Studies On Artificial Induction Cultivation For Fruiting Body Of Cordyceps Formosana

Cordyceps sensu lato fungi have very important application in medicine, agriculture,food industry and modern biotechnology with multi-functional aspects such as antibacterial,antiviral, antineoplastic, antioxidative, anti-radiation, insecticidal, calcium antagonist andimmune activities. In recent years, it is very difficult to collect valuable specimens in thefield because of destruction of natural environment and excessive indiscriminatecollections. Therefore, artificial cultivation of fruiting body becomes an effective way toprotect and utilize these biological resources. Meanwhile, they have important academicvalue in Cordyceps classification and genetic breeding. Also we can obtain medicinal andhealth food materials, and separate numerous biological active compounds.In this research，the artificial culture conditions on fruiting body of Cordycepsformosana were systematically studied including strains select, optimization of liquidculture medium, determination of optimal culture time of flask seed，effects of differentfactors and culture conditions on fruiting body formation of C. formosana. On the basis ofabove researches, culture conditions were further optimized by response surface analysis.Six strains of Hirsutella huangshanensis were screened by solid plate culture. Theresults showed that the growth of RCEF0868and RCEF5491were better than that of otherfour strains in solid media, and the colony diameter was larger; and the colony diameter ofstrain RCEF0868was the widest by14d culture. So the strain RCEF0868was selected forfurther research because of better sporulation ability and the fruiting body successfulformation in preliminary experiments.Five liquid media were screened by shake flask cultures. The results showed that dryweight of RCEF0868mycelium in liquid medium Ⅴ reached a maximum amount for15days, the size of the mycelial pellets was uniform. Liquid medium Ⅴ was selected as theliquid shake flask culture medium because of the fastest growth and pure color offermentation broth.The optimal culture time of RCEF0868liquid flask seed was determined by shakeflask culture. The results showed that the liquid shake flask seed began to grow after aboutthree days. The maximal growth rate of mycelium was at12thday, and the dry weight ofmycelium reached the maximum size at24thday and remained unchanged later. The periodwith maximal growth rate was chosen as the shake-flask seed culture and the optimal timeof liquid shaking flask is12d.Artificial solid cultivation of fruiting body of C. formosana was studied by canned bottle including the effects of different factors and culture conditions, such as solidcultivation materials, cultivation nutrient solution, pH of nutrient solution, nutrient solutionamount, inoculum size, culture temperature, illumination time, light quality on hyphalgrowth, primordium and fruiting body formation. The results showed that the optimalmaterials of artificial solid cultivation for fruiting body of C. formosana was rice; theoptimal conditions were as follows: nutrient solution Ⅴ, pH6.0nutrition solution,40mlnutrient solution amount,10ml inoculum size and25℃culture temperature, myceliumculture stage under complete dark conditions by using black plastic bags and givingillumination after primordium formation.By response surface experiment, three factors effecting obviously on artificialcultivation conditions of fruiting body of C. formosana were further optimized andanalysed. The optimum conditions on fruiting body formation of C. formosana wereestablished as follows: nutrient solution pH5.7, amount of nutrient solution40ml,inoculum size9.5ml. Under these conditions, the three parallel verification experimentswere made and the results showed that average fresh weight of fruiting body was2.32g,while theoretical model predictions was2.38g. Experimental value was close to thecalculated value. Compared with the theoretical prediction, the relative error was only2.52%.Based on above experiments, the cultivation process of fruiting body of C. formosanawas summarized and a patent was proposed. These findings provided technical support forlarge-scale industrial production.