应用原生质体融合技术培育平菇与杏鲍菇优良新菌株

为了培育出具有优良互补性状的平菇和杏鲍菇新菌株,以高产、抗杂能力强的平菇品种CCEF89和优质、适应性强的杏鲍菇品种PL7为亲本菌株,建立原生质体聚乙二醇(PEG)融合体系,得到最佳融合条件:两亲本原生质体数量按1∶1混合,30% PEG 6000,0.01 mol·L^-1Ca²⁺,32 ℃水浴30 min,融合率达0.010 1%~0.028 2%。通过拮抗反应试验和Rep-PCR分子鉴定,从515个融合再生菌株中获得12个融合菌株P1~P12;出菇试验表明,P1和P5为杏鲍菇新菌株,其他为平菇新菌株。P1和P5的产量、生物学效率均显著高于其亲本菌株PL7,并且抗杂能力显著提高;与亲本菌株CCEF89相比,平菇新菌株P4、P10和P11长满培养料和形成原基的时间缩短5.5~7.3 d,第1茬平均产量提高18.0%~24.0%,总生物学效率提高21.00%~27.67%。利用原生质体融合技术培育出了具有亲本优良互补性状的平菇和杏鲍菇新菌株,为同时进行2种不同食用菌的种质资源创新和新品种选育提供了一种新途径。

Application of protoplast fusion technology to breed new strains of Pleurotus ostreatus and Pleurotus eryngii

In order to breed new strains combining the elite complementary traits of Pleurotus ostreatus and Pleurotus eryngii, Pleurotus ostreatus variety CCEF89 with the properties of high yield and strong ability to resist contamination, and Pleurotus eryngii variety PL7 with the properties of high quality and strong adaptability were used as the parent strains to build the PEG (polyethylene glycol)protoplast fusion system.Under the optimal protoplast fusion condition of 1∶1 quantity ratio of the parent strains, 32 ℃ water bath, 30% PEG 6000, 0.01 mol·L^-1 Ca²⁺ and 30 min co-incubation, the fusion rate was up to 0.010 1%-0.028 2%. 12 protoplast fusants (P1-P12) were identified by antagonistic reaction test and Rep-PCR molecular identification from 515 protoplast fusion regeneration isolates. After the mushroom cultivation experiments, P1 and P5 were identified as new strains of P. eryngii, and the other 10 protoplast fusants were new strains of P. ostreatus. The mushroom yield and biological efficiency of new strains P1 and P5 were significantly higher than those of their parent PL7, and their ability to resist contamination were increased significantly. Comparing to the parent CCEF89, the time of medium cultivation and mushroom primordia formation of P. ostreatus new strains P4, P10 and P11 were 5.5-7.3 days earlier, the yield of the first harvest mushroom and the total biological efficiency increased by 18.0%-24.0% and 21.00%-27.67%, respectively.Using the protoplast fusion technology, we acquired P. ostreatus and P. eryngii new strains with excellent complementary properties of the parents simultaneously, and proposed a new pathway to innovate new germplasm resources and bred new varieties of two different edible fungiat the same time.