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Effects of histone methylation and deacetylation on the biosynthesis of huperzine A in the endophytic fungus Colletotrichum gloeosporioides
SHEN Peng-Yuan1，2 Kang Xin-Cong1，2 Hu Li-Qing1，2 Zhang Jia-Yin1，2 Liu Dong-Bo1，2，3，4
1.Horticulture and Landscape College，Hunan Agricultural University，Changsha，Hunan 410128，China；
2.State Key Laboratory of Subhealth Intervention Technology，Changsha，Hunan 410128，China；
3.Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization，Hunan Agricultural University，Changsha，Hunan 410128，China；
4.Hunan Co-Innovation Center for Utilization of Botanical Functional Ingredients，Changsha，Hunan 410128，China
Abstract：A histone methyltransferases(HMT)gene CgClr4 and two histone deacetylase(HDAC)gene，CgClr3 and CgSir2，were knocked out and then complemented in the endophytic fungus Colletotrichum gloeosporioides strain Cg01 in Huperziaserrata by the PEG-mediated genetic transformation system.The expression of the corresponding genes in the complemented strains were detected by RT-qPCR and huperzine A(Hup A)production in the mutant strains were analyzed by high performance liquid chromatography(HPLC).It was found that the Hup A production of the 3 gene knockout mutant strainsΔCgClr4，ΔCgClr3 andΔCgSir2 were 255μg/L，270μg/L and 244μg/L respectively，decreased by 21.3%，16.6%and 24.7%as compared with wild type(WT).Thereis no significant difference between the gene expression of the corresponding gene in the complemented strainsΔCgClr4/CgClr4，ΔCgClr3/CgClr3 andΔCgSir2/CgSir2 and that in the wild type strain，with the Hup A production to be 351.9μg/L，334.7μg/L and 331μg/L，respectively.The HupA production of complemented strains was restored to that of the wild type strain.The results showed that these three genes had the function of regulating the biosynthesis of Hup A，which provided a theoretical basis and a new insight for studying the synthetic mechanism of Hup A in endophytic fungi of Huperzia serrata.
Keywords：Histone，gene knockout，gene rescue，huperzine A，biosynthesis