Molecular phylogenetic analysis to support the identification of Cordyceps ninchukispora collected from LangBiang, Dalat, Lam Dong

Abstract

The research team from Dalat University, Lam Dong, Vietnam, collected entomopathogenic fungi samples from the Lang Biang mountain range during their routine survey. The initial morphological identification of this species indicated it as Cordyceps ninchukispora, with Beilschmiedia erythrophloia Hay. as the host. DNA was extracted using the Phenol/Chloroform method (pH = 8) and conducted PCR experiments and sequenced the target gene regions, including ITS, nrSSU, nrLSU, Tef, and Rpb1. Molecular phylogenetic trees were constructed using Neighbor-joining, Maximum likelihood, and Maximum parsimony methods to determine the relationship between Cordyceps ninchukispora and reference sequences. We successfully amplified and sequenced the target genes, resulting in a dataset containing 52, 52, 48, 39, and 52 sequences for ITS, nrSSU, nrLSU, Tef, and Rpb1. These sequences represented various genera within the families Ophiocordycipitaceae, Clavicipitaceae, Cordycipitaceae, and two sequences from the family Glomerellaceae. The results of the molecular phylogenetic analysis showed that the collected fungal sample belonged to the Cordyceps genus and formed a monophyletic group with the reference species Cordyceps ninchukispora, with high bootstrap support. In conclusion, the multigene dataset-based phylogenetic analysis strongly supported the identification of the collected sample as Cordyceps ninchukispora.