The Study Of The Effect Of Blue Light On The Growth And Quality Of Ganoderma Lucidum And The Related Regulatory Protein 14-3-3 Protein

Light is a crucial environmental signal in nature for developmental and physiological processes in various organisms,including fungi.In this paper,the blue light was utilized to culture the famous medicinal fungi Ganoderma lucidum,at the vegetative stage with dark and the reproductive stage with natural light as control,respectively.The growth rate and biomass of G.lucidum were determined.The contents of polysaccharides were determined by the phenol-sulfuric acid method and the activities of hexokinase(HK),glucose-6-phosphate dehydrogenase(G-6-PDH),?-phosphoglucomutase(?-PGM)and phosphoglucose(PGI)were measured.The correlation between the polysaccharides content and glycometabolism-related enzymes was analyzed with SPSS17.0.Genome information of G.lucidum has been obtained to cloned the key enzyme genes in four kinds of glycometabolism-related enzymes and 14-3-3 protein gene of G.lucidum,and the real-time PCR was used to understand their spatial and temporal expression.With the PDA solid culture medium,the contents of the ploysaccharides,four kinds of glycometabolism-related enzymes of G.lucidum mycelium irradiated with the blue light were significantly higher than those with dark.As to the changing trends,they both increased first and then decreased,the maximum is on the eighth.The biomass accumulation irradiated with the blue light was higher than that with dark.The content of polysaccharides had significant correlations with the activities of HK,PGI,G-6-PDH and ?-PGM in blue light.In conclusion,the blue light promote the growth rate,the accumulation of polysaccharides,the activities of glycometabolism-related enzymes of G.lucidum mycelium.HK,PGI,G-6-PDH and ?-PGM played important roles in polysaccharides biosynthesis of G.lucidum.The fruit bodies of G.lucidum were got from log cultivation,the biomass,polysaccharides and four kinds of glycometabolism-related enzymes of G.lucidum fruit body with blue light significantly higher than those with natural light.The change tendency of G.lucidum polysaccharides,HK,PGI and ?-PGM were similar,but not with that of G-6-PDH.The maximum content of polysaccharides,HK,PGI and ?-PGM appeared during the pileus formation and lowered after the first change trend.The content of polysaccharides had significant correlations with the activities of HK,PGI and ?-PGM in blue light.In conclusion,the blue light promoted the growth,polysaccharides and glycometabolism-related enzymes of G.lucidum fruit body.Through the obtained genomic information of G.lucidum,the four gens of GlPGM,GlPGI,GlG6 PDH and GlHK could be cloned.After comparison with NCBI,GlPGM,GlPGI,GlG6 PDH with Dichomitus squalens LYAD-421 SS1,GlHK with Laetiporus sulphureus 93-53 have the highest similarity.Phylogenetic analysis based on fungal PGM,PGI,G6 PDH and HK supported GlPGM,GlPGI,GlG6 PDH and GlHK were glycometabolism-related.Finally,the relative expression level of each gene of glycometabolism-related enzymes was detected by Real-time qPCR.The results showed that all of the 4 genes identified were expressed in four periods,in addition to GlPGM,the relative expression of the other 3 genes was the highest in the second period.This is consistent with the change of polysaccharides in G.lucidum.Therefore,it was speculate that the second period was the active and fast growth period and the metabolism of G.lucidum were accelerated.The GlPGM expression after blue light treatment was higher than natural light in the third and fourth period,the GlPGI and GlG6 PDH expression after blue light treatment was higher than natural light in the second,third and fourth period,the GlHK expression after blue light treatment was higher than natural light in four periods.It shows that the blue light may increase the expression of GlPGI,GlG6 PDH and GlHK,induce a series of reactions.A novel 14-3-3 gene from G.lucidum was cloned,the cDNA was 1056 bp and it contained a complete ORF which encoded 257 amino acids.Homology comparison showed that amino acids encoded by the 14-3-3 gene in G.lucidum was highly homologous with those in basidiomycetes.As the real-time PCR results showed,the gene expression by blue light was very different from that by natural light,the gene expression under the low temperature was lower than that of the control.It showed that the blue light might increase the expression of Gl14-3-3.With the fall of temperature,the growth and development of G.lucidum might be effected.