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三种鹅膏菌培养条件及所产肽类毒素的比较研究
引用本文:郭学武,汪国轮,龚建华,肖冬光.三种鹅膏菌培养条件及所产肽类毒素的比较研究[J].菌物学报,2008,27(5):745-756.
作者姓名:郭学武  汪国轮  龚建华  肖冬光
作者单位:1. 天津科技大学天津市工业微生物重点实验室,天津,300457
2. 北京生命科学研究所,北京,102206
3. 中国科学院微生物研究所,北京,100101
摘    要:以外生菌根菌鹅膏菌属三个种Amanita muscaria,A.pseudoporphyria和A.fritillaria为研究材料,以生长速率为评价指标,对其最适生长温度、pH值、光照、培养基、C及N源的利用等基本培养条件及所产肽类毒素进行了比较研究。研究结果表明,三种菌株最适生长温度有差异,A.pseudoporphyria和A.fritillaria的最适温度为28℃,A.muscaria的最适温度为22℃;A.muscaria菌丝体生长的pH值范围为5-7,另外两个菌株的pH值范围为3-6;24h光照、12h光暗交替和24h黑暗对鹅膏菌的生长速率影响不大;SPDM培养基和MMN培养基都适合三种菌株的生长,但对于A.muscaria来说PDM培养基更适合其生长。鹅膏菌能够利用比较广泛的C、N源,但三个种在利用的C、N源种类上有一定的差别。通过抑芽法实验和HPLC分析分别表明三种鹅膏菌所含肽类毒素在种类和含量上有所不同,但都对绿豆发芽有一定的抑制作用。A.pseudoporphyria和A.fritillaria菌丝体中α-amanitin的含量分别为35.56μg/gDCW(drycellweight细胞干重)和26.02μg/gDCW,不含有phalloidin和β-amanitin;A.muscaria菌丝体中没有检测到α-amanitin、β-amanitin和phalloidin。结果表明供试的三种鹅膏菌在基本培养条件及所产肽类毒素方面存在种水平上的差异。

关 键 词:毒蝇鹅膏  假褐云斑鹅膏  格纹鹅膏  生长速率  菌落特征  肽类毒素

Comparisons of culture conditions and peptide toxins in three Amanita species
Authors:GUO Xue-Wu  WANG Guo-Lun GONG Jian-Hua XIAO Dong-Guang
Institution:Tianjin University of Sciences & Technology, Tianjin Industrial Microbiology Key Lab, Tianjin 300457, China
Abstract:A comparative study was conducted to evaluate the effect of temperature,pH,light duration,C and N source uptake and culture media on growth rate,colony characteristics and peptide toxins in three Amanita species,Amanita muscaria,A. pseudoporphyria and A. fritillaria. The result indicated that the optimum growth temperature for A. muscaria,A. pseudoporphyria and A. fritillaria were 22℃,28℃,28℃,respectively. The pH range of mycelial growth was from 5 to 7 for A. muscaria and from 3 to 6 for A. pseudoporphyria and A. fritillaria. Light duration has little effect on the growth rate of these three species. SPDM (Potato 200g,Glucose 20g,KH2PO4 3.0g,(NH4)2HPO4 0.5g,CaCl2 0.05g,Wort (8°Be) 150mL,KNO3 0.1g,MgSO4·7H2O 0.5g,VB1 10mg,Distilled water 850mL pH 5.5) and MMN (CaCl2 0.05g,MgSO4 0.15g,NaCl 0.025g,FeCl3 (1%) 1.2mL,KH2PO4 0.5g,vitamin B1 100μg,(NH4)2HPO4 0.25g,Wort (8°Be) 150mL,Glucose 10g,Citric acid 0.2g,Distilled water 850mL pH 5.5) were the suitable media for the three species,while PDM (Potato 200g,Glucose 20g,K2HPO4 0.1g,Wort (8°Be) 150mL,Distilled water 1000mL pH 5.5) was better for A. muscaria. Amanitin contents in the mycelia of the three species were analyzed by HPLC and further confirmed by bud-inhibited assay. The results showed that the mycelia of A. pseudoporphyria and A. fritillaria contain α-amanitin but no β-amanitin and phalloidin. The contents of α-amanitin in mycelia of A. pseudoporphyria and A. fritillaria reached 35.56μg/g DCW (dry cell weight) and 26.02μg/g DCW,respectively. Amatoxin and phalloidin were undetected in A. muscaria. This study indicated that culture condition and peptide toxin production in three Amanita species were different.
Keywords:Amanita muscaria  Amanita pseudoporphyria  Amanita fritillaria  growth rate  colony characteristics  peptide toxins
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