Mimicking small G-proteins: an emerging theme from the bacterial virulence arsenal

The identification of the Ras superfamily of small molecular weight GTPases (G-proteins) has opened up new fields in cancer biology, immunity and infectious disease research. Because of their ubiquitous role in cellular homeostasis, small G-proteins are common targets for several pathogens, including bacteria. It is well known that pathogenic bacteria have evolved virulence factors that chemically modify GTPases or directly mimic the activities of key regulatory proteins. However, recent studies now suggest that bacterial 'effector' proteins can also mimic the activities of Ras small G-proteins despite their lack of guanine nucleotide binding or GTPase enzymatic activity. The study of these unique pathogenic strategies continues to reveal novel mechanistic insights into host cellular communication networks and the role of small G-protein signalling during human infectious disease.     

GTP-Binding Regulatory Proteins in Higher Plant Cells

The exsitence of GTP-binding regulatory proteins (for short term, often refered as G-proteins) in higher plant cells is certain. G-proteins are classified into two groups based on their molecular structures, which are the heterotrimeric G-proteins (big G-proteins) that contain three different subunits and the small G-proteins that have only one subunit (monomeric G-proteins). All G-proteins are characterized by their properties to bind with and hydrolyze GTP, by which G-proteins function as transmembrane and intracellular signalling molecules. As a distinguished participant in signal transduction, G-proteins directly and/or indirectly regulate a number of physiological processes, such as regulation of phytochrome-related physiological processes and gene expression, involvement in blue-light response, K+-channel regulation, stomatal movement, hormone regulation, protein phosphrylation dephosphorylation, etc. Although G-proteins in plant cells have not been purified, the genes for a subunit of heterotrimeric G-proteins have been cloned. More evidences for the importance of G-proteins in plant signalling processes are rapidly accumulating.     

Guanosine triphosphate-binding proteins on the plasmalemma of spinach leaf cells

The molecular mechanism of light perception through phytochrome is not well understood. This red-light photosensor has been implicated in various physiological processes, including the photoinduction of flowering. A few recent studies have shown that phytochrome initiates signal transduction chains via guanosine triphosphate (GTP)-binding proteins (G-proteins). We show here by different approaches that G-proteins exist in spinach (Spinacia oleracea L. cv. Nobel). Binding of GTP on the plasmalemma has been partially characterized and its possible regulation by red light examined by in-vitro assays. These experiments indicate a clear regulation of GTP binding by red light and also by Mastoparan. At least three G-proteins or protein subunits were found to be associated with the plasmalemma of leaf cells. The use of an antibody raised against an animal Gβ subunit confirmed the presence of heterotrimeric G-proteins. Separation of a crude membrane extract by free-flow electrophoresis also showed that some G-proteins could exist on the tonoplast.     

Plant signaling in stress: G-protein coupled receptors,heterotrimeric G-proteins and signal coupling via phospholipases

Plant growth and development are coordinalely controlled by several internal factors and environmental signals. To sense these environmental signals, the higher plants have evolved a complex signaling network, which may also cross talk with each other. Plants can respond to the signals as individual cells and as whole organisms. Various receptors including phytochromes, G-proteins coupled receptors (GPCR), kinase and hormone receptors play important role in signal transduction but very few have been characterized in plant system. The heterotrimeric G-proteins mediate the coupling of signal transduction from activated GPCR to appropriate downstream effectors and thereby play an important role in signaling. In this review we have focused on some of the recent work on G-proteins and two of the effectors, PLC and PLD, which have been shown to interact with Gα subunit and also discussed their role in abiotic stress tolerance.Key words: abiotic stress, G-protein couple receptor, heterotrimeric G-protein, phospholipases, plant receptors, signal transduction     

Signal transduction in vascular plants

We review current evidence for the presence and activity in plants of several paradigmatic components of transmembrane signal transduction systems. Components considered include the second messengers calcium, inositol 1,4,5-trisphosphate, and cyclic AMP; protein kinases and protein phosphatases; and G-proteins. At the current stage of development of the field of plant signal transduction, broad similarities between plant and the more well-studied animal systems are apparent. However, there also exist considerable differences in detail.     

Microinjection of Guanine NucleotideAnalogues into Lily Pollen Tubes Results in Isodiametric TipExpansion

Abstract: Heterotrimeric and small G-proteins aresupposed to participate in tip growth of plant cells. Quantitative changes intip growth rate after introduction of non-hydrolysable guanine nucleotideanalogues (NA) into lily pollen tubes have been interpreted as support for thehypothesis that heterotrimeric G-proteins regulate tube elongation (Ma et al.,1999 IDREF="R243-11">11). Here, we report that microinjection of guanineNA into lily pollen tubes causes loss of growth polarity, resulting inisodiametric tip swelling. Our results are compatible with current modelssuggesting an involvement of plant Rho-related small G-proteins (Rop) in themaintenance of pollen tube polarity and in tip morphogenesis.     

From plant immunity to crop disease resistance

Plant diseases caused by diverse pathogens lead to a serious reduction in crop yield and threaten food security worldwide. Genetic improvement of plant immunity is considered as the most effective and sustainable approach to control crop diseases. In the last decade, our understanding of plant immunity at both molecular and genomic levels has improved greatly. Combined with advances in biotechnologies, particularly clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9-based genome editing, we can now rapidly identify new resistance genes and engineer disease-resistance crop plants like never before. In this review, we summarize the current knowledge of plant immunity and outline existing and new strategies for disease resistance improvement in crop plants. We also discuss existing challenges in this field and suggest directions for future studies.     

The roles of ABA in plant–pathogen interactions

Defence against abiotic and biotic stresses is crucial for the fitness and survival of plants under adverse or suboptimal growth conditions. The phytohormone abscisic acid (ABA) is not only important for mediating abiotic stress responses, but also plays a multifaceted and pivotal role in plant immunity. This review presents examples demonstrating the importance of crosstalk between ABA and the key biotic stress phytohormone salicylic acid in determining the outcome of plant–pathogen interactions. We then provide an overview of how ABA influences plant defence responses against various phytopathogens with particular emphasis on the Arabidopsis–Pseudomonas syringae model pathosystem. Lastly, we discuss future directions for studies of ABA in plant immunity with emphasis on, its role in the crosstalk between biotic and abiotic stress responses, the importance of distinguishing direct and indirect effects of ABA, as well as the prospect of utilizing the recently elucidated core ABA signaling network to gain further insights into the roles of ABA in plant immunity.     

Characterization of ATDRG1, a member of a new class of GTP-binding proteins in plants

We report the initial characterization of an Arabidopsis thaliana cDNA (atdrg1), a member of a new class of GTP-binding proteins (G-proteins) in plants. The predicted ATDRG1 protein contains all five structural motifs characteristic of the G-protein superfamily. Apart from these motifs, the amino acid sequence differs substantially from all known G-proteins except for a recently discovered new family named developmentally regulated G-proteins (DRGs). Sequences closely related to atdrg1 are found in species as distant as human (80% amino acid conservation), Drosophila (74%), yeast (77%) and Caenorhabditis elegans (77%). The remarkable evolutionary conservation of these proteins suggests an important, but as yet unclear role. Phylogenetic analysis of the available homologous sequences strongly suggests a diphyletic origin of the eukaryotic DRG proteins. Northern analysis shows high levels of atdrg1 mRNA in all Arabidopsis tissues studied, and homologues of atdrg1 are present throughout the plant kingdom. In situ hybridization reveals that atdrg1 is highly expressed in actively growing tissues and reproductive organs. Southern analysis indicates the presence of either one or two copies of atdrg1 in the Arabidopsis genome. Immunolocalization studies show that the protein is present in cytoplasmic vesicles found mainly in actively growing tissues suggesting a putative role for ATDRG1 in either the regulation of vesicle transport or the regulation of enzymes involved in storage protein processing.     

Two-component elements mediate interactions between cytokinin and salicylic acid in plant immunity

Recent studies have revealed an important role for hormones in plant immunity. We are now beginning to understand the contribution of crosstalk among different hormone signaling networks to the outcome of plant-pathogen interactions. Cytokinins are plant hormones that regulate development and responses to the environment. Cytokinin signaling involves a phosphorelay circuitry similar to two-component systems used by bacteria and fungi to perceive and react to various environmental stimuli. In this study, we asked whether cytokinin and components of cytokinin signaling contribute to plant immunity. We demonstrate that cytokinin levels in Arabidopsis are important in determining the amplitude of immune responses, ultimately influencing the outcome of plant-pathogen interactions. We show that high concentrations of cytokinin lead to increased defense responses to a virulent oomycete pathogen, through a process that is dependent on salicylic acid (SA) accumulation and activation of defense gene expression. Surprisingly, treatment with lower concentrations of cytokinin results in increased susceptibility. These functions for cytokinin in plant immunity require a host phosphorelay system and are mediated in part by type-A response regulators, which act as negative regulators of basal and pathogen-induced SA-dependent gene expression. Our results support a model in which cytokinin up-regulates plant immunity via an elevation of SA-dependent defense responses and in which SA in turn feedback-inhibits cytokinin signaling. The crosstalk between cytokinin and SA signaling networks may help plants fine-tune defense responses against pathogens.     

Arabidopsis Heterotrimeric G-Proteins Play a Critical Role in Host and Nonhost Resistance against Pseudomonas syringae Pathogens

Heterotrimeric G-proteins have been proposed to be involved in many aspects of plant disease resistance but their precise role in mediating nonhost disease resistance is not well understood. We evaluated the roles of specific subunits of heterotrimeric G-proteins using knock-out mutants of Arabidopsis Gα, Gβ and Gγ subunits in response to host and nonhost Pseudomonas pathogens. Plants lacking functional Gα, Gβ and Gγ1Gγ2 proteins displayed enhanced bacterial growth and disease susceptibility in response to host and nonhost pathogens. Mutations of single Gγ subunits Gγ1, Gγ2 and Gγ3 did not alter bacterial disease resistance. Some specificity of subunit usage was observed when comparing host pathogen versus nonhost pathogen. Overexpression of both Gα and Gβ led to reduced bacterial multiplication of nonhost pathogen P. syringae pv. tabaci whereas overexpression of Gβ, but not of Gα, resulted in reduced bacterial growth of host pathogen P. syringae pv. maculicola, compared to wild-type Col-0. Moreover, the regulation of stomatal aperture by bacterial pathogens was altered in Gα and Gβ mutants but not in any of the single or double Gγ mutants. Taken together, these data substantiate the critical role of heterotrimeric G-proteins in plant innate immunity and stomatal modulation in response to P. syringae.     

Receptor kinases with leucine-rich repeats are enriched in Triton X-100 insoluble plasma membrane microdomains from plants

In mammals, signalling components at the cell surface are clustered in Triton X-100 insoluble plasma membrane microdomains. We isolated plasma membrane microdomains from Arabidopsis and mustard cotyledons and determined their protein composition by mass spectrometry. Although the protein composition of the plant vesicles differ from the composition of the animal vesicles, they are also enriched in signalling components. We identified at least seven receptor kinases with leucine-rich repeats, 10 other kinases, the β subunit of heterotrimeric G-proteins and five small GTP-binding proteins. Thus, specific signalling components are highly enriched in plant plasma membrane microdomains while others are excluded.     

An atypical heterotrimeric G-protein γ-subunit is involved in guard cell K⁺-channel regulation and morphological development in Arabidopsis thaliana

Currently, there are strong inconsistencies in our knowledge of plant heterotrimeric G-proteins that suggest the existence of additional members of the family. We have identified a new Arabidopsis G-protein γ-subunit (AGG3) that modulates morphological development and ABA-regulation of stomatal aperture. AGG3 strongly interacts with the Arabidopsis G-protein β-subunit in vivo and in vitro. Most importantly, AGG3-deficient mutants account for all but one of the 'orphan' phenotypes previously unexplained by the two known γ-subunits in Arabidopsis. AGG3 has unique characteristics never before observed in plant or animal systems, such as its size (more than twice that of canonical γ-subunits) and the presence of a C-terminal Cys-rich domain. AGG3 thus represent a novel class of G-protein γ-subunits, widely spread throughout the plant kingdom but not present in animals. Homologues of AGG3 in rice have been identified as important quantitative trait loci for grain size and yield, but due to the atypical nature of the proteins their identity as G-protein subunits was thus far unknown. Our work demonstrates a similar trend in seeds of Arabidopsis agg3 mutants, and implicates G-proteins in such a crucial agronomic trait. The discovery of this highly atypical subunit reinforces the emerging notion that plant and animal G-proteins have distinct as well as shared evolutionary pathways.     

Plant innate immunity: An updated insight into defense mechanism

Plants are invaded by an array of pathogens of which only a few succeed in causing disease. The attack by others is countered by a sophisticated immune system possessed by the plants. The plant immune system is broadly divided into two, viz. microbial-associated molecular-patterns-triggered immunity (MTI) and effector-triggered immunity (ETI). MTI confers basal resistance, while ETI confers durable resistance, often resulting in hypersensitive response. Plants also possess systemic acquired resistance (SAR), which provides long-term defense against a broad-spectrum of pathogens. Salicylic-acid-mediated systemic acquired immunity provokes the defense response throughout the plant system during pathogen infection at a particular site. Trans-generational immune priming allows the plant to heritably shield their progeny towards pathogens previously encountered. Plants circumvent the viral infection through RNA interference phenomena by utilizing small RNAs. This review summarizes the molecular mechanisms of plant immune system, and the latest breakthroughs reported in plant defense. We discuss the plant–pathogen interactions and integrated defense responses in the context of presenting an integral understanding in plant molecular immunity.     

Conventional and unconventional ubiquitination in plant immunity

Ubiquitination is one of the most abundant types of protein post‐translational modification (PTM) in plant cells. The importance of ubiquitination in the regulation of many aspects of plant immunity has been increasingly appreciated in recent years. Most of the studies linking ubiquitination to the plant immune system, however, have been focused on the E3 ubiquitin ligases and the conventional ubiquitination that leads to the degradation of the substrate proteins by the 26S proteasome. By contrast, our knowledge about the role of unconventional ubiquitination that often serves as non‐degradative, regulatory signal remains a significant gap. We discuss, in this review, the recent advances in our understanding of ubiquitination in the modulation of plant immunity, with a particular focus on the E3 ubiquitin ligases. We approach the topic from a perspective of two broadly defined types of ubiquitination in an attempt to highlight the importance, yet current scarcity, in our knowledge about the regulation of plant immunity by unconventional ubiquitination.     

Autophagy and plant innate immunity

Plant innate immunity is often associated with specialized programmed cell death at or near the site of pathogen infection. Despite the isolation of several lesion mimic mutants, the molecular mechanisms that regulate cell death during an immune response remain obscure. Recently, autophagy, an evolutionarily conserved process of bulk protein and organelle turnover, was shown to play an important role in limiting cell death initiated during plant innate immune responses. Consistent with its role in plants, several studies in animals also demonstrate that the autophagic machinery is involved in innate as well as adaptive immunities. Here, we review the role of autophagy in plant innate immunity. Because autophagy is observed in healthy and dying plant cells, we will also examine whether autophagy plays a protective or a destructive role during an immune response.     

Plant immunity: a lesson from pathogenic bacterial effector proteins

Phytopathogenic bacteria inject an array of effector proteins into host cells to alter host physiology and assist the infection process. Some of these effectors can also trigger disease resistance as a result of recognition in the plant cell by cytoplasmic immune receptors. In addition to effector-triggered immunity, plants immunity can be triggered upon the detection of Pathogen/Microbe-Associated Molecular Patterns by surface-localized immune receptors. Recent progress indicates that many bacterial effector proteins use a variety of biochemical properties to directly attack key components of PAMP-triggered immunity and effector-triggered immunity, providing new insights into the molecular basis of plant innate immunity. Emerging evidence indicate that the evolution of disease resistance in plants is intimately linked to the mechanism by which bacterial effectors promote parasitism. This review focuses on how these studies have conceptually advanced our understanding of plant–pathogen interactions.     

植物异三聚体G蛋白研究进展

异三聚体鸟嘌呤核苷结合蛋白（简称G蛋白）是真核细胞中保守的信号转导分子,通常与G蛋白偶联受体一起将细胞外信号传递到胞质中。许多研究表明植物G蛋白介导的信号转导途径在光、激素、糖等响应过程中发挥着精细的调控作用。本文重点介绍近年来植物G蛋白在复合体组成、生化特性及其工作模式等方面的研究进展。     

气孔免疫的研究进展及展望

气孔长期以来被认为是植物病原菌入侵植物体内的被动通道,而最新的研究则表明气孔作为植物先天免疫的重要环节,在限制细菌入侵方面起到主动作用。这一发现也带动了植物免疫学,即植物气孔开合调控和植物免疫学交叉学科的快速发展。基于此,本文对气孔免疫的机制研究展开综述并对其对植物水分利用效率的影响进行展望。