程東升; 鄒莉
東北林業(yè)大學(xué); 東北林業(yè)大學(xué) 哈爾濱 150040; 哈爾濱 150040

【中文摘要】 原生質(zhì)體制備是通過體細胞雜交或基因轉(zhuǎn)導(dǎo)進行種質(zhì)改良的基礎(chǔ)。探討了糙皮側(cè)耳(Pleurotus ostreatus)原生質(zhì)體制備及再生條件。用 MYG 培養(yǎng)菌絲,以甘露醇為穩(wěn)滲劑,用1.5%溶壁酶 Lywallzyme 進行酶解脫壁,在菌齡7天、酶液 pH 值4.5、溫度28℃、酶解4hr 的條件下原生質(zhì)體產(chǎn)量最高(20×106/ml,0.01g 濕菌絲),其再生率大于8%,菌絲培養(yǎng)前對接種物進行予切碎處理可進一步使產(chǎn)量提高約2倍,而再生率并不顯著下降。酶解液 pH 值為5.5和6.5時的原生質(zhì)體產(chǎn)量低于 pH4.5,但再生率有所提高。

【英文摘要】 Preparation and culture of protoplasts are basic techniques for genetic improvement of plants and fungi through somatic cell fusion or gene transfer.Conditions for preparing the protoplasts from edible fungus Pleurotus ostreahs were examined.The highest production of protoplasts with plating efficiency of more than 8% was abtained with the enzyme mixture of pH 4.5 containing 1.5% Lywallzyme,0.6mol/L mannitol as osmotic stabilizer, and incubated at 28℃,from 7-day-age mycelia cultured in MYG medium.Shredding the inocula before mycelium incubation increased by 2 times the production of protoplasts. with plating efficiency not markedly affected.The protoplast production was lower at pH5.5 or pH6.5 than at pH4.5,but plating efficiency was increased at the former two pH value.

【中文關(guān)鍵詞】 糙皮側(cè)耳; 原生質(zhì)體制備; 細胞工程
【英文關(guān)鍵詞】 Pleurotus ostreatus; Protoplast preparation; Cytological engineering
【基金】國家教委資助優(yōu)秀年輕教師基金項目

【文獻出處】 中國食用菌,Edible Fungi of China,編輯部郵箱,1993年05期 【DOI】CNKI:SUN:ZSYJ.0.1993-05-005