Optimized Artificial Cultivation Of Ophiocordyceps Sinensis Fruiting Bodies

Chinese cordyceps is a valuable biological resource epidemic to Qinghai-Tibet Plateau.In this thesis,artificial cultivation of Ophiocordyceps sinensis fruiting bodies were optimized,to increase the yields of fruiting bodies and provide the base for commercial production of this fungus.Four storage materials,such as sterile water,silica gel,mineral oil and glycerin,were used to optimize Ophiocordyceps sinensis storage methods.The effects of different sugars(glucose,maltose,and sucrose),ten plant hormones(cyclophosphine adenosine,indole acetic acid,6-benzylamino adenine,zeatin,α-naphthylacetic acid,gibberellin,indolebutyric acid,2,4-dichlorophenoxyacetic acid,tridecanol,ethephon)at three concentrations of 1μg/m L,10μg/m L,and 100μg/m L,and different concentrations of supernatants and bacterial cells of associated microbes at different introduction times on the artificial cultivation of O.sinensis fruit bodies,based on the rice and wheat medium,were determined.Supernatants of the associated microbes at different concentrations were tested to induce the budding growth and mycelial formation of O.sinensis blastospores.Of four preservation methods,silica gel was considered to be the best one for optimized preservation of O.sinensis.The dry weights of the fruiting bodies were 0.68g and 0.09g from the media containing glucose and maltose,respectively;but no fruiting bodies were obtained from the medium containing sucrose.The dry weight of fruiting bodies from the medium with glucose was 7.6 times more than that from the medium with maltose,and the mycelia and fruiting bodies from the medium with glucose appeared in two months ahead compared with the medium with maltose.No fruiting body was observed on the medium with sucrose.Significant differences were recorded among the dry weights of fruiting bodies from the media containing different plant growth regulators.No mycelia were observed from the media containing 3’,5’-cyclic AMP,triacontanol and zeatin.Mycelia but no stroma were observed from the media with 100μg/m L 6-benzyl adenine.After 6 months at 4℃,the dry weights of fruiting bodies from the media containing 100μg/m L indole-3-acetic acid,1μg/m L or 100μg/m L indole-3-butyric acid,10μg/m L gibberellin acid,1μg/m L or 10μg/m L ethephon,or 1μg/m L 2,4-dichlorophenoxyacetic acid were significantly higher than that from the control(without any plant growth regulators).Especially,the dry weights of fruiting bodies from the media containing 1μg/m L indole-3-butyric acid or 1μg/m L 2,4-dichlorophenoxyacetic acid were 15 times more than that from the control.Three associated microbes(Pseudomonas berry,P.fluorescens and Candida friedrichii)significantly influenced the artificial cultivation of O.sinensis fruiting bodies.P.berry supernatant or cells inhibited the mycelial growth.P.fluorescens supernatant and 10-fold dilution cells significantly stimulated the formation of fruiting bodies and increased the yield of fruiting bodies(1.7 and 2.8 times compared with the control).The dry weights from the culture flasks with 10-fold dilution supernatant and cells were 3.9 times and 3.5 times more than that from the control,respectively.The blastospores(10~5)were induced to budding growth when the supernatants(10~4 dilution)of unidentified Serratia and Aspergillus niger cultures were added into PM medium,with growth rates being 4 and 10 times respectively compared with the control.The blastospores(10~5)were inhibited to mycelial growth when the supernatants(10~2 dilution)of A.flavus and A.niger cultures were added into PM medium.In conclusion,the preservation method containing silica gel provide a good way for effective storage of O.sinensis.Indoleacetic acid,indolebutyric acid,and 2,4-dichlorophenoxyacetic acid stimulated the artificial cultivation of O.sinensis fruiting bodies.P.fluorescens and C.friedrichii supernatant and cells increased the dry weights of O.sinensis fruiting bodies.P.berry inhibited the growth of Ophiocordyceps sinensis mycelia.Unidentified Serratia sp.and A.niger supernatants promoted budding growth of O.sinensis blastospores.A.flavus and A.niger supernatants inhibited mycelial growth of the blastospores.The present results provide supports for optimized artificial production of fruiting bodies of O.sinensis.