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在同源和异源模型系统中,对油棕磷酸盐饥饿诱导、高亲和力磷酸盐转运蛋白基因启动子的功能进行了鉴定。

Functional characterization of the gene promoter for an Elaeis guineensis phosphate starvation-inducible, high affinity phosphate transporter in both homologous and heterologous model systems.

机构信息

Laboratory of Plantation Science and Technology, Institute of Plantation Studies, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia; Department of Plant Pathology, Rafsanjan Branch, Islamic Azad University, Rafsanjan, Iran.

Laboratory of Plantation Science and Technology, Institute of Plantation Studies, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia; Department of Agriculture Technology, Faculty of Agriculture, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia.

出版信息

Plant Physiol Biochem. 2018 Jun;127:320-335. doi: 10.1016/j.plaphy.2018.04.004. Epub 2018 Apr 9.

DOI:10.1016/j.plaphy.2018.04.004
PMID:29653435
Abstract

Oil palm is grown in tropical soils with low bioavailability of Pi. A cDNA clone specifically expressed under phosphate-starvation condition in oil palm roots was identified as a high-affinity phosphate transporter (EgPHT1). The deduced amino acid sequence has 6 transmembrane domains each at the N- and C-termini separated by a hydrophilic linker. Comparison of promoter motifs within 1500 bp upstream of ATG of 10 promoters from high- and low-affinity phosphate transporter from both dicots and monocots including EgPHT1 was performed. The EgPHT1 promoter was fused to β-glucuronidase (GUS) reporter gene and its activity was analysed by histochemical and fluorometric GUS assays in transiently transformed oil palm tissues and T homozygous transgenic Arabidopsis plants. In response to Pi-starvation, no GUS activity was detected in oil palm leaves, but a strong inducible activity was observed in the roots (1.4 times higher than the CaMV35S promoter). GUS was specifically expressed in transgenic Arabidopsis roots under Pi deficiency and starvation of the other macronutrients (N and K) did not induce GUS activity. Eight motifs including ABRERATCAL (abscisic-acid responsive), RHERPATEXPA7 (root hair-specific), SURECOREATSULTR11 (sulfur-deficiency response), LTRECOREATCOR15 (temperature-stress response), MYB2CONSENSUSAT and ACGTATERD1 (water-stress response) as well as two novel motifs, 3 (TAAAAAAA) and 26 (TTTTATGT) identified through pattern discovery, occur at significantly higher frequency (p < 0.05) in the high-than the low-affinity phosphate transporter promoters. The Pi deficiency-responsive elements in EgPHT1 includes the P1BS, W-box, E-box and the G-box. Thus, EgPHT1 is important for improving Pi uptake in oil palm with potential for engineering efficient Pi acquisition.

摘要

油棕生长在热带土壤中,磷的生物利用度较低。在油棕根中鉴定出一种在磷酸盐饥饿条件下特异性表达的 cDNA 克隆,为高亲和力磷酸盐转运体(EgPHT1)。推导的氨基酸序列在 N 和 C 末端各有 6 个跨膜结构域,由亲水接头隔开。对来自双子叶植物和单子叶植物的高亲和力和低亲和力磷酸盐转运体的 10 个启动子中 ATG 上游 1500bp 内的启动子基序进行了比较,包括 EgPHT1。将 EgPHT1 启动子与β-葡萄糖醛酸酶(GUS)报告基因融合,并在瞬时转化的油棕组织和 T 纯合转基因拟南芥植物中通过组织化学和荧光 GUS 分析其活性。在响应磷饥饿时,在油棕叶片中未检测到 GUS 活性,但在根中观察到强烈的诱导活性(比 CaMV35S 启动子高 1.4 倍)。在缺磷和其他大量营养素(氮和钾)饥饿下,GUS 特异性在转基因拟南芥根中表达,而其他大量营养素(氮和钾)饥饿不会诱导 GUS 活性。通过模式发现,发现了 8 个基序,包括 ABRERATCAL(脱落酸响应)、RHERPATEXPA7(根毛特异性)、SURECOREATSULTR11(硫缺乏响应)、LTRECOREATCOR15(温度胁迫响应)、MYB2CONSENSUSAT 和 ACGTATERD1(水分胁迫响应)以及两个新基序 3(TAAAAAAA)和 26(TTTTATGT),在高亲和力磷酸盐转运体启动子中出现的频率明显高于低亲和力磷酸盐转运体启动子(p<0.05)。EgPHT1 中的磷饥饿响应元件包括 P1BS、W-box、E-box 和 G-box。因此,EgPHT1 对提高油棕的磷吸收具有重要意义,具有提高磷吸收效率的潜力。

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