利用目标起始密码子多态性(SCoT)分子标记分析梨遗传多样性

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摘要目标起始密码子多态性(start codon targeted polymorphism, SCoT)是一种新型的遗传分子标记技术。本研究采用单因素水平设计方法，优化并建立梨的适宜SCoT分子标记体系，通过SCoT标记技术，分析了安徽省砀山县的43份梨(Pyrus spp.)的遗传多样性和亲缘关系。结果表明，优化后梨SCoT-PCR反应体系：10×Easy Taq Buffer(含2 mmol/L Mg2+)2.5 μL，模板DNA终浓度30 mg/L，上下游引物终浓度1.0 μmol/L，dNTPs 终浓度0.2 mmol/L，Taq DNA聚合酶1.0 U，总体积为25 μL。随机选取2个DNA模板进行引物筛选，最终从67条引物中筛选出16条扩增条带清晰且有差异性的引物，共扩增出145条带，其中多态性条带有126条，多态性比率为86.1%，每条引物平均扩增出9.1条带。SCoT标记的43个梨材料遗传相似性系数为0.517~0.931，平均值为0.685。聚类分析表明，在遗传相似系数为0.610的水平上，43份梨材料分为A和B两组；在遗传相似系数为0.746的水平上，A组又分为6个亚组(Ⅰ~Ⅵ)。所研究的43份梨材料具有丰富的遗传多样性，且能够被SCoT分子标记有效地检验，为进一步研究利用安徽省砀山县梨种质资源提供了依据。

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	杨君祎
	徐超
	杨芳梅
	方志
	闫冲冲
	张金云
	蔡永萍
	林毅

关键词 ：梨, 目标起始密码子多态性(SCoT), 遗传多样性, 聚类分析

Abstract：Start codon targeted polymorphism (SCoT) is a novel genetic marker technique. In this study, a single factor experiment method was designed to optimize and establish a SCoT molecular marker system for pear (Pyrus spp.). Furthermore, the diversity and genetic relationship of 43 pear cultivars, which were collected from Dangshan, Anhui Province, were analyzed with the optimized SCoT system. The results showed that the optimized SCoT-PCR in pear was as follows: A total volume of 25 μL-reaction system including 2.5 μL 10×Easy Taq buffer (with 2 mmol/L Mg2+), 30 mg/L template DNA, 1.0 μmol/L primer, 0.2 mmol/L dNTPs and 1.0 U Taq polymerase. Sixteen primer combinations were selected from 67 SCoT primers for their amplification with clear bands and significant difference. A total of 145 bands were amplified, in which 126 bands were polymorphic (86.1%) with 9.1 bands per primer on average. The optimized SCoT-PCR system was applied to 43 pear samples. The result showed abundant polymorphism in the tested pear. The value of genetic similarity coefficient was ranged from 0.517 to 0.931, with the average of 0.685. Moreover, the 43 pear varieties could be classified into 2 groups including A and B based on the similarity of 0.610 using the cluster analysis. When the genetic similarity coefficient was set at 0.746, group A could be divided into 6 subgroups (Ⅰ~Ⅵ). These results indicated that the tested 43 pear varieties had relatively high genetic diversity and could be distinguished from each other by SCoT molecular marker technology, which provide basis for further research and application on genetic resources in pear from Dangshan, Anhui province.

Key words： Pear Start codon targeted polymorphism (SCoT) Genetic diversity Cluster analysis

收稿日期: 2014-11-13 出版日期: 2015-03-17

基金资助:国家自然科学基金;国家自然科学基金;国家级大学生创新创业训练计划项目

通讯作者: 林毅 E-mail: yjsc01@ahau.edu.cn

引用本文:

杨君祎徐超杨芳梅方志闫冲冲张金云蔡永萍林毅. 利用目标起始密码子多态性(SCoT)分子标记分析梨遗传多样性[J]. 农业生物技术学报, 2015, 23(5): 588-597.

链接本文:

http://journal05.magtech.org.cn/Jwk_ny/CN/ 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2015/V23/I5/588

陈香玲, 苏伟强, 刘业强, 等. 2012. 36份菠萝种质的遗传多样性SCoT分析. 西南农业学报, 25(2): 625~629. (Chen X L, Su W Q, Liu Y Q, et al. 2012. Analysis on Genetic Diversity of 36 Pineapple Collections by SCoT Markers. Southwest China Journal of Agricultural Sciences, 25(2): 625~629.)
曹玉芬, 刘凤之, 高源, 等. 2007. 梨栽培品种SSR鉴定及遗传多样性. 园艺学报, 34(2):305~310. (Cao Y F, Liu F Z, Gao Y, et al . 2007. SSR Analysis of genetic diversity of Pear cultivars. Acta Horticulturae Sinica, 34(2):305~310.)
甘玲, 汤浩茹, 董晓莉. 2006. 梨种类和材料鉴定研究进展. 中国农学通报, 22(5): 302~307. (Gan L, Tang H R, Dong X L. 2006. An overview on identification of pear species and cultivars. Chinese Agricultural Science Bulletin, 22(5): 302~307.)
刘超, 党江波, 魏烨昕, 等. 2013. SCoT分子标记技术初步应用于烟草属部分材料的遗传分析及种间杂种的鉴定. 中国烟草学报, 19(5): 107~111. (Liu C, Dang J B, Wei Y X, et al. 2013. The application of SCoT molecular marker technology in genetic analysis and interspecific hybrid identification in genus Nicotiana. Acta Tabacaria Sinica, 19(5): 107~111.)
李正鹏, 李廷春, 樊洪泓, 等. 2011. 棉花遗传多样性SCoT和SRAP标记的研究及比较分析. 激光生物学报, 20(2): 236~244. (Li Z P, Li T C, Fan H H, et al. 2011. Comparison of SCoT and SRAP markers for genetic analysis of Gossypium hirsutum. Acta Laser Biology Sinica, 20(2): 236~244.)
路娟, 吴俊, 张绍铃, 等. 2011. 不同系统梨种质遗传多样性与分类关系的SSR分析. 南京农业大学学报, 34(2):38~46. (Lu Juan, Wu Jun, Zhang S L, et al. 2011.Genetic diversity and polygentic relationship among pears reveled by SSR analysis. Journal of Nanjing Agricultural University, 34(2):38~46.)
罗霆, 杨海霞, 岑华飞, 等. 2013. SCoT分子标记在割手密遗传图谱构建中的应用. 植物遗传资源学报, 14(4): 704~710. (Luo T, Yang H X, Cen H F, et al. 2013. Application of SCoT molecular marker in construction of molecular genetic linkage map of Saccharum spontaneum L. Journal of Plant Genetic Resources, 14(4): 704~710.)
莫文娟, 袁德义, 段经华, 等. 2009. 新高系梨9个材料SSR标记分析. 浙江林学院学报, 26(5): 639~643. (Mo W J, Yuan D Y, Duan J H, et al. 2009. Simple sequence repeat analysis on pear cultivars in Niitaka line.Journal of Zhejiang Forestry College, 26(5): 639~643.)
秦国新, 何桥, 梁国鲁, 等. 2012. 草莓属植物SCoT分析体系的建立及优化. 果树学报, 29(3): 393~397. (Qin G X, He Q, Liang G L, et al. 2012. Establishment and optimization of SCoT-PCR system in fragaria. Journal of Fruit Science, 29(3): 393~397.)
冉辛拓, 贺丽敏, 田少强, 等. 2006. 蜜梨与授粉材料亲缘关系及其花粉直感的通径分析. 河北农业科学, 10(2): 7~10. (Ran X T, He L M, Tian S Q, et al. 2006. Path analysis on genetic relationship and metaxenia effects in pears. Journal of Hebei Agricultural Sciences, 10(2): 7~10.)
沈玉英, 滕元文, 田边贤二. 2006. 部分中国砂梨和日本梨的RAPD分析. 园艺学报, 33(3): 621~624. (Shen Y Y, Teng Y W, Tanabe Kenji. 2006. RAPD Analysis for Genetic Assessment of Some Cultivars of Pyrus pyrifolia Derived from China and Japan. Acta Horticulturae Sinica, 33(3):621~624.)
宋丰顺, 朱立武, 张水明, 等. 2004. 砀山酥梨自然保护区梨种质资源RAPD分析. 安徽农业大学学报, 31(3): 358~362. (Song F S, Zhu L W, Zhang S M, et al. 2004. RAPD analysis of the pear germplasm resources in protection district of Dangshansu Pear. Journal of Anhui Agricultural University, 31(3): 358~362.)
徐国华, 吴华清, 张绍铃. 2004. 中国梨材料S基因型鉴定的初步研究. 西北植物学报, 24(10): 1861~1865. (Xu G H, Wu H Q, Zhang S L. 2004. Study on identification of S-genotypes of Chinese pear cultivars. Acta Botanica Boreali-Occidentalia Sinica, 24(10): 1861~1865.)
杨祥燕, 蔡元保, 郭凌飞, 等. 2012. 番木瓜SCoT反应体系建立及引物筛选. 热带亚热带植物学报, 20(6): 578~584. (Yang X Y, Cai Y B, Guo L F, et al. 2012. Establishment of SCoT Amplification System and Primer Selection for Carica papaya. Journal of Tropical and Subtropical Botany, 20(6): 578~584.)
张全军, 李文贵, 钟必凤, 等. 2013. 南方砂梨返花返青发生特性及其与早期落叶的关系. 西南农业学报, 26(3):1160~1163. (Zhang Q J, Li W G, Zhong B F, et al. 2013.Reflorescence and refoliation characteristics of Pyrus pyrifolia Nakai in south China and its relationship with early defoliation. Southwest China Journal of Agricultural Sciences, 26(3):1160~1163.)
赵瑞强, 高燕会, 章晓玲, 等. 2012. 铁皮石斛SCoT-PCR反应体系构建及优化. 核农学报, 26(4):648~655. (Zhao R Q, Gao Y H, Zhang X L et al. 2012. Establishment and optimization of SCoT-PCR reaction system for Dendrobium Officinale. Journal of Nuclear Agricultural Sciences, 26(4):648~655.)
朱立武, 王艳芳, 贾兵, 等. 2009. 安徽砀山酥梨自然保护区梨种质资源AFLP分析. 果树学报, 26(2): 145~150. (Zhu L W, Wang Y F, Jia Bing, et al. 2009. Study on the pear germplasm resources in Dangshan area, Anhui province by AFLP technology. Journal of Fruit Science, 26(2): 145~150.)
Bao L, Chen K S, Zhang D, et al. 2008. An assessment of genetic variability and relationships within Asian pears based on AFLP (amplified fragment length polymorphism) markers. Scientia Horticulturae, 116 (4), 374~380.
Collard B C Y, Mackill D J. 2009. Start codon targeted (SCoT) polymorphism: A simple, novel DNA marker technique for generating gene targeted markers in plants . Plant Mol Biol Rep, 27: 86~93.
Erfani J, Ebadi A, Abdollahi H, et al. 2012. Genetic diversity of some pear cultivars and genotypes using simple sequence repeat (SSR) markers. Plant Mol Biol Rep, 30: 1065~1072.
Teng Y, Tanabe K. 2004. Reconsideration on the origin of cultivated pears native to East Asia. Acta Horticulturae, 634: 175~182.
Zou Y P, Ge S. Wang X D. 2001. Molecular Marker for Systematic and Evolutionary Botany. Beijing: Science Press, 16(10): 154~158.