利用流式细胞仪鉴定槭树染色体倍性的方法探究

Research on the Method of Measuring the Ploidy of Acer L by flow cytometry

利用流式细胞仪对不同系号的槭树植物DNA的相对含量进行测定,建立一套快速、简易的鉴定槭树科植物倍性的方法。以三角枫、元宝枫等槭树类植物的叶片为材料,选取最优鉴定叶片的部位,获取最优的裂解液配方等。同时结合形态测量对4种槭树品系进行比较分析。结果表明,春季嫩叶的叶尖是最优检测时间和位置;最优裂解液是0.1mol/L的柠檬酸和0.5%的吐温20制成的混合液取200ul,再加入0.4mol/L的磷酸氢二钠600ul。经检测分析后得到,具体活细胞所占比例的关系是:三角枫1号<三角枫2号<元宝枫A号<元宝枫F号;DNA含量方面,两个三角枫品系大于两元宝枫品系。形态学比较结果上三角枫1号枝干高大挺拔,叶片肥厚宽大,与其他3个品系的槭树差异显著。

Flow cytometry was used to determine the relative DNA content of Acer L plants of different lines,and set of quick and easy methods for identifying ploidy of Acer L plants.Using the leaves of Acer buergerianum and Acer truncatum as materials,the optimal location of leaves was selected and the optimal formula of pyrolysis solution was obtained.At the same time,four kinds of Acer L lines were compared and analyzed in combination with morphological measurements.The results showed that the optimum detection time and location were the leaf tip of tender leaves in spring,and the optimum cracking solution was the mixture of 0.1 mol/L citric acid and 0.5%Tween 20 for 200 ul,followed by 0.4 mol/L disodium hydrogen phosphate 600 ul.After detection and analysis,it was found that the proportion of specific living cells was as follows:Acer buergerianum No.1<Acer buergerianum No.2<Acer truncatum No.A<Acer truncatum No.F.In terms of DNA content,the two lines of Acer buergerianum were larger than the two lines of Acer truncatum.The results showed that the branches of the Acer buergerianum No.1 were tall and straight,and the leaves were thick and wide,which was significantly different from the maple trees of the other three Acer L lines.