| 苦瓜AFLP和SRAP的PCR反应体系优化及应用比较 |
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| 引用本文： | 琚茜茜,黄如葵,黄玉辉,刘萌芽,冯诚诚.苦瓜AFLP和SRAP的PCR反应体系优化及应用比较[J].广西农业科学,2013(2):195-199. |
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| 作者姓名： | 琚茜茜 黄如葵 黄玉辉 刘萌芽 冯诚诚 |
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| 作者单位： | [1]广西大学农学院,南宁530005 [2]广西农业科学院蔬菜研究所,南宁530007 [3]广西作物遗传改良生物技术重点开放实验室,南宁530007 |
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| 基金项目： | 现代农业产业技术体系建设专项资金项目（CARS-25-G-38）;广西科技攻关项目（桂科攻1222009-3B）;广西农业科学院基本科研业务专项项目（桂农科2012YT05）;南宁市科技攻关项目（201102034B） |
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| 摘 要： | 【目的】优化苦瓜AFLP和SRAP的PCR反应体系，比较这两种分子标记在苦瓜应用中的优劣，为苦瓜突变体筛选提供参考。【方法】以8份苦瓜种质为材料，对AFLP和SRAP反应体系中各反应因素浓度进行筛选优化；分别选取5对条带清晰、重复性好的AFLP和SRAP引物对8份苦瓜材料进行扩增，比较其多态性。【结果】在AFLP反应体系中，在0-10倍稀释范围内，以连接产物稀释5倍时反应效果较好；将预扩产物稀释30倍进行选择性扩增为宜。在SRAP反应体系中，Mg^2＋浓度为2．5mmol／L、dNTP浓度为0．2mmol／L时，反应效果较好。多态性分析结果显示，5对AFLP引物共扩增出145条带，多态性条带有18条，多态性比率为12．41％；5对SRAP引物扩增条带数为109条，其中多态性条带数有9条，多态性比率为8．25％。【结论】AFLP和SRAP两种分子标记技术均可以很好反映苦瓜种质资源的遗传多样性，但AFLP扩增产物的平均多态性高于SRAP，能够比较稳定地筛选出差异条带。
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| 关 键 词： | 苦瓜 AFLP SRAP 多态性 比较 |
Optimization of PCR reaction systems of AFLP and SRAP for Momordica charantia L. and their utilization comparison |
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| Institution: | JU Xi-xi, HUANG Ru-kui, HUANG Yu-hui, LIU Meng-ya, FENG Cheng-eheng (1Agricultural College, Guangxi University, Nanning 530005, China; 2Vegetable Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007, China; 3Guangxi Crop Genetic Improvement and Biotechnology Lab, Nanning 530007, China) |
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| Abstract: | Objective]To provide references for selecting bitter gourd mutants, AFLP and SRAP reaction systems were optimized and their pros and cons were compared. Method]Screening the concentration of all the reaction factors in the method of single-factor analysis and making further optimization for AFLP and SRAP system in M.charantia, and five pairs of cleared-band and reproducible primers were selected to amplify the eight samples, and then their polymorphism was compared. Result ]It was better to dilute the connecting product for five times and pre-amplifieation product for 30 times in the AFLP reaction system and to be 2.5 mmo]/L for Mg2＋ concentration, and 0.2 mmol/L for dNTP concentration in the SRAP reaction system. The result of polymorphism analysis showed that 145 bands were amplifeid from 5 paris of AFLP primers, of which 18 were polymorphic; while 5 paris of SRAP primers produced 109 bands with 9 being polymor- phic. Furthermore, the polymorphism rate in AFLP detection （12.41%） was higher than that in SRAP （8.25%）. Conclu- sion]In screening M.cbarantia mutants, though both AFLP and SRAP could reflect the heritanee polymorphism of M. eha- rantia germplasm, the reproducibility and polymorphism of AFLP were higher than that of SRAP. |
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| Keywords: | Momordica charantia L AFLP SRAP polymorphism comparison |
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