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Highly Sensitive and Specific Monoclonal Antibody-Based Serological Methods for rice ragged stunt virus Detection in rice Plants and rice Brown Planthopper Vectors

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Journal of Integrative Agriculture 2014年第9期13卷 1943-1951页

作者： LIU Huan SONG Xi-jiao NI Yue-qun LU Li-na ZHOU Xue-ping WU Jian-xiang Institute of Biotechnology College of Agriculture & Biotechnology Zhejiang University

rice ragged stunt virus（RRSV） is a serious rice disease in Asia, causing serious yield losses on rice. The capsid protein（CP） gene of the major outer capsid protein of RRSV was expressed in Escherichia coli BL21（DE3） using the pMAL-C2 X expression vector. The recombinant protein was used as the immunogen to immunize BALB/c mice. A hybridoma cell line 8A12 secreting monoclonal antibody（MAb） against RRSV was obtained by fusing mouse myeloma cells（Sp 2/0） with spleen cells from the immunized BALB/c mice. Western blot analysis showed that the MAb 8A12 can specifically react with RRSV CP. Using the MAb, an antigen-coated-plate enzyme-linked immunosorbent assay（ACP-ELISA）, a dot enzyme-linked immunosorbent assay（dot-ELISA）, and immunocapture-RT-PCR（IC-RT-PCR） assay were developed to detect RRSV. The established ACP-ELISA, dot-blot ELISA and IC-RT-PCR methods could detect RRSV in infected rice tissue crude extracts with dilutions of 1：40 960, 1：1 280 and 1：655 360（w/v, g mL-1）, respectively. The ACP-ELISA and dot-blot ELISA methods could detect RRSV in infected insect vector crude extracts with dilutions of 1：12 800 and 1：1 600（an individual planthopper μL-1）, respectively. The field survey revealed that rice ragged stunt disease occurs on rice in Hainan, Yunnan, Guangxi, Sichuan, Guizhou, Fujian, Hunan, Jiangxi and Zhejiang in China.

关键词： rice ragged stunt virus rice brown planthopper monoclonal antibody antigen-coated-plate enzyme-linked immunosorbent assay(ACP-ELISA) dot-blot ELISA immunocapture RT-PCR

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Suppression of Jasmonic Acid-Mediated Defense by Viral-Inducible MicroRNA319 Facilitates virus Infection in rice

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Molecular Plant 2016年第9期9卷 1302-1314页

作者： Chao Zhang Zuomei Ding Kangcheng Wu Liang Yang Yang Li Zhen Yang Shan Shi Xiaojuan Liu Shanshan Zhao Zhirui Yang Yu Wang Luping Zheng Juan Wei Zhenguo Du Aihong Zhang Hongqin Miao Yi Li Zujian Wu Jianguo Wu Fujian Province Key Laboratory of Plant Virology Institute of Plant Virology Fujian Agriculture and Forestry University Fuzhou Fujian 350002 China The State Key Laboratory of Protein and Plant Gene Research College of Life Sciences Peking University Beijing 100871 China / Plant Protection Institute Hebei Academy of Agriculture and Forestry Sciences Baoding 071000 China These authors contributed equally to this article.

MicroRNAs （miRNAs） are pivotal modulators of plant development and host-virus interactions. However, the roles and action modes of specific miRNAs involved in viral infection and host susceptibility remain largely unclear. In this study, we show that rice ragged stunt virus （RRSV） infection caused increased accumulation of miR319 but decreased expression of miR319-regulated TCP （TEOSINTE BRANCHED/ CYCLOIDEA/PCF） genes, especially TCP21, in rice plants. Transgenic rice plants overexpressing miP,319 or downregulating TCP21 exhibited disease-like phenotypes and showed significantly higher susceptibility to RRSV in comparison with the wild-type plants. In contrast, only mild disease symptoms were observed in RRSV-infected lines overexpressing TCP21 and especially in the transgenic plants overexpressing miR319- resistant TCP21. Both RRSV infection and overexpression of miR319 caused the decreased endogenous jasmonic acid （JA） levels along with downregulated expression of JA biosynthesis and signaling-related genes in rice. However, treatment of rice plants with methyl jasmonate alleviated disease symptoms caused by RRSV and reduced virus accumulation. Taken together, our results suggest that the induction of miR319 by RRSV infection in rice suppresses JA-mediated defense to facilitate virus infection and symp- tom development.

关键词： miR319 rice ragged stunt virus RRSV TCP21 viral infection jasmonic acid

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