Scientia Agricultura Sinica ›› 2020, Vol. 53 ›› Issue (3): 632-641.doi: 10.3864/j.issn.0578-1752.2020.03.014

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Effects of Histone Acetylation on Ganoderma lucidum Growth, Polysaccharide and Ganoderic Acid Biosynthesis

ZHANG ZongYuan1,2,JIANG YongMei1,ZHANG WenXian1()   

  1. 1 College of Life Sciences, Fujian Normal University, Fuzhou 350108
    2 Institute of Biology Co., Ltd., Henan Academy of Sciences, Zhengzhou 450008
  • Received:2019-05-21 Accepted:2019-10-30 Online:2020-02-01 Published:2020-02-13
  • Contact: WenXian ZHANG E-mail:huzx@fjnu.edu.cn

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Abstract:

【Objective】Histone acetylation modification plays an important role in the growth, development and metabolic synthesis of fungi. Few epigenetic studies of higher medicinal fungi were reported at present. In this study, the effects of histone acetylation on the growth and development of Ganoderma lucidum (G. lucidum) and the synthesis of its main metabolites were studied by adding chemical epigenetic inhibitor octanedianiline hydroxamic acid (SAHA), which provided a theoretical basis for improving the biosynthesis of G. lucidum polysaccharides and Ganoderma acid (GA) by epigenetic means.【Method】A two-stage cultivation, liquid fermentation combined with static cultivation, was applied to culture G. lucidum. The cell was treated with different concentration of SAHA (0, 0.6, 60, 120, and 180 μmol?L -1) during the liquid static cultivation of G. lucidum. Biomass, sugar consumption, mycelial mat formation, mycelial morphology, sporulation and biosynthesis of GA and G.lucidum polysaccharides were measured or observed by conventional methods. Histone acetylation levels of G. luidum were examined by Western blot, the relative expression levels of polysaccharides biosynthesis genes (e.g. ugp, gls, and pgm), GA biosynthesis genes (e.g. hmg, sqs, se, and ls) and global regulator vet, LaeA gene were detected by qRT-PCR. 【Result】The acetylation level of histone H4 in G. lucidum treated with SAHA increased to 1.6 times as much as that under control group. SAHA inhibited the growth of G. lucidum mycelia and the production of pigments, and changed the morphology of mycelia. The formation of spores was also inhibited, and the higher the concentration of SAHA, the more obvious the inhibition degree. SAHA treatment significantly increased the yield of G. lucidum polysaccharides, up to 50%, and the biosynthesis of GA was inhibited, which decreased by 13%-27% compared with the control. The results of qRT-PCR analysis showed that the gene expression of the key enzymes in G.lucidum polysaccharides and GA synthesis were up-regulated in different degrees under SAHA treatment. The gene expression of the key enzymes in polysaccharides synthesis were increased by 1.5-3.5 times and that of the key enzymes in GA synthesis by 1.8-12.1 times. The expression of vet and LaeA genes, the global regulators, were inhibited, which was 11.3%-62.4% of the control group.【Conclusion】Histone acetylation could regulate the growth and development of G. lucidum through global regulatory factors, thus affecting the biosynthesis of GA, while histone acetylation also had an effect on G. lucidum polysaccharides biosynthesis.

Key words: Ganoderme lucidum, histone acetylation, ganoderic acid, Ganoderma lucidum polysaccharide, SAHA

Table 1

qRT-PCR primers"

基因 Gene 引物序列（5′-3′） Primer sequence (5′-3′) 参考文献 Reference
hmgr F-TCGCAGTGGCACAGGAGC R-CCCGGTGTTGGTGTTAGAAG [36]
sqs F-TGACGCTTCCTGACGAGA R-GTGGCAGTAGAGGTTGTA [36]
ls F-CTTCCGCAAGCACTACCCG R-AGCAGATGCCCCACGAGCC [36]
se F-ACTTCTGCGGGATCATATTGG R-TGTAGGATTTGCTCCTTCAGGT [34]
LaeA F-CTCCGCCGATTCTACTGG R-ACGGTCTAGCCGCTCAAA [34]
vet F-GGACACGACCCTTGGAACA R-CGGAAACGCGAACATAGCC [34]
pgm F-GGGCCTGAGGAAGAGGGTGA R-CGGTTTCG GGGGAGAAGTAG [29]
ugp F-TGGTCTCGGAACTTCTATGGG R-CAGTGCTTCTTCTCGTCCTCA [29]
gls F-TCGTTTGGG TTGGGTCTGT R-GAAGCCCTTGTCGCTCTGC [29]
18S- rRNA F-TATCGAGTTCTGACTGGGTTGT R-ATCCGTTGCTGAAAGTTGTAT [36]

Fig. 1

Effects of SAHA on the biomass and sugar consumption in the liquid static culture of G. lucidum"

Fig. 2

Effect of SAHA on the white mycelia layer growth in the liquid static culture of G. lucidum"

Fig. 3

Effect of SAHA on the level of histone acetylation of G. lucidum A: The Western blotting of histone of G. lucidum. B: Detection of the level of histone acetylation in G. lucidum by Western blot assay. Different lowercase letters indicate significant differences (P<0.05). The same as below"

Fig. 4

Effects of SAHA on aerial mycelia morphology and spore formation in liquid static culture of G. lucidum A: The mycelia morphology on liquid surface were cultured on day 6 at 0 μmol∙L-1 (Ⅰ), 120 μmol∙L-1 (Ⅱ), 180 μmol∙L-1 (Ⅲ) SAHA and the mycelia with spores on liquid surface were cultured on day 9 at 0.09% DMSO (Ⅳ) and 0 μmol∙L-1 (Ⅴ), 0.6 μmol∙L-1 (Ⅵ), 60 μmol∙L-1 (Ⅶ), 120 μmol∙L-1 (Ⅷ), 180 μmol∙L-1 (Ⅸ) SAHA, respectively. B: Sporulation of G. lucidum after treatment with 0.09% DMSO and different concentration of SAHA"

Fig. 5

Effect of SAHA on the production of ganoderic acid in G. lucidum"

Fig. 6

Effect of SAHA on total polysaccharide production of G.lucidum"

Fig. 7

Effect of SAHA on expression of ugp, gls and pgm gene"

Fig. 8

Effect of SAHA on the expression of hmgr, sqs, se and ls gene"

Fig. 9

Effect of SAHA on the expression of vet and LaeA"

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