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ARS Home » Pacific West Area » Albany, California » Plant Gene Expression Center » Research » Publications at this Location » Publication #358451
Research Project: Disease Resistance Gene Regulation through RNA Silencing for Improved Crop Protection

Location: Plant Gene Expression Center

Title: Artificial microRNA-mediated sresistance to cucumber green mottle mosiac virus in nicotiana benthamiana

Author
item LIANG, CHAIQIONG - Key Laboratory Of Textile Science & Technology
item HAO, JIANJUN - University Of Maine
item LI, JIANQIANG - Key Laboratory Of Textile Science & Technology
item Baker, Barbara
item LUO, LAIXIN - Huazhong Agricultural University

Submitted to: Frontiers in Plant Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/29/2019
Publication Date: 8/6/2019
Citation: Liang, C., Hao, J., Li, J., Baker, B.J., Luo, L. 2019. Artificial microRNA-mediated sresistance to cucumber green mottle mosiac virus in nicotiana benthamiana. Frontiers in Plant Science. 250:1591–1601. https://doi.org/10.1007/s00425-019-03252-w.
DOI: https://doi.org/10.1007/s00425-019-03252-w

Interpretive Summary: Severe economic losses to members of the squash family occur due to the virus, Cucumber green mottle mosaic virus (CGMMV). We developed a system to use artificial microRNAs to target CGMMV genes. In this study we constructed microRNAs to target the different genes in the virus including the coat protein, movement protein or replicase. The microRNAs were placed into Nicotiana benthamiana and protection from viral disease was assessed by subsequent viral infection. Our results showed that N. benthamiana expressing high levels of microRNAs to the coat protein, the movement protein or the replicase showed viral “resistance” or “tolerance” with reduced viral replication and reduced incidence of CGMMV disease compared to N. benthamiana expressing “empty” vector controls. This work demonstrates the successful application of amiRNA silencing for protection of N. benthamiana against virus replication and spread and indicates that an amiRNA strategy is a promising approach for protecting crops against CGMMV disease.

Technical Abstract: Cucumber green mottle mosaic virus (CGMMV) infects cucurbit plants, and can cause severe economic losses to crop production. RNA silencing has been employed successfully to control several viral diseases but has not yet been applied for management of CGMMV disease. We developed a CGMMV-Nicotiana benthamiana pathosystem to test if artificial microRNAs (amiRNAs) targeting CGMMV genes can protect plants from CGMMV disease. In this study we constructed six amiRNAs (amiR1-CP, amiR2-CP, amiR3-MP, amiR4-MP, amiR5-Rep and amiR6-Rep) targeting conserved sequences of CGMMV genes for coat protein (CP), movement protein (MP) or replicase (Rep) using amiRNA expression vectors based on Arabidopsis thaliana miRNA precursor backbones. The CGMMV-amiRNA vectors were introduced into N. benthamiana using Agrobacterium tumefaciens infiltration and protection from viral disease was assessed by subsequent viral infection of infiltrated plants. Our results showed that N. benthamiana expressing high levels of amiR1-CP, amiR4-MP or amiR6-Rep showed viral “resistance” or “tolerance” with reduced viral replication and reduced incidence of CGMMV disease compared to N. benthamiana expressing “empty” amiRNA vector controls. Plants expressing low levels of amiR2-CP, or moderate levels of amiR3-MP and amiR5-Rep remained susceptible to CGMMV and displayed disease symptoms. This work demonstrates the successful application of amiRNA silencing for protection of N. benthamiana against virus replication and spread and indicates that an amiRNA strategy is a promising approach for protecting crops against CGMMV disease.