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Thesis Advisor王晓军 ; 付振艳
Degree Grantor中国科学院研究生院
Place of Conferral北京
Degree Discipline有机化学
Keyword野生菌 翘鳞伞(翘鳞环锈伞) 菌种鉴定 菌丝体培养 驯化栽培
Abstract翘鳞伞(Pholiota squarrosa)为我国民间常用的食(药)用真菌。在本文中,用组织分离法分离从新疆玛纳斯林场采拾的翘鳞伞菌种,采用rDNA-ITS序列分析的方法鉴定菌种,并对菌丝体生长及人工栽培进行研究。 主要试验结果如下: 1.采用组织分离法从(疑似)翘鳞伞子实体上进行菌种分离,接种在PDA培养基,分离得到纯培养菌丝体。 2.采用rDNA-ITS序列分析的方法对(疑似)翘鳞伞菌株进行了分子鉴定。利用ITS扩增通用引物扩增了该菌株的rDNA-ITS序列,经TA克隆后将转化菌落测序,经鉴定为翘鳞环锈伞。 3.采用单因素实验筛选出了适宜菌丝生长的最适生长温度为23℃,最适PH为5。最佳碳源为麦芽糖、氮源为酵母粉。 4.对发酵培养基进行了优化:利用Plackett-Burman方法获得影响翘鳞伞生长的主要因素为K2HPO4、麦芽糖、酵母粉,通过最陡爬坡实验和正交实验确定了翘鳞伞液体生长的最优培养基配方为麦芽糖 35 g/L、酵母粉 1.5 g/L、K2HPO4 0.5 g/L、KH2PO4 0.5 g/L、MgSO4 0.5 g/L。 5.成功驯化出翘鳞伞子实体。栽培研究表明:翘鳞伞可在以木屑、棉籽壳为主料、麸皮、玉米面、石膏、磷酸二氢钾为辅料的罐头瓶栽培料上正常生长发育,一年两次栽培,接种到子实体采收约120-130天,生物学效率达53%。 关键词:野生菌,翘鳞伞(翘鳞环锈伞),菌种鉴定,菌丝体培养,驯化栽培
Other AbstractPholiota squarrosa is commonly used as edible and medical fungi. In this paper, using tissue separation method to separate mycelium of Pholiota squarrosa from Xinjiang Manas Forest.Using rDNA-ITS sequence analysis to make molecular identification.And we also made studies on mycelium cultivation and domestication of Pholiota squarrosa. The main contents include the following: 1.Mycelia were separated using tissue separation method .The tissues were inoculated to PDA medium and the pure culture mycelium was isolated. 2.Molecular identification of Pholiota squarrosa was carried out using rDNA-ITS sequence analysis. ITS sequence were amplified by universal primers. The ITS fragment was cloned to T vector, and then sequenced.The results showed that the wild mushroom is Pholiota squarrosa. 3.One-factor experimental design was used to determine that the optimum temperature was 23 ℃;The optimum pH was 5; the optimum C-source for its mycelial growth was maltose and the optimum N-source was yeast power. 4.Fermentation medium optimization was studied:Plackett-Burman design was used to evaluate the main three factors in the medium,including K2HPO4 ,maltose and yeast power. Steepest ascent path and Orthogonal experimental design were used to determine the optimal conditions:maltose 35 g/l,yeast power 1.5 g/l,K2HPO4 0.5 g/l,KH2PO4 0.5 g/l,MgSO4 0.5 g/l. 5.Artificial cultivation was made successfully.Fruiting bodies was cultivated on a substrate consisiting of 60 % sawdust,20 % cottonseed hulls,15 %wheat bran and 5% corn meal,and supplemented with 1 % gypsum and 0.2 % KH2PO4.The recorded biological efficiency was 53 %,and the duration of the cultivation cycle was 120-130 days.
Document Type学位论文
Recommended Citation
GB/T 7714
吴金平. 翘鳞伞菌丝体培养及人工栽培研究[D]. 北京. 中国科学院研究生院,2012.
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