韧皮部特异启动子CoYMV调控下CCoAOMT基因改良黄麻纤维品质的研究

项目名称： 韧皮部特异启动子CoYMV调控下CCoAOMT基因改良黄麻纤维品质的研究

项目编号： No.31471549

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 食品科学、农学基础与作物学

项目作者： 陶爱芬

作者单位： 福建农林大学

项目金额： 60万元

中文摘要： 黄麻是世界上重要的纤维作物，其纤维具有透气性好、吸湿性强、易降解等特性。然而，黄麻纤维中木质素含量较高而导致纤维粗、硬，品质不佳，限制了其在纺织等工业领域的应用。本研究拟采用同源克隆技术，克隆黄麻的木质素合成关键酶基因CCoAOMT，采用Real-time PCR技术分析其在黄麻中的表达情况，并将其转化至烟草中进行功能验证；克隆韧皮部组织特异启动子CoYMV，构建其驱动GUS基因的表达载体，转化烟草植株，分析其组织特异表达活性；构建同时包含特异启动子CoYMV和目的基因CCoAOMT的干扰表达载体，以农杆菌介导的方法将其转化至黄麻植株中，分别从DNA、RNA和蛋白质水平对转化植株进行验证；通过测定转基因黄麻植株中的木质素含量、纤维支数、纤维强力等纤维品质性状和抗倒伏、抗病性等抗逆性状，阐明在韧皮部特异启动子CoYMV作用下，CCoAOMT基因在降低黄麻木质素含量、改良黄麻纤维品质上的功能。

中文关键词： 黄麻；韧皮部特异启动子；基因克隆；纤维品质；功能验证

英文摘要： Jute is one of the most important fiber crops in the world, with the planting area and yield almost equal to that of cotton. Jute fibers exhibit a characteristically good moisture absorption performance, rapid water loss capacity and easy degradation. The quality of jute fiber is very poor mainly due to the higher lignin content, which seriously restrict the application of jute fiber in the textile industry and multi-utilization. In this study, we would isolate the full-length cDNA of the Caffeoyl CoA O-methyltransferase (CCoAOMT) gene from jute , by homologous cloning (primers were designed according to the sequence of CCoAOMT gene of other plants) and modified RACE techniques. Differential spatial and temporal expression of CCoAOMT gene would be revealed by Real-time PCR analysis. At the same time, Phloem-Specific Promoter CoYMV also would be cloned, which activity would be testified using histochcmical localization of GUS activity through trangenic tobacco plants. Moreover, the RNAi vector including both CoYMV and CCoAOMT gene would be constructed and integrated into tobacco plants to verify their function. Then, the vector would be integrated into jute plants using agrobacterium-mediated transformation technique. The transgenic plants would be screened from DNA, RNA and protein level. The characteristics as to fiber quality, such as lignin content, fiber fineness, fiber strength and anthracnose risistance would also be measured, so as to analysis the function of improving fiber quality by suppression of CCoAOMT gene through phloem-specific promoter CoYMV . The results could provide an important basis for the application of CCoAOMT gene in the improvement of jute fiber quality. This study has important significance to improve the level of jute breeding, as well as to improve economic benefits of jute planting.

英文关键词： jute;phloem-specific promoter;gene cloning;fiber quality;function verification