Development of a rapid detection method for Banana streak virus and the survey of banana streak disease on Musa spp. germplasm collection in Taiwan
Date Issued
2010
Date
2010
Author(s)
Huang, Sz-Jia
Abstract
BSV belonging to the genus Badnavirus in the family Caulimoviridae, is a double-stranded DNA virus. Part of the BSV genome usually becomes endogenous pararetroviral sequences (EPRVs) that can be found in all banana varieties containing genome derived from Musa balbisiana. It indicated an integration event happened in Musa balbisiana. EPRV does not cause symptoms on bananas, but it will sometimes become active (episomal form) during vegetative propagation such as tissue culture and cause severe symptoms on plants. Currently 6 different strains of BSV have been reported. Despite several methods have been developed for the detection of BSV including the enzyme-linked immunosorbent assay (ELISA), the immunocapture polymerase chain reaction (IC-PCR) and the multiplex IC-PCR (M-IC-PCR) etc., these methods can not detect all strains of BSV. Besides, the presence of EPRV leads to false positive results by PCR based methods to detect the episomal form of BSV. A reverse transcription PCR (RT-PCR) method has been developed for BSV strains, Mysore and OL, reported in Taiwan, to detect and to differenciate the EPRV and episomal form of BSV in our laboratory. In this study we designed several pairs of specific primer from the common regions shared by BSV strains to perfrom RT-PCR, and primer pairs were designed. One primer pair, 5747F/6720R (6S), was found to be the most effective in BSVstrain detection. After optimizing the detection conditions of 6S, we screened more than 200 of Musa spp. germplasm collected in Taiwan Banana Research Institute (TBRI), and about 11.35% of germplasm is infected by BSV. BSV strains, GF and GD, previously not reported in Taiwan were found in our survey. Some BSV positive samples (detected by RT-PCR with 6S primer pair) have been further selected for insect transmission. The result confirmed the availability of 6S primer pairs, and showed that 6S can be used to detect other BSV strains besides Mysore and OL. In addition, we also found the activation of integrated BSV in germplasm accession, including 119, 123, 222 and 232, during tissue culture propagation. The developed rapid detection method can help the management of BSV.
Subjects
Banana streak virus
EPRV
integrated form
episomal form
RT-PCR
Type
thesis
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