School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
Plant Cell. 2010 Jan;22(1):91-107. doi: 10.1105/tpc.109.071803. Epub 2010 Jan 29.
The Arabidopsis thaliana ABORTED MICROSPORES (AMS) gene encodes a basic helix-loop-helix (bHLH) transcription factor that is required for tapetal cell development and postmeiotic microspore formation. However, the regulatory role of AMS in anther and pollen development has not been fully defined. Here, we show by microarray analysis that the expression of 549 anther-expressed genes was altered in ams buds and that these genes are associated with tapetal function and pollen wall formation. We demonstrate that AMS has the ability to bind in vitro to DNA containing a 6-bp consensus motif, CANNTG. Moreover, 13 genes involved in transportation of lipids, oligopeptides, and ions, fatty acid synthesis and metabolism, flavonol accumulation, substrate oxidation, methyl-modification, and pectin dynamics were identified as direct targets of AMS by chromatin immunoprecipitation. The functional importance of the AMS regulatory pathway was further demonstrated by analysis of an insertional mutant of one of these downstream AMS targets, an ABC transporter, White-Brown Complex homolog, which fails to undergo pollen development and is male sterile. Yeast two-hybrid screens and pull-down assays revealed that AMS has the ability to interact with two bHLH proteins (AtbHLH089 and AtbHLH091) and the ATA20 protein. These results provide insight into the regulatory role of the AMS network during anther development.
拟南芥 ABORTED MICROSPORES (AMS) 基因编码一个碱性螺旋-环-螺旋(bHLH)转录因子,该基因对于绒毡层细胞发育和减数后小孢子形成是必需的。然而,AMS 在花药和花粉发育中的调控作用尚未完全确定。在这里,我们通过微阵列分析表明,在 ams 芽中改变了 549 个花药表达基因的表达,这些基因与绒毡层功能和花粉壁形成有关。我们证明 AMS 具有在体外结合含有 6 个碱基对的保守基序(CANNTG)的 DNA 的能力。此外,通过染色质免疫沉淀鉴定了 13 个与脂质、寡肽和离子运输、脂肪酸合成和代谢、黄酮醇积累、底物氧化、甲基化修饰和果胶动力学有关的基因,作为 AMS 的直接靶标。通过对其中一个下游 AMS 靶标,ABC 转运蛋白,White-Brown Complex 同源物的插入突变体的分析,进一步证明了 AMS 调控途径的功能重要性,该突变体不能进行花粉发育,是雄性不育的。酵母双杂交筛选和下拉测定表明 AMS 具有与两个 bHLH 蛋白(AtbHLH089 和 AtbHLH091)和 ATA20 蛋白相互作用的能力。这些结果为了解 AMS 网络在花药发育过程中的调控作用提供了线索。