Crucello Aline, Sforça Danilo Augusto, Horta Maria Augusta Crivelente, dos Santos Clelton Aparecido, Viana Américo José Carvalho, Beloti Lilian Luzia, de Toledo Marcelo Augusto Szymanski, Vincentz Michel, Kuroshu Reginaldo Massanobu, de Souza Anete Pereira
Centro de Biologia Molecular e Engenharia Genética (CBMEG), Universidade Estadual de Campinas, Campinas, São Paulo, Brazil.
Centro de Biologia Molecular e Engenharia Genética (CBMEG), Universidade Estadual de Campinas, Campinas, São Paulo, Brazil; Instituto de Biologia (IB), Universidade Estadual de Campinas, Departamento de Biologia Vegetal, Campinas, São Paulo, Brazil.
PLoS One. 2015 Apr 2;10(4):e0122122. doi: 10.1371/journal.pone.0122122. eCollection 2015.
Trichoderma harzianum IOC-3844 secretes high levels of cellulolytic-active enzymes and is therefore a promising strain for use in biotechnological applications in second-generation bioethanol production. However, the T. harzianum biomass degradation mechanism has not been well explored at the genetic level. The present work investigates six genomic regions (~150 kbp each) in this fungus that are enriched with genes related to biomass conversion. A BAC library consisting of 5,760 clones was constructed, with an average insert length of 90 kbp. The assembled BAC sequences revealed 232 predicted genes, 31.5% of which were related to catabolic pathways, including those involved in biomass degradation. An expression profile analysis based on RNA-Seq data demonstrated that putative regulatory elements, such as membrane transport proteins and transcription factors, are located in the same genomic regions as genes related to carbohydrate metabolism and exhibit similar expression profiles. Thus, we demonstrate a rapid and efficient tool that focuses on specific genomic regions by combining a BAC library with transcriptomic data. This is the first BAC-based structural genomic study of the cellulolytic fungus T. harzianum, and its findings provide new perspectives regarding the use of this species in biomass degradation processes.
哈茨木霉IOC-3844分泌高水平的纤维素分解活性酶，因此是用于第二代生物乙醇生产生物技术应用的有前途的菌株。然而，哈茨木霉的生物质降解机制在基因水平上尚未得到充分探索。目前的工作研究了该真菌中六个基因组区域（每个区域约150 kbp），这些区域富含与生物质转化相关的基因。构建了一个由5760个克隆组成的BAC文库，平均插入长度为90 kbp。组装后的BAC序列揭示了232个预测基因，其中31.5%与分解代谢途径相关，包括那些参与生物质降解的途径。基于RNA-Seq数据的表达谱分析表明，假定的调控元件，如膜转运蛋白和转录因子，与碳水化合物代谢相关基因位于相同的基因组区域，并表现出相似的表达谱。因此，我们展示了一种通过将BAC文库与转录组数据相结合来聚焦特定基因组区域的快速有效工具。这是对纤维素分解真菌哈茨木霉的首次基于BAC的结构基因组研究，其研究结果为该物种在生物质降解过程中的应用提供了新的视角。
