Application of polymerase chain reaction to detect Tomato leaf curl Taiwan virus (ToLCTWV) in Bemisia argentifolii feeding on virus-infected tomato

• Main Authors: Wu, Jia-Chien, 吳佳倩
• Other Authors: Chen, Tsang-Hai
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/61671004561016850611

碩士 === 國立屏東科技大學 === 植物保護系 === 94 === Tomato leaf curl Taiwan virus (ToLCTWV) is a serious infected virus of tomato in Taiwan, which causing symptoms of leaf yellowing, leaf-curling, stunting, shortening of internodes and severe reduction of leaf size on tomato. ToLCTWV is a whitefly (Bemisia argentifolii) transmitted geminivirus (Geminiviridae, Begomovirus). In this study, polymerase chain was used to detect ToLCTWV in infected whitefly. Feeding on infected tomato, ToLCTWV can be detected in female whitefly in 5 minutes and is 20 min in male. Vector feeding 24 hours on ToLCTWV-infected tomato was analyzed, amount of 5 pooled crawlers, 2 pooled nymphs, and one single pupa of whitefly shows ToLCTWV can be detected available. However, no ToLCTWV could be detected in eggs production of infected whitefly on watermelon. Detection of ToLCTWV containable study reveals the infected proportion in female was 85.8% and is higher than 70.3% in male. Samples from field collection also indicated the infectious female (33.8%) was higher than that in male (18.7%). DNA template of ToLCTWV-infected whitefly was diluted to see the detected sensitivity and the results were 2-10 in female and 2-5 in male. Analysis shown that ToLCTWV can be detected in whitefly in 30 days. Two variants each from eastern and western Taiwan were cotransmitted to tomato by whiteflies, and PCR-RFLP was used to detect the ToLCTWV variants in feeding whitefly and its feeding site of tomato. Results shown that ToLCTWV can be detected in feeding whitefly and its feeding site of plants, however, the pattern of eastern ToLCTWV were detected more than that of western pattern. In the mixed infection analysis, 75 detections were contained ToLCTWV, 32 of them revealed the coexisted ToLCTWV pattern between feeding whitefly and its feeding site, whereas 10 revealed the reverse ToLCTWV pattern between whitefly and plant. Sixteen and seventeen can be detected only in whitefly and plant, respectively. Our studies provided a simple and rapid ToLCTWV identified method in whitefly, which will apply to the ToLCTWV influence of field vector and tomato.