ISHS Acta Horticulturae 828: International Symposium on Recent Advances in Banana Crop Protection for Sustainable Production and Improved Livelihoods
DEVELOPMENT OF SIMPLE MOLECULAR PROTOCOLS TO DETECT BANANA BUNCHY TOP VIRUS WITH THE PHYTOPASS SYSTEM |
| Authors: | J.P. Busogoro, O. Dutrecq, H. Jijakli |
| Keywords: | conservation, crude extracts, infection, PCR |
| Abstract:
A new protocol was developed for the molecular detection of Banana bunchy top virus (BBTV) in banana. Fragments of banana tissue from BBTV-infected plants were collected by scraping leaves with a PhytoPASS abrasive membrane. The samples collected were stored at room temperature until tested by PCR. To detect BBTV, the PhytoPASS membrane covered by plant-tissue fragments was introduced into a 15-ml tube containing 1 ml of 4°C KAJI extraction buffer and swirled rapidly. The resultant crude suspension was diluted 100 times in distilled water prior to PCR amplification with previously described BBTV-specific primers. A specific DNA fragment was amplified suggesting that BBTV infection could be detected without purified DNA or the immunocapture step preceding the PCR reaction. Detection of BBTV in stored samples also suggests that the newly developed protocol could be used to analyse materials collected in different production areas. Because it involves the direct use of crude extracts, this simplified protocol allows quick virus detection with minimal risk of manipulation errors, misidentification, and cross-contamination when many samples are tested. Adopting this protocol would make the routine use of PCR for BBTV detection possible in all banana-growing regions. | |
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