Institute of Plant Genetics, Polish Academy of Sciences, Strzeszyńska 34, Poznań, Poland.
Folia Microbiol (Praha). 2013 Jul;58(4):325-33. doi: 10.1007/s12223-012-0214-6. Epub 2012 Nov 29.
The present study includes the molecular characteristics of Trichoderma pleurotum and Trichoderma pleuroticola isolates collected from green moulded cereal straw substrates at 47 oyster mushroom farms in Poland. The screening of the 80 Trichoderma isolates was performed by morphological observation and by using the multiplex PCR assay. This approach enabled specific detection of 47 strains of T. pleurotum and 2 strains of T. pleuroticola. Initial identifications were confirmed by sequencing the fragment of internal transcribed spacer regions 1 and 2 (ITS1 and ITS2) of the rRNA gene cluster and the fragment including the fourth and fifth introns and the last long exon of the translation-elongation factor 1-alpha (tef1) gene. ITS and tef1 sequence information was also used to establish the intra- and interspecies relationship of T. pleurotum and T. pleuroticola originating from the oyster mushroom farms in Poland and from other countries. Comparative analysis of the ITS sequences showed that all T. pleurotum isolates from Poland represent one haplotype, identical to that of T. pleurotum strains from Hungary and Romania. Sequence analysis of the tef1 locus revealed two haplotypes ("T" and "N") of Polish T. pleurotum isolates. The "T" type isolates of T. pleurotum were identical to those of strains from Hungary and Romania. The "N" type isolates possessed a unique tef1 allele. Detailed analysis of the ITS and tef1 sequences of two T. pleuroticola isolates showed their identicalness to Italian strain C.P.K. 1540.
本研究包括从波兰 47 个蚝菇农场的绿霉谷物秸秆基质中收集的侧耳木霉和侧耳木霉分离物的分子特征。通过形态观察和多重 PCR 分析对 80 株木霉分离物进行了筛选。这种方法能够特异性检测到 47 株侧耳木霉和 2 株侧耳木霉。通过对核糖体 RNA 基因簇的内部转录间隔区 1 和 2（ITS1 和 ITS2）片段和包括翻译延伸因子 1-α（tef1）基因的第四和第五内含子以及最后长外显子的片段进行测序，对初始鉴定进行了确认。ITS 和 tef1 序列信息也用于建立源自波兰和其他国家的蚝菇农场的侧耳木霉和侧耳木霉的种内和种间关系。ITS 序列的比较分析表明，波兰所有的侧耳木霉分离株代表一个单倍型，与匈牙利和罗马尼亚的侧耳木霉菌株相同。tef1 基因座序列分析显示，波兰侧耳木霉分离株存在两种单倍型（“T”和“N”）。“T”型侧耳木霉分离株与来自匈牙利和罗马尼亚的菌株相同。“N”型分离株具有独特的 tef1 等位基因。对两株侧耳木霉分离株的 ITS 和 tef1 序列的详细分析表明，它们与意大利菌株 C.P.K. 1540 的同一性。
