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標題: Fusarium solani及F. ventricosum菌株間可溶性蛋白與同功異構酵素電 泳圖譜之比較
Comparison the soluble protein and isozyme electrophoresis patterns between Fusarium solani and F. ventricosum
作者: 曾春僑
Tzeng, chun-chiao
關鍵字: Fusarium
出版社: 植物病理學系
摘要: Fusarium 屬真菌之分類主要是依據其胞子叢、大胞子及小胞子的形態而 來,然而, 其中卻存在著許多極小之差異,以致於在較細微之鑑定上難 以達到理想之效果。近幾年來,為解決形態判定上之難題,利用真菌細胞 體內抽取之各種物質進行電泳,比較彼此間之差異性的方法,已廣泛的被 應用於一些較具爭議性之真菌分類問題上。 在 Fusarium 屬中,F. sloani及 F. ventricosum 之形態構造極為相似,兩者除產胞方式有些許 不同外,其餘並無大之差異性存在,因此,這兩種真菌是否應分開或合併 成一種目前仍具許多之爭議性。本實驗利用聚丙烯醯胺凝膠電泳比較其電 泳圖譜之差異以及透過 UPGMA 分析法比較兩者間之相似係數。在所有之 分離株中均可發現到三條主要蛋白條帶,80, 60 及 50 kd 之存在;除此 之外,所有之分離株不論在條帶之數目及濃度上均有所差異。在孔雀藍染 色以及銀染上相似度最高值分別為 0.97 及 0.94,最低值分別為 0.59 及 0.56。在二十二種受測試的同功異構酵素中,十六種之結果完全相同 ,惟其中的十五種出現不同濃度之差異。所測試到的二十個等位基因中有 十四個完全相同;在反應相異的基因座上分別可觀察到一至三個的等位基 因之存在。 在一些分離株的核酸電泳中,可發現到質體 DNA 之存在。在 所測試的限制酵素中並無法找到一個適合所有菌株之內切酵素以作為限制 片段圖譜分析用。由我們所得知結果看來,此兩種真菌間存在著相當程度 之相似性,因此建議將兩種真菌併稱為 F. solani。 Taxonomy of the genus Fusarium is based primarily on morphology of theirsporodochia, microconidia and macroconidia, however, many of their morphologyis hard to identify because of the small differences between these species.Recently, gel electrophoresis extracted from various fungal components hasreceived considerable attention as a possible aid in resolving some of theproblems inherent in fungal taxonomy. Two Fusarium species, F. solani and F.ventricosum, their morphological structure were correspondly only in thedifferences of conidiophore structure. Some arguments were raised whether thetwo species are combined or not. These two Fusarium species were compared byusing SDS-polyacrylamide gel electrophoresis of total soluble proteins and thesimilarity coefficients were obtained through the unweight pair-group methodwith arithmetic average (UPGMA). Three major bands, 80, 60 and 50 kd, wereobserved in all the isolates. And the differences of total soluble proteinswere not only in their concentration but also in the location that appeared inthe gel. The highest similarity value within the Coomassie blue stainingcluster was 0.97 and silver slat staining cluster was 0.94. The lowestsimilarity values within the Coomassie blue stain cluster was 0.59 and silversalt stain cluster 0.56. The activity of twenty-two enzymes were screened inoptimum buffer system. The electrophomorphs of all isolates were the same insixteen isozyme reaction and fifteen enzymes were showed strong activity andacceptable resolution. Fourteen of the 20 alleles exhibited completehomogeneity among all isolates surveyed and six of the alleles were variableand revealed one to three putative alleles per locus. The plasmid were existedin some isolates and non of suitable restriction enzyme were found to digestthe DNA of these isolates for restriction fragment length polymorphisms. Theresults suggested that these isolates of the two Fusarium species maybecombined together and named as F. solani due to their high degree ofsimilarity in soluble proteins and isozymes.
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