Screening of Rice Genes Interacting with p5b of Rice Black-Streaked Dwarf Virus

LU Ying1, 2, YANG Jian 1, ZHANG Heng-mu1, CHEN Jian-ping1

doi:10.1016/S1672-6308(13)60128-6

Rice Science >

2013 , Vol. 20 >Issue 3: 185 - 190

DOI: https://doi.org/10.1016/S1672-6308(13)60128-6

Research Paper

Screening of Rice Genes Interacting with p5b of Rice Black-Streaked Dwarf Virus

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1State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control / Key Laboratory of Plant Protection and Biotechnology, Ministry of Agriculture / Zhejiang Provincial Key Laboratory of Plant Virology / Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China; 2College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China

Revised date: 2012-12-31

Online published: 2013-03-27

Supported by

This work was funded by the National Science and Technology Support Program of China (Grant No. 2012BAD19B03), the National High Technology Research and Development Program of China (Grant No. 2007AA10Z414), the National Basic Research Program of China (Grant No. 2010CB126203), the International Science and Technology Cooperation Project (Grant No. 2007DFB30350), the Special Fund for Agro-scientific Research in the Public Interest of China (Grant No. 201003031), the Zhejiang Provincial Science and Technology Project, China (Grant No. 2010C12027), and the Zhejiang Provincial Foundation for Natural Science, China (Grant Nos. Z305165 and Y3090657).

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Abstract

Rice black-streaked dwarf virus (RBSDV) is a recognized member of the genus Fijivirus, family Reoviridae. Its genome has ten double-stranded RNA (dsRNA) segments (S1–S10), in which the fifth genome segment (S5) contains two open reading frames (ORFs) with a partially overlapping region. The second ORF of RBSDV S5 encodes a viral nonstructural protein named p5b with unknown function. To reveal the function of p5b, its gene was ligated into the bait plasmid pGBKT7 and an expression library containing rice cDNAs was constructed using plasmid pGADT7 for yeast two-hybrid assay. The bait protein p5b was detected in yeast by western blot, and the result of an auto-activation test showed that p5b could not autonomously activate the expression of reporter genes in yeast. Then the bait protein p5b was used for screening the cDNA expression libraries of rice. Gene fragments of some pivotal enzymes involved in photosynthesis, respiration and other important metabolic processes, were identified to interact with p5b in yeast, suggesting that these interactions may play roles in symptom development in infected plants.

Key words： rice black-streaked dwarf virus; interaction protein; yeast two-hybrid

Cite this article

LU Ying1, 2, YANG Jian 1, ZHANG Heng-mu1, CHEN Jian-ping1 . Screening of Rice Genes Interacting with p5b of Rice Black-Streaked Dwarf Virus[J]. Rice Science, 2013 , 20(3) : 185 -190 . DOI: 10.1016/S1672-6308(13)60128-6

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