玉米生长素响应因子基因ZmARF10在干旱胁迫中的功能分析OA北大核心CSTPCD
Functional Analysis of Maize Auxin Response Factor Gene ZmARF10 under Drought Stress
生长素响应因子(ARF)是一类具有B3 结构域的转录因子,是调节生长素应答反应、控制基因表达的直接分子.前期通过分析转录组数据,在玉米中筛选到 1 个编码 ARF 蛋白并积极参与干旱-复水胁迫响应的基因ZmARF10.为进一步研究该基因调控玉米抗旱性的分子机制及抗旱分子育种提供新的思路,首先通过系列生物信息学分析软件对该基因进行生物信息学分析,之后利用实时荧光定量PCR(qRT-PCR)检测ZmARF10 在玉米不同组织中的表达模式,分析在高温、干旱、高盐、ABA胁迫和解除胁迫处理下的表达情况,以及在不同玉米自交系间的表达差异,最后利用CRISPR/Cas9 技术分析ZmARF10 的功能.结果表明,ZmARF10 位于玉米的第 3 号染色体,编码区全长2127 bp,编码708 个氨基酸,具有典型的B3 结构域;该基因ATG上游2kb区域内含有茉莉酸甲酯、生长素、脱落酸、低温响应等应答元件;系统发育树显示,ZmARF10 基因编码的蛋白质与高粱同源蛋白的亲缘关系最近.qRT-PCR结果表明,ZmARF10 是组成型表达基因,在玉米成熟根中的表达量最高;在高温、干旱、高盐以及ABA 等 4 种胁迫处理下,ZmARF10 基因的表达量均显著上调,干旱胁迫后上调倍数最高达8.2 倍;干旱胁迫后,ZmARF10 基因在抗旱自交系郑36 中的表达量上升幅度显著高于干旱敏感自交系B73.观察拟南芥野生型与ARF10 缺失型突变体发现,与野生型相比,干旱条件下突变体植株出现叶片萎蔫甚至干死的现象,根系发生卷曲,根系分支数减少,侧根生长发育受阻;测定生理生化指标发现,干旱胁迫后缺失型突变体的相对含水量、叶绿素含量、净光合速率显著低于野生型植株,说明敲除ARF10 基因后拟南芥抗旱能力下降.
Auxin response factor(ARF)is a class of transcription factors with B3 domain,which is a direct molecule regulating auxin response and controlling gene expression.A gene,ZmARF10,which encoded ARF protein and actively participated in drought-rehydration stress response,was previously screened in maize by analyzed tran-scriptome data.To further research the molecular mechanism of ZmARF10 regulating drought resistance of maize,and also provide a new idea for molecular breeding of drought resistance,the gene was firstly analyzed by bioinfor-matics software.Secondly,Quantitative Real-time polymerase chain reaction(qRT-PCR)was used to detect the ex-pression pattern of ZmARF10 in different tissues,under high temperature,drought,high salt,ABA and restoring treatments,and in different inbred lines.Finally,the function of ZmARF10 was analyzed using CRISPR/Cas9 technology.The results showed that ZmARF10 was located on chromosome 3 of maize,with a total length of 2127 bp,and encoded 708 amino acids with a typical B3 domain.The upstream 2 kb region of ATG of this gene con-tained response elements related to methyl jasmonate,auxin,abscisic acid and low temperature.The phylogenetic tree showed that the protein encoded by the ZmARF10 gene was closely related to sorghum.qRT-PCR results showed that ZmARF10 was a constitutive expression gene,and the expression level of ZmARF10 was the highest in mature corn roots of maize.Under high temperature,drought,high salt and ABA treatments,the expression of this gene was significantly up-regulated,and the up-regulation ratio was up to 8.2 times after drought stress.After drought stress,the expression level of ZmARF10 gene was significantly higher in the drought-resistant inbred line Zheng 36 than that in the drought-sensitive inbred line B73.Investigation of Arabidopsis wild type and ARF10-de-ficient mutants showed that,compared with wild type,the mutant plants showed leaf wilting and even dry death,roots curled,root branch number decreased,and lateral root growth and development were hindered under drought conditions.Determination of physiological and biochemical indexes showed that the relative water content,chloro-phyll content and net photosynthetic rate of the deficient mutants were significantly lower than those of the wild-type plants after drought stress,indicating that the drought resistance of Arabidopsis decreased after ARF10 gene was knocked out.
曹丽茹;叶飞宇;李伟亚;马晨晨;庞芸芸;梁小菡;张新;鲁晓民
河南省农业科学院 粮食作物研究所,河南 郑州 450002||神农种业实验室,河南 郑州 450002河南省农业科学院 粮食作物研究所,河南 郑州 450002
农业科学
玉米生长素响应因子非生物胁迫抗旱性表达模式
MaizeAuxin response factorAbiotic stressDrought toleranceExpression pattern
《华北农学报》 2024 (002)
1-10 / 10
国家自然科学基金青年科学基金项目(32201708);河南省重大科技专项子课题(221100110300);现代农业产业技术体系建设专项(CARS-02-08)
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