利用nested PCR-DGGE技术分析江苏啤酒大麦真菌群落结构

Nested PCR-DGGE Analysis of the Fungi Community of Jiangsu Local Malting Barley

为了探索真菌群落对啤酒大麦麦芽的影响,首先要了解啤酒大麦真菌群落结构。文中采用9种不同方法提取了啤酒大麦总DNA,利用嵌套聚合酶链式反应-变性梯度凝胶电泳(nested PCR-DGGE)技术,分析了啤酒大麦真菌群落结构,明确了啤酒大麦中优势真菌种属信息。不同DNA提取方法,nested PCR-DGGE分析结果有显著差异。啤酒大麦中的主要真菌为Cochliobolus sativus、Alternaria alternata、Fusarium sporotrichioides、Coniothyrium fuckelii、Aspergillus sp.和Saccharomyces cerevisiae等子囊菌门真菌以及Sporobolomyces roseus、Cryptococcus magnus和Entorrhiza fineranae等担子菌门真菌。该研究丰富了对啤酒大麦真菌群落结构的认识,同时也证明nested PCR-DGGE技术是一种能够快速有效地研究啤酒大麦真菌群落结构的技术。

The objective of this study was to investigate the fungal communities in the malting barley. In this pa- per, nine different DNA extraction methods and the nested PCR-DGGE （nested polymerase chain reaction-denaturing gradient gel electrophoresis） were used. DNA sequencing was proceeded to obtain the dominant fungal population in- formation. The results of nested PCR-DGGE analysis had a sensible difference to different DNA extraction methods. Results showed that total of nine fungi species were detected, including Cochliobolus sativus, Alternaria alternata, Fusarium sporotrichioides, Coniothyrium fuckelii, Aspergillus sp. , Saccharomyces cerevisiae, Sporobolomyces roseus, Cryptococcus magnus, and Entorrhiza fineranae. The study enriched the knowledge of fungal communities in the malting barley, and proved that nested PCR-DGGE was a powerful tool for revealing fungal diversity in the malting barley.