Souza Thiago Pereira, Marques Simone Cristina, da Silveira e Santos Débora Marques, Dias Eustáquio Souza
Department of Biology, Federal University of Lavras/ UFLA, BOX 3037, Lavras, MG, 37200-000, Brazil.
World J Microbiol Biotechnol. 2014 Sep;30(9):2419-25. doi: 10.1007/s11274-014-1667-3. Epub 2014 May 22.
The composition and genetic diversity of fungal populations during phase II of compost production for the cultivation of Agaricus subrufescens was determined using culture-dependent and -independent methods on days 3, 6, 10, 12, and 14 of phase II composting. The isolates were morphologically characterized and subsequently analyzed using repetitive extragenic palindromic sequences (rep-PCR), and the intergenic region was sequenced to genetically identify the isolates. Changes on in the filamentous fungi population were analyzed using denaturing gradient gel electrophoresis (DGGE), and the resulting bands were sequenced. The population did not significantly change from day 3 to 10 (2.55 x 10(5) -6 x 10(5) CFU g(-1)), and maximum counts on day 14 of phase II composting (6.92 log CFU g(-1)). In the morphological characterization, Scytalidium thermophilum, Thermomyces lanuginosus, and Thermomyces ibadanensis were the most abundant identified species. The 26 most abundant isolates identified by morphological analysis were characterized using rep-PCR. A significant amount of genetic diversity was detected among the isolates of all three studied species. Based on the DGGE analysis, the diversity of the fungi was reduced during phase II composting, and S. thermophilum was the predominant species identified throughout the entire process. Thus, this study presents the first report of the involvement of T. ibadanensis in the production of compost for Agaricus mushroom cultivation.
在姬松茸栽培堆肥生产的第二阶段第3、6、10、12和14天,使用依赖培养和不依赖培养的方法确定了真菌种群的组成和遗传多样性。对分离株进行形态学表征,随后使用重复外基因回文序列(rep-PCR)进行分析,并对基因间区域进行测序以从基因上鉴定分离株。使用变性梯度凝胶电泳(DGGE)分析丝状真菌种群的变化,并对产生的条带进行测序。从第3天到第10天,种群数量没有显著变化(2.55×10⁵ - 6×10⁵ CFU g⁻¹),在堆肥生产第二阶段的第14天达到最高计数(6.92 log CFU g⁻¹)。在形态学表征中,嗜热粘帚霉、疏绵状嗜热丝孢菌和伊巴丹嗜热丝孢菌是鉴定出的最丰富的物种。通过形态学分析鉴定出的26个最丰富的分离株使用rep-PCR进行表征。在所有三个研究物种的分离株中检测到大量的遗传多样性。基于DGGE分析,在堆肥生产第二阶段真菌的多样性降低,嗜热粘帚霉是整个过程中鉴定出的优势物种。因此,本研究首次报道了伊巴丹嗜热丝孢菌参与姬松茸栽培堆肥的生产。