龙眼ISSR-PCR反应体系的优化及指纹图谱初探

基金项目:

福建省农业科技重大专项(2004NE02-1);福建省财政专项资助(STIF-Y06)

Inter simple sequence repeats system optimization and fingerprinting for Longan

1.
Fujian Breeding Engineering Technology Research Center for Longan and Loquat, Fuzhou, Fujian 350013, China;
2.
Pomology Institute, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350013, China;
3.
Agricultural college, Hebei University of Engineering, Handan, Hebei 057150, China

摘要: 以立冬本龙眼DNA为模板研究了龙眼ISSR-PCR反应体系的主要成分对ISSR扩增结果的影响,并探讨了构建龙眼指纹图谱的可行性。结果表明:在25μL的PCR反应体系中Mg2+的最适浓度为2.0mmol·L-1;dNTP最适浓度为0.2mmol·L-1;引物的最适浓度为0.7μmol·L-1;模板20ng;TaqDNA聚合酶用量为1.5U。利用优化后的反应体系对福建不同地区龙眼基因型进行了ISSR扩增,扩增产物在250bp～2kb;利用丰富的多态性DNA谱带构建了12个基因型龙眼的指纹图谱。
- 龙眼 /
- ISSR-PCR /
- 优化 /
- 指纹图谱
Abstract: Factors affecting the Inter-Simple Sequence Repeats (ISSR) analysis of longan,such as annealing temperature and concentrations of template DNA,Mg2 + and dNTP were examined.The optimal ISSR-PCR reaction system (25 μL) was found to be: 20 ng template DNA,1.5 U Taq DNA polymerase,0.7 μmol·L-1 primer,2.0 mmol·L-1 Mg2+,and 0.2 mmol·L-1 dNTPs.Using the single primer (ISSR-13),fingerprints of 12 genotypes were constructed from different origins with their abundant polymorphism of DNA spectra.The results suggested that ISSR-PCR could be a suitable reaction system for studying longan genetic diversity.
- Longan /
- ISSR-PCR /
- optimization /
- fingerprint

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