茶树CsPLt基因的荧光定量PCR分析

摘要

The cDNA-AFLP was applied to identify genes expressed differentially in cold acclimatized tea plant. A cDNA fragment, TDF6 encoding a polyol transporter was isolated. The cDNA fragment had 77 % ,76 % ,75 % homology with polyol transporter genes from Populus trichocarpa ,Haiea brasiliensu and Lotus japonicus by BLAST on NCBI, and named CsPU. qRT-PCR analysis showed that CsPLt gene were up-regulated by cold, dehydration, NaCl, the maximum expression were 7.3, 12.2, 5.3 times higher than that before treatment, respectively. For different tea organs, transcriptional level of the CsPLt gene was the highest in bud, next was in flower, the third was in root and stem, and higher than in mature leaves and the lowest in seed. It was deduced that CsPU gene was very important for tea plant in response to abiotic stress and was strictly controlled in expression of different organs.%利用cDNA-AFLP技术进行茶树低温胁迫处理的基因表达差异分析,获得低温诱导后表达的差异片段TDF6(transcript derived fragment,TDF).经BLAST比对发现该片段与杨树、橡胶树、百脉根的多元醇转运子(polyol transporter)分别有77％、76％、75％的同源性,命名为CsPLt.利用实时荧光定量PCR技术对多元醇转运子基因在不同胁迫以及不同组织中的表达进行分析,结果表明,多元醇转运子基因能被低温、脱水、高盐诱导表达,最大表达量分别比处理前提高7.3、12.2、5.3倍.在芽中表达最高,其次是花蕾,嫩茎表达量与嫩根相近,均比成熟叶种的表达量高,在种子中表达最低.推测该基因在组织中的表达受到严格控制以及在响应非生物胁迫中发挥重要作用.