不同处理方法对榆属植物叶片总DNA提取效果的影响

Effect of different treatments on Ulmus plant DNA extraction from leaves

为探明榆属植物基因组DNA的有效提取方法,以春榆无性系、白榆无性系、金叶榆无性系1号和2号的不同成熟度叶片为材料,并对其做不同预处理,然后采用CTAB法提取榆树基因组DNA,结果表明：嫩叶提取的DNA得率是老叶的1.4-3.0倍,嫩叶的A260/A280值在1.82-1.87之间,老叶的A260/A280值大于1.90;鲜叶片提取的DNA得率是干叶片的1.0-2.0倍,鲜叶片的A260/A280值在1.86-1.94之间,干叶片的A260/A280值在1.90-1.98之间;用低浓度的乙醇（50%-75%）浸泡新鲜嫩叶12-24h可以有效减少DNA的降解,得率在498.62-1 295.97ng/μL,A260/A280值在1.84-1.94之间;预处理液中加入3g的PVP具有较好除酚去多糖的效果,所提取的DNA得率在862.51-1 791.44ng/μL,A260/A280值在1.92-1.98之间。

In order to explore effective extraction method of Ulmus plant genomic DNA,leaves of different maturity from Ulmus davidiana var.japonica,Ulmus pumila,gold leaf elm clones 1and 2as materials,were treated differently,with extraction of elm genomic DNA by CTAB.The results showed that the yield of DNA extracted from new leaves was 1.4to 3.0times as high as that from the old leaves,with A260/A280 of value new leaves between 1.82to1.87,and that of the old leaves more than 1.90.The yield of fresh sample with A260/A280 value between 1.86 to 1.94 was 1.0-2.0times as high as that of the dry leaves with A260/A280 value of 1.90 to 1.98.With a low concentration of alcohol（50% - 75%）,soaking fresh leaves for 12 - 24 hcould reduce DNA degradation,with the yield of DNA between498.62ng/μL and 1 295.97ng/μL,the A260/A280 value between 1.84 and 1.94.Pretreatment solution by adding 3g of PVP had a good effect on removing the phenol and the carbohydrate,with the yield of DNA between 862.51ng/μL and 1 791.44ng/μL,the A260/A280 value between 1.92 and 1.98,which proved better quality of DNA.