摘要
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1229bp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:gus基因在抗性愈伤组织和阳性转基因植株中均能表达;转基因植株在接种稻瘟病菌后12h,GUS表达量是处理前的2.7倍;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12h,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。
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 trans- genic plants were increased by 3.1- and 3.5-fold, respectively. It suggested that M. grisea, SA, and MeJA were inductive factors of OsQ 16p promoter.
出处
《作物学报》
CAS
CSCD
北大核心
2012年第6期980-987,共8页
Acta Agronomica Sinica
基金
国家高技术研究发展计划(863计划)项目(2007AA10Z188
2008AA10Z108)
国家转基因生物新品种培育项目(2009ZX08001-005B)资助
