东方百合'西伯利亚'LoNAC48基因的克隆、表达及功能分析OA北大核心CSTPCD
Cloning,Expression and Functional Analysis of LoNAC48 Gene in Lilium Oriental Hybrid 'Siberia'
国产百合(Lilium spp.)种球培育周期长、品质差,是导致我国百合种球长期依赖进口的重要因素.植物特异性转录因子NAC在调节植物生长发育、抗性建立等方面发挥着重要作用,在百合中挖掘、鉴定并研究NAC转录因子基因用于转基因技术改良植物性状,是解决上述问题的一种策略.本研究在百合转录组数据库中筛选到1个携带NAC结构域的基因转录本,基于PCR技术克隆并鉴定其为LoNAC48基因(GenBank No.PP078617).生物信息学分析表明,LoNAC48基因包含1个长度为777 bp完整的开放阅读框,编码258个氨基酸,属于不稳定的疏水性蛋白;进化树分析显示,其与岷江百合(Lilium regale)LrNAC48高度同源;亚细胞定位结果表明,LoNAC48属于核转录因子.组织表达分析表明,LoNAC48基因在鳞茎、根、花等组织器官中均有表达,其中叶片中表达量最高;不同逆境响应结果表明,LoNAC48基因的表达易受低温、高温、干旱、盐、过氧化氢胁迫的影响,在低温、盐与过氧化氢胁迫下,与对照相比,12 h时表达量均显著上调,干旱胁迫24h时表达量显著降低,且在盐胁迫与过氧化氢胁迫条件下,LoNAC48的表达模式几乎是一致的.通过转基因拟南芥(Arabidopsis thaliana)株系表型分析显示,过表达LoNAC48基因可显著促进拟南芥生长并增加叶片面积.以上结果表明,LoNAC48基因具备抗逆和调控植物生长发育双重功能.本研究为百合分子辅助育种,改良百合性状提供了基因资源.
The long cultivation cycle and poor quality of lily bulbs by produced in China is a important factors that lead to long-term dependence on imports.The plant-specific transcription factor NAC plays an important role in regulating plant growth and development,resistance establishment,etc.,and mining,identifying and studying related genes in lily is a strategy to solve the above problems.In this study,one gene transcript carrying NAC domain was screened in the lily transcriptome database,and the gene was cloned and identified as LoNAC48 gene(GenBank No.PP078617)based on PCR.Bioinformatics analysis showed that LoNAC48 gene contained a complete open reading frame with a length of 777 bp encoding 258 amino acids,which was an unstable hydrophobic protein,and phylogenetic tree analysis showed that it was highly homologous to Lilium regale LrNAC48 protein.The results of subcellular localization showed that LoNAC48 was a nuclear transcription factor,and tissue expression analysis showed that LoNAC48 gene was present in bulbs,roots,flowers and other tissues and organs expression,among which the expression level was the highest in leaves;The results of different stress responses showed that the expression of LoNAC48 gene was susceptible to low temperature,high temperature,drought,salt and hydrogen peroxide(H2O2)stresses,Under low temperature,salt,and hydrogen peroxide stress,the expression level was significantly upregulated at 12 hours compared to the control,and significantly decreased at 24 hours of drought stress,and the expression pattern of LoNAC48 was almost the same under salt stress and hydrogen peroxide stress.Phenotypic analysis of transgenic Arabidopsis thaliana lines showed that overexpression of LoNAC48 gene could significantly promote the growth of Arabidopsis thaliana and increase the leaf area.These results indicated that LoNAC48 gene had the dual functions of stress resistance and regulation of plant growth and development.This study provides genetic resources for molecular-assisted breeding of lily and improvement of lily traits.
胡晓聪;王可;孙亮
金华职业技术学院农学院,金华 321017金华职业技术学院农学院,金华 321017||浙江农林大学风景园林与建筑学院,杭州 311300
园艺学与植物营养学
百合LoNAC48基因克隆生物信息学分析功能分析
LilyLoNAC48Gene cloningBioinformatics analysisFunctional analysis
《农业生物技术学报》 2024 (006)
1297-1306 / 10
2023年度浙江省高校国内访问工程师"校企合作项目"(FG2023011)
评论