Study On Protoplast Preparation Regeneration And Fusion Of Different Pleurotus Varieties

Protoplast fusion is cell engineering breeding technology,which is developing rapidly in recent years.It can use germplasm resources across species and category.At the same time,it can shorten breeding cycle and open a new way for distant hybridization.,.Excellent protoplasted monokaryons of Pleurotus abalonus were selected based on the protoplast technology among Pleurotus cultivated species(Pleurotus ostreatus?Pleurotus geesteranus and Pleurotus abalonus)by single factor experiment.This paper will contribute to improving Pleurotus strains breeding and providing theoretical basis and technical reference.?The paper studied the preparation and regeneration of Pleurotus ostreatus on fungus age,osmotic stabilize,enzymatic hydrolysis time,biomass of mycelium and the cultured time.The results showed that the 0.03g mycelium were added into 1 mL 2%enzyme hydrolysis solution and 0.6mol/L sucrose as osmotic stabitizer which had been cultured in liquid PDA medium for 7 days.These mycelium was treated 2.5h under 30?and the number of protoplast reached to2.6×10~7/ml.The frequency of protoplast regeneration can reach up to 6%?The paper studied the preparation and regeneration of Pleurotus geesteranus on fungus age,osmotic stabilize,enzymatic hydrolysis time,biomass of mycelium and the cultured time.The results showed that the 0.04g mycelium were added into 1 mL 2%enzyme hydrolysis solution and 0.6mol/L sucrose as osmotic stabitizer which had been cultured in liquid PDA medium for 6days.These mycelium was treated 2h under 35?and the number of protoplast reached to 2.4×10~7/ml.The frequency of protoplast regeneration can reach up to0.7%.?The paper studied the preparation and regeneration of Pleurotus abalonus on fungus age,osmotic stabilize,enzymatic hydrolysis time,biomass of mycelium and the cultured time.The results showed that the 0.04g mycelium were added into 1 m L 2%enzyme hydrolysis solution and 0.6mol/L sucrose as osmotic stabitizer which had been cultured in liquid PDA medium for 12 days.These mycelium was treated 2h under 30?and the number of protoplast reached to 1.2×10~7/ml.The frequency of protoplast regeneration can reach up to 0.3%.Excellent protoplasted monokaryons of Pleurotus abalonus were selected based on the protoplast technology among Pleurotus cultivated species(Pleurotus ostreatus?Pleurotus geesteranus and Pleurotus abalonus).lt took Pleurotus ostreatus and Pleurotus abalonus monokaryontic strain as parent material,Using a fusion agent PEG4000 for fusion.Finally,this experiment acquired two strains of fusion named R1,R2.Esterase isozyme zymogram demonstrated that R1 and R2had specific electrophoretic belt of parents.The results of ISSR analysis showed that two strains of fusion contained genetic material of both parents.