Scientia Agricultura Sinica ›› 2015, Vol. 48 ›› Issue (7): 1321-1329.doi: 10.3864/j.issn.0578-1752.2015.07.07

• PLANT PROTECTION • Previous Articles     Next Articles

Verification of Verticillium dahliae Pathogenicity of Glycometabolism Related Genes by Using Host-Induced Gene Silencing Method

ZHAO Yu-lan, SU Xiao-feng, CHENG Hong-mei   

  1. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081
  • Received:2014-10-23 Online:2015-04-01 Published:2015-04-01

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Abstract: 【Objective】 The objective of this study is to screen glycometabolism related genes from Verticillium dahliae, confirm the relationship between those genes and the growth and pathogenicity of V. dahliae, and to provide a theoretical basis for the control of plant Verticillium wilt. 【Method】 The glycometabolism related genes of V. dahliae were screened using host-induced gene silencing (HIGS) method. The Tobacco rattle virus vectors containing target genes were injected into tobacco for transient expression of those genes, then V.d991 was inoculatedto those transgenic tobacco using the root-dip approach, PDS would be whitened 7-10 days later, and their disease indexes were counted after 14 days. The fluorescence quantitative PCR was used to detect the amount of V.d991 biomass and the expression of VDPDF-1, VDEG-1, VDMAN-1and VDPD-1. 【Result】 Four glycometabolism related genes from V. dahliae (VDPDF-1, VDEG-1, VDMAN-1 and VDPD-1) were selected because of the great change of their disease indexes. Transgenic tobacco disease indexes of these four genes were reduced by 35%, 30%, 20%, and 10%, respectively, compared with the negative control. After injected with the mixture of VDEG-1/VDPDF-1, VDPD-1/VDPDF-1/ VDMAN-1, VDPD-1/VDMAN-1, VDPD-1/VDPDF-1,and VDEG-1/VDPD-1, transgenic tobacco disease indexes were reduced by 45%, 45%, 40%, 40%, and 35%. In addition to VDPD-1 transgenic tobacco, the transgenic tobacco disease indexes had significant differences compared with the control. After inoculation withV.d991, the V.d991 biomass and expression of target gene were decreased, mixed gene injection had better interference effect than the single gene. The most obvious biomass reduction wereEG/PDF (VDEG-1/VDPDF-1)and EG/PD (VDEG-1/VDPD-1), reached to 0.94, and the most significant interference effect was VDPDF-1, from PD/PDF (VDPD-1/VDPDF-1)，reduced by 0.91. 【Conclusion】 Four genes were selected from V. dahliae by HIGS method. When these genes were interfered, those plants had high resistance to disease, mixed injection of multiple gene vectors were more significant than the single gene. These genes were confirmed with the glycometabolism in V.d991, probably related with V. dahliae pathogenicity.

Key words: gene silencing, Verticillium dahliae, pathogenicity, biomass, interference effect

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