| 板栗ISSR反应体系的优化及河北省板栗生态型的分析 |
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| 引用本文： | 韩继成,刘庆香,王广鹏,张新忠,孔德军.板栗ISSR反应体系的优化及河北省板栗生态型的分析[J].河北农业科学,2008,12(4):64-66. |
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| 作者姓名： | 韩继成 刘庆香 王广鹏 张新忠 孔德军 |
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| 作者单位： | 河北省农林科学院昌黎果树研究所,河北,昌黎,066600 |
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| 基金项目： | 河北省农林科学院科研项目 |
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| 摘 要： | 以早丰和燕明2个板栗品种为试材，应用编号为ISSR52的引物，对ISSR反应体系中的dNTPs、TaqDNA聚合酶、引物、模板和甲酰胺等主要影响因子进行了优化筛选。结果表明：20μLISSR反应体系各组分的最适浓度分别为1×Buffer（含2．0mmol／L的Mg^2＋），dNTPs0．2mmol／L，TaqDNA聚合酶1．0U，引物0．2μmol／L，模板25ng。加入2．0％的甲酰胺有利于减轻背景的干扰。利用该体系对9个河北省板栗品种（系）和1个山东省板栗品种进行了分析，以明确河北省板栗居群的遗传多样性。通过对53个ISSR引物的重复筛选，获得了19个稳定的ISSR引物，在10个品种（系）中共产生93条带，33．3％为多态性务带。通过聚类分析进一步明确河北省板栗可分为燕山和太行山2个生态型，并且太行山地区的板栗具有较高的遗传多样性。
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| 关 键 词： | 板栗 简单重复序列区间 反应体系 遗传多样性 |
Optimization of Chestnut Inter-simple Sequence Repeat Reaction System and its Ecotypes in Hebei Province |
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| HAN Ji-cheng,LIU Qing-xiang,WANG Guang-peng,ZHANG Xin-zhong,KONG De-jun.Optimization of Chestnut Inter-simple Sequence Repeat Reaction System and its Ecotypes in Hebei Province[J].Journal of Hebei Agricultural Sciences,2008,12(4):64-66. |
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| Authors: | HAN Ji-cheng LIU Qing-xiang WANG Guang-peng ZHANG Xin-zhong KONG De-jun |
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| Institution: | ( Changli Institute of Pomology, Hebei Academy of Agriculture and Forestry Sciences, Changli 066600, China) |
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| Abstract: | Taking two chestnut varieties （Zaofeng and Yanming2） as materials, the optimization of main influential factors of ISSR reaction system were studied. The results showed that the optimum concentrations of components such as Buffer, dNTPs, Taq DNA polymerase, primer, template DNA in 20 μL reaction system were 1 x Buffer （ with 2.0 mmol/L Mg^2＋ ）, 0. 2 mmol/L, 1.0U, 0. 2 μmol/L and 25 ng, respectively. Furthermore, the addition of 2. 0% formamide was propitious to reduce the disturbance. The genetic diversity of 9 chestnut cultivars in Hebei Province and 1 chestnut cultivar in Shandong Province was studied by ISSR method. Nineteen stable ISSR primers were obtained from 53 tested primers. Ninety three bands were produced in 10 cuhivars with 33.3 percent polymorphic. The result of cluster analysis showed that Heibei Province chestnut could be divided into Yan mountain ecotypes and Taihang mountain ecotypes. More over, the chestnut cultivars or clones in Taihangshan Mountain were more genetic diversity. |
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| Keywords: | Chestnut ISSR(Inter Simple Sequence Repeat) Reaction system Genetic diversity |
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