The CYP51F1 Gene of Leptographium qinlingensis: Sequence Characteristic, Phylogeny and Transcript Levels.

Lulu Dai,Ranran Zhang,Mingyuan Ma,Zhumei Li,Jiamin Yu,Hui Chen,Thanh Pham

doi:10.3390/ijms160612014

Lulu Dai, Ranran Zhang + Show 5 more

Open Access

https://doi.org/10.3390/ijms160612014

Copy DOI

Abstract

Leptographium qinlingensis is a fungal associate of the Chinese white pine beetle (Dendroctonus armandi) and a pathogen of the Chinese white pine (Pinus armandi) that must overcome the terpenoid oleoresin defenses of host trees. L. qinlingensis responds to monoterpene flow with abundant mechanisms that include export and the use of these compounds as a carbon source. As one of the fungal cytochrome P450 proteins (CYPs), which play important roles in general metabolism, CYP51 (lanosterol 14-α demethylase) can catalyze the biosynthesis of ergosterol and is a target for antifungal drug. We have identified an L. qinlingensis CYP51F1 gene, and the phylogenetic analysis shows the highest homology with the 14-α-demethylase sequence from Grosmannia clavigera (a fungal associate of Dendroctonus ponderosae). The transcription level of CYP51F1 following treatment with terpenes and pine phloem extracts was upregulated, while using monoterpenes as the only carbon source led to the downregulation of CYP5F1 expression. The homology modeling structure of CYP51F1 is similar to the structure of the lanosterol 14-α demethylase protein of Saccharomyces cerevisiae YJM789, which has an N-terminal membrane helix 1 (MH1) and transmembrane helix 1 (TMH1). The minimal inhibitory concentrations (MIC) of terpenoid and azole fungicides (itraconazole (ITC)) and the docking of terpenoid molecules, lanosterol and ITC in the protein structure suggested that CYP51F1 may be inhibited by terpenoid molecules by competitive binding with azole fungicides.

Highlights

Leptographium qinlingensis is a fungal associate of the Chinese white pine beetle (Dendroctonus armandi) and a pathogen of the Chinese white pine (Pinus armandi) that must overcome the terpenoid oleoresin defenses of host trees
BLAST searches indicated that cytochrome P450 proteins (CYPs) genes expressed in L. qinlingensis were similar to members of the gene family CYP51 reported in other species (Table 1)
The full-length sequence from the CYP51 gene shared the highest level of amino acid sequence identity with variants from the fungal species G. clavigera kw1407, N. crassa OR74A, N. tetrasperma FGSC 2509, M. thermophila ATCC 42464, T. terrestris NRRL 8126 and O. piceae UAMH 11346 (Table 1)

Summary

Introduction

Leptographium qinlingensis is a fungal associate of the Chinese white pine beetle (Dendroctonus armandi) and a pathogen of the Chinese white pine (Pinus armandi) that must overcome the terpenoid oleoresin defenses of host trees. Associated with the Chinese white pine beetle (Dendroctonus armandi) helps the bark beetles to overcome the resistance system of the host trees [4]. Leptographium qinlingensis is an active participant in the death of the Chinese white pine (Pinus armandi) on Qinling Mountain of China [5,6]. Inoculation with L. qinlingensis increases the concentrations of monoterpenes and sesquiterpenes in the phloem and xylem of the P. armandi seedlings [10]

Results

Discussion

Conclusion