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Pcr-based Detection and Characterization of Tomato Leaf Curl and Other Related Geminiviruses in Bangladesh

M.A.Y. Akhond
Principal Scientific Officer
Biotechnology Division, BARI

M. R. Kabir
Scientific Officer
Biotechnology Division, BARI

Abstract/ Executive Summary:

The experiment was conducted with a view to characterise geminivirus strains in different crops beginning with tomato. Samples were collected from 8 different locations of the country and DNA from the infected leaf samples along with the virus particles was isolated. PCR was performed using degenerate and gene-specific primers following a standardized protocol. Most of the samples produced the diagnostic band of Tomato Leaf Curl Virus. Further analysis with PCR-RFLP and PCR-RCA methods revealed the specific virus genotypes. Sequencing of the complete ’A’ genome of 32 isolates has been done to facilitate their phylogenetic analysis. Samples of several other crops like various cucurbits and okra have been collected and DNA extracted for further analysis.

Keywords: ToLCV, Tomato, virus

Location: Biotechnology Laboratory, BARI, Gazipur

Start Date: 01-07-2010

End Date: 30-06-2014

Program Area: Variety and Species

Commodity: Tomato

Objectives:

1. To develop strategies for the diagnosis and characterization of tomato leaf viruses prevailing in Bangladesh.

To develop strategies for diagnosis and characterization of other related geminiviruses in Bangladesh

Methodology:

Sample collection	:	Virus infected plant samples were collected from different parts of the country
Isolation of nucleic acids	:	Nucleic acids were isolated form virus particles using appropriate methods
Virus detection	:	Conventional, molecular and immunochemical techniques.
Characterization	:	PCR, CAPS and RCA-RFLP method, Sequencing

Source of Information: Annual report, Biotechnology Division 2014-2015

Article Link (Online Journal):

Funding Source:

1. World Bank	Budget: 45,00,000.00 approx.
2. Asian Development Bank	Budget: 45,00,000.00 approx.
3. Government	Budget: 45,00,000.00 approx.

Total Budget (Tk.) : 45,00,000.00 approx.

Achievement/Findings:

Survey was conducted in 8 districts of Bangladesh and a total of 46 samples showing leaf curl symptoms in tomato were collected. These 46 isolates were analyzed using PCR-RFLP and RCA-RFLP methods. The presence of leaf curl virus has been confirmed in 44 using advanced molecular techniques. Among those 44 isolates, complete sequencing of ‘A’ genomes of 32 Tomato Leaf Curl Virus (ToLCV) isolates has been done on both strands which confirmed high level of polymorphism and the presence of several virus strains.

Knowledge Management: Report/Proceedings