植物硫转运蛋白研究进展

硫转运蛋白在植物对硫酸盐的吸收和转运中起着重要的作用。已经在拟南芥、大麦和小麦等植物中分离到了40多种硫转运蛋白基因。这些基因序列与其他种类生物的硫转运蛋白基因序列有着高度的保守性。利用CLUSTAL程序建立的系统进化树将植物硫转运蛋白划分为5个亚群。使用多种拓扑预测程序推测出不同植物硫转运蛋白的共同结构特点是均含有12个跨膜域。在柱花草和大麦中,硫转运蛋白基因表达调控包括植物体内硫水平的负调控和O—乙酰丝氨酸的正调控两种方式。对硫转运蛋白的组织定位和功能研究表明,高亲和硫转运蛋白主要定位于根部,在根系硫酸盐吸收中起重要作用。     

Control of sulphate assimilation and glutathione synthesis: interaction with N and C metabolism

Sulphate assimilation is an essential pathway being a source of reduced sulphur for various cellular processes and for the synthesis of glutathione, a major factor in plant stress defence. Many reports have shown that sulphate assimilation is well co-ordinated with the assimilation of nitrate and carbon. It has long been known that, during nitrate deficiency, sulphate assimilation is reduced and that the capacity to reduce nitrate is diminished in plants starved for sulphate. Only recently, however, was it shown that adenosine 5' phosphosulphate reductase (APR), the key enzyme of sulphate assimilation, is regulated by carbohydrates. In plants treated with sucrose or glucose APR was induced, whereas the activity was strongly reduced in plants grown in CO(2)-free air. The availability of cysteine is a crucial factor in glutathione synthesis, but an adequate supply of glutamate and glycine are also important. The molecular mechanisms for the co-ordination of S, N, and C assimilation are not known. O-acetylserine, a precursor of cysteine, was proposed to be the signal regulating sulphate assimilation, but most probably is not the outgoing signal to N and C metabolism. cDNA arrays revealed the induction of genes involved in auxin synthesis upon S-starvation, pointing to a possible role of phytohormones. Clearly, despite significant progress in understanding the regulation of sulphate assimilation and glutathione synthesis, their co-ordination with N and C metabolism achieved, and several potential signal molecules identified, present knowledge is still far from being sufficient.     

Plant adenosine 5'-phosphosulphate reductase: the past, the present, and the future

The sulphate assimilation pathway provides reduced sulphur for the synthesis of the amino acids cysteine and methionine. These are the essential building blocks of proteins and further sources of reduced sulphur for the synthesis of coenzymes and various secondary compounds. Several recent reports identified the adenosine 5'-phosphosulphate reductase (APR) as the enzyme with the greatest control over the pathway. In this review, a short historical excursion into the investigations of sulphate assimilation is given with emphasis on the proposed alternative pathways to APR, via 'bound sulphite' or via PAPS reductase. The evolutionary past of APR is reviewed, based on phylogenetic analysis of APR and PAPS reductase sequences. Furthermore, recent biochemical analyses of APR that identified an iron-sulphur centre as a cofactor, proposed functions for different protein domains, and addressed the enzyme mechanism are summarized. Finally, questions that have to be addressed in order to improve understanding of the molecular mechanism and regulation of APR have been identified.     

Molecular analysis and control of cysteine biosynthesis: integration of nitrogen and sulphur metabolism

Since cysteine is the first committed molecule in plant metabolism containing both sulphur and nitrogen, the regulation of its biosynthesis is critically important. Cysteine itself is required for the production of an abundance of key metabolites in diverse pathways. Plants alter their metabolism to compensate for sulphur and nitrogen deficiencies as best as they can, but limitations in either nutrient not only curb a plant's ability to synthesize cysteine, but also restrict protein synthesis. Nutrients such as nitrate and sulphate (and carbon) act as signals; they trigger molecular mechanisms that modify biosynthetic pathways and thereby have a profound impact on metabolite fluxes. Cysteine biosynthesis is modified by regulators acting at the site of uptake and throughout the plant system. Recent data point to the existence of nutrient-specific signal transduction pathways that relay information about external and internal nutrient concentrations, resulting in alterations to cysteine biosynthesis. Progress in this field has led to the cloning of genes that play pivotal roles in nutrient-induced changes in cysteine formation.     

Physiological analysis of mutants of Saccharomyces cerevisiae impaired in sulphate assimilation.

The assimilation of sulphate in Saccharomyces cerevisiae, comprising the reduction of sulphate to sulphide and the incorporation of the sulphur atom into a four-carbon chain, requires the integrity of 13 different genes. To date, the functions of nine of these genes are still not clearly established. A set of strains, each bearing a mutation in one MET gene, was studied. Phenotypic studies and enzyme determinations showed that the products of at least five genes are needed for the synthesis of an enzymically active sulphite reductase. These genes are MET1, MET5, MET8, MET10 and MET20. Wild-type strains of S. cerevisiae can use organic metabolites such as homocysteine, cysteine, methionine and S-adenosylmethionine as sulphur sources. They are also able to use inorganic sulphur sources such as sulphate, sulphite, sulphide or thiosulphate. Here we show that both of the two sulphur atoms of thiosulphate are used by S. cerevisiae. Thiosulphate is cleaved into sulphite and sulphide prior to utilization by the sulphate assimilation pathway, as the metabolism of one sulphur atom from thiosulphate requires the presence of an active sulphite reductase.     

Respiration and nitrogen assimilation: targeting mitochondria-associated metabolism as a means to enhance nitrogen use efficiency

Considerable advances in our understanding of the control of mitochondrial metabolism and its interactions with nitrogen metabolism and associated carbon/nitrogen interactions have occurred in recent years, particularly highlighting important roles in cellular redox homeostasis. The tricarboxylic acid (TCA) cycle is a central metabolic hub for the interacting pathways of respiration, nitrogen assimilation, and photorespiration, with components that show considerable flexibility in relation to adaptations to the different functions of mitochondria in photosynthetic and non-photosynthetic cells. By comparison, the operation of the oxidative pentose phosphate pathway appears to represent a significant limitation to nitrogen assimilation in non-photosynthetic tissues. Valuable new insights have been gained concerning the roles of the different enzymes involved in the production of 2-oxoglutarate (2-OG) for ammonia assimilation, yielding an improved understanding of the crucial role of cellular energy balance as a broker of co-ordinate regulation. Taken together with new information on the mechanisms that co-ordinate the expression of genes involved in organellar functions, including energy metabolism, and the potential for exploiting the existing flexibility for NAD(P)H utilization in the respiratory electron transport chain to drive nitrogen assimilation, the evidence that mitochondrial metabolism and machinery are potential novel targets for the enhancement of nitrogen use efficiency (NUE) is explored.     

Plant responses to sulphur deficiency and the genetic manipulation of sulphate transporters to improve S-utilization efficiency

Decreased inputs of S have increased the incidence of S-deficiency in crops, resulting in decreased yields and quality. Remediation by fertilizer application is not always successful because this often results in an uneven supply of S. The ability to respond to S-deficiency stress varies between crops and this is a target for the genetic improvement of S-utilization efficiency. Improved capture of resources, the accumulation of greater reserves of S and improved mechanisms for the remobilization of these reserves are required. It is an inability to over-accumulate S and subsequently, effectively remobilize S-reserves, which restricts optimum S-use efficiency. Genetic manipulation of the transporters and their expression will contribute to overcoming these limitations. Control of gene expression limits excess uptake and activity of the assimilatory pathway: the endogenous expression of sulphate transporters is regulated by S-supply, with negative regulation from reduced S-containing compounds and positive regulation by O-acetylserine, the C/N skeleton precursor of cysteine. Constitutive expression of the transporter will remove this control and may enable the accumulation of sulphate reserves. Sulphate in the vacuole and other pools of reduced sulphur, such as glutathione or protein may be remobilized under S-limiting conditions. Low efficiencies of these remobilization processes, particularly the remobilization of vacuolar sulphate, suggest that the transporters involved in the remobilization are a target for modification. Transporters are involved in facilitating the multiple trans-membrane transport steps between uptake of sulphate from the soil solution, and delivery to the site of reduction in the chloroplast or plastid. A gene family has been identified and phylogenetic relationships based on primary sequence information indicate multiple sub-groups. Groups which are expressed in roots, in shoots and in both tissue types are postulated, however, the functional roles for these groups and the identification of transporters involved in recycling remain to be confirmed.     

Plant sulfate assimilation genes: redundancy versus specialization

Sulfur is an essential nutrient present in the amino acids cysteine and methionine, co-enzymes and vitamins. Plants and many microorganisms are able to utilize inorganic sulfate and assimilate it into these compounds. Sulfate assimilation in plants has been extensively studied because of the many functions of sulfur in plant metabolism and stress defense. The pathway is highly regulated in a demand-driven manner. A characteristic feature of this pathway is that most of its components are encoded by small multigene families. This may not be surprising, as several steps of sulfate assimilation occur in multiple cellular compartments, but the composition of the gene families is more complex than simply organellar versus cytosolic forms. Recently, several of these gene families have been investigated in a systematic manner utilizing Arabidopsis reverse genetics tools. In this review, we will assess how far the individual isoforms of sulfate assimilation enzymes possess specific functions and what level of genetic redundancy is retained. We will also compare the genomic organization of sulfate assimilation in the model plant Arabidopsis thaliana with other plant species to find common and species-specific features of the pathway.     

miSSING LINKS: miRNAs and plant development

The discovery of hundreds of plant micro RNAs (miRNAs) has triggered much speculation about their potential roles in plant development. The search for plant genes involved in miRNA processing has revealed common factors such as DICER, and new molecules, including HEN1. Progress is also being made toward identifying miRNA target genes and understanding the mechanisms of miRNA-mediated gene regulation in plants. This work has lead to a reexamination of many previously characterized mutations that are now known to affect components or targets of miRNA-mediated pathways.     

Complex regulation of ABA biosynthesis in plants.

Abscisic acid (ABA) is a plant hormone that plays important roles during many phases of the plant life cycle, including seed development and dormancy, and in plant responses to various environmental stresses. Because many of these physiological processes are correlated with endogenous ABA levels, the regulation of ABA biosynthesis is a key element facilitating the elucidation of these physiological characteristics. Recent studies on the identification of genes encoding enzymes involved in ABA biosynthesis have revealed details of the main ABA biosynthetic pathway. At the same time, the presence of gene families and their respective organ-specific expression are indicative of the complex mechanisms governing the regulation of ABA biosynthesis in response to plant organ and/or environmental conditions. There have been recent advances in the study of ABA biosynthesis and new insights into the regulation of ABA biosynthesis in relation to physiological phenomena.     

Profiles of gene family members related to carotenoid accumulation in citrus genus

Citrus fruit are an important reservoir of carotenoids. Numerous studies have been carried out to identify and profile the members of gene families involved in carotenoid biosynthetic pathway to explain the diversity of coloration in citrus fruit. It was found that gene expression analysis could not always explain the changes in carotenoid content and composition, indicating that other unknown genes and mechanisms should be operative. This review summarizes and updates the current knowledge on gene families involved in the citrus carotenoid biosynthetic pathway and their roles on the regulation of carotenoid biosynthesis, as well as provides insightful questions leading to future experimentation.     

Plant sulphate transporters: co-ordination of uptake, intracellular and long-distance transport

Proton/sulphate co-transport in the plasma membrane of root cells is the first step for the uptake of sulphate from the environment by plants. Further intracellular, cell-to-cell and long-distance transport must fulfil the requirements for sulphate assimilation and source/sink demands within the plant. A gene family of sulphate transporters, which may be subdivided into five groups, has been identified with examples from many different plant species. For at least two groups, proton/sulphate co-transport activity has been confirmed. It appears that each group represents sulphate transporters with distinct kinetic properties, patterns of expression, and cell/tissue specificity related to specific roles in the uptake and allocation of sulphate. High-affinity sulphate uptake and low-affinity vascular transport, as well as vacuolar efflux, are controlled by the nutritional status of the plant. Most notably there is an apparent increase in capacity for cellular sulphate uptake and vacuolar efflux when sulphur supply is limiting. Within the groups, the individual sulphate transporters may be further subdivided by differences in temporal, cellular and tissue expression. Many of the transporters are regulated by the nutritional status of the individual tissues, to optimize sulphate movement within and between sink and source organs.     

Nitrate transporters in plants: structure, function and regulation

Physiological studies have established that plants acquire their NO(-3) from the soil through the combined activities of a set of high- and low-affinity NO(-3) transport systems, with the influx of NO(-3) being driven by the H(+) gradient across the plasma membrane. Some of these NO(-3) transport systems are constitutively expressed, while others are NO(-3)-inducible and subject to negative feedback regulation by the products of NO(-3) assimilation. Here we review recent progress in the characterisation of the two families of NO(-3) transporters that have so far been identified in plants, their structure and their regulation, and consider the evidence for their roles in NO(-3) acquisition. We also discuss what is currently known about the genetic basis of NO(-3) induction and feedback repression of the NO(-3) transport and assimilatory pathway in higher plants.