Construction of a cDNA-AFLP reaction system of Populus euphratica for abiotic stress studies  

作　　者：MA Hong-shuang XIA Xin-li YIN Wei-lun 

机构地区：[1]National Engineering Laboratory for Tree Breeding, Beijing Forestry University, Beijing 100083, E R. China [2]Preparation Office of Taiyuan Taishan Botanical Garden, Taiyuan 030006, P. R. China

出　　处：《Forestry Studies in China》2011年第2期104-111,共8页中国林学（英文版）

基　　金：supported by the National Natural Science Foundation of China (Grant Nos. 30730077,30972339, 31070597);the "948" Project of the State Forestry Administration of China (2007-4-01);the National Key Technologies R&D Program of China(2011BAD38B01)

摘　　要：We established a cDNA-AFLP （amplified fragment length polymorphism） system for Populus euphratica, after optimizing several key factors. The results show that optimized CTAB method is suitable for the extraction of ideal RNA from P euphratica; the double-stranded cDNA （ds-cDNA） is completely digested by the combination of EcoRI and MseI for 4.5 h; the reducible result can be obtained when the preamplification products were diluted 10-fold for selective amplification templates. The amplification products resulting from this reaction system were determined by silver staining and the fragments were clear and stable. Detected by RT-PCR, the optimized differential display cDNA-AFLP technique can accurately reflect the true differences of gene expressions among plants. In addition, 42 pairs of primer arrays, producing repeatable and prolific bands, were successfully selected by this improved method. This investigation establishes a cDNA-AFLP system for P. euphratica, which forms a basis for further study on stress-response genes in P. euphratica.We established a cDNA-AFLP （amplified fragment length polymorphism） system for Populus euphratica, after optimizing several key factors. The results show that optimized CTAB method is suitable for the extraction of ideal RNA from P euphratica; the double-stranded cDNA （ds-cDNA） is completely digested by the combination of EcoRI and MseI for 4.5 h; the reducible result can be obtained when the preamplification products were diluted 10-fold for selective amplification templates. The amplification products resulting from this reaction system were determined by silver staining and the fragments were clear and stable. Detected by RT-PCR, the optimized differential display cDNA-AFLP technique can accurately reflect the true differences of gene expressions among plants. In addition, 42 pairs of primer arrays, producing repeatable and prolific bands, were successfully selected by this improved method. This investigation establishes a cDNA-AFLP system for P. euphratica, which forms a basis for further study on stress-response genes in P. euphratica.

关 键 词：Populus euphratica CDNA-AFLP abiotic stress mechanism 

分 类 号：S662.9[农业科学—果树学] Q78[农业科学—园艺学]