Development of plant virus vectors for genetic engeering in fruit frees

用于无果遗传工程的植物病毒载体的开发

• 批准号: 15380032
• 负责人: YOSHIKAWA Nobuyuki
• 金额: $ 9.92万
• 依托单位: Iwate University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15380032/
• 关键词: Virus vector; Apple latent spherical virus; virus-induced gene silencing; FT gene; Promotion of flowering in plants; RNAサイレンシグ; ダイアポジン; ウイルス誘導ジーンサイレンシグ; プロテアーゼ

Apple latent spherical virus (ALSV) has isometric virus particles, c. 25 nm in diameter, and contains two ssRNA species (RNA1 and RNA2) and three capsid proteins. The virus is what is called a latent virus that infects most host plants symptomlessly. The aims of the research are the development of plant virus vectors for expressing foreign genes in plants and the establishment of the genetic engineering system in fruit trees by the virus vectos. The results were summarized as follows.(1)Construction of the ALSV-RNA2 vectors.Infectious cDNA clones of ALSV-RNAs were successfully constructed and modified into viral vectors for stable expression of foreign genes in plants.(2)Expression of foreign genes in plants using ALSV vectors.The ALSV vector which expresses green fluorescence protein (GFP) has been constructed and used for the tracing of cell-to-cell and long distance movement of the virus in infected plants. ALSV expressing yellow and cyan fluorescent proteins (YFP and CFP) have also … More been constructed and used investigate the interaction of populations of identical, but differently tagged viruses in doubly infected plants. The results indicated that ALSV-YFP and ALSV-CFP were always distributed separately in infected leaves. The ALSV vector could also express an antifungal peptide (diapausin) from an insect in plants.(3)Use of ALSV vectors for inducing RNA silencing of a transgene and endogenous genes in plants.ALSV expressing GFP infection efficiently triggered virus-induced gene silencing of a transgene (GFP gene) in transgenic tobacco plants. We also successfully silenced the phytoene desaturase (PDS), sulfur (SU), and proliferating cell nuclear antigen (PCNA) genes in four Nicotiana species (N.tabacum, N.benthamiana, N.occidentalis, and N.glutinosa) and Arabidopsis thaliana plants by ALSV vectors.(4)Promotion of flowering in plants Infected with ALSV expressing flowering locus T (FT) gene of Arabidopsis thaliana.The infection of ALSV expressing FT resulted in the strong promotion of flowering in four Nicotiana species, petunia, and A.thaliana. Less

苹果潜伏球形病毒（ALSV）具有等轴病毒颗粒，c.直径为25 nm，含有两种ssRNA（RNA 1和RNA 2）和三种衣壳蛋白。这种病毒是一种潜伏病毒，它可以不经任何途径感染大多数寄主植物。本研究的目的是开发在植物中表达外源基因的植物病毒载体，并利用病毒载体建立果树基因工程体系。结果总结如下。（1）ALSV-RNA_2载体的构建：成功构建了ALSV-RNA_2感染性cDNA克隆，并将其改造成病毒载体，使外源基因在植物中稳定表达。（2）利用ALSV载体在植物中表达外源基因构建了表达绿色荧光蛋白（GFP）的ALSV载体，并将其用于病毒在感染植物中的细胞间和远距离移动的追踪。表达黄色和青色荧光蛋白（YFP和CFP）的ALSV也具有 ...更多信息 已构建和使用的调查群体的相同，但不同的标记病毒在双重感染的植物。结果表明，ALSV-YFP和ALSV-CFP在感病叶片中始终是分离分布的。ALSV载体还可以在植物中表达来自昆虫的抗真菌肽（diapausin）。（3）ALSV载体在植物中诱导转基因和内源基因的RNA沉默中的用途表达GFP的ALSV感染有效地引发转基因烟草植物中病毒诱导的转基因（GFP基因）的基因沉默。我们还成功地沉默八氢番茄红素去饱和酶（PDS），硫（SU），和增殖细胞核抗原（PCNA）基因在四个烟草物种（烟草，烟草本塞姆，N. octagentalis，和N. numerosa）和拟南芥植物的ALSV载体。（4）表达拟南芥开花位点T（FT）基因的ALSV对植物开花的促进作用表达FT基因的ALSV感染烟草、矮牵牛和拟南芥4种植物后，对植物开花有明显的促进作用。少

项目成果

Mapping the RNA-binding domain on the Apple chlorotic leaf spot virus movement protein

• DOI: 10.1099/vir.0.80493-0
• 发表时间: 2005-01-01
• 期刊: JOURNAL OF GENERAL VIROLOGY
• 影响因子: 3.8
• 作者: Isogai, M;Yoshikawa, N
• 通讯作者: Yoshikawa, N

A movement protein and three capsid proteins are all necessary for the cell-to-cell movement of apple latent spherical cheravirus

一个运动蛋白和三个衣壳蛋白对于苹果潜伏球形樱桃病毒的细胞间运动都是必需的

• 发表时间: 2006
• 期刊: Archives of Virology 151
• 作者: Yoshikawa;N.;Okada;K.;Asanma;K.;Watanabe;K.;et al.
• 通讯作者: et al.

C.Li, Sasaki, N., Isogai, M., Yoshikawa, N.: "Stable expression of foreign proteins in herbaceous and apple plants using Apple latent spherical virus RNA2 vectors"Archives of Virology. 149(In press). (2004)

C.Li、Sasaki, N.、Isogai, M.、Yoshikawa, N.：“使用苹果潜伏球形病毒 RNA2 载体在草本植物和苹果植物中稳定表达外源蛋白”病毒学档案。

異なる蛍光色素でタグをしたウイルス利用による混合感染と干渉作用の解析

不同荧光染料标记病毒的混合感染和干扰效应分析

• 发表时间: 2006
• 期刊: 植物ウイルス病研究会レポート(ISSN 0919-2956) 8
• 作者: 高橋 翼;吉川信幸;夏秋知英
• 通讯作者: 夏秋知英

リンゴ小球形潜在ウイルスの感染・移行とRNAサイレンシングの誘導

苹果球状潜伏病毒的感染、转移及RNA沉默的诱导

• 发表时间: 2006
• 期刊: 植物感染生理談話会論文集 42
• 作者: 吉川信幸;山蔦 翼;佐々木伸浩;五十嵐亜紀;山形広輔;八重樫 元;高橋 翼;磯貝雅道
• 通讯作者: 磯貝雅道