Camellia sinensis CsMYB4a participates in regulation of stamen growth by interaction with auxin signaling transduction repressor CsAUX/IAA4

Subgroup 4(Sg4)members of the R2R3-MYB are generally known as negative regulators of the phenylpropanoid pathway in plants.Our previous research showed that a R2R3-MYB Sg4 member from Camellia sinensis(CsMYB4a)inhibits expression of some genes in the phenylpropanoid pathway,but its physiological function in the tea plant remained unknown.Here,CsMYB4a was found to be highly expressed in anther and filaments,and participated in regulating filament growth.Transcriptome analysis and exogenous auxin treatment showed that the target of CsMYB4a might be the auxin signal pathway.Auxin/indole-3-acetic acid 4(AUX/IAA4),a repressor in auxin signal transduction,was detected from a yeast two-hybrid screen using CsMYB4a as bait.Gene silencing assays showed that both CsIAA4 and CsMYB4a regulate filament growth.Tobacco plants overexpressing CsIAA4 were insensitive to exogenous a-NAA,consistent with overexpression of CsMYB4a.Protein-protein interaction experiments revealed that CsMYB4a interacts with N-terminal of CsIAA4 to prevent CsIAA4 degradation.Knock out of the endogenous NtIAA4 gene,a CsIAA4 homolog,in tobacco alleviated filament growth inhibition and a-NAA insensitivity in plants overexpressing CsMYB4a.All results strongly suggest that CsMYB4a works synergistically with CsIAA4 and participates in regulation of the auxin pathway in stamen.

Protein post-translational modifications in auxin signaling

Protein post-translational modifications(PTMs),such as ubiquitination,phosphorylation,and small ubiquitin-like modifier(SUMO)ylation,are crucial for regulating protein stability,activity,subcellular localization,and binding with cofactors.Such modifications remarkably increase the variety and complexity of proteomes,which are essential for regulating numerous cellular and physiological processes.The regulation of auxin signaling is finely tuned in time and space to guide various plant growth and development.Accumulating evidence indicates that PTMs play critical roles in auxin signaling regulations.Thus,a thorough and systematic review of the functions of PTMs in auxin signal transduction will improve our profound comprehension of the regulation mechanism of auxin signaling and auxin-mediated various processes.This review discusses the progress of protein ubiquitination,phosphorylation,histone acetylation and methylation,SUMOylation,and S-nitrosylation in the regulation of auxin signaling.

Auxin signaling module OsSK41-OsIAA10-OsARF regulates grain yield traits in rice

Auxin is an important phytohormone in plants, and auxin signaling pathways in rice play key roles in regulating its growth, development, and productivity. To investigate how rice grain yield traits are regulated by auxin signaling pathways and to facilitate their application in rice improvement, we validated the functional relationships among regulatory genes such as OsIAA10, OsSK41, and OsARF21 that are involved in one of the auxin(OsIAA10) signaling pathways. We assessed the phenotypic effects of these genes on several grain yield traits across two environments using knockout and/or overexpression transgenic lines.Based on the results, we constructed a model that showed how grain yield traits were regulated by OsIAA10 and OsTIR1, OsAFB2, and OsSK41 and OsmiR393 in the OsSK41-OsIAA10-OsARF module and by OsARF21 in the transcriptional regulation of downstream auxin response genes in the OsSK41-OsIAA10-OsARF module. The population genomic analyses revealed rich genetic diversity and the presence of major functional alleles at most of these loci in rice populations. The strong differentiation of many major alleles between Xian/indica and Geng/japonica subspecies and/or among modern varieties and landraces suggested that they contributed to improved productivity during evolution and breeding. We identified several important aspects associated with the genetic and molecular bases of rice grain and yield traits that were regulated by auxin signaling pathways.We also suggested rice auxin response factor(OsARF) activators as candidate target genes for improving specific target traits by overexpression and/or editing subspecies-specific alleles and by searching and pyramiding the ‘best' gene allelic combinations at multiple regulatory genes in auxin signaling pathways in rice breeding programs.

Auxin signaling:Research advances over the past 30 years

Auxin,one of the first identified and most widely studied phytohormones,has been and will remain a hot topic in plant biology.After more than a century of passionate exploration,the mysteries of its synthesis,transport,signaling,and metabolism have largely been unlocked.Due to the rapid development of new technologies,new methods,and new genetic materials,the study of auxin has entered the fast lane over the past 30 years.Here,we highlight advances in understanding auxin signaling,including auxin perception,rapid auxin responses,TRANSPORT INHIBITOR RESPONSE 1 and AUXIN SIGNALING F-boxes(TIR1/AFBs)-mediated transcriptional and non-transcriptional branches,and the epigenetic regulation of auxin signaling.We also focus on feedback inhibition mechanisms that prevent the over-amplification of auxin signals.In addition,we cover the TRANSMEMBRANE KINASE-mediated non-canonical signaling,which converges with TIR1/AFBs-mediated transcriptional regulation to coordinate plant growth and development.The identification of additional auxin signaling components and their regulation will continue to open new avenues of research in this field,leading to an increasingly deeper,more comprehensive understanding of how auxin signals are interpreted at the cellular level to regulate plant growth and development.

Hydrogen Peroxide-Mediated Growth of the Root System Occurs via Auxin Signaling Modification and Variations in the Expression of Cell-Cycle Genes in Rice Seedlings Exposed to Cadmium Stress

The link between root growth, H2O2, auxin signaling, and the ceil cycle in cadmium （Cd）-stressed rice （Oryza sativa L. cv. Zhonghua No. 11） was analyzed in this study. Exposure to Cd induced a significant accumulation of Cd, but caused a decrease in zinc （Zn） content which resulted from the decreased expression of OsHMA9 and OsZIP. Analysis using a Cd-specific probe showed that Cd was mainly localized in the meristematic zone and vascular tissues. Formation and elongation of the root system were significantly promoted by 3-amino-l,2,4-triazole （AT）, but were markedly inhibited by N,N＇. dimethylthiourea （DMTU） under Cd stress. The effect of H2O2 on Cd-stressed root growth was further confirmed by examining a gain-of-function rice mutant （carrying catalasel and glutathione-S-transferase） in the presence or absence of diphenylene iodonium. DR5-GUS staining revealed close associations between H2O2 and the concentration and distribution of auxin. H2O2 affected the expression of key genes, including OsYUCCA, OsPIN, OsARF, and OslAA, in the auxin signaling pathway in Cd-treated plants. These results suggest that H2O2 functions upstream of the auxin signaling pathway. Furthermore, H2O2 modified the expression of cell-cycle genes in Cd-treated roots. The effects of H2O2 on root system growth are therefore linked to auxin signal modification and to variations in the expression of cell-cycle genes in Cd-stressed rice. A working model for the effects of H2O2 on Cd-stressed root system growth is thus proposed and discussed in this paper.

Loss of GSNOR1 Function Leads to Compromised Auxin Signaling and Polar Auxin Transport

Cross talk between phytohormones, nitric oxide （NO）, and auxin has been implicated in the control of plant growth and development. Two recent reports indicate that NO promoted auxin signaling but inhibited auxin transport probably through S-nitrosylation. However, genetic evidence for the effect of S-nitrosylation on auxin physiology has been lacking. In this study, we used a genetic approach to understand the broader role of S-nitrosylation in auxin physiology in Arabidopsis. We compared auxin signaling and transport in Col-0 and gsnorl-3, a loss-of-function GSNOR1 mutant defective in protein de-nitrosylation. Our results showed that auxin signaling was impaired in the gsnorl-3 mutant as revealed by significantly reduced DR5-GUS/ DR5-GFP accumulation and compromised degradation of AXR3NT-GUS, a useful reporter in interrogating auxin-mediated degradation of Aux/IAA by auxin receptors. In addition, polar auxin transport was compro- mised in gsnorl-3, which was correlated with universally reduced levels of PIN or GFP-PIN proteins in the roots of the mutant in a manner independent of transcription and 26S proteasome degradation. Our results suggest that S-nitrosylation and GSNORl-mediated de-nitrosylation contribute to auxin physiology, and impaired auxin signaling and compromised auxin transport are responsible for the auxin-related morpho- logical phenotypes displayed by the gsnorl-3 mutant.

Mitochondrial Perturbation Negatively Affects Auxin Signaling

Mitochondria are crucial players in the signaling and metabolic homeostasis of the plant cell. The molecular components that orchestrate the underlying processes, however, are largely unknown. Using a chemical biology approach, we exploited the responsiveness of Arabidopsis UDP-glucosyltransferase-encoding UGT74E2 towards mitochondrial per- turbation in order to look for novel mechanisms regulating mitochondria-to-nucleus communication. The most potent inducers of UGT74E2 shared a （2-furyl）acrylate （FAA） substructure that negatively affected mitochondrial function and was identified before as an auxin transcriptional inhibitor. Based on these premises, we demonstrated that perturbed mitochondria negatively affect the auxin signaling machinery. Moreover, chemical perturbation of polar auxin transport and auxin biosynthesis was sufficient to induce mitochondrial retrograde markers and their transcript abundance was constitutively elevated in the absence of the auxin transcriptional activators ARF7 and ARF19.

Interactions of Auxinic Compounds on Ca2+ Signaling and Root Growth in <i>Arabidopsis thaliana</i>

Auxinic-like compounds have been widely used as weed control agents. Over the years, the modes of action of auxinic herbicides have been elucidated, but most studies thus far have focused on their effects on later stages of plant growth. Here, we show that some select auxins and auxiniclike herbicides trigger a rapid elevation in root cytosolic calcium levels within seconds of application. Arabidopsis thaliana plants expressing the Yellow-Cameleon (YC) 3.60 calcium reporter were treated with indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), 1-naphthalene acetic acid (NAA), and two synthetic herbicides, 2,4-dichlorophenoxyacetic acid (2,4-D) and mecoprop [2-(4-chloro- 2-methylphenoxy) propanoic acid], followed by monitoring cytosolic calcium changes over a 10 minute time course. Seconds after application of compounds to roots, the Ca2+ signaling-mediated pathway was triggered, initiating the plant response to these compounds as monitored and recorded using Fluorescence Resonance Energy Transfer (FRET)-sensitized emission imaging. Each compound elicited a specific and unique cytosolic calcium signature. Also primary root development and elongation was greatly reduced or altered when exposed at two concentrations (0.10 and 1.0 μM) of each compound. Within 20 to 25 min after triggering of the Ca2+ signal, root growth inhibition could be detected. We speculate that differences in calcium signature among the tested auxins and auxinic herbicides might correlate with their variation and potency with regard to root growth inhibition.

生长素信号途径参与南瓜白粉菌胁迫的响应研究

生长素(auxin,IAA)信号途径在植物生长、生物胁迫和非生物胁迫响应中发挥着重要的作用。白粉病是南瓜普遍发生且较为严重的一种病害,为探究IAA信号途径响应白粉病胁迫的分子机制,对白粉菌处理的南瓜叶片进行了转录组测序和全基因组DNA甲基化测序分析。结果发现,在IAA信号途径中,有25个差异表达基因,53个基因发生了差异甲基化,其中16个生长素上调小RNA (SAUR)基因均发生了不同程度的甲基化,说明这些基因可能参与了白粉病胁迫响应。SAUR50(CmoCh19G007170)基因的甲基化水平降低,且甲基化区域位于基因的启动子区。在白粉病胁迫下SAUR50的表达水平显著上调,在IAA诱导下显著下调,因此该基因可能通过DNA甲基化调节其表达水平,并且通过IAA信号途径参与南瓜白粉病胁迫的调控。研究结果为IAA信号途径响应白粉病胁迫途径与抗白粉病南瓜分子育种提供了理论依据。

Calcium Signaling is Involved in Negative Phototropism of Rice Seminal Roots

Calcium ions （Ca2＋） act as an intracellular second messenger and affect nearly all aspects of cellular life. They are functioned by interacting with polar auxin transport, and the negative phototropism of plant roots is caused by the transport of auxin from the irradiated side to the shaded side of the roots. To clarify the role of calcium signaling in the modulation of rice root negative phototropism, as well as the relationship between polar auxin transport and calcium signaling, calcium signaling reagents were used to treat rice seminal roots which were cultivated in hydroculture and unilaterally illuminated at an intensity of 100-200 pmol/（m2.s） for 24 h. Negative phototropism curvature and growth rate of rice roots were both promoted by exogenous CaCI2 lower than 100 pmol/L, but inhibited by calcium channel blockers （verapamil and LaCI3）, calcineurin inhibitor （chlorpromazine, CPZ）, and polar auxin transport inhibitor （N-l-naphthylphthalamic acid, NPA）. Roots stopped growing and negative phototropism disappeared when the concentrations increased to 100 pmol/L verapamil, 12.500 ~Jmol/L LaCI3, 60 pmol/L CPZ, and 6 pmol/L NPA. Moreover, 100 pmol/L CaCI2 could relieve the inhibition of LaCI3, verapamil and NPA. The enhanced negative phototropism curvature was caused by the transportation of more auxin from the irradiated side to the shaded side in the presence of exogenous Ca2＋. Calcium signaling plays a key role as a second messenger in the process of light signal regulation of rice root growth and negative phototropism.

脱落酸调控果实成熟的分子及信号转导机制研究进展

脱落酸(abscisicacid,ABA)作为一种重要的植物激素,不仅涉及许多植物发育过程和逆境胁迫,而且在果实成熟,尤其是非呼吸跃变型果实成熟中发挥关键作用。随着植物中ABA合成、代谢和作用机制的解析及其受体识别和核心信号转导模型的建立,极大地推动了ABA在果实成熟和品质形成中的研究。一般来讲,褪绿和着色是果实成熟过程中普遍存在的现象,这一过程涉及了ABA早期信号和多种激素的协同作用并组成了复杂的网络调控机制。总之,ABA是调控果实成熟的核心机制,其中存在着乙烯依赖(呼吸跃变型)和不依赖(非呼吸跃变型)类型。综述了ABA在植物体内的合成、代谢及作用的分子机制,构建了ABA调控果实成熟的分子网络模型,为果实的品质改善和保鲜奠定理论基础。

生长素响应因子与植物的生长发育

生长素响应因子(Auxin response factor,ARF)作为一类调控生长素响应基因表达的转录因子,是生长素研究的重要内容。它可与生长素响应基因启动子区域内的生长素响应元件结合,促进或抑制基因的表达。文章介绍了植物体内ARF家族的分子生物学近年来的研究进展,同时也讨论了ARF转录因子的结构、ARF基因的表达调控、ARF在植物生长发育及信号转导中的作用以及ARF对靶基因的调控机制等内容。植物ARF成员都有一定的同源性,大多含有4个结构域,在多种组织和器官中都有表达,其表达受到转录及转录后调控,并且在介导生长素与其它激素之间相互作用方面扮演重要角色。

生长素信号转导研究进展

长素的信号转导是一个复杂的网络系统,在信号的感知上,除了存在ABP1介导的膜上感知途径外,还有其他的感知途径。G蛋白参与诱导生长素信号的胞内传递,生长素信号转导的第二信使包括离子型第二信使、磷酯酶A2、脂活化蛋白激酶、MAPK和PINOIND等。AUX/IAA蛋白的泛素化降解在生长素反应中发挥关键性作用,ARF和AUX/IAA蛋白相互作用调节生长素响应基因的转录。

植物的胚形态建成及其基因调控机制研究进展

胚形态建成是植物生长发育的基础,是单细胞发育成功能性多细胞有机体的过程。高等植物在胚形态建成过程中首先确立极性随后形成轴性结构,为后续的胚形态建成和生长发育建立了空间坐标体系。笔者以模式植物拟南芥为例介绍了植物胚形态建成过程和基因调控机制,其中包括:MPK/GRD信号途径控制的合子伸长和极性化;WOX8、WOX9、WOX2指导合子不均等分裂和基顶轴方向确定;生长素信号、WOX5调控网络指导的根端分生组织确立、WUS/CLV3、STM调控网络指导顶端分生组织确立;以子叶形成为代表的侧生器官发生和近远轴发育模式的确立需要生长素信号控制和TCP/CUC基因的表达拮抗;SCR/SHR调控途径参与胚基部径向发育过程机理等。认为对胚形态建成过程中基因调控机理的研究有助于从一个全新的角度认识植物生长发育过程。

不定根发生分子调控机制的研究进展

不定根发生问题,既是植物无性繁殖和工厂化育苗实践的核心问题,又是植物发育和形态建成等方面的重要理论问题。由于不定根发生过程的复杂性,到目前为止对其调控机制的了解还十分有限。大量研究证实,不定根发生与植物生长素类物质密切相关,因此现有的研究不仅围绕生长素及其信号传导途径展开,而且还涉及到基因表观遗传学调控水平。目前已经鉴定出一些与不定根发生相关的生长素信号传导因子,如NO、cGMP、microRNAs等。同时,还克隆到一些与不定根发生相关的基因,如OsPIN1、OsCKI1、NPK1、ARL1等。此外,发现DNA甲基化可以抑制DNA与蛋白(MeCP2) 的结合,从而抑制基因转录;microRNA可以使基因沉默来调控不定根的发生状况。本文围绕不定根发生的激素调控、不定根发生的基因调控、不定根发生的生长素信号传导机制、表观遗传调控等几个方面综述了近年来的研究进展。

生长素的运输及其在信号转导及植物发育中的作用

生长素作为一种重要的植物激素,参与调节植物生长发育的诸多过程,如器官发生、形态建成、向性反应、顶端优势及组织分化等,其作用机理长期以来备受人们关注。生长素的极性运输能使生长素积累在植物体某些特定部位,从而形成生长素浓度梯度,生长素对植物生长发育的调节主要依赖于这一特性。系统阐述生长素的运输特点、运输机理和相关生长素极性运输载体的研究进展;并对生长素信号转导途径中的重要组分及其机理进行了总结;同时较系统地对生长素参与植物体各器官发育过程及调节情况进行综述。

Ath-miR169d介导的拟南芥叶片发育的分子调控机制

【目的】探究ath-miR169d在拟南芥叶片发育过程中的作用,明确其调控的分子机制,为增强叶菜类植物的光合效率以及增加生物量和提高作物产量提供理论依据。【方法】选取ath-miR169d过表达和降低表达的拟南芥转基因株系以及野生型拟南芥,在22℃、相对湿度60%、16 h/8 h光周期条件下培养,观察不同生长阶段叶片发育表型的差异;同时构建pmiR169d::GUS表达载体,转化农杆菌EHA105,并利用蘸花法转化野生型拟南芥(Columbia,Col-0),获得转基因拟南芥,利用GUS染色对ath-miR169d在拟南芥不同组织器官中的表达模式进行研究;选取8周龄的过表达材料和野生型对照株系,对其叶片表皮细胞形态进行扫描电镜分析;对4周龄过表达材料和野生型对照株系的幼苗顶端分生组织和叶片中总IAA进行定量分析,并进行基因芯片表达谱分析,筛选差异表达基因;通过实时荧光定量PCR对生长素信号途径部分差异表达关键基因的表达情况进行分析。【结果】Ath-miR169d过表达拟南芥株系莲座叶数目较野生型少,叶片小,而ath-miR169d降低表达的拟南芥株系的莲座叶较野生型多,叶片大,且在抽薹和种子成熟之后还持续长出新叶,而野生型莲座叶则在种子成熟之后逐渐衰老干枯;扫描电镜观察到ath-miR169d过表达拟南芥株系叶片中表皮细胞小于野生型;表达模式分析表明,ath-miR169d主要在顶端分生组织(shoot apical meristem,SAM)和叶片维管系统中表达,且新叶中表达强。SAM是产生生长素的部位,IAA测定结果表明ath-miR169d过表达拟南芥株系中IAA含量显著降低,为野生型植株的38.6%,同时基因芯片表达谱分析也验证了ath-miR169d参与调控植物体内激素信号转导途径。进一步研究发现,ath-miR169d过表达株系中生长素合成关键基因YUC2和转运体蛋白基因PIN1均显著下调表达,而具有转录抑制功能的生长素响应因子1和2基因(ARF1和ARF2)表达上调;STTM miR169d低表达株系中这4个基因的表达则为相反趋势,该结果与观察到的表型相一致。【结论】Ath-miR169d通过介导生长素信号途径参与了拟南芥叶片发育的调控,ath-miR169d表达量发生变化会影响叶片的数量和大小,最终影响整个植株的生物量。

生长素信号转导途径及参与的生物学功能研究进展

生长素参与植物生长和发育诸多过程,调控众多生理反应,在植物整个生命周期中自始至终发挥着调节作用.研究生长素的作用机制,对深入认识植物生长发育的生理过程有着重要的意义.综述了与生长素信号转导途径相关的3类主要蛋白组分:生长素/吲哚乙酸蛋白(auxin/indoleacetic acids proteins,Aux/IAAs)、生长素响应因子(auxin response factors,ARFs)和SCF(SKP1-CDC53/CUL1-F-box)复合体,及相关的SGT1(suppressor of the G2 allele of skp1)基因,并对生长素相关基因表达的模式及其生物学功能进行了总结.

植物根系向地性感应的分子机理与养分吸收

植物根系向地性是决定根系空间生长趋势的主要因素之一,对于养分吸收具有重要影响。认识根系向地性感应和根系生长变化的分子机理及其与养分吸收的关系,可为遗传改良根系性状、提高植物养分吸收效率提供理论依据。本文从重力感应、信号转导和生长素非对称分布等方面总结了植物根系向地性感应的分子机理,探讨了根系在养分胁迫下(特别是磷胁迫下)向地性变化的生理基础及其与养分吸收(特别是磷吸收)的关系,最后对根系向地性研究的若干问题进行了展望。

水稻中生长素作用的分子机理研究进展

水稻的生长与发育是受多个植物激素协同调控的,其中认识最早和研究最深的是生长素。随着分子技术的进步,研究生长素的作用机制已经深入到细胞和基因水平,包括生长素的代谢、运输、信号感知和传导,并且生长素的信号传导在水稻生长发育过程中与其他激素相互交叉、协调作用。研究生长素的作用机制对植物生长发育具有重要意义。