南方根结线虫Mi-eft2基因RNAi沉默效应研究

发布时间：2018-06-16 01:22

本文选题：南方根结线虫 + Mi-eft　；参考：《热带作物学报》2016年06期

【摘要】：以南方根结线虫延长因子2基因(Mi-eft2)T克隆质粒为模板,克隆该基因的3个部分片段(各200 bp),作为体外合成ds RNA的模版,利用该ds RNA(F1ds,F2ds,F3ds)干扰南方根结线虫2龄幼虫(J2)的Mi-eft2表达。结果显示,利用real-time PCR作为检测方法,干扰处理后的J2线虫Mi-eft2的表达量(F1、F2、F3),与对照处理的J2线虫Mi-eft2的表达量(F0)相比,分别降低了21%、69%、0.02%。将经过干扰处理的J2线虫和作为对照处理的J2线虫(500头/株)分别接种5株番茄,培养45 d后观察表型。发现干扰处理后J2线虫与对照处理的J2线虫接种番茄后产生的根结数相比较,分别减少了57.09%、68.94%、7.01%。说明Mi-eft2表达的沉默效应会使南方根结线虫对番茄造成的危害程度相应降低,也说明了利用RNAi干扰该基因表达的方法有利于对南方根结线虫病害防治方法的探索。
[Abstract]:Using Mi-eft2T clone plasmid of the prolongation factor 2 gene of Rhizodendron solandii as template, three partial fragments of this gene (200bp each, used as template for synthesis of DS RNA in vitro) were used to interfere with the expression of Mi-eft2 of 2nd instar nematode larvae of Rhizodd-knot nematode. The results showed that the expression of Mi-eft2 in J2 nematode treated by interference was significantly lower than that in the control group (F0), and the expression of Mi-eft2 in J2 nematode treated with interference was decreased by 210.69% and 0.02%, respectively, compared with that of the control treated J2 nematode, the results showed that the expression of Mi-eft2 in J2 nematode treated by interference was significantly lower than that in control group. Five tomato strains were inoculated with interference treated J2 nematode and 500 J2 nematode per plant as control, and the phenotypes were observed after 45 days of culture. It was found that the number of root knots produced by J2 nematodes after inoculation with tomato decreased by 57.0968.94and 7.01, respectively. It indicated that the silencing effect of Mi-eft2 expression would reduce the damage caused by the southern root knot nematode to tomato, and that the method of interfering the gene expression with RNAi was beneficial to the exploration of the control methods of the disease of the southern root knot nematode.
【作者单位】：中国热带农业科学院热带生物技术研究所农业部热带作物生物学与遗传资源利用重点实验室;
【基金】：国家自然科学基金项目资助“沉默延长因子2基因在黄灯笼辣椒根结线虫防治中的作用研究”(No.31101210) 海南省自然科学基金项目“RNAi沉默Mi-eft2基因对南方根结线虫病害防治的研究”(No.313081) 公益性行业(农业)科研专项经费项目资助“南繁区生物安全监测预警及控制”(No.201403075)
【分类号】：S432.45

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