葫芦科作物基因功能验证平台技术VIGS体系的创建

西瓜、甜瓜和黄瓜等葫芦科作物基因组测序的完成，积累了抗病、抗逆等相关表达序列标签，奠定了葫芦科作物优异基因挖掘及基因功能研究的基础。病毒诱导的基因沉默(VIGS)技术是基因功能验证平台技术，目前亟需研究葫芦科作物的VIGS技术。黄瓜绿斑驳花叶病毒(CGMMV)寄主范围广，能系统侵染多种葫芦科作物，且基因组只有6.4Kb，易于在DNA水平进行基因重组。本研究在病毒侵染性克隆成功构建的基础上，以八氢番茄红素脱氢酶PDS基因为指示基因，以CGMMV为病毒载体，通过点突变和DNA重组等技术，创制葫芦科作物基因验证功能研究的VIGS体系。随后用西瓜、黄瓜和甜瓜上参与叶绿体发育的核基因CH42验证该VIGS载体的广泛性和适用性。该项目成果为在葫芦科作物上利用VIGS研究其基因功能提供快速、高效、高通量的平台技术，加快葫芦科作物功能基因组学的研究，促进我国葫芦科作物产业可持续发展。

The sequencing of genome of watermelon, muskmelon, cucumber , a large of sequences of resistance genes to disease, resistance to stress and other related expressed sequence tags(ESTs) lay foundation of screening of genes and study of gene function of cucurbit crops. Virus inducing gene silencing (VIGS ) is one of powerful tools of gene function research in crops, and VIGS for cucurbits are urgently needed to be explored. Cucumber green mottle mosaic virus(CGMMV) can infect several cucurbitaceous crops, the size of the CGMMV genome is only 6.4Kb, it is easy to construct the mutant at DNA level, it is very suitable for the VIGS vector construction ,. Based on the infectious clone of CGMMV, the PDS and are used to construct the VIGS vector in cucurbitaceous crop. Subsequently, to further verify the universality and currency of VIGS, the CH42 gene is studied in the project. The implementation of the project will provide a fast, efficient, high-throughput platform technology of the gene function research of the cucurbitaceous crops and accelerate the functional genomics research of cucurbit crops and promote the sustainable development of industry of cucurbit crops .