Kim Insoon, Kobayashi Ken, Cho Euna, Zambryski Patricia C
Department of Plant and Microbial Biology, 111 Koshland Hall, University of California, Berkeley, CA 94720.
Proc Natl Acad Sci U S A. 2005 Aug 16;102(33):11945-50. doi: 10.1073/pnas.0505622102. Epub 2005 Aug 8.
The axial body pattern of Arabidopsis is determined during embryogenesis by auxin signaling and differential gene expression. Here we demonstrate that another pathway, cell-to-cell communication through plasmodesmata (PD), is regulated during apical-basal pattern formation. The SHOOT MERISTEMLESS (STM) promoter was used to drive expression in the shoot apical meristem (SAM) and a subset of cells at the base of the hypocotyl of 1x,2x, and 3x soluble green fluorescent proteins (sGFPs), and the P30 movement protein of Tobacco mosaic virus (TMV) translationally fused to 1x and 2x sGFP. In the early heart stage, 2x sGFP (54 kDa) moves throughout the whole embryo, whereas 3x sGFP (81 kDa) shows more restricted movement. As the embryo develops, PD apertures are down regulated to form local subdomains allowing transport of different sized tracers. For example, movement of 2x sGFP to the cotyledon, and 3x sGFP to root tips, becomes restricted. Subdomains of cell-to-cell transport align with the apical-basal embryo body axis and correspond to the shoot apex, cotyledons, hypocotyl, and root. Studies with P30-GFP fusions reinforce the distinction between embryonic symplastic subdomains. Although P30 targets embryo cell walls as puncta (diagnostic for functional localization of P30 to PD in adult plants), P30 cannot dilate embryonic PD to overcome the barriers for transport between symplastic subdomains, suggesting that specific boundaries separate symplastic subdomains of the embryo. Thus, cell-to-cell communication via plasmodesmata conveys positional information critical to establish the axial body pattern during embryogenesis in Arabidopsis.
拟南芥的轴向身体模式在胚胎发生过程中由生长素信号传导和差异基因表达决定。在这里,我们证明了另一条途径,即通过胞间连丝(PD)进行的细胞间通讯,在顶基模式形成过程中受到调控。使用无茎尖分生组织(STM)启动子驱动在茎尖分生组织(SAM)以及下胚轴基部的一部分细胞中表达1倍、2倍和3倍可溶性绿色荧光蛋白(sGFP),以及将烟草花叶病毒(TMV)的P30运动蛋白与1倍和2倍sGFP进行翻译融合。在早期心形期,2倍sGFP(54 kDa)在整个胚胎中移动,而3倍sGFP(81 kDa)的移动则受到更多限制。随着胚胎发育,PD孔径下调以形成局部亚域,允许不同大小的示踪剂运输。例如,2倍sGFP向子叶的移动以及3倍sGFP向根尖的移动受到限制。细胞间运输的亚域与胚胎的顶基轴对齐,并对应于茎尖、子叶、下胚轴和根。对P30-GFP融合体的研究强化了胚胎共质体亚域之间的区别。尽管P30以斑点形式靶向胚胎细胞壁(这是成年植物中P30在PD上功能定位的诊断特征),但P30不能扩张胚胎PD以克服共质体亚域之间运输的障碍,这表明特定的边界将胚胎的共质体亚域分隔开。因此,通过胞间连丝进行的细胞间通讯传递了对拟南芥胚胎发生过程中建立轴向身体模式至关重要的位置信息。