应用18S rDNA同源性分析和常规微生物培养方法,对蘑菇基肥中的真菌种群进行了研究.采用纯培养技术分离基肥中的真菌,机械破壁法提取分离真菌总DNA,真菌特异引物对EF4/EF3 扩增18S rDNA,并进行测序及序列相似性比较,结果表明:纯培养技术分离自基肥中的真菌种属较少,从11株分离菌中只鉴定获得2株不同真菌Chaetomium elatum和Penicillium expansum.18S rDNA部分片段2次聚合酶链反应(PCR)扩增及温度梯度凝胶电泳(TGGE)分析结果表明:不同时间取自同一地点的原始蘑菇基肥样本中的真菌种群结构要比纯培养技术的分析结果复杂得多.研究结果表明,TGGE分析与高效自动测序技术结合将为生物技术和应用微生物生态过程提供一分子研究方法.
[1] 杭晓敏, 杨虹, Whiteby Corne. 垃圾填埋场中厌氧真菌18S rDNA的PCR扩增及鉴定 [J]. 生物工程学报 2001.doi:10.3321/j.issn:1000-3061.2001.05.009
[2] Wiegant W M. Growth characteristics of the thermophilic fungus Scytalidium thermophilum in relation to production of mushroom compost [J]. Applied and Environmental Microbiology 1992, 58.
[3] Blanc M, Beffa T, Aragno M, Marilley L. Thermophilic bacterial communities in hot composts as revealed by most probable number counts and molecular (16S rDNA) methods [J]. FEMS Microbiology Ecology 1999, 2(2).
[4] FERMOR T R, GRANT W D. Degradation of fungal and actinomycete mycelia by Agaricus bisporus [J]. Journal of General Microbiology 1985, 131.
[5] Hawksworth DL, Rossman AY. Where are all the undescribed fungi? [J]. Phytopathology 1997, 9(9).
[6] WHITE T J, BRUNS T, LEE S. Amplification and direct sequencing of fungal ribosomal RNA genes for phyologenetics [A]. San Diego Calif:Academic Press 1990.
[7] Smit E, Leeflang P, Glandorf B, Elsas JD van, Wernars K, van Elsas JD. Analysis of fungal diversity in the wheat rhizosphere by sequencing of cloned PCR-amplified genes encoding 18S rRNA and temperature gradient gel electrophoresis [J]. Applied and Environmental Microbiology 1999, 6(6).