馬愛民 ; 賀冬梅 ; 潘迎捷
上海市農(nóng)業(yè)科學(xué)院食用菌研究所 ; 農(nóng)業(yè)部食用菌遺傳育種重點實驗室 上海 201106

【中文摘要】 通過對溶壁酶組成及濃度,滲透壓穩(wěn)定劑、菌絲培養(yǎng)方法等研究,建立了有效的雙孢蘑菇原生質(zhì)體制備與再生系統(tǒng)。采用1.5％Lywallzyme、0.6M KCl為滲透壓穩(wěn)定劑,25℃酶解4小時,原生質(zhì)體的產(chǎn)量可達到107個/ml;純化后的原生質(zhì)體在0.6M蔗糖為滲透壓穩(wěn)定劑的PDMA培養(yǎng)基中,25℃恒溫培養(yǎng)5～7天,再生率達1%以上,根據(jù)原生質(zhì)體再生菌落出現(xiàn)時間的先后順序,并以菌落形態(tài)特征、菌絲生長速度。羧甲基纖維素酶(Cx酶)活性及子實體形成能力上的差異為鑒定同核體的主要標記,從兩種形態(tài)類型,不同來源的12個雙孢蘑菇菌株中均分離到同核原生質(zhì)體,同核率為2％～15%,平均為11.75％。

【英文摘要】 Based on testing of different combinations and concentration of lysozymes, osmotic stabilizers and different methods of mycelium incubation, an efficient system of protoplast preparation and regeneration in Agaricus bisporus was established in present paper. It was proved that after the mycelium being digested with 1.5% lywallzyme in 0.6M KCl for 4h at 25℃, the protoplast yield reached 107/ml, and that after the purified protoplasts being incubated in PDMA medium with 0.6M sucrose as osmotic stabilizer for 5-7 days at 25℃, the regeneration rate was over 1%. Based on different regeneration time, using the difference of colonial morphology, activity of Cx enzyme and fruiting ability as main markers for identification of homokaryotic protoplast, the homokaryotic protoplasts were isolated from protoplasts of 12 strains with a rate of 2% to 15%, averaging out at 11.75%.

【中文關(guān)鍵詞】 雙孢蘑菇; 同核原生質(zhì)體; 分離與鑒定
【英文關(guān)鍵詞】 Agaricus bisporus; Homokaryotic protoplast; Isolation and identification
【基金】上海市科委資助項目

【文獻出處】 食用菌學(xué)報,Acta Edulis Fungi,編輯部郵箱,1995年03期 【DOI】CNKI:SUN:SYJB.0.1995-03-000