|
The fruiting body of the Ganoderma lucidum has always been greatly valued
in traditional Chinese medioine and in recent years some compounds have
been found to have antitimor activities in the metabolite. However world
wide it is merely classified as a wood decaying fungi that causes white
rots of a wide variety of trees and timber. Even in many scientific papers
G. lucidum is mainly deseribed as phytopathogenic fungi.
THe taxonomy system of the Ganoderma, used for the last 100 years is based
on the morphology of the fruiting body. At present , a large variety of
Ganoderma research is being conducted such as chemical
composition,biological activity, artificial cultivation, cross-breeding,
in connection with plant protection etc., therefore a much clearer
identifieation system ofr cultures needs to be established. New parameters
are needed to differentiate between Ganoderma interspeeies and
intraspooies.
The parameters described in this thesis are as follows:
(1) macroscopic characteristics of the colony and fruiting body.
(2) microscopic characteristios of the hyphae and basidiospores.
(3) growth rate and optimum temperature.
(4) extracellular hydrolytic enzymes assay by API-ZYM kets.
(5) electrophoretic patterns of extracellular laccase isozywes.
(6) di-mon mating and incompatibility test.
(7) restrietion fragment length polymorphisms in total DNA.
Using the above parameters, the following species found in Taiwan were
identified : G. applanatum (Pers.) Pat., G. formosanum Chang & Chen, G.
fornicatum (Fr.) Pat. G. lucidum (W. Curt.: Fr.) Karst., G. microsporum
Hseu, G. neo-japonicum Imaz., G. tropicum (Jungh.) Bres., G. tsugae Murr.
Also the relationship between 10 different isolates of G. lucidum were
identified.
The above parameters can be used for different levels of the taxonomy
system. For genus identification, the morphology of the fruiting body and
basidiospore. For species identification, the most important parameters
are the restriction fragment length polymorphisms, di-mon mating and
incompatibiligy test. Microscopy of hyphae espedially strains with or
without chlamydospore, and the morphology of the colony can be used as
supporting data. For variety identification, the RFLPs, extracellular
hydrolytic enzymes assay by API-ZYM kets, laccase isozyme patterns, growth
rate of colony and optimum temperature are used. These parameters can be
used to clearly identify the relationship between isolates of the same
species.
When the morphology of different species is very similar, such as that of
the G. Lucidum and G. tsugae, two new parameters must be used for more
precise identification-the compatibility test and RFLPs results.
靈芝自古為我國滋補強壯的珍貴藥材，近年來由靈芝代謝物中發現具有抗腫瘤等生理
活性成分，而成為廣受重視的藥用真菌。但靈芝屬也是廣泛分佈的白腐型真菌，文獻
記載其中的代表種G.lucidum（W. Curt.: Fr. ）Karst. 亦為植物病原性真菌。由於
靈芝屬的分類仍以子實體的形態特徵為主，對於靈芝屬的相關研究中，無論就植病防
治、生理、藥理、化學成分，人工栽培及育種等項目皆需有明確的菌株鑑定系統，做
為人工培養菌絲體時異種間和同種內菌株類緣關係判斷的指標。
本論文中利⑴人工培養時菌落、子實體的巨觀特性，⑵菌絲體和擔孢子的微觀特性，
⑶菌落生長速率和最適生長溫度，⑷API-ZYM 分析細胞外酵素活性，⑸細胞外潻氧化
同功酵素電泳分析圖譜，⑹單一雙核體與單一單核體菌株間的親和性反應，⑺以
Random prtoe set GLH-2標記的Hind Ⅲ 限制水解基因片段多型性圖譜等結果為分
類指標時，可以明確區別自本省採集分離的G.applanatum（Pers.）Pat. G.
formosanum Chang & Chen, G. fornicatum （Fr.）Pat., G. lucidum, G.
neo-japonicum Imaz., G. tropicum （Jungh.）Bres., G. tsugae Murr. 等菌株外
，並可探討10株不同來源的G. lucidum菌株間的類緣關係。
在此分類系統中所用各項指標適用於不同的分類階層。子實體和擔孢子的形態特性為
建立靈芝屬時的依據，故可用於異屬間之區別。對於靈芝屬內異種間菌株種源關係的
判斷時，以限制水解基因片段多型性圖譜和菌株間的親和性反應結果為主要指標，
而以菌落形態和菌絲體中厚膜孢子的有無等為輔助的參考指標。在同種內的不同品系
菌株間關係，可由限制水解基因片段多型性圖譜、API-ZYM 分析胞外酵素活性結果
、漆氧化同功酵素電泳圖譜、菌落生長速率和最適生長溫度等特性做為同種菌株間
類緣關係鑑別的參考。
利用此系統的分類結果顯示，今後對於子實體外型相似的菌種如G. lucidum和G.
tsuqae等的分類鑒定時，必須配合菌絲體間親和性反應與限制水解基因片段的多型
性圖譜等結果加以確認。
|