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首页> 外文期刊>African Journal of Biotechnology >Development of a transformation method for the nematophagous fungus Dactylellina cionopaga
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Development of a transformation method for the nematophagous fungus Dactylellina cionopaga

机译：线虫真菌Dactylellina cionopaga转化方法的开发

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Dactylellina cionopaga?is a trapping fungus that produces adhesive columns and a two-dimensional network.?The factors that influence protoplast preparation and regeneration of?D. cionopaga?were analyzed, and?poly ethylene glycol (PEG)-CaCl2- or?Agrobacterium tumefaciens-mediated transformation was conducted?to develop a transformation system for the fungus and provide a tool for studying the function of nematode infection-related?genes.?The results indicate that between 4.175±1.025×106and?3.08±1.4×107,?protoplasts/ml were obtained under optimized conditions and that the protoplasts could be regenerated on?potato dextrose agar?(PDA), RA and IM regeneration media.?D. cionopaga?transformation using PEG-CaCl2?or?A. tumefaciensdisplayed?4.2 to 11 resistant colonies/μg DNA using 106?protoplasts?and?180-270 resistant colonies using 106?conidia. Molecular?analysis and microscopy of randomly selected transformants showed that the target genes were integrated into the genome of?D. cionopaga?and that green fluorescence could be detected in transformants containing pK2-BarGFP, which carried a?glufosinate ammonium?resistance gene and the enhanced green fluorescence protein gene. The methods used in this study for protoplast preparation and convenient?Agrobacterium-mediated transformation of?D. cionopaga?represent useful tools for genetic research on this nematophagous fungus. This is the first report on protoplast generation and transformation of?D. cionopaga.
机译：Dactylellina cionopaga是一种捕集真菌，会产生黏附柱和二维网络。影响原生质体制备和D再生的因素。分析了紫苏病，并进行了聚乙二醇（PEG）-CaCl 2-或根癌农杆菌介导的转化，以开发真菌的转化系统，并为研究线虫感染相关基因的功能提供了工具。结果表明，在最佳条件下可得到4.175±1.025×106〜3.08±1.4×107的原生质体/ ml，原生质体可在马铃薯葡萄糖琼脂（PDA），RA和IM再生培养基上再生。 D．使用PEG-CaCl2或A进行cionopaga转化。使用106个原生质体时，根瘤菌显示出4.2至11个耐药菌落/μgDNA，使用106个分生孢子菌显示出180-270个耐药菌落。随机选择的转化子的分子分析和显微镜检查表明，靶基因已整合到ΔD基因组中。 cionopaga？和含有pK2-BarGFP的转化子中可以检测到绿色荧光，该转化子带有草铵膦抗性基因和增强的绿色荧光蛋白基因。本研究中使用的方法用于原生质体的制备和方便的农杆菌介导的ΔD转化。 cionopaga代表了对这种线虫真菌进行遗传研究的有用工具。这是关于原生质体产生和转化的首次报道。 cionopaga。

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