Development of a 4-in-1 multiplex reverse-transcription polymerase chain reaction (RT-PCR) detection method and survey of four banana viruses in Taiwan

• Main Authors: Po-Chen Wu, 吳柏辰
• Other Authors: 洪挺軒
• Format: Others
• Language: zh-TW
• Published: 2019
• Online Access: http://ndltd.ncl.edu.tw/handle/r67m2j

碩士 === 國立臺灣大學 === 植物病理與微生物學研究所 === 107 === Banana is one of the most important fruits in the world and ranks the third of export crop in Taiwan. In addition to Panama disease caused by Fusarium oxysporum f.sp. cubense (Foc), the banana plants also face with the threat from different viruses such as cucumber mosaic virus (CMV), banana bunchy top virus (BBTV), banana streak virus (BSV) and banana bract mosaic virus (BBrMV). These four viruses cause various symptoms including stunting, foliar chlorosis or mottling, poor fruit production and decline. CMV and BBTV are the most widespread and serious in Taiwan among four viruses, but BSV and BBrMV have potential impact for several new promoting banana and plantain cultivars. The main propagative method of banana is the tissue culture in Taiwan. To avoid the banana viruses transmitted by vegetative propagation of tissue culture, Bureau of Animal and Plant Health Inspection and Quarantine has established “the certificate instruction of banana diseases for banana seedlings” to promote the pathogen-free nursery system of banana. This system needs accurate and efficient detection methods for those four viruses to verify the seedlings without virus infection. However, the whole procedure of complete virus detection is complicate and time-consuming when the four virus detections have to be conducted individually. This study is dedicated to develop a 4-in-1 multiplex RT-PCR (reverse transcription – polymerase chain reaction) to simultaneously detect four viruses in one assay for saving time and budget. Most banana-producing countries have the same problem caused by these four viruses. Thus, this 4-in-1 virus detection has the internationally commercialized potential for the quarantine and inspection. The developed multiplex RT-PCR was applied to investigate the distribution and disease-severity of four banana viruses in Taiwan. The banana samples collected from different banana-cultivated areas such as Pingtung, Kaohsiung, Tainan, Chiayi, Yunlin, Nantou, Taitung and Hualien showed that the infectious percentages of CMV and BBTV are 7.7 % and 13.9 %, respectively. BBrMV was detected only in few samples (Saba cultivar) collected from Taiwan Banana Research Institute (TBRI), one from Pingtung and one from Yunlin. The BSV-infected sample has not yet been found in the field. This devised multiplex RT-PCR technique could be successfully and efficiently applied in the investigations of four banana viruses in different banana cultivars.