A novel sensor to map auxin response and distribution at high spatio-temporal resolution

Auxin is a key plant morphogenetic signal but tools to analyse dynamically its distribution and signalling during development are still limited. Auxin perception directly triggers the degradation of Aux/IAA repressor proteins. Here we describe a novel Aux/IAA-based auxin signalling sensor termed DII-VENUS that was engineered in the model plant Arabidopsis thaliana. The VENUS fast maturing form of yellow fluorescent protein was fused in-frame to the Aux/IAA auxin-interaction domain (termed domain II; DII) and expressed under a constitutive promoter. We initially show that DII-VENUS abundance is dependent on auxin, its TIR1/AFBs co-receptors and proteasome activities. Next, we demonstrate that DII-VENUS provides a map of relative auxin distribution at cellular resolution in different tissues. DII-VENUS is also rapidly degraded in response to auxin and we used it to visualize dynamic changes in cellular auxin distribution successfully during two developmental responses, the root gravitropic response and lateral organ production at the shoot apex. Our results illustrate the value of developing response input sensors such as DII-VENUS to provide high-resolution spatio-temporal information about hormone distribution and response during plant growth and development.     

Auxin transport inhibitors block PIN1 cycling and vesicle trafficking

Polar transport of the phytohormone auxin mediates various processes in plant growth and development, such as apical dominance, tropisms, vascular patterning and axis formation. This view is based largely on the effects of polar auxin transport inhibitors. These compounds disrupt auxin efflux from the cell but their mode of action is unknown. It is thought that polar auxin flux is caused by the asymmetric distribution of efflux carriers acting at the plasma membrane. The polar localization of efflux carrier candidate PIN1 supports this model. Here we show that the seemingly static localization of PIN1 results from rapid actin-dependent cycling between the plasma membrane and endosomal compartments. Auxin transport inhibitors block PIN1 cycling and inhibit trafficking of membrane proteins that are unrelated to auxin transport. Our data suggest that PIN1 cycling is of central importance for auxin transport and that auxin transport inhibitors affect efflux by generally interfering with membrane-trafficking processes. In support of our conclusion, the vesicle-trafficking inhibitor brefeldin A mimics physiological effects of auxin transport inhibitors.     

Auxin transport is sufficient to generate a maximum and gradient guiding root growth

The plant growth regulator auxin controls cell identity, cell division and cell expansion. Auxin efflux facilitators (PINs) are associated with auxin maxima in distal regions of both shoots and roots. Here we model diffusion and PIN-facilitated auxin transport in and across cells within a structured root layout. In our model, the stable accumulation of auxin in a distal maximum emerges from the auxin flux pattern. We have experimentally tested model predictions of robustness and self-organization. Our model explains pattern formation and morphogenesis at timescales from seconds to weeks, and can be understood by conceptualizing the root as an 'auxin capacitor'. A robust auxin gradient associated with the maximum, in combination with separable roles of auxin in cell division and cell expansion, is able to explain the formation, maintenance and growth of sharply bounded meristematic and elongation zones. Directional permeability and diffusion can fully account for stable auxin maxima and gradients that can instruct morphogenesis.     

结球白菜在个体发育中苗端淀粉粒积累的变化

本文描述了结球白菜苗瑞细胞中淀粉粒的含量在个体发育过程中的变化规律,结果是: 结球白菜苗端在分化和形成叶原基的过程中,淀粉粒主要积累于肋状分生组织细胞中,且随苗端产生叶原基活动的加强而增多;随营养苗端向生殖苗端的转化,其淀粉粒的含量明显减少,转化完成后的生殖苗端细胞中,极少见到淀粉粒。由此可见,淀粉物质的积累和变化与叶原基的分化形成及白菜包心结球有着密切的关系。     

AtSNX1 defines an endosome for auxin-carrier trafficking in Arabidopsis

Polarized cellular distribution of the phytohormone auxin and its carriers is essential for normal plant growth and development. Polar auxin transport is maintained by a network of auxin influx (AUX) and efflux (PIN) carriers. Both auxin transport and PIN protein cycling between the plasma membrane and endosomes require the activity of the endosomal GNOM; however, intracellular routes taken by these carriers remain largely unknown. Here we show that Arabidopsis thaliana SORTING NEXIN 1 (AtSNX1) is involved in the auxin pathway and that PIN2, but not PIN1 or AUX1, is transported through AtSNX1-containing endosomes. We demonstrate that the snx1-null mutant exhibits multiple auxin-related defects and that loss of function of AtSNX1 severely enhances the phenotype of a weak gnom mutant. In root cells, we further show that AtSNX1 localizes to an endosomal compartment distinct from GNOM-containing endosomes, and that PIN2 accumulates in this compartment after treatment with the phosphatidylinositol-3-OH kinase inhibitor wortmannin or after a gravity stimulus. Our data reveal the existence of a novel endosomal compartment involved in PIN2 endocytic sorting and plant development.     

冬小麦节与节间发生的发育解剖学研究

本文从发育解剖学角度研究了冬小麦顶端分生组织各区的发展衍生情况.在冬小麦中,顶端分生组织应分为原分生组织、初生分生组织及位于二者之间的过渡区.节与节间由过渡区细胞分化产生。节间开始伸长及小穗原基分化均发生在起身期.     

Control of phyllotaxy by the cytokinin-inducible response regulator homologue ABPHYL1

Phyllotaxy describes the geometric pattern of leaves and flowers, and has intrigued botanists and mathematicians for centuries. How these patterns are initiated is poorly understood, and this is partly due to the paucity of mutants. Signalling by the plant hormone auxin appears to determine the site of leaf initiation; however, this observation does not explain how distinct patterns of phyllotaxy are initiated. abphyl1 (abph1) mutants of maize initiate leaves in a decussate pattern (that is, paired at 180 degrees), in contrast to the alternating or distichous phyllotaxy observed in wild-type maize and other grasses. Here we show that ABPH1 is homologous to two-component response regulators and is induced by the plant hormone cytokinin. ABPH1 is expressed in the embryonic shoot apical meristem, and its spatial expression pattern changes rapidly with cytokinin treatment. We propose that ABPH1 controls phyllotactic patterning by negatively regulating the cytokinin-induced expansion of the shoot meristem, thereby limiting the space available for primordium initiation at the apex.     

拟南芥PIN2介导的生长素极性运输调控植物根向地性

主要观察了拟南芥生长素输出载体PIN2及其介导的极性运输、生长素诱导合成对根尖生长素不对称分布和根向地性反应的影响.结果表明:拟南芥PIN2基因突变、诱导内源生长素IAA及用抑制剂NPA或TIBA抑制生长素极性运输都严重影响了根尖生长素不对称分布的形成,最终抑制植物根向地性反应,暗示PIN2介导的生长素极性运输和生长素不对称分布在根向地性反应中起着关键性调控作用.从这些研究结果可进一步理解生长素调控植物根向地性的分子机理.     

A novel putative auxin carrier family regulates intracellular auxin homeostasis in plants

The phytohormone auxin acts as a prominent signal, providing, by its local accumulation or depletion in selected cells, a spatial and temporal reference for changes in the developmental program. The distribution of auxin depends on both auxin metabolism (biosynthesis, conjugation and degradation) and cellular auxin transport. We identified in silico a novel putative auxin transport facilitator family, called PIN-LIKES (PILS). Here we illustrate that PILS proteins are required for auxin-dependent regulation of plant growth by determining the cellular sensitivity to auxin. PILS proteins regulate intracellular auxin accumulation at the endoplasmic reticulum and thus auxin availability for nuclear auxin signalling. PILS activity affects the level of endogenous auxin indole-3-acetic acid (IAA), presumably via intracellular accumulation and metabolism. Our findings reveal that the transport machinery to compartmentalize auxin within the cell is of an unexpected molecular complexity and demonstrate this compartmentalization to be functionally important for a number of developmental processes.     

Auxin inhibits endocytosis and promotes its own efflux from cells

One of the mechanisms by which signalling molecules regulate cellular behaviour is modulating subcellular protein translocation. This mode of regulation is often based on specialized vesicle trafficking, termed constitutive cycling, which consists of repeated internalization and recycling of proteins to and from the plasma membrane. No such mechanism of hormone action has been shown in plants although several proteins, including the PIN auxin efflux facilitators, exhibit constitutive cycling. Here we show that a major regulator of plant development, auxin, inhibits endocytosis. This effect is specific to biologically active auxins and requires activity of the Calossin-like protein BIG. By inhibiting the internalization step of PIN constitutive cycling, auxin increases levels of PINs at the plasma membrane. Concomitantly, auxin promotes its own efflux from cells by a vesicle-trafficking-dependent mechanism. Furthermore, asymmetric auxin translocation during gravitropism is correlated with decreased PIN internalization. Our data imply a previously undescribed mode of plant hormone action: by modulating PIN protein trafficking, auxin regulates PIN abundance and activity at the cell surface, providing a mechanism for the feedback regulation of auxin transport.     

Auxin promotes Arabidopsis root growth by modulating gibberellin response

The growth of plant organs is influenced by a stream of the phytohormone auxin that flows from the shoot apex to the tip of the root. However, until now it has not been known how auxin regulates the cell proliferation and enlargement that characterizes organ growth. Here we show that auxin controls the growth of roots by modulating cellular responses to the phytohormone gibberellin (GA). GA promotes the growth of plants by opposing the effects of nuclear DELLA protein growth repressors, one of which is Arabidopsis RGA (for repressor of gal-3). GA opposes the action of several DELLA proteins by destabilizing them, reducing both the concentration of detectable DELLA proteins and their growth-restraining effects. We also show that auxin is necessary for GA-mediated control of root growth, and that attenuation of auxin transport or signalling delays the GA-induced disappearance of RGA from root cell nuclei. Our observations indicate that the shoot apex exerts long-distance control on the growth of plant organs through the effect of auxin on GA-mediated DELLA protein destabilization.     

植物根尖分生组织干细胞调控模式

静止中心(QC)形成和干细胞区特化是植物根尖分生组织确立的标志。静止中心位于根尖分生组织中心,干细胞围绕在静止中心细胞周围。依赖于生长素的PLT途径和不依赖于生长素的SCR/SHR途径共同发挥维持静止中心细胞稳定的作用。静止中心和干细胞区的柱干细胞之间存在类似于WUS/CLV3的WOX5/ACR4/CLE40的反馈抑制调节途径,该调节途径维持着静止中心细胞和柱干细胞之间的平衡。静止中心和其他类型干细胞之间也可能存在类似的反馈抑制调节途径。生长素、细胞分裂素、赤霉素等植物激素信号在根干细胞功能发挥方面也起到重要作用,与各种基因一起组成根分生组织干细胞调控网络。     

裸燕麦不同发育时期的茎尖外植体中内源激素与体细胞胚胎发生的关系

裸燕麦的茎尖开始拉长,小穗原基开始分化和小穗形成三个不同的分化阶段的外植体在诱导体细胞胚胎发生时,随小穗原基的分化和形成过程,要求外源2.4-D浓度越来越高内源生长素(1AA)和细胞分裂素(CTK)分析结果表明三个不同分化阶段的1AA/CTK比值分别为1.36:1,l:1.02和1:2.08,而脱分化的愈伤组织和胚性愈伤组织中二者比值分别为1.81:1和5.10:1。我们推测,体细胞胚胎发生过程中内源生长素的高比值是要求大量提高外源2.4-D浓度的原因。