Abstract:

Lonicera japonica Thunb. is an important Chinese medicinal material in our country， which has many functions such as anti-inflammatory， anti-bacterial， anti-virus， anti-oxidation and anti-cancer. With the increasing contradiction between supply and demand in markets of Lonicera japonica Thunb.， it is essential to breed high yield and high quality varieties by marker-assisted selection. The amino acid sequence of Lonicera japonica Thunb. was scanned with NCBI-Blast tool， and a series of bioinformatics analyses including phylogenetic analysis， conserved domain analysis， expression pattern analysis， physical properties and chemical analysis and protein structure prediction were performed. In this study， the amino acid sequence of Lonicera japonica Thunb. was scanned with NCBI-Blast tool， and 8 candidate genes containing MADS domain were selected， based on ABE genes regulating petal in Arabidopsis. Expression pattern analysis in LjaFGD found that the expression in flower of GWHGAAZE014905 and GWHGAAZE016592 was significantly higher than that in other parts， which might positively regulate flower shape of Lonicera japonica Thunb.. GWHGAAZE014905 was a class B gene containing MADS domain that regulating the development of floral organs； GWHGAAZE016592 was an AP3 homologous gene. Bioinformatics analysis found that GWHGAAZE014905 and GWHGAAZE016592 belonged to stable hydrophilic proteins and non-secreted proteins， both including Motif1， Motif3， Motif4， Motif2， Motif6 and Motif5， 3D structure template of proteins were both 6byy.2.A and 4ox0.2.C. Subcellular localization analysis indicated that GWHGAAZE014905 was targeted to the chloroplast； GWHGAAZE016592 was targeted to the chloroplast， which belonged to membrane protein， contained a transmembrane helix region in 151~173 bp. This research provided genetic resources and molecular markers for the cultivation of top quality and high varieties in Lonicera japonica Thunb. by molecular marker-assisted selection.

Key words: Lonicera japonica Thunb., gene of flower shape, expression pattern analysis, bioinformatics analysis

摘要：

金银花作为我国重要的中药材，具有消炎、抗菌、抗病毒、抗氧化、防癌等多种功效。随着金银花市场供需矛盾日益加剧，通过分子标记辅助选择育种方法来培育高产优质品种势在必行。通过NCBI的Blast工具扫描金银花蛋白组数据发掘花形候选基因，并执行候选基因的亲缘关系分析、结构域分析、表达模式分析、理化性质分析、蛋白质结构预测等一系列生物信息学分析。依据拟南芥调控花形的ABE类基因，通过NCBI-Blast工具扫描金银花氨基酸序列，筛选出包含MADS结构域的8个调控花形的金银花候选基因。经LjaFGD表达模式分析发现，金银花的花中GWHGAAZE016592和GWHGAAZE014905表达量显著高于其他部位，可能正向调控金银花花形。GWHGAAZE014905是一个包含MADS结构域的调控花器官发育的B类基因；GWHGAAZE016592是AP3同源基因。生物信息学分析发现，GWHGAAZE016592和GWHGAAZE014905均是稳定的亲水蛋白，属于非分泌蛋白，包括Motif1、Motif3、Motif4、Motif2、Motif6和Motif5，蛋白质三级结构模板为6byy.2.A和4ox0.2.C。GWHGAAZE014905被定位到细胞核上，而GWHGAAZE016592被定位到叶绿体上，且包含1个位于151~173 bp的跨膜螺旋区域，属于膜蛋白。研究结果为分子标记辅助选择方式培育道地高产优质金银花品种提供了基因资源和分子标记。

关键词: 金银花, 花形基因, 表达模式分析, 生物信息学分析