| 杨树RAPD标记技术体系的建立与优化 |
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| 引用本文： | 张玉平. 杨树RAPD标记技术体系的建立与优化[J]. 中国农学通报, 2014, 30(13): 20-24. DOI: 10.11924/j.issn.1000-6850.2013-2814 |
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| 作者姓名： | 张玉平 |
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| 作者单位： | 北京市农林科学院林业果树研究所 |
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| 基金项目： | 科技部科技基础性工作资助项目“国家核桃板栗种质资源平台的运行服务”（2013-048）; 农业部作物种质资源保护项目“核桃板栗种质资源收集、保存与利用”（2014NWB009）; 山东省农业良种工程“果树地方优势品种发掘与种质创新利用研究”鲁科农学（2011-086） |
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| 摘 要： | 本研究主要对杨树RAPD体系进行优化，以银白杨为试材，利用L16(45)正交试验设计和2个单因素试验，研究各主要参数的适宜浓度。结果表明，杨树RAPD优化后的反应体系：20μL反应体系中，含Mg2+ 1.5 mmol/L、Taq酶2.0 U、引物0.2μmol/L、dNTPs 0.2 mmol/L、模板DNA 50 ng。扩增程序为：94℃预变性3 min、94℃变性40 s、38℃退火60 s、72℃延伸90 s，共38个循环；最后在72℃延伸7 min后4℃保温。通过1.5%的琼脂糖凝胶电泳检测其扩增产物，证明该体系稳定可靠，可以用于杨树的遗传学分析。
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| 关 键 词： | 诊断方法 诊断方法 |
| 收稿时间： | 2013-10-29 |
| 修稿时间： | 2014-01-02 |
Establishment and Optimization of RAPD Technique in Poplar |
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| Affiliation: | Jia Jianyun, Chen Xin, Liu Hongdui, Zhang Junlin, Xu Li, Zhang Lisi, Liu Qingzhong (1.Shandong Institute of Pomology, Shandong Provincial Key Laboratory of Fruit Tree Biotechnology Breeding, Tai＇an Shandong 271000; 2Shandong Academy of Agricultural Sciences, Jinan 250100; 3Landscaping Ltd of Hangzhou City, Hangzhou 310000) |
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| Abstract: | The analysis of genetic relationship of Ping＇ou hybrid hazelnut laid the foundation for the geneticmapping and the breeding of choiceness gene. 21 main cultivars were selected for the experiment. After DNAextraction, SSR molecular marker and NTSYSpc（version 2.11 a） analysis of the results, constructed adendrogram. 6 primer pairs of Corylus avellana were selected for the detection of the tested materials. 71genetic loci were detected, which were polymorphic loci in all. The percentage of polymorphic bands was100%. The SSR-derived genetic similarity was 0.34-0.82, with an average of 0.55. All the samples weredivided into 4 groups when the threshold value was 0.74. C1, C13 and C2 constituted the groupⅠ; C6 and C15constituted the groupⅡ; C7, C9, C14, C16 and C10 constituted the groupⅣ. The others constituted the groupⅢ. There were genetically abundant diversities among Ping＇ou hybrid hazelnut. The genetic relationships ofthe main cultivars were complex. Analysis system established in this study could offer reference for otherrelative researches. |
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| Keywords: | amplified procedure |
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