韓增華; 張丕奇; 戴肖東; 孔祥輝; 馬慶芳; 張介馳
黑龍江省科學院微生物研究所
【中文摘要】 利用正交試驗研究了黑木耳原生質體制備、再生的最佳條件,結果表明,原生質體制備最佳條件是酶解溫度31℃、酶濃度1.0%、菌齡5 d、酶解時間4 h,原生質體再生的最佳條件是酶解溫度27℃、酶解時間4 h、酶濃度1.5%、菌齡11 d。穩滲劑種類對黑木耳原生質體再生率的試驗結果表明,采用0.5 mol/L蔗糖為穩滲劑,各黑木耳菌株的原生質體再生率最高。HW2號菌株原生質體經核染色其單核化比率最高;在熒光顯微鏡下觀察菌絲核相,單、雙核清晰可辨。
【英文摘要】 Conditions for the preparation and regeneration of Auricularia auricula protoplasts were optimized using orthogonal testing.Highest protoplast yields were obtained from 5-day-old mycelium after treatment with 1.0%(w/v) lywallzyme for 4 h at 31 ℃,while the highest protoplast regeneration rate was obtained from 11-day-old mycelium after treatment with 1.5%(w/v) lywallzyme for 4 h at 27 ℃.Of four osmotic stabilizers tested(sucrose,mannitol,MgSO4 and KCl),the highest rate of protoplast regeneration was obtained using 0.5 mol/L sucrose.Monokaryotic and dikaryotic mycelia were readily distinguishable using fluorescence microscopy.The highest rate of monokaryotic protoplast formation was recorded with A.auricula,strain HW2.
【中文關鍵詞】 黑木耳; 原生質體; 再生; 單核體; 熒光核染色
【英文關鍵詞】 Auricularia auricula; protoplast; regeneration; monokaryotic protoplast; fluorescence staining of nuclei
【基金】黑龍江省科學院基金"應用分子生物學技術進行黑木耳單核原生質體雜交育種"(編號:03BCCAHZH08)的部分研究內容
【文獻出處】 食用菌學報,Acta Edulis Fungi,編輯部郵箱,2008年03期 【DOI】CNKI:SUN:SYJB.0.2008-03-003