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节节麦 TmHKT1;4-A1 基因启动子在转基因拟南芥中驱动胁迫诱导、发育调控和组织特异性基因表达。

Promoter of the TmHKT1;4-A1 gene of Triticum monococcum directs stress inducible, developmental regulated and organ specific gene expression in transgenic Arbidopsis thaliana.

机构信息

Biotechnology and Plant Improvement Laboratory, Centre of Biotechnology of Sfax (CBS)/University of Sfax, B. P "1177", 3018, Sfax, Tunisia.

Laboratory of Legumes, Centre of Biotechnology Bordj Cedria, University of Carthage-Tunis, BP 901, 2050, Hammam Lif, Tunisia.

出版信息

World J Microbiol Biotechnol. 2018 Jun 20;34(7):99. doi: 10.1007/s11274-018-2485-9.

DOI:10.1007/s11274-018-2485-9
PMID:29926196
Abstract

HKT transporters which mediate Na-specific transport or Na-K co-transport, play an important role in protecting plants from salinity stress by preventing Na-over-accumulation in leaves. In this work, a 1508-bp genomic fragment upstream of the TmHKT1;4-A1 translated sequence from Triticum monococcum has been isolated, cloned, and designated as the ''PrTmHKT1;4-A1'' promoter. Sequence analysis of ''PrTmHKT1;4-A1'' revealed the presence of cis-regulatory elements which could be required for abiotic stress and abscisic acid (ABA) responsiveness. The PrTmHKT1;4-A1 sequence was fused to the β-glucuronidase gene and the resulting construct was transferred into Arabidopsis plants. Histochemical assays of stably transformed Arabidopsis plants showed that PrTmHKT1;4-A1 is active in this heterologous system. Under control conditions, GUS histochemical staining was observed significantly only in leaves of 20-day-old plants. Histological sections prepared at this stage and in leaves revealed activity localized in leaf veins (phloem and bundle sheath). In flowers, GUS activity was detected only in sepals. After 3 days of challenging the plants with salt, dehydration or ABA treatments, the PrTmHKT1;4-A1 transformed Arabidopsis plants showed a substantial increase in the GUS staining in leaves, compared to untransformed plants under the same conditions. Real time qPCR expression analysis of the uidA gene, showed that GusA transcripts were up-regulated by salt, dehydration, and ABA treatments. All together, these results showed that PrTmHKT1;4-A1 is an age-dependent, abiotic-stress-inducible, organ-specific and tissue-specific promoter in a heterologous dicot system.

摘要

HKT 转运蛋白介导 Na 特异性运输或 Na-K 共转运,通过防止 Na 在叶片中过度积累,在保护植物免受盐胁迫方面发挥重要作用。在这项工作中,从小麦属(Triticum monococcum)TmHKT1;4-A1 翻译序列的上游分离、克隆并命名为 1508bp 基因组片段 ''PrTmHKT1;4-A1'' 启动子。''PrTmHKT1;4-A1'' 的序列分析表明存在顺式调控元件,这些元件可能是生物胁迫和脱落酸(ABA)响应所必需的。PrTmHKT1;4-A1 序列与β-葡萄糖醛酸酶基因融合,所得构建体被转入拟南芥植物中。稳定转化的拟南芥植物的组织化学分析表明,PrTmHKT1;4-A1 在这种异源系统中是活跃的。在对照条件下,仅在 20 天大的植物叶片中观察到 GUS 组织化学染色显著。在这个阶段和叶片中制备的组织切片显示活性定位于叶片脉(韧皮部和束鞘)中。在花朵中,仅在萼片中检测到 GUS 活性。在对植物进行盐、脱水或 ABA 处理 3 天后,与相同条件下的未转化植物相比,PrTmHKT1;4-A1 转化的拟南芥植物叶片中的 GUS 染色显著增加。uidA 基因的实时 qPCR 表达分析表明,GusA 转录物受盐、脱水和 ABA 处理上调。总之,这些结果表明,PrTmHKT1;4-A1 是一种在异源双子叶系统中具有年龄依赖性、非生物胁迫诱导、器官特异性和组织特异性的启动子。

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本文引用的文献

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Characterization of Two HKT1;4 Transporters from Triticum monococcum to Elucidate the Determinants of the Wheat Salt Tolerance Nax1 QTL.来自一粒小麦的两个HKT1;4转运蛋白的特性分析,以阐明小麦耐盐性Nax1 QTL的决定因素。
Plant Cell Physiol. 2016 Oct;57(10):2047-2057. doi: 10.1093/pcp/pcw123. Epub 2016 Jul 20.
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A comparative gene analysis with rice identified orthologous group II HKT genes and their association with Na(+) concentration in bread wheat.
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BMC Plant Biol. 2016 Jan 19;16:21. doi: 10.1186/s12870-016-0714-7.
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OsHKT1;4-mediated Na(+) transport in stems contributes to Na(+) exclusion from leaf blades of rice at the reproductive growth stage upon salt stress.在盐胁迫下,生殖生长阶段水稻茎中由OsHKT1;4介导的钠离子转运有助于叶片中钠离子的外排。
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