Development of bud blight viral disease resistant transgenic tomato
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Date
2013
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UAS Dharwad
Abstract
Tomato, an economically important crop in many countries is affected by many viral
diseases including bud blight disease caused by the Groundnut bud necrosis virus, belongs
to the family Bunyaviridae a member of group Tospovirus . Survey was carriedout to
assess the GBNV in major tomato growing regions of Dharwad, Belgaum, Haveri, Kolar
and Bangaluru rural districts during summer 2011. From the survey it is revealed that the
highest incidence (68%) of bud blight was noticed in fields of Poovandahalli village in
Kolar district, whereas least (12%) incidence of the disease was recorded at Singanalli
village of Dharwad district.
The coat protein (CP) gene of five GBNV isolates of Karnataka were cloned and
characterized. The sequence similarity index revealed, that the GBNV- To- DWD coat
protein gene sequence had 96.0 per cent homology with GBNV-To-BGM, 99.2 per cent
homology with GBNV- To- HVR, 94.9 per cent homology with GBNV- T0-KLR and 94.8
per cent homology with GBNV-To-BAN-R isolates. Highest homology of 99.2 per cent was
found between CP gene sequences of GBNV To- DWD and GBNV- To- HVR followed by
96.1 per between GBNV-To- BGM and GBNV-To-BAN-R. Therefore, GBNV- T0 -DWD
isolate was selected for plant transformation studies. The transgenic tomato (DMT-2) lines
were developed by following in planta transformation method.
PCR analysis of the genomic DNA isolated from T1 transformants demonstrated that
the transgene was integrated in the genome of the twenty four out of thirty transgenic tomato
plants.
ELISA using the antibody against the coat protein indicated that the transgenic plants
expressed the coat protein at varying levels. Bioassay results also indicated the coat protein
mediated resistance by the stability of the transgene.