玉米ARID转录因子家族鉴定及表达分析

Identification and expression analysis of ARID transcription factor family in maize

  • 摘要: 【目的】鉴定玉米ARID转录因子家族成员,并进行表达分析,为深入研究ARID转录因子生物学功能及玉米遗传育种提供理论参考。【方法】从拟南芥数据库TAIR获取7个ARID转录因子家族成员的氨基酸序列,与玉米基因组编码蛋白进行同源比对,鉴定出玉米ARID转录因子家族成员,采用生物信息学方法对其理化性质、系统发育进化、启动子区顺式作用元件、生育期组织表达模式和蛋白互作网络等进行预测分析,并用实时荧光定量PCR检测其在激素处理及低温胁迫下的表达模式。【结果】从玉米中共鉴定出12个ARID转录因子家族成员(ZmARID1~ZmARID12),编码的氨基酸数量为448~1875个,均为亲水性蛋白,除ZmARID12外,其余均为不稳定蛋白;ZmARID2、ZmARID4、ZmARID9和ZmARID10蛋白定位于叶绿体和细胞核,ZmARID12定位于叶绿体和细胞质,ZmARID1定位于叶绿体,其余成员均定位于细胞核。玉米ARID家族基因启动子区域不仅含有多种光响应元件(G-box、Sp1、GATA-motif、Box4、MRE、AE-box),还含有茉莉酸甲酯(MeJA)响应元件(CGTCA-motif)、脱落酸(ABA)响应元件(ABRE)、赤霉素(GA)响应元件(GARE-motif、TATC-box)、生长素(IAA)响应元件(TGA-element、AuxRR-core)和水杨酸(SA)响应元件(TCA-element)。ZmARID1基因在授粉后12 d(12 DAP)的胚乳中表达水平较高;除了ZmARID1ZmARID3基因外,其余成员在2~4 mm和6~8 mm穗长的原基中均有较高表达;ZmARID3基因在成熟叶片和营养分生组织中表达水平较高;ZmARID5ZmARID6ZmARID11基因在成熟叶片中表达水平较高,说明部分家族成员表达具有组织特异性。实时荧光定量PCR检测结果显示,GA处理后,ZmARID12基因的相对表达量较对照显著升高(P<0.05,下同);ABA处理后,ARID家族基因相对表达量显著升高或下降,但IAA、JA、SA和低温胁迫处理后整体呈下降趋势。【结论】玉米ARID家族成员在全生育期具有明显的组织表达特异性,且在响应多种激素和低温胁迫中发挥重要作用。

     

    Abstract: 【Objective】The purpose of the study was to identify family members of maize ARID transcription factors and perform expression analysis to provide theoretical reference for further research on the biological functions of ARID transcription factors and maize genetic breeding. 【Method】The amino acid sequences of seven ARID transcription factor family members were obtained from the Arabidopsis thaliana database TAIR, and homologous alignment was performed with the encoded proteins in the maize genome to identify the members of the maize ARID transcription factor family. Bioinformatics methods were used to predict and analyze their physicochemical properties, phylogenetic evolution, cisacting elements in the promoter region, tissue expression pattern during growth period and protein interaction networks.And real-time fluorescence quantitative PCR was used to detect its expression patterns under hormone treatment and low temperature stress. 【Result】Twelve members of ARID transcription factor family in maize were identified, named ZmARID1-ZmARID12, with 448-1875 encoded amino acids, all of which were hydrophilic proteins. Except for ZmARID12, all other proteins were unstable proteins. Except for ZmARID2, ZmARID4, ZmARID9, and ZmARID10 proteins, which were located in the chloroplast and nucleus, ZmARID12, which was located in the chloroplast and cytoplasm, and ZmARID1, which was located in the chloroplast, the rest of the members were located in the nucleus. The promoter region of the maize ARID family gene contained not only multiple light response elements(G-box, Sp1, GATAmotif, Box 4, MRE, AE-box), but also methyl jasmonate(MeJA) response elements(CGTCA-motif), abscisic acid(ABA) response elements(ABRE), gibberellin(GA) response elements(GARE-motif, TATC-box), auxin(IAA) response elements(TGA-element, AuxRR-core), and salicylic acid(SA) response elements(TCA-element). ZmARID1 gene exhibited increased expression level in the endosperm at 12 d after pollination(12 DAP). Except for ZmARID1 and ZmARID3 genes, the rest of the members displayed high expression levels in the primordia of 2-4 mm and 6-8 mm ear lengths. The expression level of ZmARID3 gene was high in mature leaves and vegetative meristem. ZmARID5, ZmARID6 and ZmARID11 genes were expressed at high levels in mature leaves, indicating that the expression of some family members were specific for tissue. The real-time fluorescence quantitative PCR detection results showed that after GA treatment, the relative expression of ZmARID12 gene was significantly up-regulated compared to the control(P<0.05,the same below). The relative expression of ARID family genes were significantly up-regulated or down-regulated under ABA treatment, but under IAA, JA, SA and low temperature stress treatments, the relative expression of ARID family genes showed an overall downward trend. 【Conclusion】The maize ARID family members exhibit obvious tissue expression specificity throughout the whole growth period and play important roles in responding to multiple hormones and low temperature stress.

     

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