【摘要】 为解析乙烯利刺激巴西橡胶树橡胶增产的分子机制,应用抑制性消减杂交技术(suppression subtractive hybridization,SSH)成功构建了乙烯利刺激条件下橡胶树胶乳与未处理橡胶树胶乳差异表达的cDNA消减文库。经蓝、白斑筛选,共得到了118个阳性克隆。菌落PCR分析结果表明,92%左右的阳性克隆含有大小200～500bp的插入片段。随机选取25个克隆进行了测序,并用基因检索工具(Genbank)对获得的EST进行了Blastx分析,结果表明,5个EST属于无任何序列线索的未知序列,6个属于未知功能的推测蛋白基因,2个属于未知功能的蛋白基因,3个与已知功能基因具有较高的同源性。应用半定量PCR技术,对编号203片段进行的表达分析结果表明,该片段为乙烯利诱导特异表达基因,在乙烯利处理后24h范围内随处理时间的增加而表达量增强,叶片中没有检测到该基因的表达。更多还原

【Abstract】 A SSH library of differently expressed cDNA was constructed employing two kinds of latex,latex pro-duced under ethephon stimulation for 12 h,and the control without any treatment,in order to illuminate the molecular mechanism underlying higher latex yield in Hevea brasiliensis stimulated by Eth.118 ESTs were ob-tained using the technique of screening the blue-white colony and PCR,in which the 25 selected positive clones were sequenced,indicating 5 ESTs fall into no-clued unknown sequences,6 for the deduced protein genes with unknown functions,2 for the unknown function protein genes,and 3 have high similarity with the known corre-sponding gene respectively,through blastx analysis in Genbank.We checked the expressing profiles of No 203 EST indicating that its transcripts were detected only in the ethphon-induced latex and no transcripts in the leaves,and the expressing intensity depended on the time after application of ethephon.更多还原