水稻稻瘟病菌诱导表达启动子OsQ16p的克隆与功能分析

qRT-PCR分析表明，日本晴OsQ16基因受稻瘟病菌诱导表达。利用PCR技术从日本晴基因组中克隆了该基因编码区5′端上游1 229 bp的启动子序列，命名为OsQ16p。用其取代pBI121中gus基因上游的CaMV35S启动子，构建重组表达载体pBIQ16p，经农杆菌介导转化日本晴，获得转基因植株。GUS组织化学染色和qRT-PCR分析表明: (1) gus基因在抗性愈伤组织和阳性转基因植株中均能表达; (2)转基因植株在接种稻瘟病菌后12 h，GUS表达量是处理前的2.7倍; (3)抗病相关信号分子水杨酸(salicylic acid，SA)和茉莉酸甲酯(methyl jasmonate，MeJA)喷施转基因植株叶面后12 h，GUS表达量分别为处理前的3.1倍和3.5倍。以上结果表明，OsQ16p启动子具有启动活性，并明显受稻瘟病菌、MeJA和SA诱导表达。

Cloning and Functional Analysis of Magnaporthe oryzae-Induced Promoter OsQ16p in Rice

The qRT-PCR analysis showed that,in Nipponbare(Oryza sativa L.ssp japonica),the expression of OsQ16 gene was induced by Magnaporthe grisea.The 1 299 bp-fragment of 5’-end of OsQ16 gene,named as OsQ16p,was amplified by PCR from Nipponbare.The plasmids pBIQ16p was constructed by replacing the CaMV35S promoter of pBI121 with the OsQ16p,and transformed into Nipponbare through Agrobacterium-mediated transformation.The analysis of GUS activity and qRT-PCR showed that gus gene could express in transgenic plants and calli.The expression of gus gene in transgenic plants was obviously enhanced by M.grisea.After treatment with the resistance-related signaling molecules SA and MeJA,the GUS activities in transgenic plants were increased by 3.1-and 3.5-fold,respectively.It suggested that M.grisea,SA,and MeJA were inductive factors of OsQ16p promoter.