| Ophiocordyceps bispora (Stifler) GH. Sung, J.M.Sung, Hywel-Jones & Spatafora was firstly known from Tanzania, Eastern Africa, where it was a specific pathogen of termite —Macrotermes natalensis (Haviland) Fuller. During 2014～2015, a large of O. bispora were collected from Ailao Mountain, Yunnan province, China. Literature review resulted that this fungus was newly recorded in Asia. Several aspects of observation were performed to research biological feature of this fungus in present paper, attempt to lay a foundation for further research and utilization. This study was included the systematic status, cultural character, active ingredients and the anti-aging effect of this species.O. bispora was belonged to the genus Ophiocordyceps, with being characterized by the clavate ascus with long stipe, but without ascus cap and the ascus only behaviour two long—fusiform ascus spores which adhere together, becoming 7～15 separation when mature. The phylogenetic analyses based on 5-gene dataset(nrSSU, nrLSU, tef-1A, rpb1 and rpb2)showed that O. bispora was clustered with Hirsutella thompsonii Fisher、 Hirsutella necatrix Minteret & Brady、Hirsutella minnesotensis Chen, Liu & Chen and Hirsutella cryptosclerotiμm Fernandez Garcia Evans & Samson into one clade. It was indicated that O. bispora had closer phylogenetic relationship with those species. Comparing with Ophiocordyceps species distributed in the Hengduan Mountains, the phylogenetic position of O. bispora was located at the base of phylogenetic tree. It was suggested that O. bispora should be an ancient species distributed in The Hengduan Mountains as its specific ascus and ascus spores.Two associated fungi of O. bispora were identified as Bionectria ochroleuca (Schwein.) Schroers & Samuels and Paecilomyces lilacinus (Thom) Samson based on morphologic observation and ITS sequences analysis.The cultural condition of O. bispora were investigated. It was resulted that the optimum carbon source was corn, the optimum nitrogen source was silkworm, the optimum temperature was 30℃, and the optimum pH was 9. The optimal medium formula was corn 3%, silkworm 2%, MgSO40.15%, and KH2PO40.1%.The polysaccharides, D-mannitol, saponin, flavonoid, triterpene of O. bispora and its mycelium cultured were determined by UV spectrophotometer. The result showed that D-mannitol of O. bispora and polysaccharides of its mycelium cultured were much higher than those of Ophiocordyceps sinensis. The contents of D-mannitol of O. bispora and polysaccharides in its mycelium cultured were 114.33mg/g and 63.04mg/g, respectively. The contents of D-mannitol and polysaccharides of O. sinensis were 89.20mg/g and 44.79mg/g, respectively. But saponin, flavoneand triterpene in O. bispora and its mycelium cultured were lower than those of O. sinensis. The contents of saponin, flavone and triterpene in O. bispora and its mycelium cultured were 14.97mg/g and 12.92mg/g,1.61mg/g and 1.29mg/g,34.65mg/g and 17.10mg/g, respectively. The contents of saponin, flavone and triterpene of O. sinensis were 34.30mg/g, 11.01mg/g and 41.67mg/g, respectively.The nucleosides of O. bispora and its mycelium cultured were determined by HPLC. Compared with O. sinensis, the contents of uracil, inosine, adenine and 2’-deoxy-adenosine in O. bispora were higher than those of O. sinensis, while the uridine, guanosine, adenosine and the thymidine were lower than those of O. sinensis. The contents of uracil,2’-deoxy-adenosineand and adenine in mycelium cultured of O. bispora were much higher than those of O. sinensis. The contents of uracil, 2’-deoxy-adenosineand and adenine in mycelium cultured of O. bispora were 819.28μg/g,785.96μg/g, and 344.57μg/g, respectively. The contents of uracil, 2’-deoxy-adenosineand and adenine of O. sinensis were75.43μg/g,24.33μg/g and 46.80μg/g, respectively. The thymidine wasn’t detected in O. bispora, and the adenosine was not detected in mycelium cultured of O. bispora.The pharmacology experiment suggested that mycelium cultured of O. bispora might have an effect of delaying senescence. The maximum life span, the average life expectancy, and half the time of death of Drosophila melanogaster Meigen were significantly extended by feeding with low concentrations(0.5%) of mycelium cultured of O. bispora.In summary, O. bispora was belonged to ophiocordyceps, being newly recorded in Asia. The active ingredients of both O. bispora and its mycelium cultured were similar to those of O. sinensis. The pharmacology experiment suggested that the O. bispora might have the effect of delaying senescence. Two associated fungi of the O. bispora were identified as B. ochroleuca and P. lilacinus. O. bispora and its associated fungi —B. ochroleuca and P. lilacinus might be a great potential of biocontrol. |
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