Construction and application of MCBL plate for facilitation of chromosome recombination in fungi.
A medium with camphor and benomyl MCBL plate was designed and constructed based on the proposed mechanism that d-camphor could induce the fusion of nuclear membrane while benomyl could induce the nondisjunctional recombination of chromosome in fungi. This so-called co-induction plate consisted of 0.1% d-camphor (W/V) and 0.5 microgram/L benomyl contained in Czapek's minimal medium. The precautions to be taken in the construction procedure of this plate was described in detail. One typical example of intergeneric fusion-cross, Aspergillus niger x Trichoderma reesei, was investigated, comparing the ratios of genotypes and phenotypes of fusant progenies produced by the co-induction of MCBL plate and by the step-by-step induction with camphor and benomyl separately. The results showed that the heterodiploid state was extremely transient and the recombinant haploid was hardly obtained when induced by the routine step-by-step method, whereas the ratios of apparent diplodization and nondisjunctional recombinant haplodization among all types of varied segregates on MCBL plate were greatly improved, compared with those on the routine plates containing single reagents, which indicated that the transient heterodiploid could be grasped and transformed further into nondisjunctional recombinant haploid when co-induced on the MCBL plate. The age of regenerated mycelium to be induced was found to have a dominant influence on the co-induction effects of MCBL plate. The mechanism of co-induction by MCBL plate and its promising applications were discussed.
Ai
Y
Y
School of Life Sciences, Zhongshan University, Guangzhou, China.
Meng
F
F
D001234:Aspergillus niger / Q000235:genetics* / Q000254:growth & development; D001542:Benomyl; D002164:Camphor; D015825:Chromosomes, Fungal; D003470:Culture Media; D005659:Fungicides, Industrial; D005818:Genetic Engineering; D005838:Genotype; D010641:Phenotype; D011995:Recombination, Genetic; D014242:Trichoderma / Q000235:genetics* / Q000254:growth & development
D003470:Culture Media; D005659:Fungicides, Industrial; D002164:Camphor; D001542:Benomyl