Establishment of flow cytometry system for DNA C-value determination of polyploid species in subgenus Yulania
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摘要: 通过对不同解离液和内标组合的筛选，采用流式细胞仪对木兰科玉兰亚属（subgen.Yulania）6种多倍体植物的DNA C-值进行测定与分析。结果显示：在解离液和标样的筛选中，WPB解离液为玉兰亚属植物的最佳选择；以星花玉兰（Magnolia stellata（Siebold&Zucc.）Maxim.）为内标时，混合样品的峰图效果最好，DNA峰的变异系数（CV）均小于5.0%。紫玉兰（M.liliiflora Desr.）、多瓣紫玉兰（M.polytepala Law，R.Z.Zhou&R.J.Zhang）、黄山玉兰（M.cylindrica E.H.Wilson）、白玉兰（M.denudata Desr.）、二乔玉兰（M.×soulangeana Soul.-Bod.）和渐尖玉兰（M.acuminata（L.）L.）的C-值大小范围在（3.92 ±0.02）pg到（5.85 ±0.03）pg之间，基因组大小从（3832.13 ±24.67）Mbp到（5964.17 ±56.06）Mbp。研究结果表明该优化的流式细胞术测定体系可以精确测定玉兰亚属多倍体植物的DNA C-值。Abstract: By screening different combinations of dissociation solutions and internal standards, the DNA C-values of six polyploid plants of subgen. Yulania (Magnoliaceae) were determined and analyzed by flow cytometry. Results showed that, in the screening of solution buffers and standard samples, woody plant buffer (WPB) was the best choice for subgen. Yulania. Furthermore, the peak pattern of mixed samples worked best when Magnolia stellata (Siebold & Zucc.) Maxim. was the internal standard and the coefficient of variation (CV) of the DNA peak was less than 5.0%. The C-values of M. liliiflora Desr., M. polytepala Law, R. Z. Zhou & R. J. Zhang, M. cylindrica E. H. Wilson, M. denudata Desr., M.×soulangeana Soul.-Bod., and M. acuminata (L.) L ranged from (3.92 ±0.02) pg to (5.85 ±0.03) pg and the genome size ranged from (3832.13 ±24.67) Mbp to (5964.17 ±56.06) Mbp. Thus, our results showed that the optimized flow cytometry system accurately determined the DNA C-value of polyploid species in the subgen. Yulania.
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Keywords:
- Flow cytometry /
- DNA C-value /
- Genome size /
- subgenus Yulania
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