Project description:M. purpureus YY-1 is widely used in food colorant production in China. In our previous study, the whole-genome information of YY-1 was clearly illustrated, which provided useful hints for evolution esearch and industrial applications. However, the presence of citrinin, which has nephrotoxic, hepatotoxic, and carcinogenic activities, attracts people′s attention to the safety of Monascus products. In order to reduce the harm of citrinin in Monascus related products, in this study, a random mutant of M. purpureus YY-1, with scarce citrinin production (designated as winter) was obtained. To analyze the biosynthesis and regulation mechanism of pigment and citrinin, transcriptomic analysis of M. purpureus YY-1 and winter was performed. Comparative transcriptomic analysis reveals pksCT, the essential gene for citrinin synthesis, showed low expression level in M. purpureus YY-1 and winter, which suggested there might be isoenzymes in M. purpureus YY-1 that were responsible for the citrinin synthesis during evolution. In addition, the expression change of transcription factors may also influence the regulatory network of citrinin synthesis pathway of in M. purpureus. Moreover, the yields of pigments produced by the winter mutant were significantly increased. Repressing central carbon metabolism and improving the acetyl-CoA pool can contribute to the high yield of pigments, and enhanced NADPH regeneration also lead the metabolic flux to pigments in M. purpureus. Investigations on biosynthesis and regulation of citrinin and pigment production in M purpureus will enhance our knowledge of the mechanisms in fungal secondary metabolite biosynthesis.
Project description:BackgroundPhytochromes are the best characterized photoreceptors that perceive Red (R)/Far-Red (FR) signals and mediate key developmental responses in plants. It is well established that photoperiodic control of flowering is regulated by PHY A (phytochrome A) gene. So far, the members of PHY A gene family remains unexplored in Lablab purpureus, and therefore, their functions are still not deciphered. PHYA3 is the homologue of phytochrome A and known to be involved in dominant suppression of flowering under long day conditions by downregulating florigens in Glycine max. The present study is the first effort to identify and characterize any photoreceptor gene (PHYA3, in this study) in Lablab purpureus and decipher its phylogeny with related legumes.ResultsPHYA3 was amplified in Lablab purpureus cv GNIB-21 (photo-insensitive and determinate) by utilizing primers designed from GmPHYA3 locus of Glycine max. This study was successful in partially characterizing PHYA3 in Lablab purpureus (LprPHYA3) which is 2 kb longer and belongs to exon 1 region of PHYA3 gene. Phylogenetic analysis of the nucleotide and protein sequences of PHYA genes through MEGA X delineated the conservation and evolution of Lablab purpureus PHYA3 (LprPHYA3) probably from PHYA genes of Vigna unguiculata, Glycine max and Vigna angularis. A conserved basic helix-loop-helix motif bHLH69 was predicted having DNA binding property. Domain analysis of GmPHYA protein and predicted partial protein sequence corresponding to exon-1 of LprPHYA3 revealed the presence of conserved domains (GAF and PAS domains) in Lablab purpureus similar to Glycine max.ConclusionPartial characterization of LprPHYA3 would facilitate the identification of complete gene in Lablab purpureus utilizing sequence information from phylogenetically related species of Fabaceae. This would allow screening of allelic variants for LprPHYA3 locus and their role in photoperiod responsive flowering. The present study could aid in modulating photoperiod responsive flowering in Lablab purpureus and other related legumes in near future through genome editing.