2021 Fiscal Year Annual Research Report
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
| Project/Area Number |
20F20392
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| Research Institution | Tokyo University of Agriculture and Technology |
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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)
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| 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
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| Project Period (FY) |
2020-11-13 – 2023-03-31
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| Keywords | potexvirus / necrosis / RNA-seq |
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| Outline of Annual Research Achievements |
To analyze mechanisms underlying necrotic symptoms, we employed Nicotiana benthamiana and plantago asiatica mosaic virus (PlAMV) pathosystem, where helicase domain of PlAMV is a determinant of necrotic symptoms caused by PlAMV. We transiently expressed helicase domain (HEL) of PlAMV by agroinfiltration and extracted total RNA from these leaf tissues at 2 days after infiltration. We sent total RNA samples for RNA-seq analysis.
We successfully obtained enough reads for bioinformatic analyses. So we next carried out bioinformatic analysis including k-mer clustering and found set of genes induced specifically in necrosis-inducing condition mediated by HEL domain. Among these genes, we found oxylipin synthesis genes and endoplasmic reticulum stress genes. We are now trying to examine the effect of these genes on systemic necrosis by virus-induced gene silencing using tobacco rattle virus vector.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
We successfully obtained enough reads for gene expression analyses by RNA-seq. We found sets of genes induced in necrosis, and some of these are consistent with the previous findings that oxylipin synthesis is involved in necrosis.
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| Strategy for Future Research Activity |
We will carry out gene-knockdown analysis to examine which genes are involved in 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.
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