柱花草种质遗传多样性的ISSR分析

草业学报 ›› 2009, Vol. 18 ›› Issue (1): 57-64.

柱花草种质遗传多样性的ISSR分析

唐燕琼¹,胡新文^1*,郭建春²,白昌军³,何华玄³

1.海南大学农学院,海南儋州571737;
2.中国热带农业科学院生物技术研究所,海南海口571101;
3.中国热带农业科学院品种资源研究所,海南儋州571737

收稿日期:2008-03-10 出版日期:2009-01-25 发布日期:2009-02-20
作者简介:唐燕琼(1968-),女,湖南衡阳人,副教授,博士。E-mail:tyq68@126.com
基金资助:
国家重点基础研究发展计划(973计划)(2007CB108903),作物遗传育种国家重点学科和海南省自然科学基金项目资助。

An analysis by ISSR of genetic diversities in Stylosanthes germplasms

TANG Yan-qiong¹, HU Xin-wen¹, GUO Jian-chun², BAI Chang-jun³, HE Hua-xuan³

1. College of Agriculture, Hainan University, Danzhou 571737, China;
2. Institute of Biotechnology and Bioscience, CATAS, Haikou 571101, China;
3. Institute of Tropical Crops Genetic Resources, CATAS, Danzhou 571737, China

Received:2008-03-10 Online:2009-01-25 Published:2009-02-20

摘要/Abstract

摘要： 应用ISSR标记,对48份柱花草种质的遗传多样性进行研究。从68个ISSR引物中筛选出14个多态性明显、反应稳定的引物,共扩增出156条谱带,平均每个引物能扩增出11.1条带,多态性条带比率达99.36%。材料间遗传相似系数为0.445~0.975,POPGENE结果分析表明,平均Shannon信息指数(I)为0.3553,平均Nei’s基因多样性(H)为0.2279,每位点平均有效等位基因数(NE)为1.3887。利用聚类分析和主成分分析表明,48份供试材料可聚为5类:有钩柱花草类、头状柱花草类、圭亚那柱花草类、西卡柱花草类和灌木柱花草类,其中,圭亚那柱花草类种内材料遗传变异较大。

Abstract: Forty-eight Stylosanthes germplasms were studied for an analysis of genetic diversity by ISSR. Fourteen effective primers selected from 68 ISSR primers were used for PCR, and 155 of the 156 DNA fragments amplified, showed polymorphisms. The average binds from each primer was 11.1 and the average percentage of polymorphic bands was 99.36%. The coefficient of genetic conformability among the germplasms was 0.445-0.975. The results from POPGENE analysis indicated that the average information index (I) of Shannon was 0.355 3, gene diversity (H) of Nei was 0.227 9, and effective number of alleles (NE) was 1.388 7. Based on analyses of cluster and principal components, the 48 Stylosanthes germplasms used in this study could be divided into 5 groups: S. hamata, S. capitata, S. scabra, S. seabrana and S. guianensis, in which the genetic differentiation of S. guianensis group was the largest.

中图分类号:

S540.32
Q943

唐燕琼,胡新文,郭建春,白昌军,何华玄. 柱花草种质遗传多样性的ISSR分析[J]. 草业学报, 2009, 18(1): 57-64.

TANG Yan-qiong, HU Xin-wen, GUO Jian-chun, BAI Chang-jun, HE Hua-xuan. An analysis by ISSR of genetic diversities in Stylosanthes germplasms[J]. Acta Prataculturae Sinica, 2009, 18(1): 57-64.

参考文献

[1] Williams R J, Reid R, Schultze-Kraft R, et al. Natural distribution of Stylosanthes[A]. In: Stace H M, Edye L A. The Biology and Agronomy of Stylosanthes[M]. Sydney, Australia: Academic Press, 1984.73-101.
[2] 蒋昌顺. 我国对柱花草的研究与利用[J]. 热带作物研究, 1995, (1): 61-65.
[3] Sousa Costa N M, Ferreira M B. Some Brazilian species of Stylosanthes[A]. In: Stace H M, Edye L A. The Biology and Agronomy of Stylosanthes[M].Sydney, Australia: Academic Press, 1984.23-48.
[4] Mannetje L T. Considerations on the taxonomy of the genus Stylosanthes[A]. In: Stace H M, Edye L A. The Biology and Agronomy of Stylosanthes[M]. Sydney, Australia: Academic Press, 1984.1-21.
[5] Glover B J, Gillies A C M, Abbott R J. Use of the polymerase chain reaction to investigate the delimitation of two agronomically important species of Stylosanthes (Aubl.) Sw[J]. Botanical Journal of Scotland, 1984, 47: 83-96.
[6] 蒋昌顺, 葛琴雅. 柱花草RAPD 反应体系的建立及其8个品种遗传多样性分析[J]. 广西植物, 2004, 24(3): 243-247.
[7] Curtis M D, Cameron D F, Manners J M. Molecular evidence that diploid Stylosanthes humilis and diploid Stylosanthes hamata are progenitors of allotetraploid Stylosanthes hamata cv ‘Verano’[J]. Genome, 1995, 38: 344-348.
[8] Jiang C S, Jia H S, Ma X R. AFLP analysis of genetic variability among Stylosanthes guianensis accessions resistant and susceptible to the Stylo[J]. Anthracnose Acta Botanica Sinica, 2004, 46(4): 480-488.
[9] Liu C J, Musial J M, Thomas D. Genetic relationship among Stylosanthes species revealed by RFLP and STS analysis[J]. Theoretical and Applied Genetics, 1999, 99: 1179-1186.
[10] Vander stappen J, Volckaert G. Molecular charaterization and classification of Stylosanthes mexicanna, S. macrocarpa, S. seabrana, and S. fruticosa by DNA sequence analysis of two chloroplast regions[J]. DNA Sequence, 1999, (3): 199-202.
[11] Vander stappen J, De Laet J, Gama-Lopez S. Phylogenetic analysis of Stylosanthes (Fabaceae) based on the internal transcribed spacer region (ITS) of nuclear ribosomal DNA[J]. Plant Systematics and Evolution, 2002, 234: 27-51.
[12] 蒋昌顺. 应用微卫星标记分析柱花草的遗传多样性[J]. 高技术通讯, 2004, (4): 25-30.
[13] Vander stappen J, Weltjens I, Volckaert G. Microsatellite markers in Stylosanthes guianensis[J]. Molecular Ecology, 1999, 8: 515-517.
[14] 蒋昌顺, 张新申. 柱花草核rDNA的ITSl序列分析[J]. 高技术通讯, 2006, (1): 73-77.
[15] Zietikiewicze, Rafalske R A, Labuda D. Genome fingerprinting by simple sequence repeat (ISSR) anchored and polymerase chain reaction amplificaion[J]. Genomica, 1994, 20: 178-183.
[16] Shen J, Ding X, Liu D, et al. Intersimple sequence repeats (ISSR) molecular fingerprinting markers for authenticating populations of Dendrobium officinale KIMURA et MIGO[J]. Biological and Pharmaceutical Bulletin, 2006, 29(3): 420-422.
[17] Amel S, Mikhtar T, Salwa Z, et al. Inter-simple sequence repeat fingerprints to assess genetic diversity in trnisian fig (Ficus carica L.) germplasm[J]. Genetic Resources and Crop Evolution, 2004, 51: 269-275.
[18] Ge X, Yu Y, Zhao N, et al. Genetic variation in the endangered inner mongolia endemic shrub Tetraena mongolica Mazim. (Zygophyllaceae)[J]. Biolgical Conservation, 2003, 111: 427-434.
[19] Wolfe A D, Xiang Q, Kephart S R. Assessing hybridization in natural populations of Penstemon (Scrophulariaceae) using hypervariable intersimple sequence repeat (ISSR) bands[J]. Milecular Ecology, 1998, 7: 1107-1125.
[20] Danilova T V, Danilov S S, Karlov G I. Assessment of genetic polmorphism in hop (Himulus lupulus L.) cultivars by ISSR-PCR analysis[J]. Russian Journal of Genetics, 2003, 39: 1252-1257.
[21] Pomper K W, Crabtree S B, Brown S P, et al. Assessment of genetic diversity of pawpaw (Asimina triloba) cultivators with intersimple sequence repeat markers[J]. Journal American Society for Horticultural Science, 2003, 128(4): 521-525.
[22] Talhinhas T, Neves-Martins J, Leitào J. AFLP, ISSR, and RAPD markers reveal high levels of genetic diversity among Lupinus ssp.[J]. Plant Breeding, 2003, 122: 507-510.
[23] 唐燕琼, 吴紫云, 郭建春, 等. 柱花草DNA提取及ISSR反应体系的正交优化[J]. 热带作物学报, 2008, 29(3): 352-357.
[24] 英彩, 赖志强, 腾少花, 等. 907柱花草的选育研究[J]. 草业科学, 1998, 15(2): 27-30.
[25] 刘国道, 白昌军, 何华玄, 等. 热研5号柱花草选育研究[J]. 草地学报, 2001, 9(1): 1-7.
[26] 范彦, 李芳, 张新全, 等. 扁穗牛鞭草种质遗传多样性的ISSR分析[J]. 草业学报, 2007, 16(4): 76-81.
[27] 刘伟, 张新全, 李芳, 等. 西南区野生狗牙根遗传多样性的ISSR标记与地理来源分析[J]. 草业学报, 2007, 16(3): 55-61.
[28] Prevost A, Wilkinson M J. A new system of comparing PCR primers applied to ISSR fingerprinting of potato cultivars[J]. Theoretical and Applied Genetics, 1999, 98: 107-112.
[29] Charters Y M, Robertson A, Wilkinson M J, et al. PCR analysis of oilseed rape cultivars (Brassica napus L. ssp. olei fera) using 5′-anchored simple sequence repeat (SSR) primers[J]. Theoretical and Applied Genetics, 1996, 92: 442-447.
[30] 邓春婷, 包满珠. 野牛草实生群体多样性的表型及ISSR分析[J]. 草业科学, 2008, 25(1): 100-105.
[31] Huang J C, Sun M. Genetic diversity and relationships of sweptotato and its wild relatives in Jpomoea series Batats (Convolvlaceae) as revealed by ISSR and restriction analysis of chloroplast DNA[J]. Theoretical and Applied Genetics, 2000, 100: 1050-1060.
[32] 隋晓青, 王堃. 克氏针茅ISSR-PCR反应体系的建立与优化[J]. 草业学报, 2008, 17(3): 71-78.