RNAi silencing of the Meloidogyne incognita Rpn7 gene reduces nematode parasitic success
RNAi silencing of the Meloidogyne incognita Rpn7 gene reduces nematode parasitic success
复制标题
RNAi 沉默南方根结线虫 Rpn7 基因可降低线虫寄生成功率
DOI：
10.1007/s10658-012-9971-y
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发表时间：
2012-09-01
影响因子：
1.8
通讯作者：
Guo, Yangdong
中科院分区：
文献类型：
--
作者：
Niu, Junhai;Jian, Heng;Guo, Yangdong
RNA干扰（RNAi）技术是植物寄生线虫（PPN）功能研究的重大突破。它提供了确定新的基本靶标并因此开发新的抗性转基因的可能性。为了验证Mi-Rpn 7作为控制根结线虫南方根结线虫的靶标的潜力，并评估我们的改良平台用于评估沉默表型的可行性，我们敲低了M.使用RNAi在体外和体内对未知的植物进行研究。在用408-bp Rpn 7 dsRNA浸泡后，前寄生第二阶段幼（J2）线虫显示特异性转录物敲低，导致在用Pluronic凝胶培养基的吸引测定中运动中断，并因此使线虫感染降低55.2%至66.5%。Rpn 7 dsRNA在转基因复合植株中的体内表达使每克根组织中的卵量减少了34%（P < 0.05），每克根组织中的卵数减少了50.8%（P < 0.05）。我们的结果表明，在M. incognita J2显著降低运动性和感染性。虽然Mi-Rpn 7的RNAi不能完全抑制毛状根中的抗性，但它对毛状根的繁殖和运动产生了显著的负面影响。无名氏此外，本文提出的改进方法为PPN基因功能分析或靶点筛选提供了技术参考。
RNA interference (RNAi) techniques provide a major breakthrough in functional analysis for plant parasitic nematodes (PPNs). It offers the possibility of identifying new essential targets and consequently developing new resistance transgenes. To validate the potential of Mi-Rpn7 as a target for controlling root knot nematode Meloidogyne incognita and to evaluate the feasibility of our modified platform for assessing silencing phenotypes, we knocked down the Rpn7 gene of M. incognita using RNAi in vitro and in vivo. After soaking with 408-bp Rpn7 dsRNA, pre-parasitic second-stage juvenile (J2) nematodes showed specific transcript knockdown, resulting in an interrupted locomotion in an attraction assay with Pluronic gel medium, and consequently in a reduction of nematode infection ranging from 55.2% to 66.5%. With in vivo expression of Rpn7 dsRNA in transformed composite plants, the amount of egg mass per gram root tissue was reduced by 34% (P < 0.05) and the number of eggs per gram root tissue was reduced by 50.8% (P < 0.05). Our results demonstrated that the silencing of the Rpn7 gene in M. incognita J2s significantly reduced motility and infectivity. Although it does not confer complete resistance, Mi-Rpn7 RNAi in hairy roots produced significant negative impacts on reproduction and motility of M. incognita. In addition, the presented modified procedure provides technique reference for PPN genes functional analysis or target screening.