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International Journal of Medicinal Mushrooms
IF: 1.423 5-Year IF: 1.525 SJR: 0.431 SNIP: 0.661 CiteScore™: 1.38

ISSN Print: 1521-9437
ISSN Online: 1940-4344

International Journal of Medicinal Mushrooms

DOI: 10.1615/IntJMedMushr.v7.i12.250
pages 263-280

Artificial Hybridization of Ganoderma lucidum and G. tsugae Murrill by Protoplast Fusion for Sustainability

Siu Wai Chiu
Department of Biology, The Chinese University of Hong Kong, Hong Kong SAR, China
Vivien Wing-Yin Luk
Department of Biology, The Chinese University of Hong Kong, Hong Kong SAR, China
Stephen Yu
Department of Biology, The Chinese University of Hong Kong, Hong Kong SAR, China
Peggy Lee
Department of Biology, The Chinese University of Hong Kong, Hong Kong SAR, China
Natalie Wai
Department of Biology, The Chinese University of Hong Kong, Hong Kong SAR, China
Polly Fu
Department of Biology, The Chinese University of Hong Kong, Hong Kong SAR, China
Karen Ka Wan Cheung
Department of Biology, The Chinese University of Hong Kong, Hong Kong SAR, China

ABSTRACT

An artificial fertile hybrid was created by protoplast fusion between a cultivar Ganoderma tsugae Murrill and a field isolate of G. lucidum (W. Curt.: Fr.) Lloyd for expanding the growth temperatures favoring production throughout the year. Actively growing mycelia were digested using 2% lywallzyme in 0.6 M mannitol to yield 228 × 103 and 178 × 103 protoplasts g−1 for G. lucidum and G. tsugae, respectively. Fusion of protoplasts was chemically induced by 30% polyethylene glycol, and the protoplast regeneration efficiency was 4.0 ± 0.2%. Positive screening was carried out by determining the survival and growth rates of the putative hybrids from 18 °C to 40 °C. After several fruiting tests, a stable hybrid was selected. Characterized by unique DNA fingerprints, it consistently produced basidiomes and basidiospores in 1.5 months using a formulated sawdust/coconut fiber substrate. Also, this artificial hybrid gained an antiproliferative property on colon cancer cells from the G. lucidum parent in an in vitro study. Oral consumption of the hybrid extract caused rapid hypoglycemic effect on BALB/c mice even after the challenge of carcinogenic streptozotocin. Also, it enhanced the recovery of cytokine expression in streptozotocin-treated mice, although there was not a horizontal transfer of an immunoprotein LZ-8-encoding gene from the parental cultivar. Thus, this study first reports the in vivo immunorecovery effect of lingzhi, demonstrates the use of protoplast fusion to cross the natural incompatibility of two biological species in the medicinal Ganoderma lucidum species complex, and demonstrates hybrid vigor for sustainable lingzhi production and commercialization.


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