濒危植物五小叶槭(Acer pentaphyllum Diels)天然种群遗传多样性的ISSR标记分析
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摘要
五小叶槭(Acer pentaphyllum Diels)是中国四川特有野生珍稀濒危植物,仅分布于雅砻江中上游干旱河谷地带;探索其遗传多样性特征可以为其遗传资源的保护和利用提供理论依据.本文采用ISSR技术对五小叶槭的5个野生种群的44份材料进行遗传多样性分析,8条引物总共检测到38个位点,其中多态性位点35个,多态位点百分率(PPL)平均为90.21%.应用分析软件Popgen32分析得出:在物种水平上,五小叶槭的香农多样性指数(I)为0.3924,Nei氏基因多样性指数(H_e)为0.2498;种群间基因分化系数(G_(st))为0.3722,种群间的基因流(N_m)为0.8433,这表明五小叶槭的遗传变异主要存在于种群内.在种群水平上,香农多样性指数(I)为0.2125, Nei氏基因多样性指数(H_e)为0.1456.结果表明木里、九龙三垭宫和康定这3个种群的遗传多样性水平较高,而雅江和九龙洛莫种群遗传多样性较低.通过Mantel检测,种群间遗传距离和地理距离之间没有显著相关性.
Acer pentaphyllum is rare and endangered plant endemic to Sichuan Province, China. This species is only distributed in the arid valley area of the upper and middle reaches of Yalong River. Exploring its genetic diversity characteristics can provide a theoretical basis for the protection and utilization of its genetic resources. In the paper, genetic diversity and genetic structure of 5 Acer pentaphyllum Diels populations from Sichuan(total about 44 individuals) were analyzed by ISSR(inter simple sequence repeat) markers. Eight highly stable and repeatable ISSR primers were used to detect a total of 38 loci, of which 35 were polymorphic sites(90.21% of the total). At the species level, the Shannon index(I) was 0.3924, Nei's gene diversity index(H_e) was 0.2498, the coefficient of gene differentiation(Gst) was 0.3722, and the gene flow(Nm) was 0.8433. The software Popgen 32 analysis of the results showed that the genetic variation mainly existed in the population. At the population Level, I and He were 0.2125 and 0.1456, respectively. All of these results showed that Muli, Sanyagong and Kangdiing population had higher levels of genetic diversity, while Yajiang and Luomo had lower genetic diversity. The mantel analysis showed no significant correlation between genetic distance and geographic distance.
Acer pentaphyllum is rare and endangered plant endemic to Sichuan Province, China. This species is only distributed in the arid valley area of the upper and middle reaches of Yalong River. Exploring its genetic diversity characteristics can provide a theoretical basis for the protection and utilization of its genetic resources. In the paper, genetic diversity and genetic structure of 5 Acer pentaphyllum Diels populations from Sichuan(total about 44 individuals) were analyzed by ISSR(inter simple sequence repeat) markers. Eight highly stable and repeatable ISSR primers were used to detect a total of 38 loci, of which 35 were polymorphic sites(90.21% of the total). At the species level, the Shannon index(I) was 0.3924, Nei's gene diversity index(H_e) was 0.2498, the coefficient of gene differentiation(Gst) was 0.3722, and the gene flow(Nm) was 0.8433. The software Popgen 32 analysis of the results showed that the genetic variation mainly existed in the population. At the population Level, I and He were 0.2125 and 0.1456, respectively. All of these results showed that Muli, Sanyagong and Kangdiing population had higher levels of genetic diversity, while Yajiang and Luomo had lower genetic diversity. The mantel analysis showed no significant correlation between genetic distance and geographic distance.
引文
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[12] Nudin N F H, Ali A M, Ngah N. ISSR marker-assisted genetic diversity analysis of Dioscorea hispida and selection of the best variety for sustainable production[J]. CR Biol, 2017, 340: 359.
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[19] 罗成, 熊宇婷, 顾蔚, 等. 华中五味子ISSR-PCR反应体系优化及引物筛选 [J]. 植物研究, 2010, 30: 588.
[20] 冯亮亮, 唐红, 李毅, 等. 甘肃红砂不同种群遗传多样性的ISSR分析 [J]. 草业学报, 2011, 20: 125.
[21] 柴胜丰, 庄雪影, 邹蓉, 等. 濒危植物毛瓣金花茶遗传多样性的ISSR分析 [J]. 西北植物学报, 2014, 34 : 93.
[22] 张云香, 胡灏禹, 何兴金. 珍稀濒危植物距瓣尾囊草遗传多样性的ISSR分析 [J]. 西北植物学报, 2013, 33: 1098.
[23] 沈浩, 刘登义. 遗传多样性概述 [J]. 生物学杂志, 2001, 18: 5.
[24] 陈宗游, 黄夕洋, 唐辉, 等. 广西甜茶种质资源遗传多样性的ISSR分析 [J]. 园艺学报, 2017, 161.
[25] 文亚峰, 韩文军, 吴顺. 植物遗传多样性及其影响因素 [J]. 中南林业科技大学学报, 2010, 30: 80.
[26] 梁婷婷. 山东省梨属野生种质资源调查与评价 [D]. 山东农业大学, 2016.
[27] 解庆, 刘志红, 李周岐. 柴松遗传多样性的RAPD分析 [J]. 西北林学院学报, 2011, 26: 112.
[28] Lewis P O, Crawford D J. Pleistocene refugium endemics exhibit greater allozymic diversity than widespread congeners in the genus Polygonella (Polygonaceae) [J]. Am J Bot, 1995, 82: 141.
[29] Zawko G, Krauss S L, Dixon K W, et al. Conservation genetics of the rare and endangered Leucopogon obtectus (Ericaceae) [J]. Mol Ecol, 2001, 10: 2389.
[30] Nybom H. Comparison of different nuclear DNA markers for estimating intraspecific genetic diversity in plants [J]. Mol Ecol, 2004, 13: 1143.
[31] Wright S. The genetical structure of populations[J]. Ann Eugenic, 1951, 15: 323.
[2] Mark S R, William A. M, Deric P. Assessment of genetic variation in Acer pentaphyllum based on amplified fragment length polymorphisms [J]. J Hortic Sci Biotech, 2008, 83:725
[3] 沙红. 五小叶槭育苗造林技术 [J] . 农业科技与信息, 2015(14): 100.
[4] 孙治宇, 李明富, 李八斤, 等. 四川雅江县珍稀濒危植物五小叶槭 [J]. 四川林业科技, 2010, 31: 86.
[5] 毕泉鑫, 金则新, 李钧敏, 等. 枫香自然种群遗传多样性的ISSR分析 [J]. 植物研究, 2010, 30: 120.
[6] 邓珍. 二倍体马铃薯组培苗耐旱性相关性状QTLs定位与分析 [D]. 北京: 中国农业科学院, 2014: 23.
[7] 金则新, 李钧敏, 顾奇萍. 云锦杜鹃自然居群遗传多样性的ISSR分析 [J]. 园艺学报, 2006, (06): 1263.
[8] 邱英雄, 傅承新. 明党参的濒危机制及其保护对策的研究 [J]. 生物多样性, 2001, 9: 151.
[9] Patel H K, Fougat R S, Kumar S, et al. Detection of genetic variation in Ocimum species using RAPD and ISSR markers [J]. Biotech, 2015, 5: 697.
[10] Thul S T, Srivastava A K, Singh S C, et al. Genetic and chemical diversity of high mucilaginous Plants of Sida complex by ISSR markers and chemical fingerprinting [J]. Mol Biotechnol, 2011, 49: 77.
[11] 王佳媛, 吴传芳, 唐亚. 基于SNP分子标记的凹叶木兰遗传多样性初步研究 [J]. 广西植物, 2012, 32: 542.
[12] Nudin N F H, Ali A M, Ngah N. ISSR marker-assisted genetic diversity analysis of Dioscorea hispida and selection of the best variety for sustainable production[J]. CR Biol, 2017, 340: 359.
[13] 王利, 邢世岩, 王正华, 等. 银杏分子系统学研究进展 [J]. 山东林业科技, 2005(6): 58.
[14] 黄琼. 新疆甜瓜果实绿色条纹性状AFLP标记的初步筛选 [D]. 乌鲁木齐: 新疆大学, 2008: 45.
[15] 曾艳玲, 谭晓风, 曾晓峰. 采用正交设计方法优化梨ISSR-PCR反应体系[J]. 湖北农业科学, 2008, 47: 1235.
[16] 张卿哲, 孟金贵, 张应华, 等. 用正交设计法优化辣椒ISSR-PCR反应体系 [J]. 西南农业学报, 2011, 24: 1034.
[17] 李瑞, 陈少瑜, 宁德鲁, 等. 油橄榄ISSR-PCR反应体系优化与引物筛选 [J]. 安徽农业科学, 2012, 40: 5797.
[18] 俞靓, 井赵斌, 魏琳, 等. 本氏针茅ISSR-PCR反应体系的建立及引物筛选 [J]. 中国草地学报, 2012, 34: 77.
[19] 罗成, 熊宇婷, 顾蔚, 等. 华中五味子ISSR-PCR反应体系优化及引物筛选 [J]. 植物研究, 2010, 30: 588.
[20] 冯亮亮, 唐红, 李毅, 等. 甘肃红砂不同种群遗传多样性的ISSR分析 [J]. 草业学报, 2011, 20: 125.
[21] 柴胜丰, 庄雪影, 邹蓉, 等. 濒危植物毛瓣金花茶遗传多样性的ISSR分析 [J]. 西北植物学报, 2014, 34 : 93.
[22] 张云香, 胡灏禹, 何兴金. 珍稀濒危植物距瓣尾囊草遗传多样性的ISSR分析 [J]. 西北植物学报, 2013, 33: 1098.
[23] 沈浩, 刘登义. 遗传多样性概述 [J]. 生物学杂志, 2001, 18: 5.
[24] 陈宗游, 黄夕洋, 唐辉, 等. 广西甜茶种质资源遗传多样性的ISSR分析 [J]. 园艺学报, 2017, 161.
[25] 文亚峰, 韩文军, 吴顺. 植物遗传多样性及其影响因素 [J]. 中南林业科技大学学报, 2010, 30: 80.
[26] 梁婷婷. 山东省梨属野生种质资源调查与评价 [D]. 山东农业大学, 2016.
[27] 解庆, 刘志红, 李周岐. 柴松遗传多样性的RAPD分析 [J]. 西北林学院学报, 2011, 26: 112.
[28] Lewis P O, Crawford D J. Pleistocene refugium endemics exhibit greater allozymic diversity than widespread congeners in the genus Polygonella (Polygonaceae) [J]. Am J Bot, 1995, 82: 141.
[29] Zawko G, Krauss S L, Dixon K W, et al. Conservation genetics of the rare and endangered Leucopogon obtectus (Ericaceae) [J]. Mol Ecol, 2001, 10: 2389.
[30] Nybom H. Comparison of different nuclear DNA markers for estimating intraspecific genetic diversity in plants [J]. Mol Ecol, 2004, 13: 1143.
[31] Wright S. The genetical structure of populations[J]. Ann Eugenic, 1951, 15: 323.