State Key Laboratory of Soil Erosion and Dryland Farming on the Loess Plateau, College of Life Sciences, Northwest A&F University, Yangling 712100, China.
New Energy Department, School of Energy and Power Engineering, Jiangsu University, Zhenjiang 212013, China.
Int J Mol Sci. 2018 Jan 8;19(1):191. doi: 10.3390/ijms19010191.
Betaine, a non-toxic osmoprotectant, is believed to accumulate considerably in plants under stress conditions to maintain the osmotic pressure and promote a variety of processes involved in growth and development. Phosphoethanolamine -methyltransferase (PEAMT), a key enzyme for betaine synthesis, is reported to be regulated by its upstream promoter. In the present investigation, by using the transgenic approach, a 1048 bp long promoter region of gene from was cloned and functionally characterized in tobacco. Computational analysis affirmed the existence of abiotic stress responsive -elements like ABRE, MYC, HST, LST etc., as well as pathogen, wound and phytohormone responsive motifs. For transformation in tobacco, four 5'-deletion constructs of 826 bp (P2), 642 bp (P3), 428 bp (P4) and 245 bp (P5) were constructed from the 1048 bp (P1) promoter fragment. The transgenic plants generated through a single event exhibited a promising expression of GUS reporter protein in the leaf tissues of treated with salt, drought, oxidative and cold stress as well as control plants. The GUS expression level progressively reduced from P1 to P5 in the leaf tissues, whereas a maximal expression was observed with the P3 construct in the leaves of control plants. The expression of GUS was noted to be higher in the leaves of osmotically- or salt-treated transgenic plants than that in the untreated (control) plants. An effective expression of GUS in the transgenic plants manifests that this promoter can be employed for both stress-inducible and constitutive expression of gene(s). Due to this characteristic, this potential promoter can be effectively used for genetic engineering of several crops.
甜菜碱是一种无毒的渗透保护剂,据信在应激条件下会在植物中大量积累,以维持渗透压并促进与生长和发育相关的各种过程。磷酸乙醇胺 -N-甲基转移酶(PEAMT)是甜菜碱合成的关键酶,据报道其受到其上游启动子的调控。在本研究中,通过转基因方法,从 中克隆并功能表征了 基因的 1048bp 长启动子区域在烟草中。计算分析证实了存在非生物胁迫响应元件,如 ABRE、MYC、HST、LST 等,以及病原体、伤口和植物激素响应元件。为了在烟草中进行转化,从 1048bp(P1)启动子片段构建了四个 5'-缺失构建体:826bp(P2)、642bp(P3)、428bp(P4)和 245bp(P5)。通过单个事件生成的转基因植物在经过盐、干旱、氧化和冷胁迫以及对照植物处理的叶片组织中表现出 GUS 报告蛋白的有希望的表达。在叶片组织中,GUS 表达水平从 P1 到 P5 逐渐降低,而在对照植物的 P3 构建体中观察到最大表达。在渗透或盐处理的转基因植物的叶片中观察到 GUS 的表达高于未处理(对照)植物的表达。GUS 在转基因植物中的有效表达表明该启动子可用于基因的胁迫诱导和组成型表达。由于这个特点,这个潜在的启动子可以有效地用于几种作物的遗传工程。