Fujisaki Koki, Iwahashi Fukumatsu, Kaido Masanori, Okuno Tetsuro, Mise Kazuyuki
Laboratory of Plant Pathology, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake-cho, Sakyo-ku, Kyoto 606-8502, Kyoto Prefecture, Japan.
Virus Res. 2009 Mar;140(1-2):103-11. doi: 10.1016/j.virusres.2008.11.007. Epub 2008 Dec 24.
Brome mosaic virus (BMV) and Spring beauty latent virus (SBLV) are closely related, tripartite RNA plant viruses. In Arabidopsis thaliana, BMV shows limited multiplication whereas SBLV efficiently multiplies. Such distinct multiplication abilities have been observed commonly in all Arabidopsis accessions tested. We used this model system to analyze the molecular mechanism of viral resistance in plants at the species level. Unlike SBLV, BMV multiplication was limited even in protoplasts and a reassortment assay indicated that at least viral RNA1 and/or RNA2 determine such distinct infectivities. By screening Arabidopsis mutants with altered defense responses, we found that BMV multiplies efficiently in cpr5-2 mutant plants. This mutation specifically enhanced BMV multiplication in protoplasts, which depended on the functions of RNA1 and RNA2. In the experiment using DNA vectors to express BMV replication proteins encoded by RNA1 and RNA2, BMV RNA3 accumulation in cpr5-2 protoplasts was similar to that in wild-type Col-0 protoplasts, despite significant reduction of accumulation levels of replication proteins, suggesting that cpr5-2 mutation could enhance BMV multiplication independently of increased accumulation, therefore enhanced translation and stabilization, of the replication proteins.
雀麦花叶病毒(BMV)和春美草潜隐病毒(SBLV)是密切相关的三分体RNA植物病毒。在拟南芥中,BMV的增殖有限,而SBLV能有效增殖。在所有测试的拟南芥种质中都普遍观察到这种明显不同的增殖能力。我们利用这个模型系统在物种水平上分析植物中病毒抗性的分子机制。与SBLV不同,即使在原生质体中BMV的增殖也有限,并且重配试验表明至少病毒RNA1和/或RNA2决定了这种明显不同的感染性。通过筛选防御反应改变的拟南芥突变体,我们发现BMV在cpr5 - 2突变体植物中能有效增殖。这种突变特异性地增强了BMV在原生质体中的增殖,这依赖于RNA1和RNA2的功能。在用DNA载体表达由RNA1和RNA2编码的BMV复制蛋白的实验中,尽管复制蛋白的积累水平显著降低,但cpr5 - 2原生质体中BMV RNA3的积累与野生型Col - 0原生质体中的相似,这表明cpr5 - 2突变可以独立于复制蛋白积累的增加、因此独立于复制蛋白翻译和稳定性的增强来增强BMV的增殖。