杜梨IRT1基因的克隆及表达分析

发布时间：2018-03-06 06:30

本文选题：杜梨　切入点：二价铁转运基因(IRT)　出处：《农业生物技术学报》2017年05期 　论文类型：期刊论文

【摘要】：二价铁转运基因(iron-regulated transporter 1 gene,IRT1)是植物缺铁时主要的二价铁转运载体,参与植物铁营养吸收过程。为克隆杜梨(Pyrus betulaefolia)IRT1基因的全长序列并研究其序列及表达特征,本实验以杜梨幼苗为材料,采用反转录PCR(reverse transcription PCR,RT-PCR)、半定量PCR和RACE技术克隆到了杜梨二价铁转运蛋白IRT1基因的c DNA全长序列,命名为PbIRT1(Gen Bank登录号:KX355331)。生物信息学分析表明,该基因全长1 369 bp,含有一个1 095 bp的开放阅读框(open reading frame,ORF),编码364个氨基酸,蛋白序列含有9个跨膜结构,并含有一个完整的锌铁转运蛋白(ZRT/IRTlike protein,ZIP)家族结构域。氨基酸序列的同源关系分析表明,杜梨PbIRT1与小金海棠(Malus xiaojinensis)Mx IRT1(Gen Bank登录号:AAO17059.1)、砀山酥梨(Py.bretschneideri)Pbr IRT1(Gen Bank登录号:XP_009357272.1)蛋白序列的亲缘关系最近,聚为同一小枝。RT-PCR和qRT-PCR表达分析表明,PbIRT1基因在叶片和茎中几乎不表达,具有较强的根组织特异性;在根系中表达量随缺铁胁迫呈逐渐上调趋势,恢复供铁又抑制其表达,正常供铁处理的表达量不随时间变化。以上研究结果表明,杜梨PbIRT1基因的表达受到缺铁胁迫的诱导,可能参与了根系对二价铁的吸收和转运过程,该结果为进一步研究IRT1基因的功能及杜梨缺铁胁迫机制提供了理论依据。
[Abstract]:The bivalent iron transport gene (IRT1) is the main bivalent iron transport vector during iron deficiency in plants. It is involved in the process of iron uptake in plants. In order to clone the full-length sequence of Pyrus betulaefolia)IRT1 gene and study its expression characteristics, the bivalent iron transport gene is a major bivalent iron transport vector in plants with iron deficiency. In this study, the full-length c DNA sequence of the divalent ferric transporter IRT1 gene was cloned by reverse transcription PCR(reverse transcription PCR RT-PCRN and RACE techniques from Pyrus pyrifolia seedlings, named PbIRT1(Gen Bank accession number: KX355331. bioinformatics analysis showed that the cDNA sequence of the IRT1 gene of Pyrus pyrifolia was identified by bioinformatics analysis. The gene is 1 369 BP in length and contains an open reading frame of 1 095 BP, encoding 364 amino acids. The protein sequence contains 9 transmembrane structures. The homology analysis of amino acid sequence of PbIRT1 and Malus xiaojinensis)Mx IRT1(Gen Bank showed that there was a close relationship between PbIRT1 and Malus xiaojinensis)Mx IRT1(Gen Bank accession number: AAO17059.1, Py. bretschneider IRT1(Gen Bank accession number: XP009357272.1). RT-PCR and qRT-PCR expression analysis showed that PbIRT1 gene was almost not expressed in leaves and stems and had strong root tissue specificity, and the expression of PbIRT1 gene in roots increased gradually with iron deficiency stress, and the expression of PbIRT1 gene recovered and inhibited the expression of PbIRT1 gene. The results showed that the expression of PbIRT1 gene was induced by iron deficiency stress and might participate in the process of absorption and transport of divalent iron by roots. The results provided a theoretical basis for further study on the function of IRT1 gene and the mechanism of iron deficiency in Pyrus.
【作者单位】：南京农业大学资源与环境科学学院/江苏省固体有机废弃物资源化高技术研究重点实验室/江苏省有机固体废弃物资源化协同创新中心/农业部长江中下游植物营养与肥料重点实验室;
【基金】：国家现代农业产业技术体系建设专项资金项目(No.CARS-29-15) 国家公益性行业(农业)科研专项(No.201203013)
【分类号】：Q943.2;S661.2

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