| 锥栗自然居群ISSR-PCR分析技术的建立 |
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| 引用本文： | 刘国彬,龚榜初,罗正荣.锥栗自然居群ISSR-PCR分析技术的建立[J].果树学报,2009,26(1). |
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| 作者姓名： | 刘国彬 龚榜初 罗正荣 |
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| 作者单位： | 刘国彬,LIU Guo-bin(中国林业科学研究院亚热带林业研究所,浙江富阳,311400;华中农业大学园艺植物生物学教育部重点实验室,武汉,430070);龚榜初,GONG Bang-chu(中国林业科学研究院亚热带林业研究所,浙江富阳,311400);罗正荣,LUO Zheng-rong(华中农业大学园艺植物生物学教育部重点实验室,武汉,430070)
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| 基金项目： | 国家农业科技成果转化资金资助项目，国家林业局重点项目 |
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| 摘 要： | 提取野生锥栗的基因组DNA作为ISSR-PCR模板,对反应体系中的模板用量、Mg2+浓度、dNTPs浓度、引物浓度和TaqDNA聚合酶用量5个主要影响因素进行梯度试验,并对扩增程序中的退火温度与循环次数进行了探讨,初步确立了适合野生锥栗的ISSR-PCR分析技术体系,即25μL反应体系中,模板DNA40或50ng、Mg2+2.25mmol.L-1、dNTPs0.2mmol.L-1、引物0.2μmol.L-1、TaqDNA聚合酶1.0U;PCR扩增程序为:94℃预变性3min;然后94℃变性45s,(Tm+2～4℃)退火45s,72℃延伸2min,共32个循环;最后72℃充分延伸5min;1.5%琼脂糖凝胶电泳分离扩增产物。
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| 关 键 词： | 锥栗 自然居群 ISSR |
Establishment of inter -simple sequence repeat reaction system in Castanea henryi |
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| LIU Guo-bin,GONG Bang-chu,LUO Zheng-rong.Establishment of inter -simple sequence repeat reaction system in Castanea henryi[J].Journal of Fruit Science,2009,26(1). |
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| Authors: | LIU Guo-bin GONG Bang-chu LUO Zheng-rong |
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| Institution: | 1Research Institute of Subtropical Forestry;Chinese Academy of Forestry;Fuyang;Zhejiang 311400 China;2Key Laboratory for Horticul- tural Plant Biology;Ministry of Education;Huazhong Agricultural University;Hubei;Wuhan 430070 China |
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| Abstract: | In order to investigate the genetic diversity and relationship within and among Castanea henryi, Zhejiang and Huangshan natural C. henryi were subjected to inter-simple sequence repeat (ISSR-PCR) analysis. Genomic DNA of C. hen- ryi was extracted from leaves as the template, the mainly influencing factors of ISSR were studied and the experiment param- eters were optimized. By adjusting the concentration of template DNA, Mg2+, dNTPs, primer and the contents of Taq DNA polymerase, the PCR amplification system... |
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| Keywords: | ISSR |
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