Studies On Cell Engineering Breeding And Semi-wild Planting Of Stropharia Rugoso-annulata

Stropharia rugoso-annulata is a kind of rare edible mushroom which is tasting delicious and being rich in nutriments.It can decomposes and makes use of angriculture and forestry waste such as straw and leaf.Stropharia rugoso-annulata protoplast preparation and regeneration was elementarily studied.The optimal conditions of protoplast preparation and regeneration was as follows: 1.5% cellulase + 1.5% snailase mixed enzyme (volume ratio was 1:1) to zymolyse hypha, 0.6 mol/L mannitol as osmotic pressure stabilizer, best cell age for 3 days, the optimal enzymolysis temperature for 30℃, 2.5h being suitable most for zymolysing .The highest yield can reach 1.70×10~7/mL under these conditions.The best osmotic pressure stabilizer for regeneration medium was mannitol and on this occasion the regeneration rate could reach to 0.89%.The best mutugenesis time was 35 seconds whose fatality rate was 73.33% for Stropharia rugoso-annulata protoplast.Exellent regeneration strain was easier obtained on this dose.A technology of protoplast combined with uv mutagenesis was used. An excellent variation strain Dq084 was obtained which was good at planting and fermentation through the speed and site on various planting material and measuring consumption ratio of reducing sugar, biomass, mycelium ball growing status, various exocellular enzyme activity in the liquid medium. Its routine planting biological efficiency was 64.43% obviously higher than the original strain.Its biomass, consumption ratio of reducing sugar and extracellular enzyme activity were higher than the original strain so it could resolve fibrin and lignin and so on. The result also showed that there was definite positive correlation between the activities of extracellular cellulose enzyme and mycelial growth rate.It was concluded that adding 0.2% xanthan gum could increase the mycelium ball density and quantities of liquid culture through the thickener experiment.The electrophoresis result was prominently different between Dq084 and the original strain by esterase isozyme measuring.This proved that genetic variation has occured to Dq084 strain and Dq084 strain was a good regetation variation strain which was excellent on erery aspects.Two kind of semi-wild planting patterns ridge cultivation and bag cultivation were searched through the new strain preparation technology of Dq084-liquid cultivation-grain nature.It was concluded that corn straw nature bag cultivation was fit for planting in winter and sping whose mycelium growed faster, growing period was less and biological conversion rate reached up to 70.41% (The contrast was 60.96%)and wheat strain nature ridge cultivation was fit for planting in summer and autumn whose biological conversion rate was 68.64%(The contrast was 57.33%).