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

doi:10.1016/S2095-3119(18)62000-3

Journal of Integrative Agriculture ›› 2018, Vol. 17 ›› Issue (11): 2492-2500.DOI: 10.1016/S2095-3119(18)62000-3

所属专题：植物病毒合辑Plant Virus

收稿日期:2017-10-10 出版日期:2018-11-01 发布日期:2018-11-01

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

Received:2017-10-10 Online:2018-11-01 Published:2018-11-01
Contact: Correspondence Jiban Kumar Kundu, E-mail: jiban@vurv.cz
About author:Pavel Cejnar, E-mail: cejnar@vurv.cz;
Supported by:
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.

摘要/Abstract

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 (WDVRepA215RBR^mut) 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 WDVRepA215RBR^mut plants to WDVRepA215 plants before significant increases in the WDV viral levels occurred. Both WDVRepA215 and WDVRepA215RBR^mut 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.

Key words: Wheat dwarf virus (WDV) , truncated Rep gene , RBR domain , qPCR screening

. [J]. Journal of Integrative Agriculture, 2018, 17(11): 2492-2500.

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

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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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