碩士 / 國立臺灣大學 / 植物病理學研究所 / 89 / 英文摘要
The viral disease of calla lily is one of the limiting factors for cultivation of the crops in Taiwan. Dasheen mosaic virus (DsMV) and Zantedeschia mosaic virus (ZaMV) which are members of the genus Potyvirus, are the most important viruses in cultivated calla lilies. In this study, we developed three kinds of detection techniques (ELISA, dot-blot hybridization and RT-PCR) for DsMV and ZaMV. Detection of DsMV and ZaMV by I-ELISA using our anti-DsMV and anti-ZaMV antiserum is a specific, sensitive and convenient method. In molecular hybridization methods, six specific DNA probes were designed which located in 3’NCR and the coat protein region of DsMV and ZaMV, respectively. The results of hybridization indicated that all the probes were highly specific and sensitive to detect the corresponding viral RNAs in crude sap extracts and purified total RNAs. In order to develop more sensitive and convenient detection methods for DsMV and ZaMV, we developed a new technique of dip-capture RT-PCR (DC-RT-PCR) which was compared with immuno-capture-RT-PCR (IC-RT-PCR) and direct-binding RT-PCR (DB-RT-PCR). The results showed that IC-RT-PCR was the best, and DC-RT-PCR was more sensitive than DB-RT-PCR. The method of DC-RT-PCR dose not need specific antibody as IC-RT-PCR, and thus is convenient and useful in detection of other viruses or new viruses. When compared the sensitivity of I-ELISA, specific DNA probe and RT-PCR for detecting DsMV and ZaMV in crude sap extracts, the results indicated that IC-RT-PCR was about 125 times more sensitive than I-ELISA, and specific DNA probe was about 4-20 times more sensitive than I-ELISA.
In order to find out the effect of ZaMV on growth and rhizome formation of calla lilies, 1st- and 2nd-generation rhizomes were used for evaluation. The experimental results indicated that ZaMV not only induced symptoms of mosaic and green island on leaves of calla lily, but also significantly reduced the weight of harvested rhizomes. In addition, the flower stems of infected plants were shortened, and the flower color and shape were abnormal. When comparing the concentrations of ZaMV as refracted in ELISA values using the antiserum to recombinant coat protein in different parts of calla lily plant, the data showed that the virion concentrations were uneven among the whole plant: the leaves had the highest, root and rhizome had the lowest levels of ZaMV.
| Identifer | oai:union.ndltd.org:TW/089NTU00364012 |
| Date | January 2001 |
| Creators | Wei-Zhou Huang, 黃偉洲 |
| Contributors | Ya-Chun Chang, 張雅君 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 83 |
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