Two mutations in the truncated Rep gene RBR domain delayed the Wheat dwarf virus infection in transgenic barley plants

Pavel Cejnar; Ludmila Ohnoutková; Jan Ripl; Tomá(s) Vl(c)ko; Jiban Kumar Kundu

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Two mutations in the truncated Rep gene RBR domain delayed the Wheat dwarf virus infection in transgenic barley plants

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Wheat dwarf virus (WDV),an important cereal pathogen,is closely related to Maize streak virus (MSV),a model virus of the Mastrevirus genus.Based on its similarity to known MSV resistance strategies,a truncated part of the WDV replicationassociated (RepA) gene (WDVRepA215) and the WDV RepA gene with a mutated retinoblastoma-related protein (RBR)interaction domain (WDVRepA215RBRmut) were cloned into the plPKb002 expression vector and transformed into immature embryos of spring barley cv.Golden Promise plants through Agrobacterium-mediated transformation.A detailed study of T1-generation plants infected by leafhoppers (Psammotettix alienus) fed on infection sources of variable strength was performed over a 5-week period encompassing the initial stages of virus infection.A DNA WDV TaqMan qPCR assay normalized using the DNA puroindoline-b SYBR Green qPCR assay for samples on a per week basis revealed an approximately 2-week delay in WDVRepA215RBRmut plants to WDVRepA215 plants before significant increases in the WDV viral levels occurred.Both WDVRepA215 and WDVRepA215RBRrnut plants showed similar levels of transgenic transcripts over the screened period;however,the transgenic plants also showed increased numbers of infected plants compared to the control plants.

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《农业科学学报（英文版）》 |2018年第11期|2492-2500|共9页
作者
Pavel Cejnar; Ludmila Ohnoutková; Jan Ripl; Tomá(s) Vl(c)ko; Jiban Kumar Kundu;
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Division of Crop Protection and Plant Health, Crop Research Institute, Prague 16106, Czech Republic;

Department of Chemical Biology and Genetics, Centre of the Region Hana for Biotechnological and Agricultural Research Palack(y)University, Olomouc 78371, Czech Republic;

Division of Crop Protection and Plant Health, Crop Research Institute, Prague 16106, Czech Republic;

Department of Chemical Biology and Genetics, Centre of the Region Hana for Biotechnological and Agricultural Research Palack(y)University, Olomouc 78371, Czech Republic;

Division of Crop Protection and Plant Health, Crop Research Institute, Prague 16106, Czech Republic;

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Two mutations in the truncated Rep gene RBR domain delayed the Wheat dwarf virus infection in transgenic barley plants

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