DETECTION OF BANANA BUNCHY TOP VIRUS IN VIRUS-INFECTED PLANTS USING POLYMERASE CHAIN REACTION

Abstract

Polymerase chain reaction (PCR) was developed for the detection of Banana bunchy top virus
(BBTV) at maximum after 210 min and at minimum after 90 min using Pc-1 and Pc-2, respectively.
PCR detection of BBTV in crude sap indicated that the freezing of banana tissue in liquid nitrogen (LN2)
before extraction was more effective than using sand as the extraction technique. BBTV was also
detected using PCR assay in 69 healthy and diseased plants using Na-PO4 buffer containing 1 % SDS.
PCR detection of BBTV in nucleic acid extracts using seven different extraction buffers to adapt the use
of PCR in routine detection in the field was studied. Results proved that BBTV was detected with high
sensitivity in nucleic acid extracts more than in infectious sap. The results also suggested the common
aetiology for the BBTV by the PCR reactions of BBTV in nucleic acid extracts from Australia, Burundi,
Egypt, France, Gabon, Philippines and Taiwan. Results also proved a positive relation between the
Egyptian-BBTV isolate and abaca bunchy top isolate from the Philippines, but there no relation was
found with the Cucumber mosaic cucumovirus (CMV) isolates from Egypt and Philippines and Banana
bract mosaic virus (BBMV) were found.