摘要
用紫外线照射稻恶苗菌(Glbberella fujikuroi)野生型分生孢子9～12 min,存活率为0.6%～23%,得到单标记营养缺陷型突变菌株5株。选出1144(ade^-)、1184(arg^-)和1175(try^-)3株,用亚硝基胍诱变处理,得到双重标记的营养缺陷型突变菌8株。将11442(ade^-,pba^-)和11844(arg^-,try^-)两菌株用原生质体融合技术使它们形成异核体。用2%蜗牛酶加2%纤维素酶以0.6mol/L NaCl 液作渗透压稳定剂制成混合酶液,裂解幼嫩菌丝1.5～3 h,获得2×10'个/g 菌丝的原生质体。经35%PEG 融合剂将等量原生质体融合,获得异核菌落230个。经过进行排除营养互养的实验;对4株异核体菌株进行子代分离的遗传学分析;发现其两种核型的比例在1:1.4～1:8.5之间;对4株异核体还进行酯酶同功酶的测定。证明这4株菌株是异核体。
Five auxotrophic mutants of G.fujikuroi were induced with UV light for 9～12 min expourse(0.6～2.3% of survival).Three of them 1144(ade^-) 1184 (arg^-)and 1175 (try^-)were rautagenized with NG and eight biauxotrophwere obtained.Protoplasts were produced by digesting10～15 hour myceliura with 2% sna- lase plus 2% celllase under the condition of 0.6 mol/L NaCl as stabilizer.Protoplasts from 11844(.arg^-,try^-)and 11442(ade^-,pba^-)were fused in the presence of PEG and CaCl_2 Four of the fusion products were tested for their heterokarysis by cross- feeding experiment isozyme analysis and progeney analysislt.proved that they we- re all heterokaryons and the ratio of two type nuclei was 1:1.4 to 1:8.5.
基金
国家自然科学基金项目之一
关键词
稻恶苗菌
原生质
营养缺陷
Gibberella fujikuroi
auxotroph
protoplast fusion
isozyme
