Development and use of an efficient DNA-based viral gene silencing vector for soybean.
Virus-induced gene silencing (VIGS) is increasingly being used as a reverse genetics tool to study functions of specific plant genes. It is especially useful for plants, such as soybean, that are recalcitrant to transformation. Previously, Bean pod mottle virus (BPMV) was shown to be an effective VIGS vector for soybean. However, the reported BPMV vector requires in vitro RNA transcription and inoculation, which is not reliable or amenable to high-throughput applications. To increase the efficiency of the BPMV vector for soybean functional genomics, a DNA-based version was developed. Reported here is the construction of a Cauliflower mosaic virus 35S promoter-driven BPMV vector that is efficient for the study of soybean gene function. The selection of a mild rather than a severe BPMV strain greatly reduced the symptom interference caused by virus infection. The DNA-based BPMV vector was used to silence soybean homologues of genes involved in plant defense, translation, and the cytoskeleton in shoots and in roots. VIGS of the Actin gene resulted in reduced numbers of Soybean mosaic virus infection foci. The results demonstrate the utility of this new vector as an efficient tool for a wide range of applications for soybean functional genomics.
Zhang
Chunquan
C
Department of Plant Pathology, Iowa State University, Ames, IA 50011, USA. czhan2@iastate.edu
Yang
Chunling
C
Whitham
Steven A
SA
Hill
John H
JH
D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics*; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D017343:Genes, Plant; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D015151:Immunoblotting; D010641:Phenotype; D018517:Plant Roots / Q000235:genetics / Q000821:virology; D018520:Plant Shoots / Q000235:genetics / Q000821:virology; D010942:Plant Viruses / Q000472:pathogenicity; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics; D013025:Glycine max / Q000235:genetics* / Q000821:virology*
D004279:DNA, Viral; D012333:RNA, Messenger; D012367:RNA, Viral