Isolation and Identification of Gomphus purpuraceus
Abstract：Gomphus purpuraceus is an important rare mycorrhizal edible fungus. It is very difficult to identify the isolates by fruiting. The tissue isolation from different parts of different developmental phases of fruiting body and mycorrhiza was made with 5 media. The results showed that mycelia from fruiting body grew very slowly and thickly, mycelia from mycorrhiza and rhizomorph grew quickly and thinly. Out of 5 media, medium D was optimal medium for tissue isolation. The isolates from different resources were appraised through DNA fingerprinting in comparison with that of dry basidiocarps of G.purpuraceus, and RAPD patterns were sharp and stable by 8 random primers. The results suggested that isolates from fruiting body had the same DNA fingerprinting patterns as that of the basidiocarps tissues (such as pileus and stipe), whose similarity coeffecient was 0.884. However, the RAPD patterns of the isolates from mycorrhiza and rhizomorph were different from the others (including the basidiocarps tissues). These results further revealed that some isolates from fruiting body were real isolates of G. purpuraceus and the isolates from mycorrhiza and rhizomorph were not real isolates of the G.purpuraceus.
Keywords：Gomphus purpuraceus; tissue isolation; DNA identification; RAPD analysis
紫陀螺菌菌株的分离及鉴定
谭爱华 马富英 罗信昌 黄亚东
摘 要：采用5种培养基配方,对紫陀螺菌子实体不同生长期的不同部位及菌根、菌索进行组织分离,结果从子实体得到的菌丝生长缓慢,菌丝致密,从菌根和菌索得到的菌丝生长快,絮状不致密.以紫陀螺菌干子实体DNA为参照样品,对得到的相关分离纯培养物进行亲菌鉴定.供试的26条随机引物中,有8条对所有供试样品可扩增出清晰、稳定的DNA指纹图谱,且从子实体分离得到的菌丝体DNA指纹与干子实体的基本一致.聚类分析结果表明,从子实体不同生长期得到的2个菌株相似系数0.979,它们与干子实体间的相似系数为0.884,从菌根和菌索得到的分离物与干子实体间的相似系数为0.389.结果表明,从子实体分离得到的2个菌株与对照物相似度最高,可以认定是紫陀螺菌的纯培养物.
关键词：紫陀螺菌; 组织分离; 亲菌鉴定; RAPD分析
CLC Number：S646
Foundation Item：supported by Committee of Science and Technology of Huibei Province and State Key Laboratory of Agricultural Microbiology
Author Resume：Tan Aihua, female, born in 1967, master. Address:Yichang Vocational and Technical College.罗信昌,Corresponding author. E-mail: xinchangluo@mail.hzau.edu.cn
Author Unit：谭爱华（华中农业大学,农业微生物国家重点实验室,武汉,430070;宜昌职业技术学院,宜昌,443003）
马富英（华中农业大学,农业微生物国家重点实验室,武汉,430070）
罗信昌（华中农业大学,农业微生物国家重点实验室,武汉,430070）
黄亚东（华中农业大学,农业微生物国家重点实验室,武汉,430070）
References：
［1］Mao X L. Macrofungi of China. Zhengzhou:Henan Science and Technology Press,2000.1～388
［2］Tan A H, Liu F Z. A primary study on ecology of Gomphus purpuraceus.Edible Fungi of China, 2002, 21(1):6
［3］Tan A H, Liu F Z,Zhang S G. Analysis of nutritive components of Gomphus purpuraceu.Acta Edulis Fungi, 2001, 8(4):15～18
［4］Luo X C. DNA fingerprinting identification of isolate in mushrooms.Edible Fungi of China, 2001, 20(3): 3～4
［5］Ma F Y, Luo X C. PCR-based restriction analysis of internal transcribed spacers of nuclear ribosomal DNA in the genus Pleurotus.Mycosystema, 2002, 21(3): 356～362
华中农业大学学报
JOURNAL OF HUAZHONG AGRICULTURAL UNIVERSITY
2004 Vol.23 No.1
