摘要
基于对樟脑（Ｃａｍｐｈｏｒ）和苯菌灵（Ｂｅｎｏｍｙｌ）可能分别诱导细胞核膜融合和染色体不分离性重组的机理认识，设计了ＭＣＢＬ共诱导平板，以察氏（Ｃｚａｐｅｋ）培养基含有１％Ｃａｍｐｈｏｒ及０５μｇ／ｍｌＢｅｎｏｍｙｌ为基础制作平板。以属间融合组合Ａｓｐｅｒｇｉｌｕｓｎｉｇｅｒ×Ｔｒｉｃｈｏｄｅｒｍａｒｅｓｅｉ为例，研究所设计的几种诱导平板处理方法对融合子代群体的基因型和表型变异的影响，结果表明常规分步诱导处理，杂合二倍体阶段缺乏或极短暂难以获得重组单倍体；而经过ＭＣＢＬ共诱导平板处理能够获得类型齐全的分离子，显著提高表观二倍化率和染色体不分离性重组单倍体的比率。显示ＭＣＢＬ共诱导平板能够有效地扩增捕捉到极短暂杂合二倍体的机率，促进不稳定异核体向重组单倍体的转化，扩增染色体不分离性重组频率。再生菌丝菌龄对共诱导效果影响显著。由此对共诱导机制和应用前景提出讨论。
A MCB L plate was designed and constructed based on the proposed mechanism that camphor could induce the nuclear membrane fusion while benomyl induce the nondisjunctional recombination of chromosome in fungi.This so called co induction plate consisted of 0.1% camphor (W/V) and 0.5×10 -6 benmyl contained in Czapek's mimual medium.The construction procedure with cautions of this plate was developed in details.One typical example of intergeneric fusion cross, Aspergillus niger×Trichoderma reesei, was investigated,comparing the ratios of genotype and phenotype of fusant progeny caused by the co induction of MCB L plate and by the step by step induction with camphor and benomyl respectively.The results showed that the heterodiploid state was extremely transient and the recombinant haploid was scarcely obtained when induced by the routine step by step method,whereas the ratios of apparent diploidization and nondisjunctional recombinant haploidization among all types of varied segregants on MCB L plate were greatly improved compared with those on the routine plates containing single reagent,which indicated that the transient heterodiploid could be grasped and transformed further into nondisjunctional recombinant haploid when co induced by MCB L plate.The age of regenerated mycelium to be induced was observed to affect dominantly on the co induction effects of MCB L plate.The mechanism of co induction of MCB L plate and its perspective applications were also discussed.
出处
《生物工程学报》
CAS
CSCD
北大核心
1998年第3期265-269,共5页
Chinese Journal of Biotechnology
基金
国家自然科学基金
中国博士后科学基金
国际科学基金
广东省自然科学基金
关键词
染色体重组
共诱导平板
真菌
构建
Chromosome recombinantion,co induction,camphor,benomyl, Aspergillus niger,Trichoderma reesei
