Isolation, identification, culture and production of adenosine and cordycepin from cicada larva infected with entomopathogenic fungi in Thailand

In this study, cicada larvae infected with entomopathogenic fungi were collected from Maha Sarakham Province in northeast Thailand. One strain of entomopathogenic fungi with cottony white cream colonies was isolated. Small subunit (SSU) rDNA, large subunit (LSU) rDNA, the elongation factor 1α (EF-1α) and the largest subunit of RNA polymerase II (rpb1) sequence analyses were used for identification of the isolate. A BLAST search in NCBI showed the sequence to be most similar to Cordyceps sp., Hirsutella sp., Ophiocordyceps longissima and O. longissima for SSU rDNA, LSU rDNA, EF-1α and rpb1, respectively. This correlated well with evidence from neighbor-joining trees, based on SSU rDNA, LSU rDNA, EF-1α and rpb1. Therefore, the isolate was classified as an anamorph strain of Ophicordyceps and assigned as Ophiocordyceps longissima isolate Cod-MK1. The O. longissima isolate Cod-MK1 was found to grow best in HCGA medium, compared with six other synthetic media. Moreover, the isolate was shown to develop numerous synnemata (stroma-like stalks) and conidia, when cultured in the applied HCGA medium at 25 to 28°C for 60 to 90 days. The content of adenosine was observed only in the extract from dried mycelia at 31.68 g/g. The content of cordycepin from dried mycelia was 335.65 g/g; lower than those of the extract from dried stroma-like stalks (366.14 g/g). Therefore, the induced culture from this study could be used for the production of adenosine and cordycepin in O. longissima isolate Cod-MK1.