基于流式细胞技术的两种槭属植物基因组大小测定

【目的】槭属植物具有重要的园林观赏价值和药用价值,开发应用前景广阔,但该属植物分子生物学及基因组学研究还相对匮乏,此次研究利用流式细胞仪技术对该属中鸡爪槭和元宝枫基因组大小进行测定,为后期槭属植物的细胞遗传学和基因组学研究奠定基础。【方法】以鸡爪槭(Acer palmatum)和元宝枫(A. truncatum)的嫩叶为材料,首次采用LB01解离液和碘化丙啶(PI)荧光染料,利用基因组大小已知的玉米(Zea mays L.)CE-777作为内标,建立适合于该两种材料的流式细胞术基因组大小测定方法。【结果】LB01解离液对测定样品与内标解离效果均较好,粒子清晰集中,无重叠峰且区分度良好,最后测得鸡爪槭的基因组1C含量为0.840 4 pg,元宝枫的基因组1C含量为0.756 0 pg。【结论】基于流式细胞仪技术,选取合适的样品处理方法,可成功测定鸡爪槭和元宝枫的基因组大小。该结果和方法可丰富槭属植物的基因库,为揭示该属植物的物种起源和进化提供重要依据,也为其后期的基因组测序工作奠定了重要基础。

Estimation of genome size of two Acer Linn. by flow cytometry

【Objective】Acer plants have potential value as ornamental plants and as a source of medicinal compounds. However, their use is hindered by a lack of molecular biology and genomic data. Presently, flow cytometry was used to examine Acer palmatum Thunb. and A. truncatum Bunge genome sizes, to provide foundational data for cytogenetics and genomics studies of Acer Linn.【Method】Fresh leaves of A. palmatum Thunb and A. truncatum were collected. LB01 and PI were used as isolation buffer. Zea mays L. CE-777 was used as the internal standard for the flow cytometry analyses. 【Result】Samples and the internal standard were better dissociated by the LB01 solution, with particles clearly distinguished and concentrated without overlapping peaks. The genome size of A. palmatum and A. truncatum was 0.840 4 and 0.756 0 pg, respectively.【Conclusion】Optimized sample processing and flow cytometry permitted the determinations of the genome sizes of A. palmatum Thunb and A. truncatum Bunge. The optimized processing and results enrich the Acer gene database and provide foundational data for investigations of origin and evolution of species, and genome sequencing.