Key Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, China.
Int J Mol Sci. 2018 Jul 3;19(7):1945. doi: 10.3390/ijms19071945.
Drought stress hinders plant growth and development, and abscisic acid (ABA) stimulates plants to respond to drought. Here, to increase plant tolerance to drought, we designed three synthetic promoters (Ap, Dp, ANDp) to determine transcription activity and drought stress resistance in plants resulting from combinations of (1) synthetic promoters and (2) the functional genes (cytosolic ABA receptor kinase 1) and (regulatory components of ABA receptor 11). Transient expression of and the dual-luciferase assay demonstrated that the basal transcriptional activities of Ap and ANDp were present at low levels under normal conditions, while the synthetic promoters were apparently induced upon either treatment of exogenous ABA or co-transformation with effector DREB2A (dehydration-responsive element binding protein 2A). Analysis of the transgenic plants (Ap:, Dp:, ANDp:, and Dp:-Ap:) showed that the synthetic promoters Ap, Dp, and ANDp increased the expression of exogenous genes in transgenic plants upon treatment of ABA or d-mannitol. ANDp: and Dp:-Ap: transgenic plants were sensitive to ABA and d-mannitol during cotyledon greening and root growth. A drought tolerance assay revealed that ANDp: and Dp:-Ap: exhibited a higher survival rate than others upon drought stress. These results indicate that the combinations ANDp: and Dp:-Ap: can be used to generate drought stress resistance in plants.
干旱胁迫会阻碍植物的生长和发育,而脱落酸(ABA)会刺激植物对干旱作出响应。在这里,为了提高植物对干旱的耐受性,我们设计了三个合成启动子(Ap、Dp 和 ANDp),以确定组合(1)合成启动子和(2)功能基因(细胞质 ABA 受体激酶 1 和 ABA 受体 11 的调节成分)对植物转录活性和干旱胁迫抗性的影响。瞬时表达和双荧光素酶测定表明,在正常条件下,Ap 和 ANDp 的基础转录活性水平较低,而在施加外源 ABA 或与效应物 DREB2A(脱水响应元件结合蛋白 2A)共转化时,合成启动子明显被诱导。对转基因植物(Ap:、Dp:、ANDp:和 Dp:-Ap:)的分析表明,在 ABA 或 d-甘露醇处理后,合成启动子 Ap、Dp 和 ANDp 增加了转基因植物中外源基因的表达。ANDp:和 Dp:-Ap:转基因植物在子叶变绿和根生长过程中对 ABA 和 d-甘露醇敏感。干旱胁迫试验表明,ANDp:和 Dp:-Ap:在干旱胁迫下的存活率高于其他株系。这些结果表明,ANDp:和 Dp:-Ap:的组合可用于提高植物的抗旱性。