马缨杜鹃查尔酮异构酶基因RdCHI1的克隆与功能解析  

Cloning and Function Analysis of Chalcone Isomerase Gene RdCHI1 in Rhododendron delavayi

作　　者：孙威 孙世宇 陈一然 王聿晗 张艳 鞠志刚 乙引 SUN Wei;SUN Shiyu;CHEN Yiran;WANG Yuhan;ZHANG Yan;JU Zhigang;YI Yin(Key Laboratory of Plant Physiology and Development Regulation,Guizhou province School of Life Sciences,Guizhou Normal University,Guiyang 550025,China;Pharmacy College,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China)

机构地区：[1]贵州师范大学生命科学学院,贵州省植物生理与发育调控重点实验室,贵阳550025 [2]贵州中医药大学药学院,贵阳550025

出　　处：《园艺学报》2022年第11期2407-2418,共12页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(31760076);贵州省教育厅自然科学研究项目(黔教合KY字[2021]059);贵州省科技计划项目(黔科合平台人才[2017]5726号);贵州省科技厅基础研究项目(黔科合基础[2018]1011)。

摘　　要：为了研究马缨杜鹃(Rhododendrondelavayi)查尔酮异构酶(CHI)的功能,以其花朵为材料,克隆获得了1个Rd CHI1基因。通过多序列比对、系统进化分析、q RT-PCR、拟南芥的遗传转化和抗逆试验,探究Rd CHI1参与花色苷合成与抗逆的作用。结果表明,Rd CHI1的CDS序列长663 bp,编码220个氨基酸,预测其蛋白分子量为23.728k D。多序列比对分析显示,Rd CHI1与拟南芥、苜蓿CHI的相似性分别为56.52%和56.07%。开花过程中Rd CHI1的表达水平先上升后下降,在雌蕊中表达量最高,花瓣和根中相对较低。拟南芥转化试验表明,Rd CHI1可以恢复CHI突变体(tt5)缺失的花色苷与黄酮醇,使其幼苗子叶与下胚轴颜色由绿色恢复为紫色。逆境表明,Rd CHI1过表达拟南芥植株在氯化钠和甘露醇胁迫下比野生型具有更好的适应力与存活率,证明Rd CHI1在植物中具有抵抗逆境的潜在作用。In order to investigate the function of Chalcone Isomerase(CHI)from Rhododendron delavayi,the flower of R.delavayi was used as study material to clone RdCHI1 gene.Multiple sequence alignment,phylogenetic analysis,q RT-PCR,genetic transformation of Arabidopsis thaliana and stress assay were employed to study the function of RdCHI1 on anthocyanin synthesis and stress resistance.The results showed that CDS sequence length of RdCHI1 is 663 bp,encoding 220 amino acids,and the predicted molecular weight of the protein is 23.78 k D.Analysis of multiple sequence alignment revealed that RdCHI1 shares 56.52%and 56.07%similarity with CHI of Arabidopsis thaliana and Medicago.The expression levels of RdCHI1 showed a trend of rising first and then decreasing during flower development,and the transcripts were the highest in pistil,but relatively lower in petal and root.Transformation of Arabidopsis found RdCHI1 could restore the biosynthesis of anthocyanins and flavonols which missed in CHI mutant(tt5),and turn the color of cotyledons and hypocotyls of seedlings from green back to purple.Meanwhile,stress assay indicated that Arabidopsis overexpressed RdCHI1 showed better adaptability and survival rate than the wild type when under Na Cl and mannitol stresses,suggesting RdCHI1 playing a potential role to stress resistance in plants.

关 键 词：马缨杜鹃 查尔酮异构酶 花色 花色苷 功能解析 

分 类 号：S685.21[农业科学—观赏园艺]