Optimization of Protoplast Preparation and Regeneration of a Medicinal Fungus

Overview

Journal Mycobiology

Date 2020 Feb 4

PMID 32010470

Citations 11

Authors

Jyun-De Wu

Jyh-Ching Chou

Affiliations

Soon will be listed here.

Abstract

is a unique medicinal fungus in Taiwan. It has been found rich in some pharmacologically active compounds for anti-cancer, hangover, and immune regulation etc. With the in-depth study of these components, it would be interesting and important to establish a molecular system for basic studies of . Thus, we would like to set up a foundation for this purpose by studying the protoplast preparation and regeneration. Firstly, we studied the optimization method of protoplast preparation of , and found various factors that may affect the yield during protoplast preparation, such as mycelial ages, pH values, and osmotic stabilizers. Secondly, in the regeneration of protoplasts, we explored the effects of various conditions on the regeneration of protoplasts, including different media and osmotic pressure. In addition, we found that citrate buffer with pH value around 3 dramatically increased the regeneration of protoplasts of , and provided a set of regeneration methodology for .

Citing Articles

Optimization of Protoplast Preparation Conditions in and Transcriptomic Analysis Throughout the Process.

Li X, Qin Y, Kong Y, Karunarathna S, Liang Y, Xu J J Fungi (Basel). 2024; 10(12).

PMID: 39728382 PMC: 11678723. DOI: 10.3390/jof10120886.

Improved Protoplast Production Protocol for Fungal Transformations Mediated by CRISPR/Cas9 in Non-Sporulating Isolates.

Coca-Ruiz V, Cabrera-Gomez N, Collado I, Aleu J Plants (Basel). 2024; 13(13).

PMID: 38999594 PMC: 11244380. DOI: 10.3390/plants13131754.

Genome analysis of Cephalotrichum gorgonifer and identification of the biosynthetic pathway for rasfonin, an inhibitor of KRAS dependent cancer.

Schuller A, Studt-Reinhold L, Berger H, Silvestrini L, Labuda R, Guldener U Fungal Biol Biotechnol. 2023; 10(1):13.

PMID: 37355668 PMC: 10290801. DOI: 10.1186/s40694-023-00158-x.

An Overview of Mycoviral Curing Strategies Used in Evaluating Fungal Host Fitness.

Khan H, Baig D, Bhatti M Mol Biotechnol. 2023; 65(10):1547-1564.

PMID: 36841858 PMC: 9963364. DOI: 10.1007/s12033-023-00695-1.

Stable overexpression and targeted gene deletion of the causative agent of ash dieback Hymenoscyphus fraxineus.

Lutz T, Hadeler B, Jaeckel M, Schulz B, Heinze C Fungal Biol Biotechnol. 2023; 10(1):1.

PMID: 36639657 PMC: 9840287. DOI: 10.1186/s40694-023-00149-y.

References

Geethangili M, Tzeng Y . Review of Pharmacological Effects of Antrodia camphorata and Its Bioactive Compounds. Evid Based Complement Alternat Med. 2009; 2011:212641. PMC: 3095428. DOI: 10.1093/ecam/nep108. View

Lu M, El-Shazly M, Wu T, Du Y, Chang T, Chen C . Recent research and development of Antrodia cinnamomea. Pharmacol Ther. 2013; 139(2):124-56. DOI: 10.1016/j.pharmthera.2013.04.001. View

Gow N, Latge J, Munro C . The Fungal Cell Wall: Structure, Biosynthesis, and Function. Microbiol Spectr. 2017; 5(3). PMC: 11687499. DOI: 10.1128/microbiolspec.FUNK-0035-2016. View

Komoda K, Naito S, Ishikawa M . Replication of plant RNA virus genomes in a cell-free extract of evacuolated plant protoplasts. Proc Natl Acad Sci U S A. 2004; 101(7):1863-7. PMC: 357018. DOI: 10.1073/pnas.0307131101. View

Eeckhaut T, Lakshmanan P, Deryckere D, Van Bockstaele E, Van Huylenbroeck J . Progress in plant protoplast research. Planta. 2013; 238(6):991-1003. DOI: 10.1007/s00425-013-1936-7. View

Biot-Pelletier D, Martin V . Evolutionary engineering by genome shuffling. Appl Microbiol Biotechnol. 2014; 98(9):3877-87. DOI: 10.1007/s00253-014-5616-8. View

Yu P, Chang Y, Liou R, Lee T, Tzean S . pks63787, a Polyketide Synthase Gene Responsible for the Biosynthesis of Benzenoids in the Medicinal Mushroom Antrodia cinnamomea. J Nat Prod. 2016; 79(6):1485-91. DOI: 10.1021/acs.jnatprod.5b00798. View

Takeshita N . Coordinated process of polarized growth in filamentous fungi. Biosci Biotechnol Biochem. 2016; 80(9):1693-9. DOI: 10.1080/09168451.2016.1179092. View

Song T, Yen G . Antioxidant properties of Antrodia camphorata in submerged culture. J Agric Food Chem. 2002; 50(11):3322-7. DOI: 10.1021/jf011671z. View

10.

Peberdy J . Fungal protoplasts: isolation, reversion, and fusion. Annu Rev Microbiol. 1979; 33:21-39. DOI: 10.1146/annurev.mi.33.100179.000321. View

11.

Huang R, Chen C, Chen H, Hsu W, Lu M . Antrodia camphorata polysaccharides exhibit anti-hepatitis B virus effects. FEMS Microbiol Lett. 2002; 209(1):63-7. DOI: 10.1111/j.1574-6968.2002.tb11110.x. View

12.

Chen C, Yang S, Shen Y . New steroid acids from Antrodia cinnamomea, a fungal parasite of Cinnamomum micranthum. J Nat Prod. 1995; 58(11):1655-61. DOI: 10.1021/np50125a002. View

13.

Veses V, Richards A, Gow N . Vacuoles and fungal biology. Curr Opin Microbiol. 2008; 11(6):503-10. DOI: 10.1016/j.mib.2008.09.017. View

14.

Geissenhoner A, Sievers N, Brock M, Fischer R . Aspergillus nidulans DigA, a potential homolog of Saccharomyces cerevisiae Pep3 (Vps18), is required for nuclear migration, mitochondrial morphology and polarized growth. Mol Genet Genomics. 2002; 266(4):672-85. DOI: 10.1007/s00438-001-0589-6. View

15.

Hseu Y, Chang W, Hseu Y, Lee C, Yech Y, Chen P . Protection of oxidative damage by aqueous extract from Antrodia camphorata mycelia in normal human erythrocytes. Life Sci. 2002; 71(4):469-82. DOI: 10.1016/s0024-3205(02)01686-7. View

16.

Rehman L, Su X, Guo H, Qi X, Cheng H . Protoplast transformation as a potential platform for exploring gene function in Verticillium dahliae. BMC Biotechnol. 2016; 16(1):57. PMC: 4960691. DOI: 10.1186/s12896-016-0287-4. View

17.

Fuchino K, Flardh K, Dyson P, Ausmees N . Cell-Biological Studies of Osmotic Shock Response in Streptomyces spp. J Bacteriol. 2016; 199(1). PMC: 5165099. DOI: 10.1128/JB.00465-16. View

18.

Du Y, Wu T, Chang F, Lin W, Hsu Y, Cheng F . Chemical profiling of the cytotoxic triterpenoid-concentrating fraction and characterization of ergostane stereo-isomer ingredients from Antrodia camphorata. J Pharm Biomed Anal. 2011; 58:182-92. DOI: 10.1016/j.jpba.2011.09.007. View

19.

Chu Y, Yang R, Chang T, Chou J . Fructification of Antrodia cinnamomea was strain dependent in malt extract media and involved specific gene expression. J Agric Food Chem. 2009; 58(1):257-61. DOI: 10.1021/jf902793e. View