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Establishment of Broad Spectrum Resistance Strategy by Switching of Biocencer

通过生物中心切换建立广谱耐药策略

基本信息

批准号：
09556071
负责人：
NAMBA Shigetou
金额：
$ 9.6万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09556071/
关键词：
high resistance broad spectrum resistance infectious transcript different virus species broad spectrum transgenic strategy 広域スペクトラムな抵抗性インフェクシャストランスクリプ不活性ミュータント polymerase cell-to-cell movement protein RNase serine-type protease P

项目摘要

For improving the research, we performed until now, by malting use of plant virus gene, mechanisms of resistance of virus infection were introduced and technique for detection was built. Within plant viruses known, more than 90% are RNA viruses. For gaining high resistance or broad spectrum resistance to these viruses, we performed our research and established related techniques. In this research project, based on our results attained these two years, gene function of Citrus tatter leaf capillovirus and several potyviruses were analyzed. Infectious transcripts were generated by using viral genes or non-coding regions of viral genomes with some modifications. The pathogenicity was analyzed when these infectious transcripts were inoculated to plants. We also constructed plant transformation vector. Several plant virus genes were engeneered into expression vector and then transformed into Ti plasmid. By transformingplant tissues by these constracts, transgenic plants were generated after differentiating and culturing the transformed plant cell. Each trangenic plant line was used for inoculation test with the same virus species or different species. And the level of resistance of each pair of plant line and inoculated virus was analyzed in detail. As a result, several interesting resistance lines were found. To improve broad spectrum resistance of transgenic strategy, more research must be done and might be profound.

为了加强这方面的研究，我们利用植物病毒基因，介绍了植物对病毒侵染的抗性机制，并建立了检测技术。在已知的植物病毒中，90%以上是RNA病毒。为了获得对这些病毒的高抗性或广谱抗性，我们进行了研究并建立了相关技术。本研究在两年研究成果的基础上，对柑橘碎叶毛细管病毒和几种马铃薯Y病毒的基因功能进行了分析。感染性转录本是通过使用病毒基因或病毒基因组的非编码区经过一些修改而产生的。将这些侵染转录物接种到植物上，分析其致病力。我们还构建了植物转化载体。将多个植物病毒基因克隆到表达载体上，转化到Ti载体上。通过这些结构转化植物组织，在转化的植物细胞分化和培养后产生转基因植株。每株转基因植株分别接种同一种或不同种的病毒。并对每对株系及接种病毒后的抗性水平进行了详细分析。结果，发现了几个有趣的抗性品系。为了提高转基因策略的广谱抗性，还需要做更多的研究，而且可能是深入的。