Regeneration and Mating Type Determination of Protoplast of Pleurotus ferulae

【Objective】 To obtain a new strain of Pleurotus ferulae with excellent traits by the protoplast monokaryotation technology in the hope of laying the theoretical and technical foundation for the selection and industrial production of new varieties of wild P. ferulae in Xinjiang. 【Method】 The protoplasmic preparation technique was used to obtain the mononuclear strain of P. ferulae and determine its mating type. 【Result】 At 3.5 h after enzymolysis, the protoplast release of the 3 wild P. ferulaeparent strains PL01, PL163 and PL176 was the maximum, with the concentration as high as 20.6 × 10⁵ /mL. Through optimized protoplast monokaryonization, 50 mononuclear strains were finally obtained from the 3 parent strains. The protoplast monokaryonization rates of parent strains PL163, PL01 and PL176 were 25.68%, 12.8% and 10.8%, respectively, with a low acquisition rate. Then, the mating type was determined, and mononuclear strains with 2 mating types were obtained from each parent strain. Among them, the acquisition ratio of the 2 mating types from PL01 and PL163 strains was 1∶3, while that of PL176 was 1∶4. Differences were found in mycelial growth rate and colony morphology among each parent strain and the obtained mononuclear strains. Subsequently, pairing was performed using mononuclear strains PL01-P16, PL001-P49, PL163-7, PL163-24, PL176-23 and PL176-39 with different mating types as hybrid breeding materials, and 12 hybrid strains with excellent traits were obtained. The antagonistic reaction showed that the 12 hybrid strains formed an obvious uplifted antagonistic band between their parents, and were significantly different from their parents. 【Conclusion】 The 3 wild P. ferulae strains present rich diversity in protoplast regeneration, mononuclear acquisition rate, growth rate and morphology. The obtained hybrid strains lay a foundation for the conservation of new varieties of P.