The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
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Optimization of a Virus-Induced Gene Silencing System with Soybean yellow common mosaic virus for Gene Function Studies in Soybeans

  • Kim, Kil Hyun (National Institute of Crop Science, Rural Development Administration) ;
  • Lim, Seungmo (Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kang, Yang Jae (Department of Plant Science and Research Institute for Agriculture and Life Sciences, Seoul National University) ;
  • Yoon, Min Young (Department of Plant Science and Research Institute for Agriculture and Life Sciences, Seoul National University) ;
  • Nam, Moon (School of Applied Biosciences, Kyungpook National University) ;
  • Jun, Tae Hwan (Department of Plant Bioscience, College of Natural Resources & Life Science, Pusan National University) ;
  • Seo, Min-Jung (National Institute of Crop Science, Rural Development Administration) ;
  • Baek, Seong-Bum (National Institute of Crop Science, Rural Development Administration) ;
  • Lee, Jeom-Ho (National Institute of Crop Science, Rural Development Administration) ;
  • Moon, Jung-Kyung (National Institute of Crop Science, Rural Development Administration) ;
  • Lee, Suk-Ha (Department of Plant Science and Research Institute for Agriculture and Life Sciences, Seoul National University) ;
  • Lee, Su-Heon (School of Applied Biosciences, Kyungpook National University) ;
  • Lim, Hyoun-Sub (Department of Applied Biology, Chungnam National University) ;
  • Moon, Jae Sun (Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Park, Chang-Hwan (National Institute of Crop Science, Rural Development Administration)
  • Received : 2015.04.16
  • Accepted : 2015.12.02
  • Published : 2016.04.01
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Abstract

Virus-induced gene silencing (VIGS) is an effective tool for the study of soybean gene function. Successful VIGS depends on the interaction between virus spread and plant growth, which can be influenced by environmental conditions. Recently, we developed a new VIGS system derived from the Soybean yellow common mosaic virus (SYCMV). Here, we investigated several environmental and developmental factors to improve the efficiency of a SYCMV-based VIGS system to optimize the functional analysis of the soybean. Following SYCMV: Glycine max-phytoene desaturase (GmPDS) infiltration, we investigated the effect of photoperiod, inoculation time, concentration of Agrobacterium inoculm, and growth temperature on VIGS efficiency. In addition, the relative expression of GmPDS between non-silenced and silenced plants was measured by qRT-PCR. We found that gene silencing efficiency was highest at a photoperiod of 16/8 h (light/dark) at a growth temperature of approximately $27^{\circ}C$ following syringe infiltration to unrolled unifoliolate leaves in cotyledon stage with a final SYCMV:GmPDS optimal density $(OD)_{600}$ of 2.0. Using this optimized protocol, we achieved high efficiency of GmPDS-silencing in various soybean germplasms including cultivated and wild soybeans. We also confirmed that VIGS occurred in the entire plant, including the root, stem, leaves, and flowers, and could transmit GmPDS to other soybean germplasms via mechanical inoculation. This optimized protocol using a SYCMV-based VIGS system in the soybean should provide a fast and effective method to elucidate gene functions and for use in large-scale screening experiments.

Keywords

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