| 宁琳.番石榴ISSR-PCR最适反应体系的建立与验证[J].中国南方果树,2017,46(4): |
| 番石榴ISSR-PCR最适反应体系的建立与验证 |
| Establishment and validation of the ISSR-PCR optimum reaction system in Psidium guajava L． |
| 投稿时间：2016-08-31 修订日期：2016-09-18 |
| DOI： |
| 中文关键词: 番石榴 ISSR-PCR 最适体系 |
| 英文关键词:guava ISSR-PCR Optimum system |
| 基金项目:农业部热带作物种质资源保护项目“番石榴（杨桃）种质资源保护”（编号：16RZZY-25） |
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| 中文摘要: |
| 为更好的将ISSR标记应用于番石榴种质研究，本研究以“维邦2号”番石榴DNA为筛选体系试验材料，采用单因子试验对ISSR反应中Mg2+浓度、dNTPs浓度、引物浓度、Taq酶浓度、DNA浓度进行了优化。实验结果表明，番石榴20µl最适反应体系为：2.0µl 10×Buffer，1.0mmol/L Mg2+，0.25mmol/L dNTPs，0.8mmol/L引物，0.2U Taq酶，10ng的DNA。利用该反应体系，选用引物UBC810对9份番石榴种质进行PCR反应，再选取“南宁本地番石榴实生2号”DNA作为模板对8条ISSR引物进行PCR反应，对所确立的扩增体系进行验证。结果显示扩增产物条带多态性丰富，且特异性强、重复性好，表明本研究所确定的反应体系适用于番石榴的ISSR分子标记。 |
| 英文摘要: |
| For better applying ISSR markers to guava germplasm research, in this study, “weibang NO.2” guava DNA being test materials for screening system, using the single-factors experiment to optimize the Mg2+ concentration, dNTPs concentration, primer concentration, Taq DNA polymerase concentration and DNA concentration in the ISSR reaction. Experimental results show that the optimal reaction system of guava 20µl is: 2.0µl 10×Buffer,1.0mmol/L Mg2+,0.25mmol/L dNTPs,0.8mmol/L primer,0.2U Taq DNA polymerase,10ng DNA. Using this reaction system for PCR reaction of 9 species of guava germplasm with primer UBC810,than using “Nanning local seedling guava No.2”guava DNA for PCR reaction with 8 ISSR primers, to validation of the established amplification system. The results showed that the amplified polymorphic bands were polymorphic, and the specificity and reproducibility were good. The results indicated that the reaction system is suitable for the ISSR molecular markers of the guava. |
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