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.
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Creator |
YOGESH EKANATHRAO THORAT
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Contributor |
Anil Sirohi
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Subject |
null
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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 |
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Date |
2018-10-09T10:01:30Z
2018-10-09T10:01:30Z 2017 |
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Type |
Thesis
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Identifier |
http://krishikosh.egranth.ac.in/handle/1/5810078135
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Language |
en_US
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Format |
application/pdf
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Publisher |
DIVISION OF NEMATOLOGY ICAR-INDIAN AGRICULTURAL RESEARCH INSTITUTE NEW DELHI
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