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从马蔺中鉴定一个新型 LmSAP 基因启动子:组织特异性和环境胁迫响应。

Characterization of a novel LmSAP gene promoter from Lobularia maritima: Tissue specificity and environmental stress responsiveness.

机构信息

Biotechnology and Plant Improvement Laboratory, Centre of Biotechnology of Sfax, University of Sfax, Sfax, Tunisia.

Plant Production Department, College of Food and Agricultural Sciences, King Saud University, Riyadh, Saudi Arabia.

出版信息

PLoS One. 2020 Jul 31;15(7):e0236943. doi: 10.1371/journal.pone.0236943. eCollection 2020.

Abstract

Halophyte Lobularia maritima LmSAP encodes an A20AN1 zinc-finger stress-associated protein which expression is up-regulated by abiotic stresses and heavy metals in transgenic tobacco. To deepen our understanding of LmSAP function, we isolated a 1,147 bp genomic fragment upstream of LmSAP coding sequence designated as PrLmSAP. In silico analyses of PrLmSAP revealed the presence of consensus CAAT and TATA boxes and cis-regulatory elements required for abiotic stress, phytohormones, pathogen, and wound responses, and also for tissue-specific expression. The PrLmSAP sequence was fused to the β-glucuronidase (gusA) reporter gene and transferred to rice. Histochemical GUS staining showed a pattern of tissue-specific expression in transgenic rice, with staining observed in roots, coleoptiles, leaves, stems and floral organs but not in seeds or in the root elongation zone. Wounding strongly stimulated GUS accumulation in leaves and stems. Interestingly, we observed a high stimulation of the promoter activity when rice seedlings were exposed to NaCl, PEG, ABA, MeJA, GA, cold, and heavy metals (Al3+, Cd2+, Cu2+ and Zn2+). These results suggest that the LmSAP promoter can be a convenient tool for stress-inducible gene expression and is a potential candidate for crop genetic engineering.

摘要

盐生植物蔓花生 LmSAP 编码一个 A20AN1 锌指应激相关蛋白,其表达受非生物胁迫和重金属在转基因烟草中上调。为了深入了解 LmSAP 的功能,我们分离了 LmSAP 编码序列上游的 1147bp 基因组片段,命名为 PrLmSAP。PrLmSAP 的计算机分析显示存在 CAAT 和 TATA 盒的共识以及非生物胁迫、植物激素、病原体和伤口反应以及组织特异性表达所需的顺式调控元件。PrLmSAP 序列与β-葡萄糖醛酸酶(gusA)报告基因融合并转移到水稻中。组织化学 GUS 染色显示转基因水稻具有组织特异性表达模式,在根、幼叶、叶片、茎和花器官中观察到染色,但在种子或根伸长区中未观察到染色。受伤强烈刺激叶片和茎中的 GUS 积累。有趣的是,当水稻幼苗暴露于 NaCl、PEG、ABA、MeJA、GA、冷和重金属(Al3+、Cd2+、Cu2+和 Zn2+)时,我们观察到启动子活性的高度刺激。这些结果表明,LmSAP 启动子可以成为诱导基因表达的便利工具,是作物遗传工程的潜在候选者。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8181/7394455/82c8076d86ea/pone.0236943.g001.jpg

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