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非洲菊(Gerbera hybrida)已成为研究复杂花序进化与发育的模式植物。以非洲菊无菌组培苗叶片为材料,分析不同浓度纤维素酶及离析酶配比对非洲菊原生质体提取效率的影响,建立了稳定、高效的非洲菊原生质体提取与瞬时转化体系。结果表明,以2.0%纤维素酶+0.3%离析酶组合,在0.4 mol·L~(–1)甘露醇溶液中,酶解4小时,酶解温度为25°C,得到产量高达2.2×10~7个·g~(–1) FW及活力较高的原生质体,可直接用于植物蛋白亚细胞定位和蛋白间相互作用实验,转化效率达80%。该研究建立了非洲菊原生质体制备和瞬时转化体系,为非洲菊基因功能研究奠定重要的技术基础。
Gerbera hybrida has become a model plant for studying the evolution and development of complex inflorescences. Gerbera jamesonii germplasm was used as material to analyze the effect of different concentration of cellulase and isozyme on the extraction efficiency of the germplasm of Gerbera jamesonii and the stable and efficient germplasm extraction and transient transformation system of Gerbera jamesonii was established. The results showed that the enzymolysis temperature was 25 ° C in 0.4 mol·L ~ (-1) mannitol solution with 2.0% cellulase + 0.3% isozyme for 4 hours, yielding up to 2.2 × 10 ~ 7 · g ~ (-1) FW and high viability of protoplasts could be directly used in plant protein subcellular localization and protein interaction experiments, the transformation efficiency of 80%. The research established the germplasm preparation and transient transformation system of Gerbera jamesonii and lay an important technical foundation for the study of the gene function of Gerbera jamesonii.