Rapid Nucleic Acid Extraction and PCR Detection of Three Significant Banana Viruses in the Philippines
Paul D. Kohnen, Maura Luisa S. Gabriel, Marissa I. Atis, and Miriam E. Pascua
College of Agriculture, Food and Sustainable Development,
Mariano Marcos State University, Ilocos Norte, Philippines
corresponding author: This email address is being protected from spambots. You need JavaScript enabled to view it.
ABSTRACT
Whole nucleic acid was rapidly isolated from leaf tissue of banana cultivars ‘Cardaba’ and ‘Lacatan’ (Musa spp.) for use in polymerase chain reactions (PCR) designed to detect three viruses commonly found in the Philippines; banana bunchy top nanovirus (BBTV), banana streak badnavirus (BSV), and banana bract mosaic potyvirus (BBrMV). Reliable sequence-specific amplification (~1100 bp, 220 bp, and 324 bp PCR products for BBTV, BSV, and BBrMV, respectively) was observed. Furthermore, BBTV and BSV could be detected simultaneously using two primer sets in the same PCR reaction. Use of these modified techniques could aid in future banana virus detection endeavors.
INTRODUCTION
Bananas (Musa spp.) are the number one export commodity in the Philippines today, making the banana industry the nation’s top dollar earner. The country ranks number five in the world in terms of production volume, in part because the country is blessed with a favorable climate well-suited for year-round banana cultivation. In addition, various cultivars including ‘Cavendish’, ‘Saba’, ‘Cardaba’, ‘Lacatan’, ‘Bungulan’, ‘Latundan’, and ‘Senorita’, are accepted in both the domestic and export markets, either as fresh or processed products (Faylon et al. 2004).
In 2002, however, the alarming spread of banana bunchy top virus (BBTV) and fusarium wilt in the northern part of Luzon culminated in an epidemic that resulted in substantial losses to growers. The research and development efforts that followed focused on the management of BBTV as well as industry rehabilitation. Virus disease surveillance was subsequently expanded to include not only BBTV, but also banana bract mosaic virus (BBrMV) and banana streak virus (BSV), especially in materials selected for micropropagation. Ongoing efforts to reduce disease incidence continue but are sometimes hampered by the use of less reliable diagnostic methods such as symptomatology and some serological assays. . . .
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