Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

A novel breeding strategy for new strains of Hypsizygus marmoreus and Grifola frondosa based on ligninolytic enzymes

  • 482 Accesses

  • 5 Citations


A novel breeding strategy for new strains of Hypsizygus marmoreus and Grifola frondosa using ligninolytic enzymes as markers was evaluated with the detection and analysis of activities and composition of 15 edible fungi. The results showed that the activity and composition of ligninolytic enzyme system varied in response to changes of fungal strains. By analyzing the growth rate of mycelia and their ability to produce ligninolytic enzymes, H. marmoreus and P. geesteranus, G. frondosa and P. sajor-caju were screened for further study. Three colonies of 26 regenerated colonies of H. marmoreus and P. geesteranus protoplast fusion and one colony of 48 regenerated colonies of G. frondosa and P. sajor-caju were selected respectively. At the same time, these four strains were identified using RAPD and ISSR molecular markers. The results showed that the strains HM5G1and PS7F1are new strains and have low similarity to parental strains H. marmoreus and G. frondosa. These results are supported by the results of antagonism tests. These two fusants were significantly higher in their ligninolytic enzyme activity than H. marmoreus and G. frondosa. The growth rates of strains HM5G1and PS7F1 were also noticeably higher than those of H. marmoreus and G. frondosa, by 1.36 and 1.5 times respectively. The biological efficiency of the strain HM5G1 was 11.5 % higher than that of the parental strain H. marmoreus. This work suggests that it is an efficient way of breeding new strains to use the decolorization of ligninolytic enzymes as a preliminary screening marker.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7


  1. Baldrian P (2006) Fungal laccases—occurrence and properties. FEMS Microbiol Rev 30:215–242. doi:10.1111/j.1574-4976.2005.00010.x

  2. Bourbonnais R, Paice MG (1990) Oxidation of non-phenolic substrates. An expanded role for laccase in lignin biodegradation. FEBS Lett 267:99–102. doi:10.1016/0014-5793(90)80298-W

  3. Glenn JK, Akileswaran L, Gold MH (1986) Mn(II) oxidation is the principal function of the extracellular Mn-peroxidase from Phanerochaete chrysosporium. Arch Biochem Biophys 251:688–696. doi:10.1016/0003-9861(86)90378-4

  4. Hammel KE, Tardone PJ, Moen MA, Price LA (1989) Biomimetic oxidation of nonphenolic lignin models by Mn(III): new observations on the oxidizability of guaiacyl and syringyl substructures. Arch Biochem Biophys 270:404–409. doi:10.1016/0003-9861(89)90044-1

  5. Hatakka A (1994) Lignin-modifying enzymes from selected white-rot fungi: production and role from in lignin degradation. FEMS Microbiol Rev 13:125–135. doi:10.1111/j.1574-6976.1994.tb00039.x

  6. Hu KH, Chu XP (2008) Effect of several factors on mycelial characteristics and fruiting quality and biological efficiency of Hypsizygus marmoreus. Edible Fungi China 27:16–18

  7. Hu KH, Huang GY, Yan S, Zhuang YH, Wei XF, Zhu HX (2009) Differential proteomics and enzyme changes of Hypsizigus marmoreus mycelium under cold stress. Mycosystema 28:584–590

  8. Kodama N, Mizuno S, Nanba H, Saito N (2010) Potential antitumor activity of a low-molecular-weight protein fraction from Grifola frondosa through enhancement of cytokine production. J Med Food 13:20–30. doi:10.1089/jmf 2009.1029

  9. Kubo K, Nanba H (1997) Anti-hyperliposis effect of maitake fruit body (Grifola frondosa). Biol Pharm Bull 20:781–785. doi:10.1111/j.1600-0625.2008.00831.x

  10. Lee J, Kang HW, Kim SW, Lee CY, Ro HS (2011) Breeding of new strains of mushroom by basidiospore chemical mutagenesis. Mycobiology 39:272–277. doi:10.5941/MYCO.2011.39.4.272

  11. Leonowicz A, Cho NS, Luterek J, Wilkolazka A, Wojtas-Wasilewska M, Matuszewska A, Hofrichter M, Wesenberg D, Rogalski J (2001) Fungal laccase: properties and activity on lignin. J Basic Microbiol 41:185–227. doi:10.1002/1521-4028(200107)41:3/43.0.CO;2-T

  12. Levin L, Melignani E, Ramos AM (2010) Effect of nitrogen sources and vitamins on ligninolytic enzyme production by some white-rot fungi. Dye decolorization by selected culture filtrates. Bioresour Technol 101:4554–4563. doi:10.1016/j.biortech.2010.01.102

  13. Niladevi KN, Sukumaran RK, Jacob N, Anisha GS, Prema P (2009) Optimization of laccase production from a novel strain-Streptomyces psammoticus using response surface methodology. Res Microbiol 164:105–113. doi:10.1016/j.micres.2006.10.006

  14. Nishiwaki T, Hayashi K (2001) Purification and characterization of an aminopeptidase from the edible basidiomycete Grifola frondosa. Biosci Biotechnol Biochem 65:424–427. doi:10.1271/bbb.65.424

  15. Obi N, Hayashi K, Miyahara T, Shimada Y, Terasawa K, Watanabe M, Takeyama M, Obi R, Ochiai H (2008) Inhibitory effect of TNF-alpha produced by macrophages stimulated with Grifola frondosa extract (ME) on the growth of influenza A/Aichi/2/68 virus in MDCK cells. Am J Chinese Med 36:1171–1183. doi:10.1142/S0192415X08006508

  16. Osma JF, Toca-Herrera JL, Rodriguez-Couto S (2010) Transformation pathway of Remazol Brilliant Blue R by immobilised laccase. Bioresour Technol 101:8509–8514. doi:10.1016/j.biortech.2010.06.074

  17. Rodriguez Couto S, Sanroman M, Gubitz GM (2005) Influence of redox mediators and metal ions on synthetic acid dye decolourization by crude laccase from Trametes hirsuta. Chemosphere 58:417–422. doi:10.1016/j.chemosphere.2004.09.033

  18. Sedlak M, Ho NW (2004) Production of ethanol from cellulosic biomass hydrolysates using genetically engineered Saccharomyces yeast capable of cofermenting glucose and xylose. Biotechnol Appl Biochem 113–116:403–416. doi:10.1016/j.biortech.2010.11.034

  19. Shervedani RK, Amini A (2012) Direct electrochemistry of dopamine on gold-Agaricus bisporus laccase enzyme electrode: characterization and quantitative detection. Bioelectrochemistry 84:25–31. doi:10.1016/j.bioelechem.2011.10.004

  20. Sun SJ, Gao W, Lin SQ, Zhu J, Xie BG, Lin ZB (2006) Analysis of genetic diversity in Ganoderma population with a novel molecular marker SRAP. Appl Microbiol Biotechnol 72:537–543. doi:10.1007/s00253-005-0299-9

  21. Sun SJ, Liu JZ, Hu KH, Zhu HX (2011) The level of secreted laccase activity in the edible fungi and their growing cycles are closely related. Curr Microbiol 62:871–875. doi:10.1007/s00284-010-9794-z

  22. Yeh JY, Hsieh LH, Wu KT, Tsai CF (2011) Antioxidant properties and antioxidant compounds of various extracts from the edible basidiomycete Grifola frondosa (Maitake). Molecules 16:3197–3211. doi:10.3390/molecules16043197

Download references


This work was financially supported by the Natural Science Foundation of China (No. 31201669), General Program of the Provincial Education Department of Fujian Province of China (No. JA11079), Special Project of Fujian Biological Industry Technology (No. 2011878), “Wu Xin” Project of Agriculture of Fujian Provincial Development and Reform Commission of China ([2012]931), and Research Funds for Distinguished Young Scientists in Fujian Agriculture and Forestry University (xjq201202 and xjq201209).

Author information

Correspondence to Kaihui Hu.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Sun, S., Li, X., Ruan, L. et al. A novel breeding strategy for new strains of Hypsizygus marmoreus and Grifola frondosa based on ligninolytic enzymes. World J Microbiol Biotechnol 30, 2005–2013 (2014). https://doi.org/10.1007/s11274-014-1624-1

Download citation


  • Hypsizygus marmoreus
  • Grifola frondosa
  • Ligninolytic enzymes
  • Breeding
  • Molecular markers