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Journal of Integrative Agriculture  2018, Vol. 17 Issue (11): 2492-2500    DOI: 10.1016/S2095-3119(18)62000-3
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Two mutations in the truncated Rep gene RBR domain delayed the Wheat dwarf virus infection in transgenic barley plants
Pavel Cejnar1, Ludmila Ohnoutková2, Jan Ripl1, Tomáš Vl?ko2, Jiban Kumar Kundu1
 
1 Division of Crop Protection and Plant Health, Crop Research Institute, Prague 16106, Czech Republic
2 Department of Chemical Biology and Genetics, Centre of the Region Hana for Biotechnological and Agricultural Research Palacký University, Olomouc 78371, Czech Republic
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Abstract  
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 replication-associated (RepA) gene (WDVRepA215) and the WDV RepA gene with a mutated retinoblastoma-related protein (RBR) interaction domain (WDVRepA215RBRmut) were cloned into the pIPKb002 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 WDVRepA215RBRmut 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. 
 
Keywords:  Wheat dwarf virus (WDV)        truncated Rep gene        RBR domain        qPCR screening  
Received: 10 October 2017   Accepted: 01 November 2018
Fund: This work was equally supported by the Czech Ministry of Education, Youth and Sports funding programme LH12161, and the Czech Ministry of Agriculture funding programme MZE RO0417.
Corresponding Authors:  Correspondence Jiban Kumar Kundu, E-mail: jiban@vurv.cz   
About author:  Pavel Cejnar, E-mail: cejnar@vurv.cz;

Cite this article: 

Pavel Cejnar, Ludmila Ohnoutková, Jan Ripl, Tomá? Vl?ko, Jiban Kumar Kundu. 2018. Two mutations in the truncated Rep gene RBR domain delayed the Wheat dwarf virus infection in transgenic barley plants. Journal of Integrative Agriculture, 17(11): 2492-2500.

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