The Expression of CARK1 or RCAR11 Driven by Synthetic Promoters Increases Drought Tolerance in Arabidopsis thaliana.

The Expression of CARK1 or RCAR11 Driven by Synthetic Promoters Increases Drought Tolerance in Arabidopsis thaliana.
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合成启动子驱动的 CARK1 或 RCAR11 表达提高拟南芥的耐旱性

DOI:
10.3390/ijms19071945
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发表时间:
2018-07-03
影响因子:
5.6
通讯作者:
Yang Y
Yang Y
中科院分区:
生物学2区
文献类型:
--
作者:
Ge H;Li X;Chen S;Zhang M;Liu Z;Wang J;Li X;Yang Y

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干旱胁迫会阻碍植物的生长发育,而脱落酸(ABA)会刺激植物对干旱的反应。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、ANDp)来确定植物中的转录活性和干旱胁迫抗性,这些启动子由(1)合成启动子和(2)功能基因CARK1(胞质ABA受体激酶1)和RCAR11(ABA受体11的调节成分)的组合产生。 eGFP 的瞬时表达和双荧光素酶测定表明,正常条件下 Ap 和 ANDp 的基础转录活性处于低水平,而合成启动子在外源 ABA 处理或与效应子 DREB2A(脱水反应元件结合蛋白 2A)共转化后明显被诱导。对转基因植物(Ap:CARK1、Dp:CARK1、ANDp:CARK1 和 Dp:RCAR11-Ap:CARK1)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:CARK1 和 Dp:RCAR11-Ap:CARK1 转基因植物在子叶绿化和根生长过程中对 ABA 和 d-甘露醇敏感。耐旱性测定显示,ANDp:CARK1 和 Dp:RCAR11-Ap:CARK1 在干旱胁迫下表现出比其他模型更高的存活率。这些结果表明ANDp:CARK1和Dp:RCAR11-Ap:CARK1的组合可用于在植物中产生干旱胁迫抗性。
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 CARK1 (cytosolic ABA receptor kinase 1) and RCAR11 (regulatory components of ABA receptor 11). Transient expression of eGFP 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:CARK1, Dp:CARK1, ANDp:CARK1, and Dp:RCAR11-Ap:CARK1) 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:CARK1 and Dp:RCAR11-Ap:CARK1 transgenic plants were sensitive to ABA and d-mannitol during cotyledon greening and root growth. A drought tolerance assay revealed that ANDp:CARK1 and Dp:RCAR11-Ap:CARK1 exhibited a higher survival rate than others upon drought stress. These results indicate that the combinations ANDp:CARK1 and Dp:RCAR11-Ap:CARK1 can be used to generate drought stress resistance in plants.
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