A high-throughput virus-induced gene silencing protocol identifies genes involved in multi-stress tolerance

BMC Plant Biology

Methodology Article

Makarla Udayakumar¹  Kirankumar S Mysore²  Venkategowda Ramegowda³  Muthappa Senthil-kumar⁴ 

[1] Department of Crop Physiology, University of Agricultural Sciences, GKVK, 560 065, Bangalore, Karnataka, India;Plant Biology Division, The Samuel Roberts Noble Foundation, 2510 Sam Noble Pkwy., 73402, Ardmore, OK, USA;Plant Biology Division, The Samuel Roberts Noble Foundation, 2510 Sam Noble Pkwy., 73402, Ardmore, OK, USA;Department of Crop Physiology, University of Agricultural Sciences, GKVK, 560 065, Bangalore, Karnataka, India;VR: Department of Crop, Soil and Environmental Sciences, MS: National Institute of Plant Genome Research, University of Arkansas, Aruna Asaf Ali Marg, 72701, 110 067, Fayetteville, New Delhi, AR, USA, India;Plant Biology Division, The Samuel Roberts Noble Foundation, 2510 Sam Noble Pkwy., 73402, Ardmore, OK, USA;VR: Department of Crop, Soil and Environmental Sciences, MS: National Institute of Plant Genome Research, University of Arkansas, Aruna Asaf Ali Marg, 72701, 110 067, Fayetteville, New Delhi, AR, USA, India;

关键词: Stress tolerance;    Drought;    Salinity;    Temperature stress;    Nonhost resistance;    Bacterial pathogens;    VIGS;    PTGS;    Translational genomics;   

DOI  :  10.1186/1471-2229-13-193

 received in 2013-07-17, accepted in 2013-11-21,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundUnderstanding the function of a particular gene under various stresses is important for engineering plants for broad-spectrum stress tolerance. Although virus-induced gene silencing (VIGS) has been used to characterize genes involved in abiotic stress tolerance, currently available gene silencing and stress imposition methodology at the whole plant level is not suitable for high-throughput functional analyses of genes. This demands a robust and reliable methodology for characterizing genes involved in abiotic and multi-stress tolerance.ResultsOur methodology employs VIGS-based gene silencing in leaf disks combined with simple stress imposition and effect quantification methodologies for easy and faster characterization of genes involved in abiotic and multi-stress tolerance. By subjecting leaf disks from gene-silenced plants to various abiotic stresses and inoculating silenced plants with various pathogens, we show the involvement of several genes for multi-stress tolerance. In addition, we demonstrate that VIGS can be used to characterize genes involved in thermotolerance. Our results also showed the functional relevance of NtEDS1 in abiotic stress, NbRBX1 and NbCTR1 in oxidative stress; NtRAR1 and NtNPR1 in salinity stress; NbSOS1 and NbHSP101 in biotic stress; and NtEDS1, NbETR1, NbWRKY2 and NbMYC2 in thermotolerance.ConclusionsIn addition to widening the application of VIGS, we developed a robust, easy and high-throughput methodology for functional characterization of genes involved in multi-stress tolerance.

【 授权许可】

Unknown   
© Ramegowda et al.; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.