以抗生素抗性为选择标记的毛壳菌种间原生质体融合

以抗生素抗性为选择标记进行角毛壳菌和球毛壳菌种间原生质体融合。角毛壳菌CH21-I-402菌株对潮霉素有抗性、对G418敏感,球毛壳菌CH08对潮霉素和G418敏感。根癌农杆菌介导的新霉素磷酸转移酶基因转化球毛壳菌,得到成功转化新霉素磷酸转移酶基因的球毛壳菌菌株,转化子对G418抗性提高3~4倍,对潮霉素仍然比较敏感。以耐潮霉素、不耐G418的角毛壳菌和不耐潮霉素、耐G418的球毛壳菌为出发菌株,以PEG6000为助融剂进行原生质体融合,用G418和潮霉素抗性进行筛选,获得再生菌株。经AFLP分析表明:再生菌株PF1、PF26为融合菌株,融合菌株菌落形态均与亲本存在明显差异。

Interspecies Protoplast Fusion Between Chaetomium cupreum and C. globosum with Antibiotic Resistance Markers

A selection system for hybrids by protoplast fusion between Chaetomium cupreum and C.globosum using dominant selective drug resistance markers were developed.C.cupreum was sensitive tc G418 (G~s) and resistant to hygromycin B(H~r).C.globosum was sensitive to both G418 (G~s) andhygromycin B (H~s).Agrobacterium tumefaciens mediated neomycin phosphotransferase Ⅱ gene transformation of C.globosum (G~s, H~s) acquired resistance to G418 strains (G~r, H~s).Protoplasts of C.cupreum (H~r, G~s) and C.globosum (H~s, G~r) were prepared, mixed in the presence of polyethylene glycol 6000 and incubated on regeneration agar plates.Two regenerated colonies showing H~r and G~r were regarded as hybrids by amplified fragment length polymorphisms analysis of the genomic DNA and they displayed a significant exchange of taxonomic characteristics between the parental strains.