Generation of nuclear hybrids overcoming the natural barrier of incompatibility: Transfer of nuclei fromLentinula edodes into protoplasts ofCoriolus versicolor

Abstract

Heterokaryotic nuclear hybrids overcoming the natural barriers of incompatibility have been studied in basidiomycetes. To produce these nuclear hybrids between incompatible mushrooms, which have several potent pharmacological effects, nuclear transfer was performed between Lentinula edodes and Coriolus versicolor. Nuclei from serine auxotrophs of Lentinula edodes, LE207 (Ser-) were transferred into the protoplasts of arginine auxotrophs of Coriolus versicolor, CV17 (Arg-), using 30% polyethylene glycol 4000 in 10 mM CaCl2-glycine solution (pH 8.0). Nuclear transfer progenies were selected by nutritional complementation on minimal media supplemented with 0.6 M sucrose. The progenies were classified based on colony morphology to L. edodes-like, C. versicolor-like and non-parental type. Most of the progenies grew slower than either parent. The number of nuclei per cell was similar but the DNA content varied between progenies. The isozyme patterns of nuclear hybrids resembled either of the parent profiles or showed a mixed profile.

Ganoderma Fusions with High Yield of Ergothioneine and Comparative Analysis of Its Genomics

“…This investigation revealed varying DNA contents among the hybrids, while the number of nuclei per cell remained consistent. Notably, certain hybrids exhibited over three times the DNA content of the parent strains, substantiating the existence of nuclear hybridization [48]. Dudits et al [49] investigated the phenomenon of gene transfer after the fusion of parsley and carrot.…”

Ultrastructural Process of Protoplast Fusion BetweenLentinula edodesandCoriolus versicolor

“…Protoplasts were prepared from L. edodes and C. versicolor as previously described by Kim et al (Kim and Kim, 1997a;Kim et al , 1997cKim et al , , 2000. Briefly, the mycelia were treated with 10 mglml Novozyme 234 (Novo, Bagsvaerd, Denmark) and 10 mglml Cellulase Onozuka (Yakult, Japan) at 30oC.…”

“…versicolor) and to generate the strains havi ng combined or synergistic pharmacological effects, we pe1formed protoplast fusion between L. edodes and C. versicolor (Kim et al, 1997b, c;2000). Our success in the production of hybrids between L. edodes and C. versicolor opened up possibilities for generati ng hybrids between genetical ly unrel ated orders as well as understanding the genetic mechani sm of fungi.…”