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蟲生真菌Nomuraea viridulus之生物學探討
Biological Characteristics of the Entomopathogenic Fungus, Nomuraea viridulus
摘要
本論文探討一種蟲生真菌Nomuraea viridulus之生物特性，本菌在溫度15~35℃範圍內皆可生長，最適生長條件為25℃下以SMA+Y(Sabouraud''''s maltose agar with yeast extract)培養基培養，其產孢量及發芽率為最高，液體震盪培養以pH7其菌絲乾重為最重，碳素源中以麥芽糖(maltose)其產孢量為最高、氮素源以蛋白腖(peptone)其產孢量為最高。於25℃下培養可同時產生蛋白酶(protease)、脂解酶(lipase)、幾丁質酶(chitinase)及澱粉酶(amylase)等酵素；另以API ZYM系統檢測胞外酵素活性，結果顯示並未偵測到脂肪分解酶(lipase (C14))、胰蛋白酶(trypsin)、胰凝乳蛋白酶(chymotrypsin)、β-葡萄糖苷酸酶(β-glucuronidase)、α-葡萄糖苷酶(α-glucosidase)及α-岩藻糖苷酶(α-fucosidase)，其他13種酶皆有活性呈現，表示本菌能分泌多種不同的胞外酵素。本菌經殺蟲劑及殺菌劑處理18天後，結果顯示殺菌劑免賴得(benomyl) 50% WP、腈硫醌(dithianon)22.7%SC、撲克拉錳(prochloraz Mn) 50% WP及多寧(dodine) 65% WP皆會抑制其生長；十一種殺蟲劑中僅加保利(carbaryl) 85% WP與培丹(cartap) 50% WP其毒性極強會抑制本菌生長，納乃得(methomyl) 90% WP、益達胺(imidacloprid) 9.6% SL、芬普尼(fipronil) 4.95% FP及窄域油(narrow range oil)99% EC則會降低產孢量，阿巴汀(abamectin) 2% EC、加保扶(carbofuran) 40.64% SC、賽洛寧(cyhalothrin) 2.8% EC、丁基加保扶(carbosulfan) 40% WP及大滅松(dimethoate) 44% EC等殺蟲劑加入仍有2×10^6 conidia/mL以上產孢量。以十一種昆蟲測試致病性之結果顯示僅有紅脈熊蟬(Crytotympana atrata)以爬行於真菌固體培養基的方式接種才受感染，感染率25%，顯示本菌的寄主範圍相當狹小。
並列摘要
This study investigated the biological characteristics of entomopathogenic fungus, Nomuraea viridulus. The mycelial growth of N. viridulus range from 15 to 35℃ and the optimum temperature for sporulation and germination was found at 25℃. The optimum growth of this fungus was when it was cultured on SMA+Y medium; at pH 7, with maltose as the carbon source, and peptone as the nitrogen source, respectively. Hydrolyte enzymes such as protease, lipase, chitinase and amylase, were present when tested with a specific culture media at 25℃. However, the activities of lipase (C14), trypsin, chymotrypsin, β-glucuronidase, α-glucosidase and α-fucosidase were not detected using the API ZYM assay. Nomuraea viridulus did not grow on media containing fungicides such as 50% benomyl WP, 22.7% dithianon SC, 50% prochloraz Mn WP or 65% Dodine WP, or those containing insecticides such as 85% carbaryl WP or 50% cartap WP. However, when inoculated with more than 2×10^6 conidia/mL, the fungus appeared on media containing the insecticides, e.g., 2% abamectin EC, 40.64% carbofuran SC, 2.8% cyhalothrin EC, 40% carbosulfan WP or 44% dimethoate EC, at 12 days after inoculation. When cultured on media containing 90% methomyl WP, 9.6% imidacloprid SL, 4.95% fipronil FP, or 99% narrow range oil EC resulted in a poor production of conidia. Only one cicada, Cryptotympana atrata, among 11 insects assayed was infected with N. viridulus causing ca. 25% mortality, indicating that N. viridulus has a narrow host range.