Shinzato Naoya, Namihira Tomoyuki, Tamaki Yasutomo, Tsukahara Masatoshi, Matsui Toru
Center of Molecular Biosciences, University of the Ryukyus, 1 Senbaru, Nishihara-cho, Okinawa 903-0213, Japan.
Appl Microbiol Biotechnol. 2009 Apr;82(6):1187-93. doi: 10.1007/s00253-009-1937-4. Epub 2009 Mar 13.
Monascus fungi are commonly used for a variety of food products in Asia, and are also known to produce some biologically active compounds. Since the use of Monascus is expected to increase in food industries, strain-level identification and management of Monascus will be needed in the near future. In the present study, random amplified polymorphic DNA (RAPD) analysis coupled with microchip electrophoresis was applied for this purpose. Evaluations of the analysis stability revealed that reproducible results could be obtained, although template DNA fragmentation could influence the resulting RAPD pattern. RAPD analysis using 15 Monascus strains consisting of four species, M. ruber, M. pilosus, M. purpureus, and M. kaoliang showed that each strain generated a unique RAPD pattern, which allows strain-level identification of Monascus. In addition, the phylogenetic tree constructed from RAPD patterns reflected M. ruber-M. pilosus and M. purpureus-M. kaoliang clusters inferred from both ITS and beta-tubulin gene sequences, which indicated that the RAPD pattern could reflect their phylogenetic traits to a certain extent. On the other hand, RAPD analysis did not support the monophyletic clustering of the four Monascus species used in this study, which suggests the necessity of reexamination of species boundaries in Monascus.
红曲霉菌在亚洲常用于多种食品，并且已知能产生一些生物活性化合物。由于预计红曲霉菌在食品工业中的使用将会增加，因此在不久的将来需要对红曲霉菌进行菌株水平的鉴定和管理。在本研究中，为此应用了随机扩增多态性DNA（RAPD）分析结合微芯片电泳技术。对分析稳定性的评估表明，尽管模板DNA片段化可能会影响最终的RAPD图谱，但仍可获得可重复的结果。对由红曲红、丛毛红曲、紫色红曲和高粱红曲这四个物种组成的15株红曲霉菌进行RAPD分析，结果表明每个菌株都产生了独特的RAPD图谱，这使得能够对红曲霉菌进行菌株水平的鉴定。此外，根据RAPD图谱构建的系统发育树反映了从ITS和β-微管蛋白基因序列推断出的红曲红-丛毛红曲以及紫色红曲-高粱红曲聚类，这表明RAPD图谱在一定程度上能够反映它们的系统发育特征。另一方面，RAPD分析并不支持本研究中所使用的四种红曲霉菌的单系聚类，这表明有必要重新审视红曲霉菌的物种界限。
