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Title 

Complete genome sequence and construction of infectious full-length cDNA clones of tobacco ringspot Nepovirus, a viral pathogen causing bud blight in soybean

Authors 

F ZhaoU S HwangSeungmo LimRan Hee YooD IgoriS H LeeH S LimJae Sun Moon

Publisher 

Springer Verlag (Germany)

Issue Date 

2015

Citation 

Virus Genes, vol. 51, no. 1, pp. 163-166

Keywords 

AgroinfiltrationBud blightcDNA infectious clonesTobacco ringspot virusWestern blot

Abstract 

Tobacco ringspot virus (TRSV, genus Nepovirus), causes severe diseases in soybean and tobacco plants. TRSV-induced bud blight disease significantly reduced both the yield and quality of soybeans. The function of the encoded viral gene product involved in TRSV infection was unclear due to the limitation of reverse genetics studies on the viral genome. Here, we represent the successful construction of infectious full-length cDNA clones of TRSV genome (RNA1 and RNA2). The cDNAs of TRSV RNA1 and RNA2 were cloned into the binary vector pPZP211 immediately downstream of a double cauliflower mosaic virus 35S promoter and upstream of the nopaline synthase terminator. Seven days after agrobacterium-mediated co-inoculation of these two constructs, Nicotiana benthamiana plants developed a systemic infection with necrotic ringspot symptoms and weak stunting of the leaves, similar to that induced by natural TRSV. The systemic infection was confirmed by transmission electron microscopy and Western blot analysis. Simultaneously, soybean, tomato, and Arabidopsis ecotype Estland were mechanically inoculated with sap prepared from TRSV-agroinfiltrated N. benthamiana leaves, showing typical symptoms of bud blight, necrotic spots, and lethal systemic necrosis, respectively. The system developed herein will be an appealing way to determine TRSV viral gene functions and study host?TRSV interactions.

ISSN 

0920-8569

Link 

http://dx.doi.org/10.1007/s11262-015-1221-x

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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