The Effect Of Change Of Nutritional Factors On The Fruiting Of Hypsizygus Marmoreus

Hypsizygus marmoreus is a edible mushroom with high nutritional and medical value,which has great market potentiality. Currently, the industrial cultivation cycle of H.marmoreus is quite long. The yield and quality of fruit body have a close relationship with the components of the medium. However, the complexity of the substrates complicated the performance of physiological studies on the fruiting process. At present, the study on H. marmoreus is mainly focused on the choose of culture conditions, the basic biology, the active ingredient analysis and pharmacological research, the research about their genetics is still weak, the research on the molecular mechanism of fruit body formation of H.marmoreus is even blank.In this study,32kinds of different liquid synthetic media were designed according to the physiological and biochemical characteristics of strain SIEF3133. The growth of mycelium, formation of primordium and development of fruit body of H.marmoreus on different media were observed, then we obtained a medium which was best for the development of fruit body; the biomass and main nutritional factors during the development of H.marmoreus were determined; then the effect of the level of nutritional factor(glucose) in medium on the formation of fruit body was investigated; SAGE was applied to analyse the gene expression profiles of H.marmoreus vegetative mycelium and primordium from media with different glucose level.The main results were as follows:1.32kinds of different liquid synthetic media were designed according to the physiological and biochemical characteristics of strain SIEF3133. The effect of carbon source(including glucose, CMC, starch, maltose, fructose and lactose) and nitrogen(L-sodium glutamate, yeast extract, tryptone, casein acid hydrolysates and ammonium nitrate) on the fruiting of H.marmoreus was investigated. The development of H.marmoreus on different media were compared and a liquid medium which was suit for the fruit body development was determined. The main components of medium were as follows(g/L):glucose,20g; L-sodium glutamate,2g; yeast extract,5g; tryptone,3g; casein acid hydrolysates,2g; KH2PO4,2g; MgSO4·7H2O,1.5g; mineral solution(SolutionA),10mL; trace element(SolutionB), solution1mL; vitamin solution(SolutionC),1mL. H.marmoreus was able to form mature fruit body with fertile basidiospores on this medium.2. Several main physiological indicators(biomass,reducing sugur, total sugar, soluble protein and free amino acid) were determined. The results revealed that the mycelium biomass reach to0.65g at the end of cultivation. The content of reducing sugar decreased from2g to0.3g and the content of free amino acid decreased from0.33mg to0.15mg, however, the change of total sugar and protein were complicate. By comparing the change of above physiological indicators, we can find that the change of nutritional factors has a close relationship with development of H.marmoreus.3. The level of glucose in medium was regulated(the level of glucose in medium was10g/L,20g/L,30g/L and40g/L, seperately) to investigate the effect of the content of the glucose on the development of H.marmoreus. The results revealed that the content of glucose in the medium affect both the speed of primordium formation and the shape of primordium; when the content of glucose in the medium reach to40g/L, the primordium were clustered.4. SAGE was applied to study the genes expression of H.marmoreus mycelium and primordium produced on media with different glucose content(10g/L and40g/L).665084,3713976,3528430and3783886tags were extracted from four SAGE libraries, and176028,185985,186925and180610tags were unambiguous tags, seperately. Due to that there is no annotated genome database for H. marmoreus, we compared all of the SAGE tags with the annotated genes of basidiomycete genome information on NCBI.237,252,260,241unambiguous tags of10-ml,10-m2,40-m3,40-m4showed homologies to basidiomycete genome database, seperately. The difference of expression profiles between mycelium and primordium and between two media with different glucose content indicates that fruit body development needs a set of genes.