忍冬属黄酮合成酶基因FNSII的SNP分析

SNP analysis of flavone synthase Ⅱ gene(FNSII) in Lonicera species

为挖掘鉴别金银花与山银花的黄酮合成酶基因(flavone synthase II gene,FNSII)SNP位点,以基源确定的5个忍冬属(忍冬、灰毡毛忍冬、红腺忍冬、黄褐毛忍冬、华南忍冬)共计12个样本为研究对象,采用同源克隆法从各忍冬属植物中克隆FNSII基因,分析不同忍冬属植物中的SNP位点。结果显示:克隆分离到的各忍冬属FNSII基因的同源性和相似性高达95%以上;成功克隆获得红腺忍冬、黄褐毛忍冬和华南忍冬的FNSII序列;系统进化分析表明,获得的忍冬属序列与基源鉴定结果一致;通过序列比对,从12个样本中克隆得到的20条FNSII序列中鉴定到38个SNP位点,其中19个SNP位点可用于区分各种属,依据各种属间SNP的差异进行分型,获得5种基因型。

Potential single nucleotide polymorphisms（SNPs） of FNSII gene is helpful to identify Lonicera japonica and Lonicera confuse. Previously, 12 samples had been identified into five Lonicera species（Lonicera japonica Thunb., Lonicera macranthoides Hand.–Mazz., Lonicera hypoglauca Miq., Lonicera fulvotomentosa, Lonicera confusa） in the base of base resource. Here, we have performed further study on these 12 samples. First, homology–based cloning was employed to clone their FNSII genes, and then SNP analysis. To our knowledge, our study is the first time to obtain the FNSII gene sequences from Lonicera hypoglauca Miq., Lonicera fulvotomentosa and Lonicera confusa. The similarity of all the cloned 20 FNSII gene fragments is over 95%, the phylogenetic analysis shows that genetic relationship of these Lonicera L. is consistent with the identification results. 38 SNPs were identified from 20 sequences in 12 samples via sequence alignment. According to the difference of SNPs, all samples can be classed into 5 genotypes according to the total 38 SNPs, which were identified from 20 sequences cloned in 12 samples and 19 of which can be used for the species indenfication.