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Title: | 臺灣產南方靈芝複合種之分類學研究 Taxonomic study of Ganoderma australe complex in Taiwan |
Authors: | Zeng-Yung Yeh 葉增勇 |
Publication Year : | 1990 |
Degree: | 碩士 |
Abstract: | 自臺灣各地區闊葉樹採獲南方靈芝Ganoderma australe (Fr.) Pat.之標本十四個。並自菌體組織獲得純培養。經雜木屑人工子實體之栽培,成功分離出菌株TAI-01、TAI-02、TAI-03、TAI-04之單孢系。由單孢系自身交配,顯示本種交配系統?異絲型、多因數四極性。 本種在形態上極易與樹舌靈芝G. applanatum (Pers. ex S. F. Gray) Pat.發生混淆。本研究參考歐美標本,及購自美國、荷蘭菌種中心樹舌靈芝的純培養菌株,就形態特徵,純培養性狀,並以南方靈芝TAI-01單核體菌株與供試雙核體菌株進行單—雙核交配試驗,三?類的薄層色層分析(TLC),以及DNA被限制酵素切割圖譜(RFLP),進行南方靈芝複合種分類學的探討。結果顯示,台灣產的南方靈芝尚可區分二群,第一群最適生長溫度範圍是28-32℃,平均生長速率?每日6.2mm,單一雙核交配成親和反應。第二群最適生長溫度範圍是24-28℃,平均生長速率?每日4.0mm,單一雙核交配成不親和反應,三?類TLC掃描儀分析光譜比第一群於Rf值0.43位置多一個波長254nm吸光波峰,DNA被限制酵素Bgl Ⅱ,Pvu Ⅱ切割圖譜與第一群也呈差異性,推斷台灣的南方靈芝己存在有二個「互不孕性群」。 南方靈芝(含兩群)明顯與樹舌靈芝有差異,單一雙核交配成不親和反應,前者之孢子大小?7.6-11.2×4.8-7.2μm,三?類TLC掃描儀分析光譜在Rf值0.26,0.37位置有波長254nm吸光波峰。後者之孢子大小?5.6-9.2×4.0-7.2μm,生長最適溫度?20-24,或24-28℃,平均生長速率每日小於3.8mm,三?類TLC掃描儀分析光譜則在Rf值0.30, 0.40, 0.46位置有波長254nm吸光波峰。兩種之DNA被限制酵素EcoR I,Bgl II,及Pvu II切割圖譜明顯呈現差異。 來自美國菌種中心ATCC 32585樹舌靈芝菌株,分離自印度亞熱帶地區的闊葉樹,能和台灣產的南方靈芝TAI-01單孢系菌株成親和反應,並且最適生長溫度及生長速率均類似於第一群,人工子實體三?類TLC掃描儀分析光譜,10% H2SO4呈色點位置,以及DNA被三種限制酵素切割之圖譜均明顯相同。由以上結果顯示ATCC 32585樹舌靈芝和台灣南方靈芝同?一種,其學名應更正?G. australe (Fr.) Pat.。 另檢查澤田兼吉於日據時代在台灣所採集的G. applanatum標本,重新檢視其形態特徵,以及依照近代菌類學家對樹舌靈芝的地理分佈侷限在北溫帶的觀念,澤田的G. applanatum之標本應更正?G. australe (Fr.) Pat.較?合理。 Fresh basidiocarps of Ganoderma. subgen. Elfvingia (Karst.) Imazeki were collected from hardwoods at different sites in Taiwan, and pure cultures were obtained. Based on the morphology of basidiocarps, they were identified as Ganoderma australe (Fr.) Pat. Monocaryotic cultures of this species were obtained from basidiospores of artificially cultivated fruit-bodies of isolates TAI- 01, TAI-02, TAI-03, and TAI-04 of G. australe. Mating system of this species proved to be heterothallic and tetrapolar multiallelic. As the results of different approaches of confirmation, i.e., comparison with type specimen and some authentic reference specimens from Europe and America, cultural identification and growth temperature kinetics, compatibility by di-mon mating reactions, patterns of triterpenoids by thin layer chromatography (TLC), the restriction fragment length polymorphism (RFLP) of ribosomal DNA (rDNA) repeating unit, the isolates of G. australe in Taiwan could be separated into two groups. Isolates of Group 1 had an average growth rate of 6.2 mm/day at its optimum temperature range of 28-32℃. The di-mon matings within this group were compatible. Isolates of Group 2 had an average growth rate of 4.0 mm/day at its optimum temperature range of 24-28℃. The di-mon matings with monocaryons of group 1 were incompatible. The absorbance spectrum of 254 nm by TLC-scanner showed a little difference at Rf 0.43. The RFLP of rDNA repeating unit by restriction enzyme EcoR Ⅰ and Bgl Ⅱ are slightly different. Therefore, it is concluded that up to date, there are two intersterility group of G. australe in Taiwan. Morphologically, G. australe and G. applanatum (Pers. ex S. F. Gray) Pat. (=G. lipsiense (Batsch) Atk.) are very similar and difficult to distinguish. The di-mon matings by monocaryotic isolates of G. australe TAI- 01 with cultures of G. applantum proved to be incompatible. G. australe had a larger spore size, namely, 7.6-11.2×4.8-7.2 μm. The absorbance spectrum of 254 nm by TLC-scanner for triterpenoids showed two different peaks at Rf 0.26, and 0.37. While G. applanatum had a smaller spore size, namely, 5.6-9.2×4.0-7.2 μm. The optimum growth temperature ranged from 20 to 24 or from 24 to 28℃. The absorbance spectrum of 254 nm by TLC-scanner for triterpenoids showed three different peaks at Rf 0.30, 0.40, 0.46. The RFLP of rDNA repeating unit of two species by restriction enzyme EcoR Ⅰ, Bgl Ⅱ and Pvu Ⅱ are also different. Four representative monocaryotic isolates of G. australe TAI-01 were mated with cultures of G. applanatum obtained from ATCC or CBS. The isolate ATCC 32585, which was isolated from a tropical hardwood in India, was compatible with monokaryotic isolates of G. australe TAI-01, and had a similar growth temperature requirement of Group 1 of G. australe from Taiwan. They had the same patterns of triterpenoids by TLC-scanner, and the same RFLP of rDNA repeating unit. It suggests that G. applanatum ATCC 32585 is identical to G. australe from Taiwan. The scientific name, therefore, should be revised as G. australe (Fr.) Pat. The specimens of G. applanatum collected by K. Sawada during 1929-1941 in Taiwan are also indentical to the Group 1 of G. australe. According to the recent species concept for G. applanatum, the scientific name of G. applanatum sensu Sawada from Taiwan should be revised as G. australe. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75738 |
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Appears in Collections: | 植物科學研究所 |
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