Wang Wei, Wang Xutong, Liu Xiaoyu, Wang Yating, Li Yingying, Hussain Saddam, Jing Xiaoxiao, Chen Siyu, Wang Shucai
Laboratory of Plant Molecular Genetics and Crop Gene Editing, School of Life Sciences, Linyi University, Linyi 276000, China.
Heilongjiang Provincial Key Laboratory of Environmental Microbiology and Recycling of Argo-Waste in Cold Region, College of Life Science and Biotechnology, Heilongjiang Bayi Agricultural University, Daqing 163319, China.
Plants (Basel). 2024 Nov 22;13(23):3282. doi: 10.3390/plants13233282.
The abscisic acid (ABA) signaling pathway is crucial for regulating downstream ABA-responsive genes, which influence plant responses to ABA and abiotic stresses. However, many ABA-responsive genes remain poorly characterized. This study reports on the identification and characterization of ABA up-regulated EAR motif-containing proteins (), a novel family of EAR motif-containing proteins in . From a previous transcriptome dataset, were identified as a family of unknown-function ABA-response genes with only five members, and the up-regulation of by ABA was further confirmed by quantitative RT-PCR (qRT PCR). All contain at least one LxLxL EAR motif and can repress reporter gene expression in Arabidopsis protoplasts. We generated CRISPR/Cas9 gene-edited , and single, () double, and () triple mutants, as well as transgenic plants overexpressing , and examined their ABA sensitivity. We found that the single and double mutants displayed wild-type responses to ABA treatment, while the triple mutants showed increased sensitivity in seed germination and cotyledon greening assays but decreased sensitivity to ABA treatment in root elongation assays. Conversely, the showed decreased sensitivity in seed germination and cotyledon greening assays but increased sensitivity to ABA treatment in root elongation assays. The qRT PCR results show that the expression level of was increased in the mutants and decreased in the plants. These findings suggest that function redundantly to regulate ABA responses in Arabidopsis, likely by modulating the expression of key regulatory genes in ABA-signaling pathway.
脱落酸（ABA）信号通路对于调控下游ABA应答基因至关重要，这些基因影响植物对ABA和非生物胁迫的反应。然而，许多ABA应答基因的特征仍不清楚。本研究报道了ABA上调的含EAR基序蛋白（）的鉴定和特征，这是中一个新的含EAR基序蛋白家族。从先前的转录组数据集中，被鉴定为一个功能未知的ABA应答基因家族，仅包含五个成员，并且通过定量RT-PCR（qRT-PCR）进一步证实了ABA对其的上调作用。所有都至少包含一个LxLxL EAR基序，并且能够在拟南芥原生质体中抑制报告基因的表达。我们构建了CRISPR/Cas9基因编辑的、和单突变体，（）双突变体，以及（）三突变体，还有过表达的转基因植物，并检测了它们对ABA的敏感性。我们发现单突变体和双突变体对ABA处理表现出野生型反应，而三突变体在种子萌发和子叶绿化试验中对ABA处理的敏感性增加，但在根伸长试验中对ABA处理的敏感性降低。相反，在种子萌发和子叶绿化试验中对ABA处理的敏感性降低，但在根伸长试验中对ABA处理的敏感性增加。qRT-PCR结果表明，在突变体中的表达水平升高，而在过表达植株中的表达水平降低。这些发现表明，在拟南芥中可能通过调节ABA信号通路中关键调控基因的表达来冗余调控ABA反应。
