| Summary: | 碩士 === 國立嘉義大學 === 生物科技研究所 === 92 === Abstract
Several fungal pathogens, including Fusarium oxysporum f. sp. lilii, Phytophthora parasitica, Ph. cinnamomi, Sclerotium rolfsii, Rhizotonia solani, Pythium sp., Botrytis elliptica could unpredictably affect the quantity and quality of lily. The identification and detection of the pathogenic fungi become important for plant health inspection and quarantine control of lily. The objective of the present study is to develop a specific and precise molecular diagnosis system for the detection of pathogenic fungi of lily bulbs. The internal transcribed spacer (ITS) regions of the rRNA were amplified, cloned, and sequenced. Based on the sequences of ITS regions, five pairs of specific PCR primers (FRs-f/FRs-r, FSr-f/FSr-r, FPp-f/FPp-r, FPy-f/FPy-r, FFo-f/FFo-r) were designed for the identification and detection for tested pathogenic fungi. Moreover, a multiplex PCR was performed to simultaneously amplify predicted-size fragments as 350 bp, 386 bp, 467 bp, 559 bp and 673 bp from the DNA of F. oxysporum f. sp. lilii, R. solani, S. rolfsii, Pythium sp., and Ph. parasitica, respectively. On the other hand, using the 18S and ITS fragment as a probe, dot-blot hybridization and biochip (Lily chip) were able to successfully identify and detect fungal pathogens of lily, which were classed as different genus or species. Furthermore, fungal pathogens were also successfully identified and detected by Lily chip from the DNA of inoculated lily bulb scale tissue. The results in the present study showed that multiplex PCR and Lily chip could provide a rapid, simple, and reliable alternative to conventional methods for detection of fungal pathogens of lily. The potential applications for plant pathogens of different crops were also discussed.
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