植物RNAウイルスの感染個体内におけるゲノム多様化と病徴発現の分子機構

2020 Fiscal Year Annual Research Report

• Project/Area Number: 20F20392
• Research Institution: Tokyo University of Agriculture and Technology
• Principal Investigator: 小松 健 東京農工大学, (連合)農学研究科(研究院), 准教授 (60451837)
• Co-Investigator(Kenkyū-buntansha): HAMIM ISLAM 東京農工大学, (連合)農学研究科(研究院), 外国人特別研究員
• Project Period (FY): 2020-11-13 – 2023-03-31
• Keywords: potexvirus / necrosis

Outline of Annual Research Achievements

We prepared inoculum of PlAMV by local lesion isolations and propagation in test host plants for the inoculation test in the next season. However, a local lesion isolate from ornamental lily plants \could not cause infection to leaves of cut flowers of ornamental lilies. In contrast, we successfully detected PlAMV by RT-qPCR from lily plants inoculated with bulk lily leaves showing symptoms. To analyze mechanisms underlying necrotic symptoms, we employed Nicotiana benthamiana and PlAMV pathosystem. In this pathosystem, helicase domain is a determinant of necrotic symptoms caused by PlAMV (Komatsu et al., 2011). We transiently expressed helicase domain of PlAMV by agroinfiltration and extracted total RNA from these leaf tissues at 2 days after infiltration for RNA-seq analysis.

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

Even though we need a start-up time of the researcher during COVID-19 situation, we could perform RNA-seq analysis and prepared for amplicon-sequencing.

Strategy for Future Research Activity

We will analyse the data of RNA-seq analysis to see whether some transcriptome changes are observed in the necrosis caused by the helicase domain of PlAMV in Nicotiana benthamiana. In parallel, we will perform amplicon-sequencing of PlAMV replicase region RT-PCR-amplified from lily plants. By bioinformatic analyses of this amplicon-sequencing, we would like to identify what kind of variants in the lily plant enhance infectivity.