Liu Chenggang, Mehdy Mona C
Section of Molecular, Cell, and Developmental Biology, University of Texas, Austin, Texas 78712, USA.
Plant Physiol. 2007 Nov;145(3):863-74. doi: 10.1104/pp.107.102657. Epub 2007 Sep 20.
In response to wounding and pathogens, jasmonate (JA) serves as a signal molecule for both induction and repression of gene expression. To examine defense-regulated gene repression in Arabidopsis (Arabidopsis thaliana), we have identified a nonclassical arabinogalactan protein (AGP) gene, AGP31, and show that its mRNA decreased to about 30% of its original level within 8 h in response to methyl JA (MeJA) treatment of whole 7-d-old seedlings. Wounding and abscisic acid treatment had similar effects. MeJA suppression primarily depends on the action of the JA-signaling protein, COI1, as shown by much lower MeJA suppression in coi1-1 mutant plants. The main mechanism of mRNA suppression by MeJA is repression of transcription, as shown by nuclear run-on experiments. The AGP31 protein shares features with several known and putative nonclassical AGPs from other species: a putative signal peptide, a histidine-rich region near the N terminus followed by a repetitive proline-rich domain, and a cysteine-rich C-terminal PAC (for proline-rich protein and AGP, containing cysteine) domain. Positive Yariv reagent interaction demonstrated that the protein is an AGP. Monosaccharide analysis of purified AGP31 indicated it is a galactose-rich AGP. Expression of an AGP31-enhanced green fluorescent protein fusion protein in transgenic cells revealed that the AGP31 protein was localized to the cell wall. AGP31 promoter-beta-glucuronidase reporter gene analysis showed expression in the vascular bundle throughout the plant, except in the flower. In the flower, beta-glucuronidase staining occurred throughout the pistil, except in the stigma. The strong preferential expression in vascular tissues suggests that AGP31 may be involved in vascular tissue function during both the defense response and development.
作为对伤口和病原体的响应,茉莉酸(JA)作为一种信号分子,既可以诱导基因表达,也可以抑制基因表达。为了研究拟南芥中防御调节的基因抑制作用,我们鉴定了一个非经典阿拉伯半乳聚糖蛋白(AGP)基因AGP31,并发现,用甲基茉莉酸(MeJA)处理7日龄的完整幼苗后,其mRNA在8小时内降至原来水平的约30%。创伤和脱落酸处理也有类似效果。如在coi1-1突变体植株中MeJA抑制作用低得多所示,MeJA抑制主要取决于JA信号蛋白COI1的作用。如核延伸实验所示,MeJA抑制mRNA的主要机制是转录抑制。AGP31蛋白与来自其他物种的几个已知和推测的非经典AGP具有共同特征:一个推测的信号肽、靠近N端的富含组氨酸区域,随后是富含脯氨酸的重复结构域,以及富含半胱氨酸的C端PAC(富含脯氨酸蛋白和AGP,含半胱氨酸)结构域。阳性的Yariv试剂相互作用表明该蛋白是一种AGP。对纯化的AGP31进行单糖分析表明它是一种富含半乳糖的AGP。在转基因细胞中表达AGP31增强型绿色荧光蛋白融合蛋白表明AGP31蛋白定位于细胞壁。AGP31启动子-β-葡萄糖醛酸酶报告基因分析表明,除了在花中,该基因在整个植株的维管束中均有表达。在花中,β-葡萄糖醛酸酶染色出现在整个雌蕊中,但柱头除外。在维管组织中的强烈优先表达表明,AGP31可能在防御反应和发育过程中参与维管组织功能。