College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, People's Republic of China.
College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, People's Republic of China; Gansu Key Laboratory of Crop Genetic and Germplasm Enhancement, Gansu Provincial Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Lanzhou 730070, People's Republic of China.
Plant Physiol Biochem. 2018 Apr;125:63-73. doi: 10.1016/j.plaphy.2018.01.027. Epub 2018 Feb 2.
The DWARF4 (DWF4) gene encodes a C-22 hydroxylase which is pivotal for brassinosteroids (BRs) biosynthesis. In this research, aimed to understand the molecular mechanism of DWF4 on regulation of potatoes tolerance to salt stress, DWF4 was cloned from potato, named as StDWF4. Its 1476 bp open reading frame encodes a protein of 491 amino acids. The StDWF4-overexpressing (OE) and interference-expressing (RNAi) transgenic potato plants were acquired using Agrobacterium-mediated transformation, respectively. Tissue specific analysis using Quantitative real-time polymerase chain reaction (qRT-PCR) demonstrated that the StDWF4 gene expressed in the leaves, stems and roots of the transgenic and un-transgenic (NT) plants, with specially increased (StDWF4-OE)/reduced (StDWF4-RNAi) expression in the roots. The content of malondialdehyde (MDA) in StDWF4-OE potato plants was lower than that of NT, and proline content was higher than that of NT. MDA and proline content in StDWF4-OE and NT under salt-stress was significantly higher than that of the control and was increased at different sampling times. The content of soluble protein, soluble sugar and the activities of superoxide dismutase (SOD), peroxidase (POD) and ascorbate peroxidase (APX) was higher in the StDWF4-OE plantlets at varied salt treatment time than in the NT potatoes. Reduction of HO content in the StDWF4-OE plants was observed. All above plant physiology indicators in the StDWF4-RNAi potatoes showed opposite variation trends. The results proved that the overexpressing of StDWF4 in potato plantlets can enhance the salt resistance by alleviating the negative effects of salt-stress. However, its interference expression in potato plants depresses the salt resistance. The results lay the groundwork for intensive study of BRs regulation in potato growth and development, and will help us to reveal the molecular mechanisms of how the BRs signaling regulate potato salt tolerance.
DWARF4(DWF4)基因编码一个 C-22 羟化酶,该酶对油菜素内酯(BRs)生物合成至关重要。在这项研究中,旨在了解 DWF4 对调控马铃薯耐盐性的分子机制,从马铃薯中克隆了 DWF4,命名为 StDWF4。其 1476 bp 的开放阅读框编码一个 491 个氨基酸的蛋白质。通过农杆菌介导的转化,分别获得了 StDWF4 过表达(OE)和干扰表达(RNAi)的转基因马铃薯植株。利用定量实时聚合酶链反应(qRT-PCR)进行组织特异性分析表明,StDWF4 基因在转基因和非转基因(NT)植株的叶片、茎和根中表达,特别是在根中表达增加(StDWF4-OE)/减少(StDWF4-RNAi)。StDWF4-OE 马铃薯植株中的丙二醛(MDA)含量低于 NT,脯氨酸含量高于 NT。StDWF4-OE 和 NT 在盐胁迫下的 MDA 和脯氨酸含量均显著高于对照,并在不同采样时间增加。在不同盐处理时间下,StDWF4-OE 植株的可溶性蛋白、可溶性糖含量以及超氧化物歧化酶(SOD)、过氧化物酶(POD)和抗坏血酸过氧化物酶(APX)的活性均高于 NT 马铃薯。在 StDWF4-OE 植株中观察到 HO 含量降低。StDWF4-RNAi 马铃薯的所有上述植物生理指标均表现出相反的变化趋势。结果证明,在马铃薯植株中过表达 StDWF4 可以通过减轻盐胁迫的负面影响来增强耐盐性。然而,其在马铃薯植株中的干扰表达会降低其耐盐性。该结果为深入研究 BRs 对马铃薯生长发育的调控奠定了基础,并将有助于揭示 BRs 信号转导调节马铃薯耐盐性的分子机制。