Record Details

Screening of a multi-virus resistant RNAi construct in cowpea through transient vacuum infiltration method

KRISHI: Publication and Data Inventory Repository

View Archive Info
 
 
Field Value
 
Title Screening of a multi-virus resistant RNAi construct in cowpea through transient vacuum infiltration method
Not Available
 
Creator K. Prasad Babu, Manamohan Maligeppagol, R. Asokan and M. Krishna Reddy
 
Subject Agroinfiltration, Tospovirus, Cowpea, GBNV
 
Description Not Available
Plant viruses are the most devastating pathogens
causing substantial economic losses in many crops. Current
viral disease management relies on prophylactics, roguing
and insect vector control, since in most crops resistant gene
pools for resistance breeding are unavailable. RNA inter-
ference, a sequence dependent gene silencing mechanism
holds great potential in imparting virus resistance. In this
study, the efficacy of a RNAi gene construct developed
against four viruses commonly infesting tomato and chilli
viz., capsicum chlorosis virus, groundnut bud necrosis
virus, cucumber mosaic virus and chilli veinal mottle virus
was evaluated. A 3546 bp dsRNA-forming construct
comprising sense-intron-antisense fragments in binary
vector pBI121 (hpRNAi-MVR) was mobilized into
Agrobacterium tumefaciens
. Cowpea (
Vigna unguiculata
)
was used as an indicator plant for GBNV agroinfiltration to
evaluate the efficacy of hpRNAi-MVR construct in con-
ferring GBNV resistance. The type of agroinfiltration,
bacterial concentration and incubation-temperatures were
optimized. Vacuum infiltration of three pulses of 20–30 s
at 66.66 kPa were effective than syringe infiltration. Of the
five Agrobacterial concentrations, OD
600
0.5 was more
efficient. Incubation temperature of 31
±
1
°
C was favor-
able for development of disease symptoms than 20
±
1
°
C
and 26
±
1
°
C. ELISA revealed a 35% decline in viral
load in hpRNAi-MVR infiltrated plants compared to vector
control plants. Quantitative real time PCR results have
shown a viral gene silencing to the extent of 930–990 folds
in hpRNAi-MVR infiltrated plants compared to vector
control. This approach is simple, rapid and efficient to
screen the efficacy of RNAi constructs developed for the
RNAi mediated plant virus management.
Not Available
 
Date 2019-03-19T03:41:04Z
2019-03-19T03:41:04Z
2019-01-22
 
Type Article
 
Identifier Prasad Babu, K., Maligeppagol, M., Asokan, R. et al. VirusDis. (2019). https://doi.org/10.1007/s13337-018-00509-y
Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/17426
 
Language English
 
Relation Not Available;
 
Publisher Springer