Application and Optimization of VIGS Experimental Technology System in Rosa Hybrida

doi:10.11924/j.issn.1000-6850.casb17090139

Abstract

Abstract: [Objective] The aim of this study is to establish an efficient and fast VIGS experimental system in Rosa hybrida, so that can improve the efficiency of infection by VIGS which is relatively low for Rosa hybrida and other woody plants and provide an experimental basis for further functional study of gene.[Method]The coding sequence of RhPDS was amplified from Rosa hybrida using RT-PCR method.Through double digested,pTRV2-RhPDS vectors were constructed.The test result was validated by gene sequencing and double digested,and then the VIGS expression vectors were transformed into Agrobacterium strain GV3101 by eletroporation.The whole plant was sprayed connected to a portable air compressor.The vacuum infiltration method was used for rose cuttings and tissue culture plant.The expression level of RhPDS gene was detected by RT-qPCR.[Result]The results showed that the recombinant vector pTRV2-RhPDS, which was verified by double enzyme digestion, showed a single band and was in good agreement with the expected target. In the method and material of infection, it is the most rapid and efficient way to infect tissue culture seedlings by vacuum osmosis. [Conclusion]In this research,we obtain an optimized technical system for VIGS which is improve the efficiency of infection by VIGS in Rosa hybrida and other woody plants and provide an solid experimental basis for further functional study of plant gene.

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