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Analysis of molecular interactions between a novel bromovirus and Arabidopsis thaliana

新型雀麦病毒与拟南芥之间的分子相互作用分析

基本信息

批准号：
13660049
负责人：
MISE Kazuyuki
金额：
$ 2.24万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13660049/
关键词：
Plant virus Bromovirus Arabidopsis thaliana Full-length cDNA Mutant Cell-to-cell movement Symptom Chromosome ウイルス全身感染 in vitro転写 Tissue-printing

项目摘要

Brome mosaic virus (BMV) is one of the most-studied plant virues, but does not infect model plants such as Arabidopsis thaliana. We have recently found that a related bromovirus, Spring beauty latnet virus (SBLV) systemically and efficiently infects A. thaliana. To establish a system for the anlysis of molecular interactions between SBLV and A. thaliana, we first constructed biologically active cDNA clones of SBLV genomic RNAs and determined their complete nucleotide sequences. To identify plant factor(s) involved in SBLV infection, (1) We inoculated 2,231 M2 (Col-0 background) plants mutagenized with EMS treament and identified one mutart line in which SBLV infection was delayed The SBLV infection in this line was shownto be affected at the step of the cell-to-cell movement Genetic analysis demonstrated that slower SBLVspread in this line was recessive. (2) We inoculated 67 Arabidopsis accesions with SBLV, and found anaccession, Pla-O in which SBLV spread was delayed at least in incculated leaves. The SBLV infection in Pla-0 was shown to be affected At the step of the cell-to-cell raovement rather than maltiplication within a single (cell.Genetic analysis demonstrated that slower SBLV spread in Pla-0 was recessive or semidominant trait. (3) Symptom development was examined in A. thaliana accenssions infected with SBLV. Of 85 Arabidopsis accessions tested, four accessions including an accession S96 were Scnsitive to SBLV infection, while the otner accessions including Col-0 were symptomless or developed only mild symptoms. Genetic study suggested that the symptom phenotype of S96 is controlled by a single semidominant lolcus. We bave designated this locus SSBI. Infectivity of SBLV in plants and protoplasts were compalable between Col-0and S96, indicating that variation of symptom phenotype was not due to the different infectivity of SBLV By using genetic markers, SSBI was mapped between ATMB3R and JM142 on chromosome IV.

雀麦花叶病毒(BMV)是研究最多的植物病毒之一，但不侵染拟南芥等模式植物。我们最近发现了一种与之相关的雀巢病毒--春美拉特病毒(Spring Beauty Latnet Virus，SBLV)，它系统而有效地侵染拟南星天牛。为了建立SBLV与拟南芥分子互作分析系统，我们首先构建了具有生物学活性的SBLV基因组DNA克隆，并测定了它们的全核苷酸序列。为了确定SBLV感染的植物因子(S)，(1)我们用EMS诱变处理的2231株M2(Col-0背景)植株，鉴定出1个SBLV感染延迟的突变株系。遗传分析表明，该突变株的SBLV感染是在细胞间运动的阶段受到影响的，遗传分析表明，SBLV在该品系中的缓慢传播是隐性的。(2)将SBLV接种到67个拟南芥植株上，发现了一个SBLV至少在嵌合叶中延缓传播的接穗Pla-O。SBLV在Pla-0中的感染主要发生在细胞间的分裂阶段，而不是在单个细胞内的增殖阶段。遗传分析表明，在Pla-0中传播较慢的SBLV是隐性或半显性的。(3)对感染SBLV的拟南星天牛进行了症状发展观察。在供试的85份拟南芥材料中，包括S96在内的4份材料对SBLV感染呈阳性反应，而包括Col-0在内的OTNER材料无症状或仅表现出轻微的症状。遗传研究表明，S96的症状表型受单个半显性棒状病毒控制。我们已经将这个位置命名为SSBI。利用遗传标记将SBLV在植株和原生质体中的侵染性与Col-0和S96进行比较，表明症状表型的差异不是由于SBLV的侵染性不同所致，SSBI被定位在第4染色体上的ATMB3R和JM142之间。