【作者】 罗升辉；

【导师】 姚方杰；

【作者基本信息】 吉林农业大学 ， 蔬菜学， 2007， 硕士

【摘要】 亚侧耳[Hohenbuehelia serotina（Perts：Fr．）Sing．]，菌肉肥厚，细嫩清香，营养丰富，是东北著名的食用菌，我国已经初步栽培成功，但是目前生产中均是采用野生菌株、凭经验进行栽培管理，品质与产量很不稳定。关于菌丝培养参数、酶活参数在菌丝生长过程中的变化以及菌株选育等系统研究尚未见报道。因此，本试验借鉴其他食用菌的研究成果，以亚侧耳为供试材料，利用改良系统选择法进行品种选育；利用纯培养方法进行环境条件参数和营养条件参数的研究；研究不同培养基配方对酶活参数及优质高产参数。旨在为亚侧耳的标准化栽培提供优良品种及优质高产参数。试验结果如下：（1）采用改良系统选择法（多孢杂交），配制100个多孢杂交组合，有19个形成锁状联合；根据生理特性（拮抗）和酯酶同工酶鉴定结果从中筛选出4个遗传特性不同的杂交菌株；再通过菌丝培养特性研究，最终筛选出生长速率和生长势均强的1个优良菌株“HE-5”。（2）环境条件参数的单因子试验表明，HE-5最适生长温度为25℃，菌丝生长量达到4.77cm，高于30℃和15℃时菌丝生长不良；最适pH为6.5，菌丝生长量达到5.68cm，大于pH8.5和小于pH4.5时菌丝生长不良。（3）营养条件参数的单因子试验表明，以果糖、麦芽糖、葡萄糖、蔗糖为碳源的菌丝生长量分别达到7.05cm、6.03cm、5.46cm、6.2cm，显著高于乳糖、可溶性淀粉和白糖；以蛋白胨为氮源的菌丝生长量达到5.97cm，显著高于NaNO₃、NH4NO₃、KNO₃和麸皮液；C／N为35：1的菌丝生长量达到5.27cm，显著高于10：1和40：1的处理；MgSO₄为最佳无机盐，显著高于K₂SO₄和ZnSO₄，菌丝生长量达到5.76cm。（4）从环境条件参数和营养条件参数的单因子试验中选出最佳温度、碳源、氮源和无机盐，做四因子三水平L₉（3⁴）正交试验。结果表明，HE-5纯培养的最适宜温度为25℃，最适培养基为牛肉膏5g、麦芽糖20g、CaCI₂1g、H₂O 1L、琼脂20g。（5）不同培养基对HE-5的纤维素酶、半纤维素酶、木质素酶及淀粉酶等胞外酶活性影响不同。但是随着培养时间的延长，各种酶活参数变化趋势差异不大。其中CMC酶活性随着子实体成熟达到最高峰，而后逐渐降低；半纤维素酶、淀粉酶活性与CMC的变化趋势相反，以配方E变化幅度最大，配方C最小；Lip酶、漆酶在菌丝生长阶段一直很高，原基形成后逐渐下降。滤纸纤维素酶变化趋势的规律性不强。（6）不同培养基配方对HE-5产量及品质的影响不同。配方E（硬杂木屑50％、麸皮15％、玉米芯30％、石膏5％）显著高于其它配方，为本试验最适栽培培养基。其子实体直径达到10.1cm，颜色浅黄，畸形菇少，商品性较好，生物学效率达到105％。综上所述，本试验选育出1个亚侧耳优良菌株，明确了培养条件参数和酶活参数，提出了纯培养最佳配方和栽培料最适配方。更多还原

【Abstract】 Hohenbuehelia serotina（Perts: Fr.） Sing. is a famous native product which has flesh, good smell and plenty of nutrition. Pleurots serotinus has cultivated successfully in China but not reported on the changes of cultural characters and enzyme activity during the growth of mycelium and the selection of Pleurots serotinus strains. In this paper, Improvement system chosen method（multispore hybridization）was used to breeding from Hohenbuehelia serotina on the base of results in others edible fungus; study of nutrition and growth conditions by pure culture;Different culture mediums were investigated to choose the best, aiming to research the change of activities of the enzyme and difference of quantity and quality of mature fruitbodies. The results are as follows:（1） 100 combinations were formulas by multispore hybridization. 19 were formed catenation. Received four hybridization strains which had different inheritable characters according to physiological characteristic and results of esterase isoenzyme.As a tesult,filtering a best strain"HE-5"which had better growth potential and rate of growth.（2） The optimum temperature is 25℃and reach the maximum of 4.77cm, next is 20℃, Pleurots serotinus grows slowly under the condition of 30℃and get the minimum of 2.626cm. The most suitable pH is 6.5 and reaching the most value of 5.68cm, The worst growth is at>pH7.5.and<PH4.5.（3） The appropriate carbon sources are fructose, maltose, glucose and sucrose, the maximum is respective 7.05cm, 6.03cm, 5.46cm and 6.2cm, the minimal is 2.32cm of sucrose; The most suitable nitrogen is peptone, the maximum is 5.97cm, was higher than NaNO₃,NH₄NO₃,KNO₃ and bran. In the study of C/N, 35:1 was greater than 10:1 and 40:1. In the experiment of mineral salt, MgSO₄ was the best greater than K₂SO₄ and ZnSO4,the growth of mycelium was 5.76cm.（4）The optimum temperature, carbon, nitrogen and mineral salt chose through the test of environment and nutrition condition were used to make L9（34） orthogonal experiment. The results showed that the suitable culture medium parameters are: 25℃, 5g beef extract, 20g maltose, 21gCaCl, H₂O1L.（5）The research on the mycelium growth of No. 5 strain of the extracellular enzyme activities on different culture mediums showed that different chemicals could influence the activities of extracellular enzymes（cellulase, hemicellulase, ligninases, amylase）. The change trend of CMC enzyme is raising before the fruitbody maturing and reaching the maximum then falling gradually; hemicellulase and amylase were reverse to the CMC enzyme, E formula has biggest change and C was smallest; The content of ligninases and laccase was always high during the mycelium growth and then descends after the anlage formed.（6）The culture mediums of saw dust: 50% wheat bran, 15% corn cob30% and 5% plaster is the best parameters in this study. Its fruiting body is 8.9cm,yellow,and abnormality shortly and better selling.Its efficiency of biology is 105%.So, In this paper we breeding a better strain of Hohenbuehelia serotina, making definite the culture condition and enzyme activity, proposing the best and optimum formula.更多还原