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

M.A.Y. Akhond
Senior Scientific Officer
Botechnology Division, BARI

M. R. Kabir
Scientific Officer
Botechnology Division, BARI

A. Muqit
Senior Scientific Officer
Plant Pathology Division, BARI

M. Al-Amin
Principal Scientific Officer

Abstract/ Executive Summary:

Nine different symptom types were identified among the infected tomato plants on the basis of visual observation. These symptoms will be used as a reference to correlate with the virus strains. Leaf samples from virus infected tomato plants were collected from seven districts of the country in the year 2006-7 and 2007-8. DNA was extracted from a total of 38 samples showing symptoms along with one healthy control (grown in greenhouse) and stored for further analysis.

Keywords: ToLCV, Tomato, lea fcurl, virus

Location: Biotechnology Laboratory, BARI

Start Date: 01-10-2006

End Date: 31-07-2009

Program Area: Variety and Species

Commodity: Tomato

Objectives:

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

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

Methodology:

Sample collection: A field survey was carried out at the experimental fields of BARI, Gazipur farm and virus infected tomato plants were assessed for symptom expression. All the symptoms were photographed for future reference. Infected leaf samples were collected from 7 districts representing the whole country with special emphasis to border areas. Samples were preserved on ice just after collection.

Virus detection: DNA from the infected leaf samples along with the virus particles were isolated and preserved for future analysis. Degenerate primers were designed on the basis of published virus sequences. PCR was performed according to a standardized protocol

Source of Information: Annual report Biotechnology Division 2009-2010

Article Link (Online Journal):

Funding Source:

1. Government

Budget: 2,00,000.00

Total Budget (Tk.) : 2,00,000.00

Achievement/Findings:

Preliminary results show that the designed virus specific primers are able to detect the virus in almost all the infected samples. Thirty six samples out of 38 produced virus specific bands upon PCR analysis. Polymorphism was detected among the samples using PCR-RFLP method. Further analyses and sequencing will be carried out to detect virus strains at the molecular level. It was also evident from the results that morphological symptoms observed in infected tomato plants are not associated with virus strains.

Knowledge Management: Report/Proceedings