茶树冷诱导基因的AFLP筛选及其表达分析

Screening Genes Related to Cold Stress from Tea Plant Using AFLP

以中小叶茶树良种舒茶早和云南大叶种73-11为材料,于4℃低温处理4 d后,采用AFLP技术筛选出冷诱导下茶树差异表达基因并进行分析.结果显示,实验共获得41个差异片段(TDFs),其中舒茶早产生19条差异条带,73-11产生38条差异条带,两品种共有条带16条.所获得的差异片段按功能可划分为4类,即信号传导蛋白、转录因子、基础代谢相关蛋白、抗逆蛋白质,此外还有一些假设蛋白质以及未知蛋白.利用qRT-PCR对Kh1、Kh3和Kh11差异片段进行验证,结果显示这3个片段均被低温诱导,表达量上升.研究表明,茶树在低温胁迫下的逆境反应非常复杂,涉及多种代谢过程中的众多基因,而且大叶种比中小叶种对低温反应更为敏感.

The cold regulated genes were screened from both Yunnan large-leaved cv.＇73-11＇ and small-medium leaved cv.＇Shuchazao＇under low temperature stress for 4 days using cDNA-AFLP.The results showed that 41 Transcript-Derived Fragments（TDFs） were obtained,including 38 TDFs from cv.＇73-11＇,19 TDFs from cv.＇Shuchazao＇,and 16 TDFs shared by the two varieties.According to blasting results,these TDFs were classified into four groups：signal transduction genes,transcription factors,primary metabolism genes and proteins related to stress.In addition,there were some genes for predicted protein and unknown protein.The results of qRT-PCR for Kh1,kh3 and kh11,showed that expression of these TDFs increased obviously under low temperature stress.It had been shown that response of tea plant to low-temperature stress is very complex with many genes of several metabolism involved,and cv.＇73-11＇ is more sensitive to the low temperature stress than cv.＇Shuchazao＇.