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

紫陀螺菌菌株的分離及鑒定

譚愛華　 馬富英　 羅信昌　 黃亞東　

摘　要：采用5種培養(yǎng)基配方,對(duì)紫陀螺菌子實(shí)體不同生長期的不同部位及菌根、菌索進(jìn)行組織分離,結(jié)果從子實(shí)體得到的菌絲生長緩慢,菌絲致密,從菌根和菌索得到的菌絲生長快,絮狀不致密.以紫陀螺菌干子實(shí)體DNA為參照樣品,對(duì)得到的相關(guān)分離純培養(yǎng)物進(jìn)行親菌鑒定.供試的26條隨機(jī)引物中,有8條對(duì)所有供試樣品可擴(kuò)增出清晰、穩(wěn)定的DNA指紋圖譜,且從子實(shí)體分離得到的菌絲體DNA指紋與干子實(shí)體的基本一致.聚類分析結(jié)果表明,從子實(shí)體不同生長期得到的2個(gè)菌株相似系數(shù)0.979,它們與干子實(shí)體間的相似系數(shù)為0.884,從菌根和菌索得到的分離物與干子實(shí)體間的相似系數(shù)為0.389.結(jié)果表明,從子實(shí)體分離得到的2個(gè)菌株與對(duì)照物相似度最高,可以認(rèn)定是紫陀螺菌的純培養(yǎng)物.

關(guān)鍵詞：紫陀螺菌; 組織分離; 親菌鑒定; 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.羅信昌,Corresponding author. E-mail: xinchangluo@mail.hzau.edu.cn

Author Unit：譚愛華（華中農(nóng)業(yè)大學(xué),農(nóng)業(yè)微生物國家重點(diǎn)實(shí)驗(yàn)室,武漢,430070;宜昌職業(yè)技術(shù)學(xué)院,宜昌,443003）　

　　　　　馬富英（華中農(nóng)業(yè)大學(xué),農(nóng)業(yè)微生物國家重點(diǎn)實(shí)驗(yàn)室,武漢,430070）　

　　　　　羅信昌（華中農(nóng)業(yè)大學(xué),農(nóng)業(yè)微生物國家重點(diǎn)實(shí)驗(yàn)室,武漢,430070）　

　　　　　黃亞東（華中農(nóng)業(yè)大學(xué),農(nóng)業(yè)微生物國家重點(diǎn)實(shí)驗(yàn)室,武漢,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

華中農(nóng)業(yè)大學(xué)學(xué)報(bào)

JOURNAL OF HUAZHONG AGRICULTURAL UNIVERSITY

2004 Vol.23 No.1