Cloning And Analysis Of The Mating Type Gene Of Monascus

With three single ascospore isolates of Monascus purpureus SM005, M. ruber GM011 and M. pilosus 3.4649 individual cultivation experiments, we found that single ascospore isolates can formed perithecia and ascospores, the sexual reproduction mode of Monascus was confirmed as homothallism. Comparing the mating type gene sequences and protien sequences of other strains in NCBI database, several primer pairs were designed to clone MAT-1 and MAT-2 sequences of Monascus. Used online software Conserved Domains to analyze Monascus MAT sequences and found that MAT-1 homologous sequence including a alpha-box domain, and the MAT-2 homologous sequence including a HMG-box domain. Used different strains MAT sequences of ascomycetes in NCBI databases and three Monascus MAT sequences to establish phylogenetic tree, reflecting the genetic relationship of different species.Thirty-three industrial Monascus strains with different metabolic traits were classified by three methods. By use of classical morphological classification, the Monascus species were identified as 21 strains of M. purpureus, 10 strains of M. ruber, M. pilosus and M. albidulus. The phylogenetic tree were established based on ITS genes, which can identify the two species of M. purpureus and M. ruber. The phylogenetic tree were established based on MAT genes, and the dendrogram is consistent with the classical morphological classification and ITS dendrogram. But the mating type genes showed a larger difference in interspecific and intraspecific, especially the phylogenetic tree based on MAT-1, M. pilosus has obvious genetic distance with M. ruber. Therefore, MAT sequence analysis can be an effective auxiliary tool in Monascus classification. the combination with other methods of classification, can more accurately classification and reveal the evolutionary relationships in interspecific and intraspecific.Based on the research of Monascus metabolism characters, found high-yielding pigment strains are almost M. purpureus, strain M31 has the highest yield of 4510 u/g. High-yielding monacolin K strains are almost M. ruber, strain M23 has the highest yield of 3.93 mg/g. After researching, there were no significant relationship between MAT phylogenetic tree and Monascus metabolism characters.