Development of oligonucleotide array for simultaneous identification of multiple crucial forest fungal pathogens

• Main Authors: Po-Yao Shu, 許博堯
• Other Authors: Shean-Shong Tzean
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/83975390954320582414

碩士 === 國立臺灣大學 === 植物病理與微生物學研究所 === 97 === Plant fungal pathogens can cause various types of diseases in agronomical, horticultural and forestry plants, which can lead to widespread plant mortality and famine. Therefore, use of time-saving, accurate, and sensitive diagnosis methods coupled with appropriate and effective control measures, is essential in reducing economic loss and enhancing public well-being. In recent years, due to climate change, an outbreak of brown root disease among community greenery street trees, artificial forests, and windbreak plants has caused serious losses, but has also forced the public to take a serious view of fungal pathogens. Attempt to rapid and accurate diagnosis, totally 67 species of wood rot fungal pathogens were collected, including many crucial pathogens, i.e. Phellinus noxius, P. pini and P. werii, Armillaria mellea, Armillaria ostyae, and several Ganoderma species. These pathogens, which have also been reported in the United States, Canada, New Zealand, Australia and Europe, have infected many coniferous and broadleaf forests, resulting in the growth decline; defoliating, early flowering, decay, wilting, lodging and other symptoms, and become a great limiting factor and a potentially serious threat in forest management. The oligonucleotide microchip developed for simultaneous rapid identification of the 67 crucial forest pathogens was based on the DIG or biotin-labeled specific probes derived forms ribosomal DNA gens (ITS1-5.8S-ITS2) by using reverse-dot hybridization. These chips can precisely and accurately identify and diagnose many Armillaria, Antrodia, Antrodiella, Heterobasidium, and Phellinus species, including A. mellea, A. ostoyae, H. annosum, P. noxius, and P. weirii, etc. with a sensitivity of 1 pg DNA/μl on nylon membrane chip, and 100 fg DNA/μl on plastic chip, respectively. And the verification and identification of forest Phellinus pathogens in authentic samples or voucher specimens can be accomplished within 7 hrs. The chip can be applied in inspection and quarantine, also for healthy free, seedling and nursery certification, and in ecology and silviculture management, as well.