荷花品种不同部位DNA提取的比较研究

Comparative Study on Extraction of DNA from Different Parts of Nelumbo nucifera Gaertn.

采用改良的3×CTAB法提取荷花品种笛女、国庆红、红苔莲的叶片、叶柄和花不同部位的DNA,并对提取的DNA进行琼脂糖凝胶电泳检测、纯度检测和ISSR分析。结果表明,除叶柄中提取的DNA效果不理想外,其余从叶片、花中提取的DNA纯度高、得率高,OD260/280比值在1.80左右,OD260/230比值在2.01左右。以红苔莲的花为试材,对改良的3×CTAB法提取基因组DNA的参数进行正交试验,选出提取DNA的最优组合。通过荷花基因组DNA的ISSR分析,完全满足实验要求。

Genomic DNA of Nelumbo nucifera Gaertn.was extracted from different parts of three cultivars of Nelumbo nucifera Gaertn.,Dinv,Guoqinghong,and Hongtailian.Yield and purity of total genomic DNA was detected by agarose gel electrophoresis and UV-spectrophotomoter.Results showed that,extraction effect of DNA from the petiole was not ideal,the purity of DNA extracted from the leaves and flowers were high.OD260/280 was 1.80 or so.OD260/230 was 2.01 or so.Taking the flowers of cultivars Hhongtailian as material,orthogonal experiment was carried out on the parameters of genome DNA extracted by the modified 3×CTAB method,and the optimal combination of DNA extraction was selected.When tested through the DNA-ISSR analysis,experiment requirement was met perfectly.