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Develop Root-Knot Nematode, Meloidogyne incognita Specific Gene Expression System in Tomato, Solanum lycopersicum L.

KrishiKosh

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Title Develop Root-Knot Nematode, Meloidogyne incognita Specific Gene Expression System in Tomato, Solanum lycopersicum L.
 
Creator YOGESH EKANATHRAO THORAT
 
Contributor Anil Sirohi
 
Subject null
 
Description t-9789
Root-knot nematode (Meloidogyne incognita) is a major constraint to successful
tomato production in India and the world. Engineering genetic resistance can be
achieved in a more selective and robust manner by delivery of dsRNA of the specific
nematode target genes through host delivered RNAi technology. Limiting the
expression of the desired gene to the roots or to the zone of nematode feeding is the best
option while developing transgenic management strategies against the root-knot
nematodes. In the present study, tomato (cv. Pusa Ruby) was transformed with two
nematode responsive root specific promoters of Arabidopsis origin, 3P and 2L, with
GUS as a reporter gene by Agrobacterium mediated transformation. Upon nematode
infection, the 3P transgenic tomato lines showed strong expression of GUS activity at
nematode infection site starting from 10 days and sustained up to 14 days of nematode
inoculation and beyond that, the GUS activity was found to be down regulated while the
2L transgenic tomato lines showed a robust GUS expression upon nematode infection
starting from 10 days and extended up to 30 days of nematode inoculation. In both the
promoters, the GUS expression was found to be restricted around the feeding nematode
and at nematode feeding site (NFS). Quantitative PCR analysis of both promoters
confirmed that the GUS transcript was up regulated upon nematode infection in tomato.
Having done this, the transgenic tomato harboured with nematode responsive root
specific promoter (2L) expressing dsRNAs of M. incognita house-keeping genes
(Splicing factor and Integrase) were generated and challenged with M. incognita
infection. Quantitative PCR analysis confirmed that the transcript of the both the
house−keeping genes got up regulated on nematode infection from 14 to 28 days.
Overall, 50% to 72% and 52% to 71% reduction in nematode galls over the susceptible
control tomato plant was observed in Splicing factor and Integrase transgenic tomato
plants; respectively. The study proves that the expression of the transgenic gene of
interest can be limited to the root/site of nematode infection and can considerably
reduce nematode parasitism
 
Date 2018-10-09T10:01:30Z
2018-10-09T10:01:30Z
2017
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/5810078135
 
Language en_US
 
Format application/pdf
 
Publisher DIVISION OF NEMATOLOGY ICAR-INDIAN AGRICULTURAL RESEARCH INSTITUTE NEW DELHI