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转双 RNA 特异性核糖核酸酶基因 PAC1 大豆提高了对多种病毒的抗性。

Increased multiple virus resistance in transgenic soybean overexpressing the double-strand RNA-specific ribonuclease gene PAC1.

机构信息

Jilin Provincial Key Laboratory of Agricultural Biotechnology, Jilin Academy of Agricultural Sciences, Changchun, 130033, China.

出版信息

Transgenic Res. 2019 Feb;28(1):129-140. doi: 10.1007/s11248-018-0108-8. Epub 2018 Dec 1.

Abstract

Viruses constitute a major constraint to soybean production worldwide and are responsible for significant yield losses every year. Although varying degrees of resistance to specific viral strains has been identified in some soybean genetic sources, the high rate of mutation in viral genomes and mixed infections of different viruses or strains under field conditions usually hinder the effective control of viral diseases. In the present study, we generated transgenic soybean lines constitutively expressing the double-strand RNA specific ribonuclease gene PAC1 from Schizosaccharomyces pombe to evaluate their resistance responses to multiple soybean-infecting virus strains and isolates. Resistance evaluation over three consecutive years showed that the transgenic lines displayed significantly lower levels of disease severity in field conditions when challenged with soybean mosaic virus (SMV) SC3, a prevalent SMV strain in soybean-growing regions of China, compared to the non-transformed (NT) plants. After inoculation with four additional SMV strains (SC7, SC15, SC18, and SMV-R), and three isolates of bean common mosaic virus (BCMV), watermelon mosaic virus (WMV), and bean pod mottle virus (BPMV), the transgenic plants exhibited less severe symptoms and enhanced resistance to virus infections relative to NT plants. Consistent with these results, the accumulation of each virus isolate was significantly inhibited in transgenic plants as confirmed by quantitative real-time PCR and double antibody sandwich enzyme-linked immunosorbent assays. Collectively, our results showed that overexpression of PAC1 can increase multiple virus resistance in transgenic soybean, and thus provide an efficient control strategy against RNA viruses such as SMV, BCMV, WMV, and BPMV.

摘要

病毒是全球大豆生产的主要制约因素,每年都会导致显著的产量损失。尽管在一些大豆遗传资源中已经鉴定出对特定病毒株系的不同程度的抗性,但病毒基因组的高突变率和田间条件下不同病毒或株系的混合感染通常阻碍了病毒病的有效控制。在本研究中,我们生成了组成型表达来自裂殖酵母的双链 RNA 特异性核糖核酸酶基因 PAC1 的转基因大豆品系,以评估它们对多种大豆侵染病毒株系和分离物的抗性反应。连续三年的抗性评估表明,与非转化(NT)植株相比,转 PAC1 基因的植株在田间条件下受到中国大豆种植区流行的大豆花叶病毒(SMV)SC3 挑战时,病情严重程度显著降低。在接种了另外四个 SMV 株系(SC7、SC15、SC18 和 SMV-R)和三个菜豆普通花叶病毒(BCMV)、西瓜花叶病毒(WMV)和豆荚斑驳病毒(BPMV)分离物后,转基因植株表现出较严重的症状和增强对病毒感染的抗性,与 NT 植株相比。与这些结果一致的是,通过定量实时 PCR 和双抗体夹心酶联免疫吸附测定证实,每个病毒分离物在转基因植株中的积累都明显受到抑制。总之,我们的结果表明,PAC1 的过表达可以提高转基因大豆对多种病毒的抗性,从而为防治 SMV、BCMV、WMV 和 BPMV 等 RNA 病毒提供了一种有效的控制策略。

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