Bhuria Monika, Goel Parul, Kumar Sanjay, Singh Anil K
Council of Scientific and Industrial Research - Institute of Himalayan Bioresource TechnologyPalampur, India; Academy of Scientific and Innovative ResearchNew Delhi, India.
Front Plant Sci. 2016 Dec 26;7:1957. doi: 10.3389/fpls.2016.01957. eCollection 2016.
Universal stress proteins (USPs) are known to be expressed in response to various abiotic stresses in a wide variety of organisms, such as bacteria, archaebacteria, protists, algae, fungi, plants, and animals. However, in plants, biological function of most of the USPs still remains obscure. In the present study, USP gene () showed induction in response to abscisic acid (ABA) and various abiotic stresses . heat, dehydration, salt, osmotic, and cold stresses. Additionally, analysis of promoter identified several -elements responsive to phytohormones and abiotic stresses such as ABRE, ERE, DRE, and HSE, etc. To functionally validate the promoter, the 1115 bp region of promoter was characterized under phytohormone and abiotic stress treatments. Deletion analysis of promoter was carried out by cloning the full length promoter (D0) and its three 5' deletion derivatives, D1 (964 bp), D2 (660 bp), and D3 (503 bp) upstream of the β-glucuronidase (GUS) reporter gene, which were then stably transformed in plants. The promoter (D0) showed minimal activity under non-stress conditions which was enhanced in response to phytohormone treatments (ABA and ACC) and abiotic stresses such as dehydration, heat, cold, salt, and osmotic stresses. The seedlings harboring D1 and D2 deletion fragments showed constitutive GUS expression even under control condition with increased activity almost under all the treatments. However, D3 seedlings exhibited complete loss of activity under control condition with induction under ACC treatment, dehydration, heat, oxidative, salt, and osmotic stresses. Thus, present study clearly showed that promoter is highly inducible by phytohormones and multiple abiotic stresses and it can be exploited as stress inducible promoter to generate multi-stress tolerant crops with minimal effects on their other important traits.
已知通用应激蛋白(USPs)在多种生物体中,如细菌、古细菌、原生生物、藻类、真菌、植物和动物,会响应各种非生物胁迫而表达。然而,在植物中,大多数通用应激蛋白的生物学功能仍不清楚。在本研究中,USP基因()显示出对脱落酸(ABA)和各种非生物胁迫(如热、脱水、盐、渗透和冷胁迫)的诱导。此外,对该基因启动子的分析鉴定出了几个对植物激素和非生物胁迫有响应的顺式作用元件,如ABRE、ERE、DRE和HSE等。为了从功能上验证该基因的启动子,在植物激素和非生物胁迫处理下对启动子的1115 bp区域进行了表征。通过将全长启动子(D0)及其三个5'端缺失衍生物D1(964 bp)、D2(660 bp)和D3(503 bp)克隆到β-葡萄糖醛酸酶(GUS)报告基因上游,进行启动子缺失分析,然后将其稳定转化到植物中。该基因的启动子(D0)在非胁迫条件下显示出最小活性,而在植物激素处理(ABA和ACC)以及脱水、热、冷、盐和渗透胁迫等非生物胁迫下活性增强。携带D1和D2缺失片段的幼苗即使在对照条件下也表现出组成型GUS表达,且在几乎所有处理下活性都有所增加。然而,D3幼苗在对照条件下表现出完全丧失活性,而在ACC处理、脱水、热、氧化、盐和渗透胁迫下有诱导表达。因此,本研究清楚地表明,该基因启动子对植物激素和多种非生物胁迫具有高度诱导性,可作为胁迫诱导启动子用于培育对多种胁迫具有耐受性且对其他重要性状影响最小的作物。