Plant Metabolic Pathways Research Articles

An expanding role for purine uptake permease-like transporters in plant secondary metabolism

For the past decade, our understanding of the plant purine uptake permease (PUP) transporter family was primarily oriented on purine nucleobase substrates and their tissue-specific expression patterns in Arabidopsis. However, a tobacco PUP-like homolog demonstrating nicotine uptake permease activity was recently shown to affect both nicotine metabolism and root cell growth. These new findings expand the physiological role for PUP-like transporters to include plant secondary metabolism. Molecular evolution analyses of PUP-like transporters indicate they are distinct group within an ancient super family of drug and metabolite transporters (DMTs). The PUP-like family originated during terrestrial plant evolution sometime between the bryophytes and the lycophytes. A phylogenetic analysis indicates that the PUP-like transporters were likely derived from a pre-existing nucleotide-sugar transporter family within the DMT super family. Within the lycophyte Selaginella, there are three paralogous groups of PUP-like transporters. One of the three PUP-like paralogous groups showed an extensive pattern of gene duplication and diversification within the angiosperm lineage, whereas the more ancestral PUP-like paralogous groups did not. Biochemical characterization of four closely related PUP-like paralogs together with model-based phylogenetic analyses indicate both subfunctionalization and neofunctionalization during the molecular evolution of angiosperm PUP-like transporters. These findings suggest that members of the PUP-like family of DMT transporters are likely involved in diverse primary and secondary plant metabolic pathways.

Ustilago takes control

Ustilago maydis Cmu1 is a virulence factor that can move through plant cells, redirecting plant metabolic pathways to favour fungal infection.

A tiling microarray for global analysis of chloroplast genome expression in cucumber and other plants.

Plastids are small organelles equipped with their own genomes (plastomes). Although these organelles are involved in numerous plant metabolic pathways, current knowledge about the transcriptional activity of plastomes is limited. To solve this problem, we constructed a plastid tiling microarray (PlasTi-microarray) consisting of 1629 oligonucleotide probes. The oligonucleotides were designed based on the cucumber chloroplast genomic sequence and targeted both strands of the plastome in a non-contiguous arrangement. Up to 4 specific probes were designed for each gene/exon, and the intergenic regions were covered regularly, with 70-nt intervals. We also developed a protocol for direct chemical labeling and hybridization of as little as 2 micrograms of chloroplast RNA. We used this protocol for profiling the expression of the cucumber chloroplast plastome on the PlasTi-microarray. Owing to the high sequence similarity of plant plastomes, the newly constructed microarray can be used to study plants other than cucumber. Comparative hybridization of chloroplast transcriptomes from cucumber, Arabidopsis, tomato and spinach showed that the PlasTi-microarray is highly versatile.

AtIPD: A Curated Database of Arabidopsis Isoprenoid Pathway Models and Genes for Isoprenoid Network Analysis

Isoprenoid biosynthesis is one of the essential metabolic pathways in plants and other organisms. Despite the importance of isoprenoids for plant functions, not much is known about the regulation of isoprenoid synthesis. Quantitative technologies and systems approaches are now increasingly used to investigate the regulation of metabolic pathways and networks. Prerequisite for systems approaches is the knowledge of network elements and topologies. Information that can be extracted from the public metabolic pathway databases such as AraCyc and KEGG is often not sufficiently comprehensive and current. Therefore we have built a database of manually curated isoprenoid pathway models and genes, the Arabidopsis thaliana Isoprenoid Pathway Database (AtIPD; http://www.atipd.ethz.ch). The database was compiled using information on pathways and pathway genes from BioPathAt (Lange and Ghassemian, 2003, 2005), KEGG (http://www.genome.jp/kegg), AraCyc (http://www.arabidopsis.org/biocyc), SUBA (http://suba.plantenergy.uwa.edu.au), and from the literature. AtIPD can be searched or browsed to extract data and external links related to isoprenoid pathway models, enzyme activities or subcellular enzyme localizations. To display quantitative gene-related data on curated pathway models, we created image annotation and mapping files for integrated use with the MapMan tool (http://mapman.gabipd.org/web/guest/mapman). Additionally, we built SBML XML files of the isoprenoid pathway images using the Cell DesignerTM tool (http://www.celldesigner.org). Users can download all image and annotation files for customization, e.g., adding pathway structural and regulatory network elements or modifying pathway images to visualize other quantitative protein or metabolite data. AtIPD therefore represents a valuable resource for isoprenoid network analysis.

Identification of Genes in the Phenylalanine Metabolic Pathway by Ectopic Expression of a MYB Transcription Factor in Tomato Fruit

Altering expression of transcription factors can be an effective means to coordinately modulate entire metabolic pathways in plants. It can also provide useful information concerning the identities of genes that constitute metabolic networks. Here, we used ectopic expression of a MYB transcription factor, Petunia hybrida ODORANT1, to alter Phe and phenylpropanoid metabolism in tomato (Solanum lycopersicum) fruits. Despite the importance of Phe and phenylpropanoids to plant and human health, the pathway for Phe synthesis has not been unambiguously determined. Microarray analysis of ripening fruits from transgenic and control plants permitted identification of a suite of coregulated genes involved in synthesis and further metabolism of Phe. The pattern of coregulated gene expression facilitated discovery of the tomato gene encoding prephenate aminotransferase, which converts prephenate to arogenate. The expression and biochemical data establish an arogenate pathway for Phe synthesis in tomato fruits. Metabolic profiling and ¹³C flux analysis of ripe fruits further revealed large increases in the levels of a specific subset of phenylpropanoid compounds. However, while increased levels of these human nutrition-related phenylpropanoids may be desirable, there were no increases in levels of Phe-derived flavor volatiles.

Databases on Food Phytochemicals and Their Health-Promoting Effects

Considerable information on the chemistry and biological properties of dietary phytochemicals has accumulated over the past three decades. The scattering of the data in tens of thousands publications and the diversity of experimental approaches and reporting formats all make the exploitation of this information very difficult. Some of the data have been collected and stored in electronic databases so that they can be automatically updated and retrieved. These databases will be particularly important in the evaluation of the effects on health of phytochemicals and in facilitating the exploitation of nutrigenomic data. The content of over 50 databases on chemical structures, spectra, metabolic pathways in plants, occurrence and concentrations in foods, metabolism in humans and animals, biological properties, and effects on health or surrogate markers of health is reviewed. Limits of these databases are emphasized, and needs and recommendations for future developments are underscored. More investments in the construction of databases on phytochemicals and their effects on health are clearly needed. They should greatly contribute to the success of future research in this field.

From hormones to secondary metabolism: the emergence of metabolic gene clusters in plants

Gene clusters for the synthesis of secondary metabolites are a common feature of microbial genomes. Well-known examples include clusters for the synthesis of antibiotics in actinomycetes, and also for the synthesis of antibiotics and toxins in filamentous fungi. Until recently it was thought that genes for plant metabolic pathways were not clustered, and this is certainly true in many cases; however, five plant secondary metabolic gene clusters have now been discovered, all of them implicated in synthesis of defence compounds. An obvious assumption might be that these eukaryotic gene clusters have arisen by horizontal gene transfer from microbes, but there is compelling evidence to indicate that this is not the case. This raises intriguing questions about how widespread such clusters are, what the significance of clustering is, why genes for some metabolic pathways are clustered and those for others are not, and how these clusters form. In answering these questions we may hope to learn more about mechanisms of genome plasticity and adaptive evolution in plants. It is noteworthy that for the five plant secondary metabolic gene clusters reported so far, the enzymes for the first committed steps all appear to have been recruited directly or indirectly from primary metabolic pathways involved in hormone synthesis. This may or may not turn out to be a common feature of plant secondary metabolic gene clusters as new clusters emerge.

Capillary electrophoresis-mass spectrometry analysis of trehalose-6-phosphate in Arabidopsis thaliana seedlings

Trehalose-6-phosphate (T6P) is an intermediate in the plant metabolic pathway that results in trehalose production. T6P has been shown to inhibit the sucrose nonfermenting-1-related protein kinase 1, which is a major regulator of metabolism. The quantitation of T6P has proven difficult due to the complexity of the plant matrix and the low abundance of T6P in plant tissues. The aim of this work was to develop a quantitation method for T6P present in Arabidopsis tissues, with capillary electrophoresis (CE) coupled to electrospray ionization-mass spectrometry (MS) with a sheath liquid (SL) interface. The CE-MS method was first optimized with respect to T6P signal intensity and separation of isomers by studying the composition of the background electrolyte (BGE) and SL. The use of triethylamine (TEA) in the BGE was favorable, providing separation of T6P from sucrose-6-phosphate and minimizing ionization suppression. Replacing ammonium acetate with TEA enhanced T6P signal intensities more than four times. The optimized method allowed quantification of T6P in plant extracts with good linearity (r 2 > 0.99) within a biologically relevant concentration range. The limit of quantification was 80 nM in Arabidopsis extracts, corresponding to 33 pmol/g plant fresh weight. The CE-MS method was applied to the determination of T6P in seedlings from wild type (WT) Arabidopsis and mutants lacking the trehalase AtTRE1, tre1-1, challenged with trehalose or sorbitol. T6P accumulation in tre1-1 plants grown on sorbitol was about twice the level of T6P found in WT. CE-MS is shown to be a fast and reliable technique to analyze phosphodisaccharides for seedling extracts. The low sample volume requirement of CE and its direct MS coupling makes it an attractive alternative for anion-exchange liquid chromatography–MS.

Transcription factors as tools to engineer enhanced drought stress tolerance in plants

Plant growth and productivity are greatly affected by abiotic stresses such as drought, salinity, and temperature. Drought stress is one of the major limitations to crop productivity worldwide due to its multigene nature, making the production of transgenic crops a challenging prospect. To develop crop plant with enhanced tolerance of drought stress, a basic understanding of physiological, biochemical, and gene regulatory networks is essential. In the signal transduction network that leads from the perception of stress signals to the expression of stress-responsive genes, transcription factors (TFs) play an essential role. Because TFs, as opposed to most structural genes, tend to control multiple pathways steps, they have emerged as powerful tools for the manipulation of complex metabolic pathways in plants. One such class of TFs is DREB/CBF that binds to drought responsive cis-acting elements. Transgenic plants have been developed with enhanced stress tolerance by manipulating the expression of DREB/CBF. Recently the functions of an increasing number of plant TFs are being elucidated and increased understanding of these factors in controlling drought stress response has lead to practical approaches for engineering stress tolerance in plants. The utility of the various TFs in plant stress research we review is illustrated by several published examples. The manipulation of native plant regularity networks therefore represents a new era for genetically modified crops. This review focuses on the recent understanding, latest advancements related to TFs and present status of their deployment in developing stress tolerant transgenic plants.

Photorespiratory and Respiratory Carbon Isotope Fractionation in Leaves

The photorespiratory pathway in plants is initiated with the uptake of O2 instead of CO2 by the photosynthetic enzyme Rubisco. This generates a compound in the leaf that must then be recycled through enzymatic reactions distributed over the leaf chloroplasts, peroxisomes, and mitochondria, resulting in a net loss of CO2. The pathway was thought to be well understood, but recent studies using mutants lacking expression of the genes coding for peroxisomal malate dehydrogenase (PMDH) have suggested that there are additional mechanisms that alter the stoichiometry of photorespiratory CO2 release to O2 uptake. Any changes in this stoichiometry will likely cause a change in the isotopic composition of the CO2 being released since it would involve additional enzymes or carbon sources. The isotopic composition of the CO2 being released was measured under both photorespiratory and non-photorespiratory conditions, as well as under different isotopic conditions. Measurements were taken using a tunable diode laser spectroscopy system, which allows high frequency in vivo gas exchange measurements of different CO2 isotopologues. Comparing the PIB isotopic signature of wild type Arabidopsis to that of PMDH mutants, may improve our understanding of alternate metabolic pathways in plants. The decarboxylation fractionation seen is significantly higher in PMDH mutant plants relative to wild-type, which can be partially explained by higher rates of photorespiratory decarboxylation seen in previous studies. The photorespiratory fractionation also varied significantly between wild-type and mutant plants.

Performance of chick pea under the influence of gibberellic acid and oxygenated peptone during germination

The experiments were carried out at the Post Graduate Research Center, to study the influence of Gibberellic Acid (50 ppm) and Oxygenated Peptone (1% aqueous solution) on chick pea (Cicer arietinum L. cv. Vijay) during germination by giving pre-sowing soaking treatment for 6 hours using petriplate method. Both the treatments enhanced the germination process. GA treatment was useful to increase shoot length, mobilization efficiency, emergence index, speed of germination and co-efficient of germination while oxygenated peptone showed an upper hand in root length, shoot/root ratio, biomass and vigour index. GA led to comparatively more synthesis of nucleic acids while oxygenated peptone showed more increase in total carbohydrates and soluble protein content. However, the activity of enzymes like amylase, catalase and protease showed upper hand with oxygenated peptone as compared to GA. In fact GA is costlier and can not be used in organic farming as it enters metabolic pathways of plant and alters them. Hence the use of oxygenated peptone is recommended being less expensive and usable under organic farming condition as it does not enter the plant metabolic pathways and yet brings about significant positive effect.

Phenolics and Plant Allelopathy

Phenolic compounds arise from the shikimic and acetic acid (polyketide) metabolic pathways in plants. They are but one category of the many secondary metabolites implicated in plant allelopathy. Phenolic allelochemicals have been observed in both natural and managed ecosystems, where they cause a number of ecological and economic problems, such as declines in crop yield due to soil sickness, regeneration failure of natural forests, and replanting problems in orchards. Phenolic allelochemical structures and modes of action are diverse and may offer potential lead compounds for the development of future herbicides or pesticides. This article reviews allelopathic effects, analysis methods, and allelopathic mechanisms underlying the activity of plant phenolic compounds. Additionally, the currently debated topic in plant allelopathy of whether catechin and 8-hydroxyquinoline play an important role in Centaurea maculata and Centaurea diffusa invasion success is discussed. Overall, the main purpose of this review is to highlight the allelopacthic potential of phenolic compounds to provide us with methods to solve various ecology problems, especially in regard to the sustainable development of agriculture, forestry, nature resources and environment conservation.

Effects of molybdenum deficiency and defects in molybdate transporter MOT1 on transcript accumulation and nitrogen/sulphur metabolism in Arabidopsis thaliana

Molybdenum (Mo) is a micronutrient essential for plant growth, as several key enzymes of plant metabolic pathways contain Mo cofactor in their catalytic centres. Mo-containing oxidoreductases include nitrate reductase, sulphite oxidase, xanthine dehydrogenase, and aldehyde oxidase. These are involved in nitrate assimilation, sulphite detoxification, purine metabolism or the synthesis of abscisic acid, auxin and glucosinolates in plants. To understand the effects of Mo deficiency and a mutation in a molybdate transporter, MOT1, on nitrogen and sulphur metabolism in Arabidopsis thaliana, transcript and metabolite profiling of the mutant lacking MOT1 was conducted in the presence or absence of Mo. Transcriptome analysis revealed that Mo deficiency had impacts on genes involved in metabolisms, transport, stress responses, and signal transductions. The transcript level of a nitrate reductase NR1 was highly induced under Mo deficiency in mot1-1. The metabolite profiles were analysed further by using gas chromatography time-of-flight mass spectrometry, capillary electrophoresis time-of-flight mass spectrometry, and ultra high performance liquid chromatography. The levels of amino acids, sugars, organic acids, and purine metabolites were altered significantly in the Mo-deficient plants. These results are the first investigation of the global effect of Mo nutrition and MOT1 on plant gene expressions and metabolism.

Application of genetic engineering in potato breeding

Potato breeding programmes worldwide are undergoing a period of rapid change. In order to be successful, breeders must adapt and incorporate the newest up-to-date techniques as they become available. Recent advances in biotechnology make it possible to develop and cultivate more and more sophisticated transgenic crops with multiple modified traits. Gene transfer methods can be used for a wide range of fundamental studies, contributing to a better understanding of the mechanisms of plant/pathogen interactions and the metabolic pathways in plants. Transgenic potato plants are being generated worldwide to investigate the impact of transgene expression on parameters as complex as yield. Historically, potato was one of the first successfully transformed crop plants. Nowadays, transgenic potatoes have been introduced into the food chain of people and animals in several countries. Some of the genetic modifications give potato plants increased resistance to biotic and abiotic environmental factors, while others lead to improved nutritional value, or cause the plants to produce proteins of the immune system of humans or animals or substances that may be used as vaccines in humans or veterinary medicine. The trend today is towards the generation of crops with output traits, e.g. modified starch or carotenoids, or the production of pharmaceuticals in tubers, whereas the early targets were input traits, e.g. herbicide resistance, pest or virus resistance. This review provides a summary of examples illustrating the versatility and applicability of transgenic biology in potato improvement.

From sunlight to phytomass: on the potential efficiency of converting solar radiation to phyto‐energy

The relationship between solar radiation capture and potential plant growth is of theoretical and practical importance. The key processes constraining the transduction of solar radiation into phyto-energy (i.e. free energy in phytomass) were reviewed to estimate potential solar-energy-use efficiency. Specifically, the out-put:input stoichiometries of photosynthesis and photorespiration in C(3) and C(4) systems, mobilization and translocation of photosynthate, and biosynthesis of major plant biochemical constituents were evaluated. The maintenance requirement, an area of important uncertainty, was also considered. For a hypothetical C(3) grain crop with a full canopy at 30°C and 350 ppm atmospheric [CO(2) ], theoretically potential efficiencies (based on extant plant metabolic reactions and pathways) were estimated at c. 0.041 J J(-1) incident total solar radiation, and c. 0.092 J J(-1) absorbed photosynthetically active radiation (PAR). At 20°C, the calculated potential efficiencies increased to 0.053 and 0.118 J J(-1) (incident total radiation and absorbed PAR, respectively). Estimates for a hypothetical C(4) cereal were c. 0.051 and c. 0.114 J J(-1), respectively. These values, which cannot be considered as precise, are less than some previous estimates, and the reasons for the differences are considered. Field-based data indicate that exceptional crops may attain a significant fraction of potential efficiency.

Molecular Evolution of the Endosperm Starch Synthesis Pathway Genes in Rice (Oryza sativa L.) and Its Wild Ancestor, O. rufipogon L.

The evolution of metabolic pathways is a fundamental but poorly understood aspect of evolutionary change. One approach for understanding the complexity of pathway evolution is to examine the molecular evolution of genes that together comprise an integrated metabolic pathway. The rice endosperm starch biosynthetic pathway is one of the most thoroughly characterized metabolic pathways in plants, and starch is a trait that has evolved in response to strong selection during rice domestication. In this study, we have examined six key genes (AGPL2, AGPS2b, SSIIa, SBEIIb, GBSSI, ISA1) in the rice endosperm starch biosynthesis pathway to investigate the evolution of these genes before and after rice domestication. Genome-wide sequence tagged sites data were used as a neutral reference to overcome the problems of detecting selection in species with complex demographic histories such as rice. Five variety groups of Oryza sativa (aus, indica, tropical japonica, temperate japonica, aromatic) and its wild ancestor (O. rufipogon) were sampled. Our results showed evidence of purifying selection at AGPL2 in O. rufipogon and strong evidence of positive selection at GBSSI in temperate japonica and tropical japonica varieties and at GBSSI and SBEIIb in aromatic varieties. All the other genes showed a pattern consistent with neutral evolution in both cultivated rice and its wild ancestor. These results indicate the important role of positive selection in the evolution of starch genes during rice domestication. We discuss the role of SBEIIb and GBSSI in the evolution of starch quality during rice domestication and the power and limitation of detecting selection using genome-wide data as a neutral reference.

식물의 탄소대사공학 연구동향

Yield productivity of staple crops must be increased at least 50% by 2050, in order to feed the world population which is expected to reach 90 billions. Photosynthetic carbon assimilation and carbohydrate metabolism leading to the production of starch would be the final frontier to quest for new sources of technology enabling such a drastic increase of crop productivity. In this review, attempts to genetically engineer plant photosynthetic carbon reduction cycle and metabolic pathways to increase starch production are introduced.

Using the Arabidopsis information resource (TAIR) to find information about Arabidopsis genes.

The Arabidopsis Information Resource (TAIR; http://arabidopsis.org) is a comprehensive Web resource of Arabidopsis biology for plant scientists. TAIR curates and integrates information about genes, proteins, gene function, gene expression, mutant phenotypes, biological materials such as clones and seed stocks, genetic markers, genetic and physical maps, biochemical pathways, genome organization, images of mutant plants, protein sub-cellular localizations, publications, and the research community. The various data types are extensively interconnected and can be accessed through a variety of Web-based search and display tools. This unit primarily focuses on some basic methods for searching, browsing, visualizing, and analyzing information about Arabidopsis genes and describes several new tools such as a new TAIR genome browser (GBrowse), and the TAIR synteny viewer (GBrowse_syn). We also describe how to use AraCyc for mining plant metabolic pathways.

ChemInform Abstract: Benzoxazinoid Biosynthesis, a Model for Evolution of Secondary Metabolic Pathways in Plants.

AbstractChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.

Lipid biochemists salute the genome

The biochemistry of plant metabolic pathways has been studied for many generations; nevertheless, numerous new enzymes and metabolic products have been discovered in the last 5-10 years. More importantly, many intriguing questions remain in all areas of metabolism. In this review, we consider these issues with respect to several pathways of lipid metabolism and the contributions made by the Arabidopsis genome sequence and the tools that it has spawned. These tools have allowed identification of enzymes and transporters required for the mobilization of seed storage lipids, as well as transporters that facilitate movement of lipids from the endoplasmic reticulum to the chloroplast in green leaf cells. Genomic tools were important in recognition of novel components of the cutin and suberin polymers that form water-impermeable barriers in plants. The waxes that also contribute to these barriers are exported from cells of the epidermis by transporters that are now being identified. Biochemical and genetic knowledge from yeast and animals has permitted successful homology-based searches of the Arabidopsis genome for genes encoding enzymes involved in the elongation of fatty acids and the synthesis of sphingolipids. Knowledge of the genome has identified novel enzymes for the biosynthesis of the seed storage lipid, triacylglycerol, and provided a refined understanding of how the pathways of fatty acid and triacylglycerol synthesis are integrated into overall carbon metabolism in developing seeds.