Analysis of the mitochondrial and nuclear DNA of basidiomycete heterokaryon.

对影响菌丝体原生质体分离及原生质体再生和回复的一些因素进行了研究。auricula-judae和A. polytricha进行了调查。酶系和菌龄对原生质体分离有影响。在Novozym 234 1%和几丁质酶0.1%的组合中，以培养4天的菌丝体为材料，原生质体产量最高。黑木耳（auricula-judae）和6日龄黑木耳（A.用培养4天的毛木耳菌丝体分离的原生质体，获得了较高的原生质体回复率。黑木耳（auricula-judae）和6日龄黑木耳（A.为了通过原生质体融合选择杂种细胞，用两种不同的代谢抑制剂碘乙酸钠盐（TAS）和焦碳酸二乙酯（DP）处理两个单核菌株的原生质体。从原生质体的破碎率和回复率的结果来看，每一单核菌株（Mo A 11和Mo P 16）的IAS和DP浓度均为4%，原生质体融合率最高的条件是10 /. tsee，275次，间隔约1 see，场强为3 μ m/em，对电融合获得的融合子进行了研究，以阐明其特性（Mo A 11和Mo P 16）和5个融合子，所有融合子对Mo A 11和Mo P 16均形成拮抗线。经同工酶分析，具有单核菌株的两条特异性带。通过限制性内切酶分析，确定Fu 9线粒体DNA的分子大小为85.8kbp。5株融合子均具有线粒体质粒样DNA，脉冲场电泳结果表明，亲本、单核株和融合子均具有4条染色体DNA，说明5株融合子均为异核体。

Some factors affecting the isolation of protoplasts from mycelia and the regeneration and reversion of isolated protoplasts for A. auricula-judae and A. polytricha were investigated. Protoplast isolation was affected by enzyme system and mycelial age. Maximum yield of protoplasts was obtained by a combination of Novozym 234 1%, and chitinase 0.1%, and by using mycelia of 4-day-old culture for A. auricula-judae and that of 6-day-old culture for A. 'polytricha.High protoplast reversion was obtained by using protoplasts isolated from mycelia of 4-day-old culture for A. auricula-judae and that of 6-day-old culture for A. polytricha.In order to select hybrid cells by protoplast fusion, the protoplasts of both monokaryotic strains were treated with two different metabolic inhibitors, iodoacetic acid sodium salt(TAS)and diethylpy rocarbonate(DP). From the results of broken and reversion frequencies of protoplasts, the concentrations of IAS and DP were found to be 4% for each monokaryotic strain(Mo A 11 and Mo P 16).The condition which gave the largest fusion frequency of protoplasts was a DC pulse of 10 /. tsee, 275 times with intervals of approximately I see, and a field strength 3 kv/em.Fusants obtained by electrical protoplast fusion, were investigated in order to clarify their characteristics.In paired cultures between monokaryotic strains(Mo A 11 and Mo P 16)and 5 fusants, all the fusants formed antagonistic lines against both Mo A 11 and Mo P 16.All 5 fusants were found, by analyzing the electrophoretic isozyme, to possess both specific bands which were present in the monokaryotic strains. By using restriction endonuclease analysis, the molecular size of mitochondrial DNA of Fu 9 was confirmed to be 85.8 kbp. All 5 fusants had a mitochondrial plasmid-like DNA.The result of pulsed field electrophoresis indicated that parent, monokaryotic, and fusant strains had 4 chromosomal DNA.From these evidences, it is concluded that all 5 fusants are heterokaryon.