香菇金针菇远缘亲本原生质体融合及融合子检测

PROTOPLAST FUSION BETWEEN LENTINUS EDODES AND FLAMMULINA VELUTIPES AND FUSANT IDENTIFICATION

采用１５ｇ／Ｌ溶壁酶处理获得了香菇单核体Ｌ３８－Ｓ０３、Ｌ５２－ａｄｅ和金针菇单核体ＦＶ１１－Ｓ２３的原生质体。１ｇ／Ｌ焦磷酸二乙酯灭活的Ｌ３８－Ｓ０３和腺嘌呤缺陷型Ｌ５２－ａｄｅ的原生质分别与８ｇ／Ｌ碘代乙酰胺灭活的ＦＶ１１－Ｓ２３的原生质体进行ＰＥＧ－Ｃａ２＋促融合，在ＲＣＭ、ＩＲＭＭ培养基上再生，得到１４２个融合子。其中１７个菌株能结实出菇。筛选出Ｆ０１、Ｆ０２和Ｆ０３３个融合子菌株，并从培养特征、显微结构、拮抗反应、同工酶谱分析和出菇特征等方面与亲本菌株Ｌ３８、Ｌ５２及ＦＶ１１进行了比较，结果显示融合子既有双亲菌株的特征，又有别于双亲菌株。

Lywallzyme 15 g/L was used to obtain protoplast from Lentinus edodes monocaryons L 38 S 03 and L 52 ade and Flammulina velutipes monocaryon F V11 S 23 . Purified protoplasts were inactivated by metabolic inhibitors. 8 g/L iodoacetamide at 22 ℃ for 5 min and 1 g/L dithy myrocarbonate at 4 ℃ for 30 min were found to be satisfactory for inactivating F V11 S 23 and L 38 S 03 protoplasts. Protoplast fusion was conducted with PEG Ca 2+ system. Regeneration of fusants from L 38 S 03 ×F V11 S 23 was carried out on double layer RCM medium and L 52 ade ×F V11 S 23 on double layer IRMM medium. 17 out of 142 fusants formed fruit bodies on saw dust medium.