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An efficient virus‐induced gene silencing vector for maize functional genomics research
Author(s) -
Wang Rong,
Yang Xinxin,
Wang Nian,
Liu Xuedong,
Nelson Richard S.,
Li Weimin,
Fan Zaifeng,
Zhou Tao
Publication year - 2016
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/tpj.13142
Subject(s) - biology , functional genomics , gene silencing , gene , agrobacterium , viral vector , cucumber mosaic virus , transformation (genetics) , genetics , vector (molecular biology) , cultivar , genome , virus , function (biology) , virology , computational biology , genomics , botany , rna , recombinant dna
Summary Maize is a major crop whose rich genetic diversity provides an advanced resource for genetic research. However, a tool for rapid transient gene function analysis in maize that may be utilized in most maize cultivars has been lacking, resulting in reliance on time‐consuming stable transformation and mutation studies to obtain answers. We developed an efficient virus‐induced gene silencing ( VIGS ) vector for maize based on a naturally maize‐infecting cucumber mosaic virus ( CMV ) strain, ZMBJ ‐ CMV . An infectious clone of ZMBJ ‐ CMV was constructed, and a vascular puncture inoculation method utilizing Agrobacterium was optimized to improve its utility for CMV infection of maize. ZMBJ ‐ CMV was then modified to function as a VIGS vector. The ZMBJ ‐ CMV vector induced mild to moderate symptoms in many maize lines, making it useful for gene function studies in critically important maize cultivars, such as the sequenced reference inbred line B73. Using this CMV VIGS system, expression of two endogenous genes, Zm PDS and ZmIspH , was found to be decreased by 75% and 78%, respectively, compared with non‐silenced tissue. Inserts with lengths of 100–300 bp produced the most complete transcriptional and visual silencing phenotypes. Moreover, genes related to autophagy, Zm ATG 3 and Zm ATG 8a , were also silenced, and it was found that they function in leaf starch degradation. These results indicate that our ZMBJ ‐ CMV VIGS vector provides a tool for rapid and efficient gene function studies in maize.