【作者】 邢蕾；

【导师】 李明；

【作者基本信息】 河北农业大学 ， 蔬菜学， 2008， 硕士

【摘要】 平菇（Pleurotus ostreatus）是一类重要的食用菌,其适应性强,栽培原料来源广,产量高,效益好,近年来发展迅猛,已成为城乡人民大众化的菌类蔬菜。但是,目前平菇产业发展面临着品种由于长期无性繁殖而退化明显、产量不稳、育种手段单一等实际问题,为了提高育种的效果和加速品种更新换代,本实验采用原生质体再生技术进行平菇新菌株的选育,对平菇原生质体制备及再生条件进行优化,将得到的再生菌株进行初步筛选的一系列试验,旨从细胞水平上探索一条食用菌育种新途径,为发展和完善食用菌新菌株选育提供依据。1.本试验对影响平菇原生质体制备及再生条件的几大因素综合酶种类、酶浓度、菌龄、酶解温度、酶解时间、稳渗剂及培养基等条件进行了研究,结果表明,以PDA液体综合培养基培养5d的幼嫩菌丝体,单一酶以2%浓度的溶壁酶或混合酶以1%的纤维素酶和1%的蜗牛酶,在30℃下作用3h,以0.6mol/L的MgSO₄为渗透压稳定剂是制备平菇原生质体的最佳条件;以菌龄为3d,酶解时间为1h,0.6mol/L的MgSO₄为渗透压稳定剂的PDA再生综合培养基为原生质体再生的最佳条件,再生率最高达到4.35%。2.采用优化的条件进行原生质体的制备与再生,最终再生出98个再生菌株,对再生植株菌丝体进行镜检,获得47个具有锁状联合的菌株。将再生菌株分别与出发菌株进行拮抗试验,获得39个再生菌株菌丝与其亲本有明显的拮抗线。3.通过对39个再生菌株与出发菌株的菌丝颜色、密度、生长势等方面的比较试验,得到各方面优于出发菌株的再生植株。在平板中进行菌丝长速测定发现,各菌株间长速有差异性,7、11、13、17、26、36、37高于出发菌株,生长速度与生长势有相关性。对栽培种进行长速测定发现,各菌株差异显著,而且和平板中菌丝长速相关,平板中长速快的,栽培种的长速也快。4.抗绿霉试验获得在抗杂能力优于出发菌株的再生菌株,通过抗绿霉试验,发现1、3、7、14、18、24、25、26、30、33、37、38这几个再生菌株与绿霉的拮抗线宽于出发菌株,说明其抗杂能力比出发菌株高,长势好的菌株抗杂能力也强。可作为进一步筛选的依据。更多还原

【Abstract】 Pleurotus ostreatus is an important type of edible fungus,it is strong adaptability, widely cultivated sources of raw materials,high yield and efficiency with rapid development in recent years,it has become a popular vegetable fungi for the urban and rural people.However,due to long-term asexual breeding,the Pleurotus ostreatus industry development is faced the apparent degradation,lack of resources,output is low and unstable,a single breeding of practical issues.In order to enhance the effectiveness and speed of the breeding new strains,the experimental breed the new strain of Pleurotus ostreatus by renewable protoplast technology,protoplast preparation and regeneration condition,renewable strains of screening,such as a series of re-screening test,purport from the cellular level to explore a new way Mushroom breeding for the development and improvement of food Selection of new strains provide the basis.Protoplast preparation and regeneration condition,renewable strains of screening,such as a series of re-screening test, purport from the cellular level to explore a new way mushroom breeding for the development and improvement of food selection of new strains provide the basis.1.We have studied the condition of the preparation of Pleurotus ostreatus protoplast in many details and it shows that using the mycelia cultrued in PDA medium at the age of 5 days,2%lywallzyme or the mixture of 1%cellulas and 1%snailas under the condition as 30℃,3h enzyme action with 0.6mol/L MgSO₄ as osmotic stabitizor,pH is 6.5 is the suitable condition of preparing protoplast ofPleurotus ostreatus.The Pleurotus ostreatus the age of hyphe was 3 days,the digesting time wasl hour,the osmotic stabilizer was 0.6 mol/L MgSO₄.the frequency of regeneration was the best and resch up 4.35%.2.Under the best condition of separation and regeneration,we get 98 regenerative strains,.Observed under the microscope,47 strains have clampconnection.The result of antagonist response shows that 39 strains are different from original strains.3.Through mycelial color,mycelial growth,mycelial density compared experiment,from 39 regenerative strains we get some strains such as 7,11,13,17,26,36,37 have stronger growing trend,biger density than original strains and they grow more quickly.Different cloture spawn has different growing state,and they have positive correlation with mother cultures stains.4.Compared with contaminated rate between regenerative strains and original strains,1,3,7,14,18,24,25,26,30,33,37,38 have stronger ability of anti-bacteria capacity.It can be used as the basis for further selection.更多还原