Cloning and characterization of coat protein gene of Tomato leaf curl virus infecting tomato and its phylogenetic relationship with other members of geminiviridae (Record no. 290256)
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| 000 -LEADER | |
|---|---|
| fixed length control field | 03208nam a22001577a 4500 |
| 082 ## - DEWEY DECIMAL CLASSIFICATION NUMBER | |
| Classification number | 660.6 |
| Item number | ATH/CL PG |
| 100 ## - MAIN ENTRY--PERSONAL NAME | |
| Personal name | Athira S M |
| 245 ## - TITLE STATEMENT | |
| Title | Cloning and characterization of coat protein gene of Tomato leaf curl virus infecting tomato and its phylogenetic relationship with other members of geminiviridae |
| 260 ## - PUBLICATION, DISTRIBUTION, ETC. (IMPRINT) | |
| Place of publication, distribution, etc | Vellayani |
| Name of publisher, distributor, etc | Department of Plant Biotechnology, College of Agriculture |
| Date of publication, distribution, etc | 2022 |
| 300 ## - PHYSICAL DESCRIPTION | |
| Extent | 74p. |
| 502 ## - DISSERTATION NOTE | |
| Dissertation note | BSc - MSc(Int.) |
| 520 ## - SUMMARY, ETC. | |
| Abstract | The study entitled ‘Cloning and characterization of coat protein gene of Tomato leaf curl virus infecting tomato and its phylogenetic relationship with other members of Geminiviridae’ was carried out at College of Agriculture, Vellayani during the year 2021- 2022. The objective of the study was molecular characterization and cloning of coat protein gene of Tomato leaf curl virus (ToLCV) infecting tomato and its phylogenetic analysis with other members of Geminiviridae. Symptomatology of virus infected tomato plants was studied. Infected plants were collected from different regions of Vellayani campus, Kerala Agricultural University and the virus was maintained by graft inoculation. The virus was serologically characterized using DAS-ELISA (Double Antibody Sandwich-ELISA) and DIBA (Dot Immuno Binding Assay) using ToLCNDV (Tomato leaf curl New Delhi virus) antisera and higher viral titer was shown by plants with severe reduction in leaf size (8-fold absorbance value). Genomic DNA was extracted from the infected samples, and coat protein (CP) gene-specific primers were used for molecular detection. PCR yielded expected amplicon size of 500bp and 600bp and were sequenced. The BLAST analysis of the sequence revealed similarities with Tomato leaf curl Palampur Virus (ToLCPMV) and ToLCNDV of 95% and 94%, respectively. Both bipartite virus and monopartite virus with a satellite DNA were detected by rolling circle amplification (RCA), which was followed by Restriction Fragment Length Polymorphism (RFLP). PCR was done using RCA fragments as template with DNA A specific primers and the amplicons obtained were cloned. Sequencing of cloned genes followed by BLAST analysis showed 98.61 per cent similarities with ToLCPMV isolates. Phylogenetic analysis of partial DNA A gene of Vellayani isolate with other strains of ToLCV showed close relation to ToLCPMV infecting cucurbits. Comparitive analysis of partial DNA A sequence with other viruses in genera Begomovirus showed closest relation with Melon leaf curl virus and Cotton leaf curl virus from Pakistan. Comparison of partial amino acid sequence of CP of ToLCV Vellayani isolates with other mono and bipartite begomoviruses revealed a maximum of 99.11 per cent similarity with pre coat protein genes of ToLCPMV. According to the current investigation, the Begomovirus that infects tomatoes in the Vellayani region is bipartite as well as monopartite with a satellite DNA. The CP and DNA A genome phylogenetic analyses revealed a strong relationship between the Vellayani isolate and the ToLCPMV isolates infecting cucurbits. |
| 650 ## - SUBJECT ADDED ENTRY--TOPICAL TERM | |
| Topical term or geographic name as entry element | Plant Biotechnology |
| 700 ## - ADDED ENTRY--PERSONAL NAME | |
| Personal name | Ayisha R (Guide) |
| 942 ## - ADDED ENTRY ELEMENTS (KOHA) | |
| Source of classification or shelving scheme | |
| Item type | Theses |
| Not for loan | Collection code | Permanent location | Current location | Shelving location | Date acquired | Full call number | Barcode | Date last seen | Koha item type |
|---|---|---|---|---|---|---|---|---|---|
| Not For Loan | Reference Book | KAU Central Library, Thrissur | KAU Central Library, Thrissur | Theses | 2023-05-05 | 660.6 ATH/CL PG | 175689 | 2023-05-05 | Theses |