| Lentinula Edodes industry,one of the most important pillar in Chinese edible fungus,plays an significant role in adjusting rural economic and industry stucture and in promoting rural economy development.Although China has produced a large number of the Lentinus Edodes,but commercial process is very low.Lagging of breeding work has a certain restriction on the development of Lentinula Edodes industry.Therefore,breeding work is imminent.This paper,researching work starts on the YJ-01 and thestrains which from laboratory.In order to obtain high yield and quality Lentinula Edodes new strains,using protoplast mutagenesis and single spore hybridization technology.The main findings are as follows:1.The optimum conditions for the protoplast preparation of Lentinus Edodes strains YJ-01 were settled: pH=5.0,0.5 mol L-1 MgSO4 as stabilizing agent,mixed enzyme(1% snail enzyme +1% cellulase)to enzyme hydrolysis of 5th-day mycelium for 2 h at 27.9?,obtain the highest protoplast production.2.The optimum conditions for protoplast regeneration of Lentinus edodes strains YJ-01 were settled: pH=5.5,0.5 mol L-1 sucorse as stabilizing agent,using mixed enzyme(1% snail enzyme +1% cellulase)to enzyme hydrolysis of 6.5th-day mycelium for 2.9 h at 27.8?,obtain the highest protoplast regeneration rate.3.547 regenerated strains were obtained by UV mutagenesis of protoplast.(1)17 strains of fine regenerated strains were obtained by screening with different cultivated materials.Strain Y209 is most suitable for the apple sawdust,the growth index(79.88)is 3.56 times higher than that of YJ-01;Strain Y72 is most suitable for willow wood sawdust,the growth index(70.63)is 3.67 times higher than that of YJ-01;Strain Y369 is most suitable for cleyera sawdust,the material connecting quantity(338.24 g)with daily average connecting quantity 7.87 g d-1,2.76 times higher than that of YJ-01.(2)Continuous subculture of 17 excellent regenerated strains and test if fruitbodies could carry out in liquid medium.The results showed that strain Y150 has the most stable mycelial physiological properties,and strains Y440,Y365,Y314,Y243 and Y283 could form the fruitbodies in the liquid medium.(3)Using isozyme and RAPD to identify these 17 of excellent regenerated strains,results showed that strains Y263,Y440,Y314,Y454,Y381,Y317,Y211,Y150 and Y463 were variant strains.By comprehensive comparison,the mutant strains Y440 and Y314 were identified as excellent mutants.4.23 species were collected and 1166 strains were isolated from 18 strains and 1367 strains of hybrid strain were obtained according to hybridization(1)The physiological properties of 238 hybrid strains of 9608 and 808 strain were screened.18 strains of dominant strains were obtained(B80D111,B80D20,B46D111,B10D175,B6D178,B80D51,B70D21,B6D125,B60D111,B81D111,B60D99,B10D167,B9D176,B76D20,B6D149,B27D21,B81D99 and B46D20).Among which,strain B60D111 is the best in cleyera sawdust,the material connection quantity(261.54 g),with daily average connecting quantity 6.08 g d-1,is 1.29 times of the parents 9608-1,1.6 times of 808-1.B70D184 performed optimally on PDA tablets,the growth index(32.73)was 2.42 times and 3.32 times than that of parents 808-1 and 9608-1.(2)Continuous subculture of 18 excellent regenerated strains and test if fruitbodies could carry out in liquid medium.The results showed that strain B6D178 has the most stable mycelial physiological properties,and strains B10D167 and B10D175 could form the fruitbodies in the liquid medium.(3)Using isozyme and RAPD to identify these 18 of excellent regenerated strains,results showed that strains B60D99,B6D149,B60D111,B27D21,B10D175,B10D167,B80D111,B80D51 and B70D20 were variant strains.By comprehensive comparison,B10D175 was identified as excellent hybrid strain.(4)Screening of the remaining 1129 strains for high temperature resistance.The results showed that after high temperature treatment,133 strains of hybrid strains were able to restore growth,of which F149B92?F`47B90?A59D97 and other 20 strains can grow under 35?.After high temperature treatment and 6 d 25? recovery,the growth of 26 hybrid strains is still good,of which F149B92 shows best,the growing rate relative to the processing before the decrese 65.99%,after the F203B92(38.42%),F98B92(64.19%)and F133B92(49.08%).F203B92?F149B92?F133B92?F98B92 and F128B66 were determined to be high temperature resistant strains. |
PDF Full Text Request