| The production cycle of Hypsizygus marmoreus is usually 100-120 days,which is about twice the production cycle of Flammulina velutipes and Pleurotus eryngii,and the production efficiency is relatively low.The previous research of this group showed that the addition of hydrogen-rich water,saponin and kojic acid could not only effectively increase the yield of the fruit body,but also shorten the post-ripening stage of the hyphae and promote the germination of the primordium.In this study,the effects of exogenous vitamin C addition on the yield,antioxidant content,enzyme activity,antioxidant enzyme gene and the related gene expression in the signal pathway were studied.The mechanism of exogenous vitamin C affecting the yield of H.marmoreus was explored.The research content and results are as follows:(1)In this study,we first added different concentrations of vitamin C by exogenous sources.It was found that four different concentrations of vitamin C can effectively increase the yield of fruit body by 1.8%-9.7%,in which the concentration of 400 mg/Lshowed the best.Then the effect of vitamin C with the best concentration of 400 mg/L on the yield of H.marmoreus was explored at different culture time.It was found that the yield of the fruiting body of the treated group at 60 days was higher than that of the control group at 80 days,indicating that vitamin C can shorten the production cycle of H.marmoreus.(2)The physiological and biochemical indexes(MDA,H2O2,anti-superoxide anion,Vc,GSH,GSSG)and antioxidant enzyme activities(SOD,CAT,GR,GPX,TrxR,TPX)of mycelium in four key periods of growth and development of H.marmoreus were studied.And the relative expression level of its enzyme gene.Exogenous vitamin C can reduce the content of MDA and the content of H2O2 in cells.In the hyphae recovery period,color transition period and primordial phase,the content of MDA in the treatment group was lower than that in the control group.In the period of hyphae recovery and color change,the content of H2O2 in the treatment group was lower than that in the control group and the difference in H2O2 content between the control group and the treatment group was not significant.The vitamin C treatment group can maintain the anti-superoxide anion activity at a high level in the hyphae recovery period,the primordial phase and the fruit body phase,effectively removing the superoxide anion in the cells,thereby reducing the degree of membrane lipid oxidation of the H.marmoreus cell.However,the anti-superoxide anion activity in the treatment group was lower than that in the control group.The content of Vc in the exogenous vitamin C treatment group showed a decreasing trend,whereas the control group increased first and then decreased.During the recovery period of mycelium,the content of Vc in the treatment group was higher than that in the control group(P<0.05).It was lower in the color-changing phase and the primordial treatment group than in the control group;the fruiting-phase treatment group was higher than the control group.But the difference is not significant.The GSH content and GSSG content in the control and treatment groups increased first and then decreased.In the four critical periods of growth and development of H.marmoreus,the SOD and CAT activities in exogenous vitamin C treatment group were significantly higher in the fruiting stage than in the control group,and were lower in the color change period than in the control group,and the changes in the other two periods were not significant.The reaction mechanism of SOD and CAT two antioxidant enzymes after exogenous addition of vitamin C is similar.The activity of GR enzyme was higher in the fruiting stage treatment group than in the control group,and there was no significant difference between the control group and the treatment group in the hyphae recovery period,and it was lower in the color-changing stage and the primordial-stage treatment group than in the control group.In the color-changing period,the TPX activity of the exogenous vitamin C treatment group was lower than that of the control group,and there was no difference in the TPX activity between the mycelial recovery period,the primordial phase and the fruiting stage control group and the exogenous vitamin C treatment group.These four enzymes play a minor role in the antioxidant mechanism regulated by vitamin C.In the hyphae recovery period and color change period,the GPX activity in the treatment group was higher than that in the control group,especially in the color-changing period,with significant difference(P<0.05),but the difference in GPX activity between the control group and the treatment group in the primordial and fruiting stages was not significantly;TrxR activity in the hyphae recovery,color change,primordial and fruiting stage treatments was higher than that of the control group.The relative expression of sod gene in exogenous vitamin C treatment group was higher than that in the control group.The relative expression of cat and gr in the vitamin C treatment group was lower than that in the control group,and the relative expression was increased in other periods.The relative expression levels of gpx,trxr and tpx were up-regulated during the recovery period.(3)The relative expression of ROS signaling pathway,Ca2+signaling pathway and MAPK signaling pathway related genes in vitamin C growth and development was studied.It was found that the relative expression levels of pka,pkc and ste7 were up-regulated during hyphal recovery,color transition and primordial phase.The relative expression levels of cpe and ste20 genes were higher in the hyphae recovery and primordial treatment groups than in the control group,and lower in the color-changing and fruit-child phases than in the control group.The relative expression levels of noxa and noxb genes in the treatment group were higher in the hyphae recovery and fruiting stage than in the control group,and were lower in the color-changing phase and the primordial phase than in the control group;in the hyphal recovery period and in the primordial phase,the vitamin C treatment group noxr gene was relative.The expression level was several times that of the control group.There was no significant difference in the gene expression between the treatment group and the control group.The mapk gene in the treatment group was down-regulated during the hyphal recovery period and the color change period,and was up-regulated in the primordial and fruiting stages.In summary,this study shows that the addition of exogenous vitamin C after sputum can shorten the time of mycelial recovery and regeneration,which is conducive to the proliferation and differentiation of mycelium.The addition of exogenous vitamin C can reduce the content of malondialdehyde and reactive oxygen species in the cells,increase the expression of antioxidant enzymes,and activate the expression of cac,pka,pkc,ste7,cpe,ste20,noxa,noxb and noxr genes in signaling pathways.In this study,the mechanism of exogenous vitamin C on the formation of the fruit body and its yield increase was revealed and the direction was initially explored. |
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