薰衣草非特异性脂质转移蛋白基因nsLTP2-1和nsLTP2-2的克隆与功能分析

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中文摘要：非特异性脂质转移蛋白(nsLTP,non-specific lipid transfer proteins)在植物脂质转运和分泌中发挥重要作用.本研究从薰衣草(Lavandula angustifolia)中克隆到2个Ⅱ型nsLTP基因,命名为nsLTP2-1和nsLTP2-2,并对其进行功能分析.生信分析表明,nsLTP2-1和nsLTP2-2分别编码119个和117个氨基酸,具有脂转移蛋白(LTP,lipid transfer proteins)保守结构域和8个高度保守的半胱氨酸残基;系统进化分析显示它们处于两个分支,与同科的紫苏(Perilla frutescens)相似性最高.基因表达分析显示2个基因均在花蕾中高表达,在叶片、茎和花瓣中几乎不表达,在花萼中的表达存在差异,nsLTP2-1和nsLTP2-2分别在成熟花萼和幼嫩花萼中表达量更高;2个基因在花蕾和叶片中的表达均受到强光诱导,且在花蕾中的表达均受脱落酸诱导,而叶片中nsLTP2-1和nsLTP2-2的表达分别受茉莉酸甲酯和乙烯诱导.亚细胞定位显示2个nsLTPs均定位在细胞膜和细胞壁上,可能与次生代谢物的转运有关.过表达nsLTP2-1和nsLTP2-2烟草叶片经尼罗红染色后,经485～543 nm激发光激发,叶片腺毛头部的荧光显示多于野生型,说明本研究中的nsLTPs可能在脂类的合成和转运中起重要作用.这些结果为明确薰衣草脂转移蛋白在脂类及萜类转运中的功能研究提供了参考.

外文标题：Cloning and Functional Analysis of Nonspecific Lipid Transfer Protein Genes nsLTP2-1 and nsLTP2-2 in Lavender

外文摘要：Non-specific lipid transfer proteins(nsLTPs)play a crucial role in the transport and secretion of lipids in plants.In this study,two type Ⅱ nsLTPs were cloned,namely nsLTP2-1 and nsLTP2-2,and analyzes it with functional analysis from Lavandula angustifolia.The analysis of sequence characteristics revealed that the nsLTP2-1 and nsLTP2-2 genes putatively encode 119 and 117 amino acids,respectively,which exhibit conserved lipid transfer proteins(LTP)domains and 8 highly conserved cysteine residues.The phylogenetic analysis revealed that these genes are located in separate branches and share the closest genetic relationship with Perilla frutescens,a member of the Labiatae family.The analysis of gene expression showed that both genes exhibit high expression levels in flower buds,with minimal expression observed in leaves,stems,and petals.However,notable disparities in expression were observed in the calyx,with nsLTP2-1 and nsLTP2-2 demonstrating higher expression levels in mature and young calyxes,respectively.The expression of these genes in flower buds and leaves was found to be stimulated by heavy light,while their expression in flower buds was also induced by abscisic acid.Additionally,the expression of nsLTP2-1 and nsLTP2-2 in leaves was found be triggered by methyl jasmonate and ethylene,respectively.The subcellular localization of the yellow fluorescent protein(EYFP)fusion protein demonstrated that both nsLTPs were localized on the cell membrane and cell wall,indicating they may be related to the transport of secondary metabolites.Following the overexpression of nsLTP2-1 and nsLTP2-2,tobacco leaves were subjected to Nile red staining,after 485-543 nm excitation light excitation,the fluorescence of leaf glandular hair was more than that of wild type.This observation suggests a potential significance of the nsLTPs investigated in this study with regards to lipid synthesis and transport.These findings provide a fundamental basis for elucidating the role of lavender lipid transfer protein in lipid and terpenoid transportation.

外文关键词：

Lavandula angustifolianon-specific lipid transfer proteinshormone treatmentabiotic stressgene expressionsubcellular localizationNile red staining

作者：

张夏夏、陈凌娜、杨扬、赵旌汝、陈永坤

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作者单位：

新疆师范大学生命科学学院/新疆特殊环境物种保护与调控生物学实验室/干旱区植物逆境生物学实验室,乌鲁木齐 830054

关键词：

薰衣草非特异性脂质转移蛋白激素处理非生物胁迫基因表达亚细胞定位尼罗红染色

基金：

新疆维吾尔自治区自然科学基金新疆维吾尔自治区自然科学基金新疆师范大学博士科研启动基金

项目编号：

2022D01A2162022D01A215XJNUBS2109

出版年：

2024

DOI：

10.13430/j.cnki.jpgr.20231220007

植物遗传资源学报

中国农业科学院作物科学研究所　中国农学会

植物遗传资源学报

CSTPCD北大核心

影响因子：1.415

ISSN：1672-1810

年,卷(期)：2024.25(5)