小花草玉梅正常和自然变异植株的AP3-3基因研究  

Cloning and Sequence Analysis of AP3-3 Gene in Normal Plant and Natural Variant from Anemone rivularis var.flore-minore

作　　者：张婷[1] 邢妮[1] 王超[1] 刘虎岐[1] 

机构地区：[1]西北农林科技大学生命科学学院,陕西杨陵712100

出　　处：《西北植物学报》2016年第2期231-240,共10页Acta Botanica Boreali-Occidentalia Sinica

基　　金：中医药行业科研专项(201207002);中医药公共卫生专项(财社[2011]76号)

摘　　要：野生小花草玉梅(Anemone rivularis var.flore-minore)正常植株和花被片自然变异植株的外观形态差异很大,该研究以二者为材料,利用常规PCR和高效热不对称PCR(Hi-Tail PCR)技术从其正常和变异植株的基因组中各分离得到1个B类基因。序列分析证明,二者隶属于B类MADS-box基因AP3家族的旁系同源基因AP3-3分枝,分别命名为NArAP3-3(正常植株)和VArAP3-3(变异植株)。NArAP3-3基因全长3 795bp,VArAP3-3基因全长3 898bp,二者均含有1个666bp的开放阅读框(ORF),可编码221个氨基酸,具有典型的植物MADS-box基因结构,其编码肽链包含了MADS区、K区、Ⅰ区和C区。对比NArAP3-3和VArAP3-3基因的全长序列,发现VArAP3-3基因比NArAP3-3多了1段49bp的插入,且在ORF序列与NArAP3-3基因相比有4个碱基突变。对二者的全长序列、所编码的221个氨基酸及插入序列的生物信息学分析显示,二者在基因启动子、蛋白质基本性质、结构功能域、二级三级预测结构等方面均有差异,推测这些差异可能是花被片变异产生的原因之一。该研究结果为进一步探索其变异机制奠定了基础。The study takes normal plant and tepal natural variant which have big differences in appearance as materials from wild Anemone rivularis var.flore-minore.The class B genes were isolated from genomes of normal plant and natural variant by using conventional and Hi-Tail PCR techniques.They were proved to be AP3-3branch of paralogs which belong to AP3 gene family from B class MADS-box genes by sequence analysis and named NArAP3-3（in normal plant）and VArAP3-3（in natural variant）,respectively.The full length of NArAP3-3gene was 3 795 bp whereas 3 898 bp in VArAP3-3,they both have a 666 bp open reading frame（ORF）and can encode 221 amino acids.The protein showed a typical MADS-box gene structure containing MADS domain,K domain,Ⅰ region and C terminus.The comparison result of full length sequences between NArAP3-3and VArAP3-3genes was found to have a 49 bp insertion in VArAP3-3gene,and there are also 4base mutations in ORF sequence compared with the NArAP3-3gene.By bioinformatics analysis of the two’s full length sequences,221 amino acids and insertion sequence,we can find that there are differences in all aspects of gene promoter,basic properties of protein,structure function domain,advanced prediction structure and so on.These differences may be one of the causes of tepals variation,and this study lays the foundation for the further exploration of its variation mechanism.

关 键 词：小花草玉梅 花被片变异 Hi-Tail PCR MADS-BOX基因 NArAP3-3和VArAP3-3基因 

分 类 号：Q754[生物学—分子生物学]