Genetic manipulation in the genome of Soybean yellow mottle mosaic virus (SYMMV) and infectivity analysis for exploring its potentiality as a transient gene delivery system
| dc.contributor.advisor | Anirban Roy | |
| dc.contributor.author | Saurabh Kumar Dubey | |
| dc.date.accessioned | 2017-11-18T08:52:15Z | |
| dc.date.available | 2017-11-18T08:52:15Z | |
| dc.date.issued | 2016 | |
| dc.description | T-9529 | en_US |
| dc.description.abstract | Genetic engineering with the genome of plant viruses resulted in the development of vectors that could be utilized for expression of foreign proteins, silencing of host genes and delivery of other genetic constructs into plants. Virus-derived transient vector system also provided an alternative to transgenic approach. Though many viruses have been utilized for such purposes in many crops, relatively less attention was given to pulse crops particularly those grown in tropical or sub-tropical climates. In the present investigation, an infectious clone of a pulse infecting carmovirus, Soybean yellow mottle mosaic virus was manipulated using inverse PCR and overlapping extension PCR methods and two different mutant amplicons were generated. Among these mutants, the coat protein deleted mutant was not stable after cloning. However, the other mutant, where a multiple cloning site (MCS) with four restriction sites (Kpn2I, Bst1107I, KspAI, and RruI) was inserted successfully at the beginning of the coat protein ORF of SYMMV genome, was found to be stable in E. coli. This MCS-added full-length mutant successfully reproduced the disease symptoms upon agroinoculation. However, compared to the wild type, the mutant produced milder symptoms and took longer time for expression of symptoms in the different legume species (black gram, mung bean, guar bean, French bean, and soybean). The mutant could produce virion after inoculation and could move systemically as evidenced by electron microscopy, ELISA, and RT-PCR. The real-time estimation of transcript abundance showed that the replication of this mutant is markedly reduced in the legume host species. The approach for inserting an MCS into the infectious construct has been standardized and the same strategy will be helpful for introducing more manipulation in the SYMMV genome in future to enhance the efficacy of the construct so that it could be used as a vector for gene delivery in the plant | en_US |
| dc.identifier.uri | http://krishikosh.egranth.ac.in/handle/1/5810035988 | |
| dc.keywords | Genetic manipulation in the genome of Soybean yellow mottle mosaic virus (SYMMV) and infectivity analysis for exploring its potentiality as a transient gene delivery system | en_US |
| dc.language.iso | en_US | en_US |
| dc.publisher | Division of Plant Pathology Indian Agricultural Research Institute New Delhi –1 | en_US |
| dc.sub | Plant Pathology | en_US |
| dc.subject | null | en_US |
| dc.theme | Genetic manipulation in the genome of Soybean yellow mottle mosaic virus (SYMMV) and infectivity analysis for exploring its potentiality as a transient gene delivery system Saurabh Kumar Dubey | en_US |
| dc.these.type | M.Sc | en_US |
| dc.title | Genetic manipulation in the genome of Soybean yellow mottle mosaic virus (SYMMV) and infectivity analysis for exploring its potentiality as a transient gene delivery system | en_US |
| dc.type | Thesis | en_US |