7种植物花瓣ACO基因的克隆与分析

为获得通用的适用于植物花瓣ACO基因克隆与检测的引物，根据Genbank中收录的双子叶木本植物的ACO基因序列，设计1对简并引物，并用其对随机选择的7种植物(紫罗兰、多头菊、冬樱、迎春、云南杜鹃、垂丝海棠、山茶)的花瓣的RNA进行PCR扩增。结果发现：从7种植物花瓣中都能扩增出约800 bp的特异片段，登录Genbank发现均未被注册，将序列提交Genbank数据库，获得7种植物花瓣ACO基因的登录号(JX503067、JX503068、JX503069、JX503070、KC112390、KC112392、KC112393)；在NCBI上进行blast比对，发现所克隆的7个ACO基因均与其他植物的ACO基因有一定的序列相似性，其中冬樱ACO基因与千叶桃花ACO基因同源性最高，为98%，迎春ACO基因与中华猕猴桃ACO6基因的同源性最低，为88%，将7个ACO基因的片段进行多序列比较分析，发现两两之间序列平均相似性为77%。

Cloning and analysis of ACO gene from petals in seven plants

To obtain general primers for detection and cloning of ACO gene of plant petal, a pair of degenerate prmiers was designed based on the published sequences of ACO genes of dicotyledonous woody plants in Genbank, which were used to clone ACO genes from petals in 7 plants, namely, Matthiola incana, Dendranthema morifolium, Prunus majestica Koehhe., Jasminum nudiflorum, Rhododendron yunnanense Franch., Malus halliana, Camellia japonica. The results indicated that specific fragments of about 800 bp could be amplified from all of the 7 plants and the sequences have not been registered on Genbank database. The sequences were submitted to Genbank database and the accessing numbers were JX503067, JX503068, JX503069, JX503070, KC112390, KC112392 and KC112393. The 7 sequences show high sequence identities with ACO genes of other plants through NCBI blast analysis, the highest homology of nucleotide sequence existed between Prunus majestica Koehhe. ACO gene and Prunus persica Batsch. ACO gene, which is 98%, and the lowest homology of nucleotide sequence existed between Jasminum nudiflorum ACO gene and Actinidia chinensis ACO6, which is 88%. It was found that the average sequence identity through comparison between any two samples was 77% by multiple sequence alignments.