發(fā)布時(shí)間：2018-03-06 06:30

  本文選題：杜梨　切入點(diǎn)：二價(jià)鐵轉(zhuǎn)運(yùn)基因(IRT)　出處：《農(nóng)業(yè)生物技術(shù)學(xué)報(bào)》2017年05期 　論文類型：期刊論文

【摘要】：二價(jià)鐵轉(zhuǎn)運(yùn)基因(iron-regulated transporter 1 gene,IRT1)是植物缺鐵時(shí)主要的二價(jià)鐵轉(zhuǎn)運(yùn)載體,參與植物鐵營養(yǎng)吸收過程。為克隆杜梨(Pyrus betulaefolia)IRT1基因的全長(zhǎng)序列并研究其序列及表達(dá)特征,本實(shí)驗(yàn)以杜梨幼苗為材料,采用反轉(zhuǎn)錄PCR(reverse transcription PCR,RT-PCR)、半定量PCR和RACE技術(shù)克隆到了杜梨二價(jià)鐵轉(zhuǎn)運(yùn)蛋白IRT1基因的c DNA全長(zhǎng)序列,命名為PbIRT1(Gen Bank登錄號(hào):KX355331)。生物信息學(xué)分析表明,該基因全長(zhǎng)1 369 bp,含有一個(gè)1 095 bp的開放閱讀框(open reading frame,ORF),編碼364個(gè)氨基酸,蛋白序列含有9個(gè)跨膜結(jié)構(gòu),并含有一個(gè)完整的鋅鐵轉(zhuǎn)運(yùn)蛋白(ZRT/IRTlike protein,ZIP)家族結(jié)構(gòu)域。氨基酸序列的同源關(guān)系分析表明,杜梨PbIRT1與小金海棠(Malus xiaojinensis)Mx IRT1(Gen Bank登錄號(hào):AAO17059.1)、碭山酥梨(Py.bretschneideri)Pbr IRT1(Gen Bank登錄號(hào):XP_009357272.1)蛋白序列的親緣關(guān)系最近,聚為同一小枝。RT-PCR和qRT-PCR表達(dá)分析表明,PbIRT1基因在葉片和莖中幾乎不表達(dá),具有較強(qiáng)的根組織特異性;在根系中表達(dá)量隨缺鐵脅迫呈逐漸上調(diào)趨勢(shì),恢復(fù)供鐵又抑制其表達(dá),正常供鐵處理的表達(dá)量不隨時(shí)間變化。以上研究結(jié)果表明,杜梨PbIRT1基因的表達(dá)受到缺鐵脅迫的誘導(dǎo),可能參與了根系對(duì)二價(jià)鐵的吸收和轉(zhuǎn)運(yùn)過程,該結(jié)果為進(jìn)一步研究IRT1基因的功能及杜梨缺鐵脅迫機(jī)制提供了理論依據(jù)。
[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.
【作者單位】： 南京農(nóng)業(yè)大學(xué)資源與環(huán)境科學(xué)學(xué)院/江蘇省固體有機(jī)廢棄物資源化高技術(shù)研究重點(diǎn)實(shí)驗(yàn)室/江蘇省有機(jī)固體廢棄物資源化協(xié)同創(chuàng)新中心/農(nóng)業(yè)部長(zhǎng)江中下游植物營養(yǎng)與肥料重點(diǎn)實(shí)驗(yàn)室;
【基金】：國家現(xiàn)代農(nóng)業(yè)產(chǎn)業(yè)技術(shù)體系建設(shè)專項(xiàng)資金項(xiàng)目(No.CARS-29-15) 國家公益性行業(yè)(農(nóng)業(yè))科研專項(xiàng)(No.201203013)
【分類號(hào)】：Q943.2;S661.2

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1 孫靜文;趙世誠;范洪黎;周衛(wèi);;莧菜IRT1基因克隆、序列及表達(dá)分析[J];生物技術(shù)通報(bào);2012年02期

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本文編號(hào)：1573701