Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (12): 60-70.doi: 10.13560/j.cnki.biotech.bull.1985.2021-0141

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Bioinformatics,Subcellular Localization and Expression Analysis of erg4 in Penicillium expansum

HAN Zhan-hong1(), ZONG Yuan-yuan1, ZHANG Xue-mei1, WANG Bin1, PRUSKY Dov2, BI Yang1()   

  1. 1. College of Food Science and Engineering,Gansu Agricultural University,Lanzhou 730070
    2. Department of Postharvest Science of Fresh Produce,Agricultural Research Organization,Volcani Center,Bet Dagan Israel 50250,Israel
  • Received:2021-02-04 Online:2021-12-26 Published:2022-01-19
  • Contact: BI Yang E-mail:hanzhanhong009@163.com;biyang@gsau.edu.cn

Abstract:

Ergosterol,a fungus-specific component in cell plasma membranes,plays an essential role in fungal growth and development. The erg4 gene encodes the catalytic enzyme for the last step of ergosterol synthesis,whereas,the function of this gene in Penicillium expansum is unknown. In this study,the full-length CDSs of erg4A,erg4B and erg4C in P. expansum were cloned by RT-PCR,and their genes’ structures,transmembrane domain and hydrophobicity of the erg4 encoded proteins were analyzed via bioinformatics. The subcellular localizations of 3 proteins were also analyzed by fusion green fiuorescent protein method. In addition,the expression differences of 3 genes in different growth stages,different medium conditions,and dark and blue light conditions were measured. The full-length CDSs of erg4A,erg4B and erg4C were 1 476 bp,1 491 bp and 1 596 bp,respectively,and the number of encoded amino acids were 491,496 and 531,respectively. The encoded proteins were all transmembrane proteins and presented hydrophobicity. The co-localization results of green fluorescent protein and endoplasmic reticulum fluorescent probe showed that Erg4A,Erg4B and Erg4C were all located in endoplasmic reticulum. There were significant differences in the expressions of erg4A,erg4B and erg4C in spore,spore germination and mature mycelium stages. The expression of erg4A had no significant variation in the three stages,while the expressions of erg4B and erg4C were significantly up-regulated,and that of erg4B was the most obvious. There was no significant variation in the expression of erg4A under CY liquid and solid culture conditions. The expressions of erg4B and erg4C under CY liquid culture conditions were significantly higher than those under solid culture conditions,and the up-regulation of erg4B was the most obvious. The expressions of erg4A and erg4B under blue light were significantly higher than those under dark conditions,and the up-regulation of erg4B was the most obvious. erg4C was not sensitive to blue light condition. In conclusion,Erg4A,Erg4B and Erg4C were all located in the endoplasmic reticulum,and the expressions of erg4A,erg4B and erg4C vaied significantly in different growth stages,solid and liquid cultural conditions,and dark and blue light culture conditions,among which erg4B was the most active.

Key words: Penicillium expansum erg4, bioinformatics analysis, subcellular localization, expression differences