Biocontrol of Plant Diseases Laboratory, Agricultural Research Service, U.S. Department of Agriculture, Beltsville, Maryland 20705.
Appl Environ Microbiol. 1990 Oct;56(10):3052-6. doi: 10.1128/aem.56.10.3052-3056.1990.
Methodology was developed to isolate and regenerate protoplasts from the biocontrol fungus Gliocladium virens and to transform them to benomyl resistance with a Neurospora crassa beta-tubulin gene. Southern blots demonstrated that multiple copies of the vector integrated into the chromosomal DNA of stable biotypes but not of abortive transformants. Analysis of nuclear condition in vegetative and asexual structures demonstrated that no structure of G. virens is dependably uninucleate and thus preferentially suitable for transformation.
方法学被开发出来以从生防真菌木霉中分离和再生原生质体，并通过脉孢菌 β-微管蛋白基因将其转化为苯并咪唑抗性。Southern 印迹表明，载体的多个拷贝整合到稳定生物型的染色体 DNA 中，但不整合到流产转化体中。对营养和无性结构中核条件的分析表明，木霉没有一种结构是可靠的单核的，因此不优先适合转化。
