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用原生质体融合法获得稻恶苗菌异核体

HETEROKARYON FORMATION BY PROTOPLAST FUSION OF AUXOTROPH IN GIBBERELLA FUJIKUROI

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【作者】 颜耀祖李秀玉

【Author】 Yan Yaozu Li Xiuyu (College of Biology)

【机构】 北京农业大学生物学院北京农业大学生物学院

【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。

【Abstract】 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 CaCl2 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
  • 【分类号】Q78
  • 【被引频次】1
  • 【下载频次】22
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