primers,87 discernible DNA fragments in all weighted from 200-3 000 bp were cloned,and 68 in those were polymorphic loci,with percentage of polymorphic loci at the species level,PPL=78.16%,Nei's index H=0.291 3,and Shannon's Information Index I=0.431 0.The A.luodianensis grew on soil surface(TM)possessed the highest level of genetic diversity(PPL=47.13%,H=0.195 3 and I=0.283 4),while the A.luodianensis in stone crevice exhibited the lowest levels of genetic diversity(PPL=39.08%,H=0.147 4 and I=0.219 2).Coefficient of genetic differentiation(G_(st)) was 0.415 1(from the POPGENE32),and the gene flows(N_m) was 0.704 5.Among the 5 microhabitats,genetic identity varied from 0.777 3-0.999 4,genetic distance was different from 0.000 6-0.251 9,and the mean of them were 0.865 6-0.148 3severally.Genetic diversity in niches at mean value was with PPL of 43.91%.Coefficient of genetic variation in niches was with G_(st) of 0.4054.The results indicated that differences of microhabitats increased the genetic diversity of A.luodianensis.
By random amplified polymorphic DNA(RAPD) markers analysis,5 kinds of karst microhabitats of Ampelocalamus luodianensis was conducted to study genetic diversity and genetic structure.Through 8 RAPD primers,87 discernible DNA fragments in all weighted from 200-3 000 bp were cloned,and 68 in those were polymorphic loci,with percentage of polymorphic loci at the species level,PPL=78.16%,Nei's index H=0.291 3,and Shannon's Information Index I=0.431 0.The A.luodianensis grew on soil surface(TM)possessed the highest level of genetic diversity(PPL=47.13%,H=0.195 3 and I=0.283 4),while the A.luodianensis in stone crevice exhibited the lowest levels of genetic diversity(PPL=39.08%,H=0.147 4 and I=0.219 2).Coefficient of genetic differentiation(G_(st)) was 0.415 1(from the POPGENE32),and the gene flows(N_m) was 0.704 5.Among the 5 microhabitats,genetic identity varied from 0.777 3-0.999 4,genetic distance was different from 0.000 6-0.251 9,and the mean of them were 0.865 6-0.148 3severally.Genetic diversity in niches at mean value was with PPL of 43.91%.Coefficient of genetic variation in niches was with G_(st) of 0.4054.The results indicated that differences of microhabitats increased the genetic diversity of A.luodianensis.
引文
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2.金祺,刘济明.不同生境条件对小蓬竹生长的影响[J].中南林业调查规划,2014,33(4):52-56,66.Jin Q,Liu J M.Study on the influence of different habitats for the growth of Drepanostachyum luodianense[J].Central South Forest Inventory and Planning,2014,33(4):52-56,66.
3.Welsh J,Mcclelland M.Fingerprinting genomes using PCRwith arbitrary primer[J].Nucleic Acids Research,1990,18(24):7213-7218.
4.瞿印权,徐雯,何天友,等.DNA分子标记技术在竹亚科植物遗传多样性研究中的应用[J].安徽农业科学,2017,45(34):141-143.Qu Y Q,Xu W,He T Y,et al.Application of DNA molecular marker in the study of genetic diversity of bambsoideae[J].Journal of Anhui Agricultural Sciences,2017,45(34):141-143.
5.刘济明,王敏,闫国华,等.濒危竹种小蓬竹(Drepanostachyum luodianense)RAPD-PCR反应体系优化[J].浙江农业学报,2014,26(3):667-674.Liu J M,Wang M,Yan G H,et al.Optimization of RAPD-PCR conditions for a rare and endangered bamboo of Drepanostachyum luodianense[J].Acta Agriculturae Zhejiangensis,2014,26(3):667-674.
6.张志毅.林木遗传学基础:2版[M].北京:中国林业出版社,2012.Zhang Z Y.Genetic basis of forest trees:2nd ed[M].Beijing:China Forestry Publishing House,2012.
7.卢永飞.退化喀斯特植被恢复与生境协变关系研究---以花江地区为例[D].贵阳:贵州大学,2007:6-13.Lu Y F.A Study on covariance between plants and environment in natural restoration process of degraded karst forests:a case study at Huajiang of Guizhou[D].Guiyang:Guizhou Univercity,2007:6-13.
8.郝久程,贾鑫,穆晓红,等.大连地区岛屿与大陆玉竹种群遗传多样性的ISSR分析[J].植物研究,2017,37(5):709-714.Hao J C,Jia X,Mu X H,et al.Genetic diversity between island and mainland natural populations of Polygonatum odoratum in Dalian Area by ISSR[J].Bulletin of Botanical Research,2017,37(5):709-714.
9.邱月群,董文渊,王逸之,等.筇竹天然居群遗传多样性研究[J].西北林学院学报,2017,32(2):155-160.Qui Y Q,Dong W Y,Wang Y Z,et al.Genetic diversity in natural populations of Qiongzhuea tumidinoda[J].Journal of Northwest Forestry University,2017,32(2):155-160.
10.张安世,张中海,齐秀娟,等.猕猴桃SCo T遗传多样性分析及指纹图谱的构建[J].植物研究,2017,37(2):259-265.Zhang A S,Zhang Z H,Qi X J,et al.Genetic diversity and fingerprints with SCo T markers in Actinidia[J].Bulletin of Botanical Research,2017,37(2):259-265.
11.余鸽.巴山木竹克隆结构及遗传多样性研究[D].咸阳:西北农林科技大学,2016.Yu G.Clonal structure and genetic diversity of Bashania fargesii[D].Xianyang:Northwest A&F University,2016.
12.王波,高素萍,陈其兵.珍稀濒危观赏竹种筇竹遗传多样性研究[J].竹子研究汇刊,2016,35(1):13-16,21.Wang B,Gao S P,Chen Q B.Genetic diversity of the rare and endangered species Qiongzhuea tumidinoda[J].Journal of Bamboo Research,2016,35(1):13-16,21.
13.Abeysinghe P D,Triest L,De Greef B,et al.Genetic and geographic variation of the mangrove tree Bruguiera in Sri Lanka[J].Aquatic Botany,2000,67(2):131-144.
14.徐静静,赵冰,张良英,等.西藏地区雪层杜鹃遗传多样性的AFLP分析[J].植物研究,2017,37(1):88-95.Xu J J,Zhao B,Zhang L Y,et al.Assessment of genetic diversity of Rhododendron nivale in Tibetan,China using AFLP markers[J].Bulletin of Botanical Research,2017,37(1):88-95.
15.施建敏,叶学华,陈伏生,等.竹类植物对异质生境的适应---表型可塑性[J].生态学报,2014,34(20):5687-5695.Shi J M,Ye X H,Chen F S,et al.Adaptation of bamboo to heterogeneous habitat:phenotypic plasticity[J].Acta Ecologica Sinica,2014,34(20):5687-5695.
16.Amomt,Tikendra L,Rahaman H,et al.Evaluation of genetic relationship between 15 bamboo species of North-EastIndia based on ISSR marker analysis[J].Molecular Biology Research Communications,2018,7(1):7-15.
17.王敏,刘济明,徐雪娇,等.濒危竹种小蓬竹群落不同小生境的小气候变化特征[J].贵州农业科学,2013,41(6):78-82.Wang M,Liu J M,Xu X J,et al.Microclimate characteristics of different microhabitats of a rare and endangered bamboo of Drepanostachyum luodianense communities[J].Guizhou Agricultural Sciences,2013,41(6):78-82.
18.Doylej J,Doyle J L.A rapid DNA isolation procedure for small quantities of freshleaf tissue[J].Phytochemical Bulletin,1987,19:11-15.
19.李雪平,牟少华,彭镇华,等.毛竹等28个刚竹属竹种的AFLP分析[J].安徽农业大学学报,2014,41(2):179-182.Li X P,Mou S H,Peng Z H,et al.AFLP analysis of 28Phyllostachys bamboo species including Mosobamboo[J].Journal of Anhui Agricultural University,2014,41(2):179-182.
20.Shapcott A.The patterns of genetic diversity in Carpentaria acuminata(Arecaceae),and rainforest history in northern Australia[J].Molecular Ecology,1998,7(7):833-847.
21.刘济明,王敏,张东凯,等.小蓬竹不同小生境土壤的理化性质[J].贵州农业科学,2012,40(6):177-180.Liu J M,Wang M,Zhang D K,et al.Soil Physico-chemical Properties of Drepanostachyum luodianense in Different Microhabitats[J].Guizhou Agricultural Sciences,2012,40(6):177-180.
22.马乃训,陈光才.我国特产珍稀濒危竹种及标准探讨[C].//中国林学会首届竹业学术大会论文集.浙江:中国林学会竹子分会,2004:45-50.Ma N X,Chen G C.Discussion on rare and endangered bamboo species and their standard in China[C].//Proceedings of the first bamboo academic conference of China forestry society.Zhejiang:Bamboo Branch of China Forestry Society,2004:45-50.
23.葛颂.植物群体遗传结构研究的回顾和展望[C].//李承森.植物科学进展(第1卷).北京:高等教育出版社,1998:1-30.Ge S.Review and prospect of studies on genetic structure of plant population[C].//Li C S.Advances in plant science(vol.1).Beijing:Advanced Education Press,1998:1-30.