收稿日期: 2014-04-22
修回日期: 2014-05-07
网络出版日期: 2015-03-15
基金资助
国家自然科学基金(31370281)和转基因生物新品种培育重大专项(2009ZX08009-130B)资助
Construction of RNAi and root-specific expression vectors of genes related to low Cd accumulation in rice
Received date: 2014-04-22
Revised date: 2014-05-07
Online published: 2015-03-15
Cd极易被水稻等作物吸收并伴随食物链在人体内累积, 严重威胁人类健康.基于水稻Cd累积机制的现有研究, 克隆与水稻Cd累积相关的基因OsLCD和OsHMA 3, 并克隆一个维管组织特异性表达的启动子OsMTP 11 P, 利用Gateway 技术及传统酶切、连接方法, 成功构建适用于水稻等单子叶植物农杆菌转化的RNA 干扰载体pRI-M-LCD和根特异性表达载体pCB2022-H.期望通过这2个载体共转化水稻, 使大量的Cd扣留在根细胞的液泡中, 限制Cd向地上部分及籽粒中的转运, 从而实现Cd在水稻籽粒中的低累积.
陈煜娴 , 冯珊珊 , 柴兴苹 , 柴团耀 . 水稻低Cd累积相关基因RNA干扰载体及根特异性表达载体的构建[J]. 中国科学院大学学报, 2015 , 32(2) : 185 -191 . DOI: 10.7523/j.issn.2095-6134.2015.02.006
Cd accumulated in soil can be easily absorbed by rice and enriched in the human body through food chain. Investigation of the mechanism for the low Cd accumulation in rice has an important practical value for standard rice breeding and safety of food crops. Multiple genes are involved in the process of the absorption, transport, and accumulation of Cd in rice. The technique of multi-gene co-transformation facilitates the genetic control of low Cd accumulation in rice grain. We cloned two genes that are related to this process, i.e., OsLCD and OsHMA 3 , and a vascular-specific promoter OsMTP 11 P. The RNAi vector pRI-M-LCD and the root-specific expression vector pCB2022-H were constructed by using the gateway technology and traditional enzyme digestion and ligation methods. Through Agrobacterium tumefaciens-mediated transformation, the transgenic rice plants containing both the two transgenes are expected to sequester more Cd into the root vacuoles and to reduce the root-to-shoot translocation of Cd, resulting in low Cd accumulation in grains. The successful construction of the two vectors will be useful in molecular regulation of low Cd accumulation in rice and will lay the foundation for breeding new varieties of rice that accumulates lower Cd in grains.
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