Turnage Michael A, Muangsan Nooduan, Peele Charles G, Robertson Dominique
Department of Botany, North Carolina State University, Raleigh, NC 27695, USA.
Plant J. 2002 Apr;30(1):107-14. doi: 10.1046/j.1365-313x.2002.01261.x.
Gene silencing, or RNA interference, is a powerful tool for elucidating gene function in Caenorhabditis elegans and Drosophila melanogaster. The vast genetic, developmental and sequence information available for Arabidopsis thaliana makes this an attractive organism in which to develop reliable gene-silencing tools for the plant world. We have developed a system based on the bipartite geminivirus cabbage leaf curl virus (CbLCV) that allows silencing of endogenous genes singly or in combinations in Arabidopsis. Two vectors were tested: a gene-replacement vector derived from the A component; and an insertion vector derived from the B component. Extensive silencing was produced in new growth from the A component vectors, while only minimal silencing and symptoms were seen in the B component vector. Two endogenous genes were silenced simultaneously from the A component vector and silencing of the genes was maintained throughout new growth. Because the CbLCV vectors are DNA vectors they can be inoculated directly from plasmid DNA. Introduction of these vectors into intact plants bypasses transformation and extends the kinds of silencing studies that can be carried out in Arabidopsis.
基因沉默，即RNA干扰，是一种用于阐明秀丽隐杆线虫和黑腹果蝇基因功能的强大工具。拟南芥拥有丰富的遗传、发育和序列信息，这使其成为开发适用于植物界的可靠基因沉默工具的理想生物体。我们基于双分体双生病毒卷心菜叶卷曲病毒（CbLCV）开发了一种系统，该系统能够在拟南芥中单独或组合沉默内源基因。测试了两种载体：一种是源自A组分的基因置换载体；另一种是源自B组分的插入载体。A组分载体在新长出的组织中产生了广泛的沉默，而B组分载体仅出现了最小程度的沉默和症状。从A组分载体中同时沉默了两个内源基因，并且在整个新长出的组织中基因沉默得以维持。由于CbLCV载体是DNA载体，它们可以直接从质粒DNA进行接种。将这些载体导入完整植株无需进行转化，从而扩展了可在拟南芥中开展的沉默研究类型。
