Signaling Molecule In Plants Research Articles

Signalling molecules responsive to ozone-induced oxidative stress in Salvia officinalis

Tropospheric ozone (O3) is the most important gaseous pollutant and induces a mass of negative impacts on vegetation at functional and genic levels. The aim of the present study was to investigate the role of reactive oxygen species and signalling molecules in sage plants exposed to O3 (200 ppb, 5 h). Ozone exposure induced only a transient oxidative burst, as confirmed by the rapid peak of anion superoxide during the first hours of exposure (+16% compared to controls). The spontaneous reaction of O3 with membrane fatty acids stimulates peroxidative processes, as demonstrated by the rise of thiobarbituric acid reactive substances concentration starting after 1 h of exposure (+25%). The formation of lipid-based signalling molecules (e.g. jasmonic acid) may be regarded as a sort of O3-perception. The concomitant accumulation of salicylic acid suggests that sage responds early to O3 by inducing cellular antioxidants mechanisms in order to minimize O3-oxidative burst. The transient increase of abscisic acid (+25% at the end of the treatment) twinned with the maximal ethylene emission (about two-fold higher than controls) could be interpreted as a first attempt by plants to regulate the signalling responses induced by O3. In order to investigate the involvement of transcription factors in managing oxidative protection, BLASTX analysis against the Salvia miltiorrhiza sequence genome was carried out using Arabidopsis thaliana WRKY sequences as queries. Six gene sequences were identified for sage WRKYs and their relative gene expression analyses were characterized. WRKY4, WRKY5, WRKY11 and WRKY46 were up-regulated by O3 at 2 and 5 h of exposure and they showed similarity with AtWRKY48, AtWRKY22 and AtWRKY53 in A. thaliana. These results suggest that WRKYs could play a pivotal role in the signalling mechanisms during the responses of plants to O3.

Molecular Dynamics Simulations Reveal Differentiated Context-Dependent Conformational Dynamics of Two Proteins of the Same Family.

The Arabidopsis pyrabactin resistant 1 (PYR1)-like family of proteins (PYLs) are receptors of abscisic acid (ABA), an essential small signaling molecule in plants. Here, we report a comparative molecular dynamics (MD) study on two PYL members, PYR1 and PYL10, which, despite their highly similar sequences and structures, have been suggested to belong to two different subclasses of PYLs, one being dimeric and relying on binding to ABA to inhibit downstream type 2C protein phosphatases (PP2Cs) and the other being monomeric and able to constitutively inhibit downstream PP2Cs without ABA. MD simulations have been carried out on these proteins in various monomeric or complexation states. Analyses of the simulations unambiguously confirm that ABA has large effects on the conformational dynamics of PYR1 but not PYL10, whereas a downstream PP2C has much larger effects on PYL10 than on PYR1. The differentiated effects are consistent with the functional differences between the two proteins. Potential of mean forces (PMFs) calculated by umbrella sampling showed that binding to ABA strengthens the PYR1-PP2C complex, increasing the PMF change for dissociation from 7.5 to 12.0 kcal mol-1. On the other hand, the same PMF change for an apo-PYL10-PP2C complex was computed to be 9.5 kcal mol-1, suggesting stronger binding in apo-PYL10-PP2C than in apo-PYR1-PP2C. Several specific sequence features that may contribute to the functional differentiation between PYR1 and PYL10 are suggested based on the intersubunit residue-residue contacts occurred in the simulations.

Hydrogen Sulfide Disturbs Actin Polymerization via S-Sulfhydration Resulting in Stunted Root Hair Growth.

Hydrogen sulfide (H2S) is an important signaling molecule in plants. Our previous report suggested that H2S signaling affects the actin cytoskeleton and root hair growth. However, the underlying mechanisms of its effects are not understood. S-Sulfhydration of proteins is regulated directly by H2S, which converts the thiol groups of cysteine (Cys) residues to persulfides and alters protein function. In this work, we studied the effects of S-sulfhydration on actin dynamics in Arabidopsis (Arabidopsis thaliana). We generated transgenic plants overexpressing the H2S biosynthesis-related genes l-CYSTEINE DESULFHYDRASE (LCD) and d-CYSTEINE DESULFHYDRASE in the O-acetylserine(thiol)lyase isoform a1 (oasa1) mutant and Columbia-0 backgrounds. The H2S content increased significantly in overexpressing LCD/oasa1 plants. The density of filamentous actin (F-actin) bundles and the F-actin/globular actin ratio decreased in overexpressing LCD/oasa1 plants. S-Sulfhydration also was enhanced in overexpressing LCD/oasa1 plants. An analysis of actin dynamics suggested that S-sulfhydration inhibited actin polymerization. We also found that ACTIN2 (ACT2) was S-sulfhydrated at Cys-287. Cys-287 is adjacent to the D-loop, which acts as a central region for hydrophobic and electrostatic interactions and stabilizes F-actin filaments. Overaccumulation of H2S caused the depolymerization of F-actin bundles and inhibited root hair growth. Introduction of ACT2 carrying a Cys-287-to-Ser mutation into an act2-1 mutant partially suppressed H2S-dependent inhibition of root hair growth. We conclude that H2S regulates actin dynamics and affects root hair growth.

Nitric oxide alleviates cell death through protein S-nitrosylation and transcriptional regulation during the ageing of elm seeds

Seed ageing is a major problem in the conservation of germplasm resources. The involvement of possible signalling molecules during seed deterioration needs to be identified. In this study, we confirmed that nitric oxide (NO), a key signalling molecule in plants, plays a positive role in the resistance of elm seeds to deterioration. To explore which metabolic pathways were affected by NO, an untargeted metabolomic analysis was conducted, and 163 metabolites could respond to both NO and the ageing treatment. The primary altered pathways include glutathione, methionine, and carbohydrate metabolism. The genes involved in glutathione and methionine metabolism were up-regulated by NO at the transcriptional level. Using a biotin switch method, proteins with an NO-dependent post-translational modification were screened during seed deterioration, and 82 putative S-nitrosylated proteins were identified. Eleven of these proteins were involved in carbohydrate metabolism, and the activities of the three enzymes were regulated by NO. In combination, the results of the metabolomic and S-nitrosoproteomic studies demonstrated that NO could activate glycolysis and inhibit the pentose phosphate pathway. In summary, the combination of these results demonstrated that NO could modulate carbohydrate metabolism at the post-translational level and regulate glutathione and methionine metabolism at the transcriptional level. It provides initial insights into the regulatory mechanisms of NO in seed deterioration.

NAD+ Biosynthesis and Signaling in Plants

Many metabolic processes that occur in living cells involve oxidation-reduction (redox) chemistry underpinned by compounds, such as glutathione, ascorbate, and/or pyridine nucleotides. Among these redox carriers, nicotinamide adenine dinucleotide (NAD+) represents one of the cornerstones of cellular oxidation and is essential for plant growth and development. In addition to its redox role, compelling evidence of a role for NAD+ as a signal molecule in plants is emerging. In non-plant systems, NAD+ biosynthesis can be the product of two independent pathways—the de novo and the salvage pathway. Despite the fact that homologous plant genes exist for the composite enzymes of most, if not all, of these pathways research into plant NAD+ biosynthesis per se has lagged behind that of microbial and mammalian systems. Indeed, much plant work has been focused on additional roles of the composite enzymes in processes, such as DNA repair or post-translational modification of proteins. That said, in recent years, a number of studies, mostly in Arabidopsis, have been carried out in order to fill this knowledge gap. Therefore, in this review, we intend to present a synthesis of such molecular genetic studies as well as biochemical analyses that have begun to elucidate the regulatory hierarchies both of NAD+ biosynthesis and the parallel activities of the enzymes that are involved in these pathways.

The hydrogen sulfide, a downstream signaling molecule of hydrogen peroxide and nitric oxide, involves spermidine-regulated transcription factors and antioxidant defense in white clover in response to dehydration

Hydrogen sulfide (H2S) nowadays is regarded as a novel gaseous signaling molecule in plants during environmental stress response. The spermidine (Spd), an important plant growth regulator, plays critical roles in regulating stress tolerance in plant species. Objectives of this study were to examine whether Spd could act as a H2S signaling activator to regulate stress-responsive transcription factors (TFs) and antioxidant defense and to further explore the possible crosstalk among Spd-induced hydrogen peroxide (H2O2), nitric oxide (NO), and H2S signaling in leaves of white clover. Results showed that dehydration or exogenous application of Spd could significantly increase L/D-cysteine desulfhydrate (L/DCD) activities and then cause a quick accumulation of H2S, followed by significant improvement of antioxidant enzyme (SOD, CAT, GPOX, APX, GR, DHAR, and MDHAR) activities and transcript levels of TFs (bZIP37, bZIP107, DREB2, DREB4, and WRKY108715) and genes encoding antioxidant enzymes. The inhibitors (AOA and NH2OH) of L/DCD activities could effectively attenuate dehydration- or Spd-induced these effects in leaves. Additionally, dehydration-induced increase in H2S accumulation, L/DCD and antioxidant enzyme activities, and genes expression were remarkably abolished by the application of Spd biosynthesis inhibitor (DCHA), whereas exogenous Spd significantly alleviate the inhibitory effect of DCHA on these parameters under dehydration condition. This indicated that H2S which was derived from L/DCD pathway might act as a downstream signaling of Spd, and Spd-activated H2S signaling was involved in dehydration-regulated TFs and antioxidant defense associated with dehydration tolerance. Further analyses found that Spd-induced H2O2 production could not be attenuated by scavengers of NO and H2S, but the scavenger of H2O2 effectively inhibited Spd-induced increases in NO and H2S. The Spd-induced H2S signaling was also substantially inhibited by the application of the NO scavenger. The result implied that Spd-induced H2O2 could be an upstream signal molecule of NO and H2S, and the Spd-induced H2S might act as the downstream signaling of NO in leaves. These obtained results provide the basic evidence to understand Spd-regulated signal transduction in white clover.

Melatonin significantly influences seed germination and seedling growth of Stevia rebaudiana Bertoni.

BackgroundMelatonin (MEL) is a signaling molecule in plants that affects developmental processes during vegetative and reproductive growth. Investigations have proved that exogenously applied MEL also has the potential to improve seed germination and plant development.MethodsIn the present study, seeds of stevia, a species with a very low germination rate, were germinated on an agar gel (AG) containing MEL at various concentrations (5, 20, 100, and 500 µM) in light. Seeds germinated on AG without MEL were used as controls. For the first 24 or 48 h of germination, the seeds were maintained in darkness as a pre-incubation step. Some seeds were not exposed to this pre-incubation step.ResultsAt concentrations of 20 and 5 µM, MEL significantly improved germination, but only in seeds pre-incubated in darkness for 24 h (p < 0.001). At concentrations of 100 and 500 µM, MEL had an inhibitory effect on germination, regardless of the pre-incubation time. Melatonin also affected plantlet properties. At a concentration of 20 µM, MEL increased plantlet fresh weight and leaf numbers. At a concentration of 5 µM, it promoted plantlet height. Regarding root development, the most favorable MEL concentration was 500 µM. Biochemical analysis revealed that MEL promoted higher pigment concentrations but hampered superoxide dismutase activity. On the other hand, the concentrations of sugars and phenolics, as well as the activities of catalase and peroxidase, increased at a MEL concentration of 500 µM.DiscussionThe results suggest that MEL can improve germination of positively photoblastic stevia seeds and that it can play a role in plantlet development. However, the effects observed in the present study depended on the quantity of MEL that was applied.

Cytokinin Signaling in Mycobacterium tuberculosis.

ABSTRACTIt was recently reported that the human-exclusive pathogen Mycobacterium tuberculosis secretes cytokinins, which had only been known as plant hormones. While cytokinins are well-established, adenine-based signaling molecules in plants, they have never been shown to participate in signal transduction in other kingdoms of life. M. tuberculosis is not known to interact with plants. Therefore, we tested the hypothesis that cytokinins trigger transcriptional changes within this bacterial species. Here, we show cytokinins induced the strong expression of the M. tuberculosis gene Rv0077c. We found that Rv0077c expression is repressed by a TetR-like transcriptional repressor, Rv0078. Strikingly, cytokinin-induced expression of Rv0077c resulted in a loss of acid-fast staining of M. tuberculosis. While acid-fast staining is thought to be associated with changes in the bacterial cell envelope and virulence, Rv0077c-induced loss of acid-fastness did not affect antibiotic susceptibility or attenuate bacterial growth in mice, consistent with an unaltered mycolic acid profile of Rv0077c-expressing cells. Collectively, these findings show cytokinins signal transcriptional changes that can affect M. tuberculosis acid-fastness and that cytokinin signaling is no longer limited to the kingdom Plantae.

Exogenous nitric oxide donor and arginine provide protection againstshort-term drought stress in wheat seedlings.

Nitric oxide (NO) is an important plant signaling molecule that has a vital role in abiotic stress tolerance. In the present study, we assesseddrought-induced (15 and 30% PEG, polyethylene glycol) damage in wheat (Triticum aestivum L. cv. Prodip) seedlings and mitigation by the synergistic effect of exogenous Arg (0.5mM l-Arginine) and an NO donor (0.5mM sodium nitroprusside, SNP). Drought stress sharply decreased the leaf relative water content (RWC) but markedly increased the proline (Pro) content in wheat seedlings. Drought stress caused overproduction of reactive oxygen species (ROS) and methylglyoxal (MG) due to the inefficiency of antioxidant enzymes, theglyoxalase system, and theascorbate-glutathione pool. However, supplementation with theNO donor and Arg enhanced the antioxidant defense system (both non-enzymatic and enzymatic components) in drought-stressed seedlings. Application of theNO donor and Arg also enhanced the glyoxalase system and reduced the MG content by increasing the activities of the glyoxalase system enzymes (Gly I and Gly II), which restored the leaf RWC and further increased the Pro content under drought stress conditions. Exogenous NO donor and Arg application enhanced the endogenous NO content, which positively regulated the antioxidant system and reduced ROS production. Thus, the present study reveals the crucial roles of Arg and NO in enhancing drought stress tolerance in wheat seedlings by upgrading their water status and reducing oxidative stress and MG toxicity.

Isoprenoid-derived plant signaling molecules: biosynthesis and biological importance.

The present review summarizes current knowledge of the biosynthesis and biological importance of isoprenoid-derived plant signaling compounds. Cellular organisms use chemical signals for intercellular communication to coordinate their growth, development, and responses to environmental cues. The skeletons of majority of plant signaling molecules, mediators of plant intercellular 'broadcasting', are built from C5 units of isoprene and therefore belong to a huge and diverse group of natural substances called isoprenoids (terpenoids). They fill many important roles in nature. This review summarizes current knowledge of the biosynthesis and biological importance of a group of isoprenoid-derived plant signaling compounds.

Quantitative assessment of chemotropism in pollen tubes using microslit channel filters.

We present a semi-in vitro chemotropism assay that can be used to evaluate the chemoattractant effect of diffusible plant signaling molecules on growing pollen tubes. We constructed an array of microslit channels in a microfluidic device that prevented the passage of randomly growing pollen tubes but permitted ones that are responsive to the chemoattractant. Depending on the microslit channel size, 80%–100% of the randomly growing Torenia fournieri pollen tubes were excluded from reaching the source of the attractant. Thus, the selection of pollen tubes that are capable of responding to chemoattractants from a mixed population can be realized using this platform.

Novel roles of ascorbate in plants: induction of cytosolic Ca2+ signals and efflux from cells via anion channels.

Ascorbate is not often considered as a signalling molecule in plants. This study demonstrates that, in Arabidopsis roots, exogenous l-ascorbic acid triggers a transient increase of the cytosolic free calcium activity ([Ca2+]cyt.) that is central to plant signalling. Exogenous copper and iron stimulate the ascorbate-induced [Ca2+]cyt. elevation, while cation channel blockers, free radical scavengers, low extracellular [Ca2+], transition metal chelators, and removal of the cell wall inhibit this reaction. These data show that apoplastic redox-active transition metals are involved in the ascorbate-induced [Ca2+]cyt. elevation. Exogenous ascorbate also induces a moderate increase in programmed cell death symptoms in intact roots, but it does not activate Ca2+ influx currents in patch-clamped root protoplasts. Intriguingly, the replacement of gluconate with ascorbate in the patch-clamp pipette reveals a large ascorbate efflux current, which shows sensitivity to the anion channel blocker, anthracene-9-carboxylic acid (A9C), indicative of the ascorbate release via anion channels. EPR spectroscopy measurements demonstrate that salinity (NaCl) triggers the accumulation of root apoplastic ascorbyl radicals in an A9C-dependent manner, confirming that l-ascorbate leaks through anion channels under depolarization. This mechanism may underlie ascorbate release, signalling phenomena, apoplastic redox reactions, iron acquisition, and control the ionic and electrical equilibrium (together with K+ efflux via GORK channels).

Metabolomic "Dark Matter" Dependent on Peroxisomal β-Oxidation in Caenorhabditis elegans.

Peroxisomal β-oxidation (pβo) is a highly conserved fat metabolism pathway involved in the biosynthesis of diverse signaling molecules in animals and plants. In Caenorhabditis elegans, pβo is required for the biosynthesis of the ascarosides, signaling molecules that control development, lifespan, and behavior in this model organism. Via comparative mass spectrometric analysis of pβo mutants and wildtype, we show that pβo in C.elegans and the satellite model P.pacificus contributes to life stage-specific biosynthesis of several hundred previously unknown metabolites. The pβo-dependent portion of the metabolome is unexpectedly diverse, e.g., intersecting with nucleoside and neurotransmitter metabolism. Cell type-specific restoration of pβo in pβo-defective mutants further revealed that pβo-dependent submetabolomes differ between tissues. These results suggest that interactions of fat, nucleoside, and other primary metabolism pathways can generate structural diversity reminiscent of that arising from combinatorial strategies in microbial natural product biosynthesis.

Biosynthesis and in vitro enzymatic synthesis of the isoleucine conjugate of 12-oxo-phytodienoic acid from the isoleucine conjugate of α-linolenic acid

The isoleucine conjugate of 12-oxo-phytodienoic acid (OPDA-Ile), a new member of the jasmonate family, was recently identified in Arabidopsis thaliana and might be a signaling molecule in plants. However, the biosynthesis and function of OPDA-Ile remains elusive. This study reports an in vitro enzymatic method for synthesizing OPDA-Ile, which is catalyzed by reactions of lipoxygenase (LOX), allene oxide synthase (AOS), and allene oxide cyclase (AOC) using isoleucine conjugates of α -linolenic acid (LA-Ile) as the substrate. A. thaliana fed LA-Ile exhibited a marked increase in the OPDA-Ile concentration. LA-Ile was also detected in A. thaliana. Furthermore, stable isotope labelled LA-Ile was incorporated into OPDA-Ile. Thus, OPDA-Ile is biosynthesized via the cyclization of LA-Ile in A. thaliana.

Sorption, plant uptake and metabolism of benzodiazepines

Reuse of treated wastewater for irrigation of crops is growing in arid and semi-arid regions, whilst increasing amounts of biosolids are being applied to fields to improve agricultural outputs. Due to incomplete removal in the wastewater treatment processes, pharmaceuticals present in treated wastewater and biosolids can contaminate soil systems. Benzodiazepines are a widely used class of pharmaceuticals that are released following wastewater treatment. Benzodiazepines are represented by a class of compounds with a range of physicochemical properties and this study was therefore designed to evaluate the influence of soil properties on the sorption behaviour and subsequent uptake of seven benzodiazepines (chlordiazepoxide, clonazepam, diazepam, flurazepam, oxazepam, temazepam and triazolam) in two plant species. The sorption and desorption behaviour of benzodiazepines was strongly influenced by soil type and hydrophobicity of the chemical. The partitioning behaviour of these chemicals in soil was a key controller of the uptake and accumulation of benzodiazepines by radish (Raphanus sativus) and silverbeet (Beta vulgaris). Benzodiazepines such as oxazepam that were neutral, had low sorption coefficients (Kd) or had pH-adjusted log octanol-water partition coefficients (log Dow, pH6.3) values close to 2 had the greatest extent of uptake. Conversely, benzodiazepines such as flurazepam that had an ionised functional groups and greater Kd values had comparatively limited accumulation in the selected plant species. Results also revealed active in-plant metabolism of benzodiazepines, potentially analogous to the known metabolic transformation pathway of benzodiazepines in humans. Along with this observed biological transformation of benzodiazepines in exposed plants, previously work has established the widespread presence of the plant signalling molecule γ-amino butyric acid (GABA), which is specifically modulated by benzodiazepines in humans. This highlights the need for further assessment of the potential for biological activity of benzodiazepines following their plant uptake.

Hydrogen Sulfide Increases Production of NADPH Oxidase-Dependent Hydrogen Peroxide and Phospholipase D-Derived Phosphatidic Acid in Guard Cell Signaling.

Hydrogen sulfide (H2S) is an important gaseous signaling molecule in plants that participates in stress responses and development. l-Cys desulfhydrase 1, one of the enzymatic sources of H2S in plants, participates in abscisic acid-induced stomatal closure. We combined pharmacological and genetic approaches to elucidate the involvement of H2S in stomatal closure and the interplay between H2S and other second messengers of the guard cell signaling network, such as hydrogen peroxide (H2O2) and phospholipase D (PLD)-derived phosphatidic acid in Arabidopsis (Arabidopsis thaliana). Both NADPH oxidase isoforms, respiratory burst oxidase homolog (RBOH)D and RBOHF, were required for H2S-induced stomatal closure. In vivo imaging using the cytosolic ratiometric fluorescent biosensor roGFP2-Orp1 revealed that H2S stimulates H2O2 production in Arabidopsis guard cells. Additionally, we observed an interplay between H2S and PLD activity in the regulation of reactive oxygen species production and stomatal movement. The PLDα1 and PLDδ isoforms were required for H2S-induced stomatal closure, and most of the H2S-dependent H2O2 production required the activity of PLDα1. Finally, we showed that H2S induced increases in the PLDδ-derived phosphatidic acid levels in guard cells. Our results revealed the involvement of H2S in the signaling network that controls stomatal closure, and suggest that H2S regulates NADPH oxidase and PLD activity in guard cells.

Correction: Hydrogen sulfide alleviates postharvest ripening and senescence of banana by antagonizing the effect of ethylene.

[This corrects the article DOI: 10.1371/journal.pone.0180113.].

UDP-Glucose: A Potential Signaling Molecule in Plants?

This perspective paper focuses on the most recent results suggesting a potential role for UDP-Glucose as a signaling molecule in plants. In animals, UDP-Glucose is well-established as an extracellular signaling molecule that is sensed by G-protein coupled receptors, activating several downstream defense mechanisms. Recent studies have shown that abnormal growth occurred in both vegetative and reproductive tissue of plants with reduced UDP-Glucose levels, and this could be rescued by exogenous UDP-Glucose. In plants with increased biomass accumulation, the genes involved in UDP-Glucose production were up-regulated. However, excessive endogenous accumulation of UDP-Glucose induced programmed cell death (PCD), and this could also be obtained by exogenous UDP-Glucose application. Plants with decreased UDP-glucose were insensitive to pathogen induced PCD. We speculate that UDP-Glucose acts as an extracellular signaling molecule in plants, and that it may be perceived as a damage-associated molecular pattern.

The effects of putrescine are partly overlapping with osmotic stress processes in wheat

Polyamine metabolism is in relation with several metabolic pathways and linked with plant hormones or signalling molecules; in addition polyamines may modulate the up- or down-regulation of gene expression. However the precise mechanism by which polyamines act at the transcription level is still unclear. In the present study the modifying effect of putrescine pre-treatment has been investigated using the microarray transcriptome profile analysis under the conditions where exogenous putrescine alleviated osmotic stress in wheat plants. Pre-treatment with putrescine induced the unique expression of various general stress-related genes. Although there were obvious differences between the effects of putrescine and polyethylene glycol treatments, there was also a remarkable overlap between the effects of putrescine and osmotic stress responses in wheat plants, suggesting that putrescine has already induced acclimation processes under control conditions. The fatty acid composition in certain lipid fractions and the antioxidant enzyme activities have also been specifically changed under osmotic stress conditions or after treatment with putrescine.

Identification, characterization, and expression of the SWEET gene family in Phalaenopsis equestris and Dendrobium officinale

Sugars are important molecules that function not only as primary metabolites, but also as nutrients and signal molecules in plants. The sugar transport protein genes family SWEET has been recently identified. The availability of the Dendrobium officinale and Phalaenopsis equestris genome sequences offered the opportunity to study the SWEET gene family in this two orchid species. We identified 22 and 16 putative SWEET genes, respectively, in the genomes of D. officinale and P. equestris using comprehensive bioinformatics analysis. Based on phylogenetic comparisons with SWEET proteins from Arabidopsis and rice, the DoSWEET and PeSWEET proteins could be divided into four clades; among these, clade II specifically lacked PeSWEETs and clade IV specifically lacked DoSWEETs, and there were orthologs present between D. officinale and P. equestris. Protein sequence alignments suggest that there is a predicted serine phosphorylation site in each of the highly conserved MtN3/saliva domain regions. Gene expression analysis in four tissues showed that three PeSWEET genes were most highly expressed in the flower, leaf, stem, and root, suggesting that these genes might play important roles in growth and development in P. equestris. Analysis of gene expression in different floral organs showed that five PeSWEET genes were highly expressed in the column (gynostemium), implying their possible involvement in reproductive development in this species. The expression patterns of seven PeSWEETs in response to different abiotic stresses showed that three genes were upregulated significantly in response to high temperature and two genes were differently expressed at low temperature. The results of this study lay the foundation for further functional analysis of SWEET genes in orchids.