RESUMO
Nearly 90% of flowering plants depend on animals for reproduction. One of the main rewards plants offer to pollinators for visitation is nectar. Nesocodon mauritianus (Campanulaceae) produces a blood-red nectar that has been proposed to serve as a visual attractant for pollinator visitation. Here, we show that the nectar's red color is derived from a previously undescribed alkaloid termed nesocodin. The first nectar produced is acidic and pale yellow in color, but slowly becomes alkaline before taking on its characteristic red color. Three enzymes secreted into the nectar are either necessary or sufficient for pigment production, including a carbonic anhydrase that increases nectar pH, an aryl-alcohol oxidase that produces a pigment precursor, and a ferritin-like catalase that protects the pigment from degradation by hydrogen peroxide. Our findings demonstrate how these three enzymatic activities allow for the condensation of sinapaldehyde and proline to form a pigment with a stable imine bond. We subsequently verified that synthetic nesocodin is indeed attractive to Phelsuma geckos, the most likely pollinators of Nesocodon We also identify nesocodin in the red nectar of the distantly related and hummingbird-visited Jaltomata herrerae and provide molecular evidence for convergent evolution of this trait. This work cumulatively identifies a convergently evolved trait in two vertebrate-pollinated species, suggesting that the red pigment is selectively favored and that only a limited number of compounds are likely to underlie this type of adaptation.
Assuntos
Flores/metabolismo , Magnoliopsida/metabolismo , Pigmentação/fisiologia , Néctar de Plantas/metabolismo , Pólen/metabolismo , Adaptação Fisiológica/fisiologia , Animais , Aves/fisiologia , Lagartos/fisiologia , Polinização/fisiologia , Reprodução/fisiologiaRESUMO
A few Capsicum (pepper) species produce yellow-colored floral nectar, but the chemical identity and biological function of the yellow pigment are unknown. A combination of analytical biochemistry techniques was used to identify the pigment that gives Capsicum baccatum and Capsicum pubescens nectars their yellow color. Microbial growth assays, visual modeling, and honey bee preference tests for artificial nectars containing riboflavin were used to assess potential biological roles for the nectar pigment. High concentrations of riboflavin (vitamin B2) give the nectars their intense yellow color. Nectars containing riboflavin generate reactive oxygen species when exposed to light and reduce microbial growth. Visual modeling also indicates that the yellow color is highly conspicuous to bees within the context of the flower. Lastly, field experiments demonstrate that honey bees prefer artificial nectars containing riboflavin. Some Capsicum nectars contain a yellow-colored vitamin that appears to play roles in (1) limiting microbial growth, (2) the visual attraction of bees, and (3) as a reward to nectar-feeding flower visitors (potential pollinators), which is especially interesting since riboflavin is an essential nutrient for brood rearing in insects. These results cumulatively suggest that the riboflavin found in some Capsicum nectars has several functions.
Assuntos
Capsicum , Néctar de Plantas , Riboflavina , Capsicum/fisiologia , Capsicum/crescimento & desenvolvimento , Riboflavina/metabolismo , Néctar de Plantas/química , Abelhas/fisiologia , Animais , Flores/fisiologia , Espécies Reativas de Oxigênio/metabolismo , CorRESUMO
The black nectar produced by Melianthus flowers is thought to serve as a visual attractant to bird pollinators, but the chemical identity and synthesis of the black pigment are unknown. A combination of analytical biochemistry, transcriptomics, proteomics, and enzyme assays was used to identify the pigment that gives Melianthus nectar its black color and how it is synthesized. Visual modeling of pollinators was also used to infer a potential function of the black coloration. High concentrations of ellagic acid and iron give the nectar its dark black color, which can be recapitulated through synthetic solutions containing only ellagic acid and iron(iii). The nectar also contains a peroxidase that oxidizes gallic acid to form ellagic acid. In vitro reactions containing the nectar peroxidase, gallic acid, hydrogen peroxide, and iron(iii) fully recreate the black color of the nectar. Visual modeling indicates that the black color is highly conspicuous to avian pollinators within the context of the flower. Melianthus nectar contains a natural analog of iron-gall ink, which humans have used since at least medieval times. This pigment is derived from an ellagic acid-Fe complex synthesized in the nectar and is likely involved in the attraction of passerine pollinators endemic to southern Africa.
Assuntos
Magnoliopsida , Néctar de Plantas , Humanos , Ácido Elágico , Compostos Férricos , Tinta , Flores , Peroxidases , PolinizaçãoRESUMO
Nectar volume and sugar composition are key determinants of the strength of plant-pollinator mutualisms. The main nectar sugars are sucrose, glucose and fructose, which can vary widely in ratio and concentration across species. Brassica spp. produce a hexose-dominant nectar (high in the monosaccharides glucose and fructose) with very low levels of the disaccharide sucrose. Cell wall invertases (CWINVs) catalyze the irreversible hydrolysis of sucrose into glucose and fructose in the apoplast. We found that BrCWINV4A is highly expressed in the nectaries of Brassica rapa. Moreover, a brcwinv4a null mutant: (i) has greatly reduced CWINV activity in the nectaries; (ii) produces a sucrose-rich nectar; but (iii) with significantly less volume. These results definitively demonstrate that CWINV activity is not only essential for the production of a hexose-rich nectar, but also support a hypothetical model of nectar secretion in which its hydrolase activity is required for maintaining a high intracellular-to-extracellular sucrose ratio that facilitates the continuous export of sucrose into the nectary apoplast. The extracellular hydrolysis of each sucrose into two hexoses by BrCWINV4A also likely creates the osmotic potential required for nectar droplet formation. These results cumulatively indicate that modulation of CWINV activity can at least partially account for naturally occurring differences in nectar volume and sugar composition. Finally, honeybees prefer nectars with some sucrose, but wild-type B. rapa flowers were much more heavily visited than flowers of brcwinv4a, suggesting that the potentially attractive sucrose-rich nectar of brcwinv4a could not compensate for its low volume.
Assuntos
Brassica rapa/citologia , Brassica rapa/metabolismo , Néctar de Plantas/fisiologia , Açúcares/metabolismo , beta-Frutofuranosidase/metabolismo , Animais , Abelhas , Brassica rapa/genética , Parede Celular/enzimologia , Regulação da Expressão Gênica de Plantas , Hidrólise , Mutação , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Polinização , Sementes/genética , Sementes/fisiologia , Açúcares/química , beta-Frutofuranosidase/genéticaRESUMO
Floral nectar is a sugary solution produced by nectaries to attract and reward pollinators. Nectar metabolites, such as sugars, are synthesized within the nectary during secretion from both pre-stored and direct phloem-derived precursors. In addition to sugars, nectars contain nitrogenous compounds such as amino acids; however, little is known about the role(s) of nitrogen (N) compounds in nectary function. In this study, we investigated N metabolism in Cucurbita pepo (squash) floral nectaries in order to understand how various N-containing compounds are produced and determine the role of N metabolism in nectar secretion. The expression and activity of key enzymes involved in primary N assimilation, including nitrate reductase (NR) and alanine aminotransferase (AlaAT), were induced during secretion in C. pepo nectaries. Alanine (Ala) accumulated to about 35% of total amino acids in nectaries and nectar during peak secretion; however, alteration of vascular nitrate supply had no impact on Ala accumulation during secretion, suggesting that nectar(y) amino acids are produced by precursors other than nitrate. In addition, nitric oxide (NO) is produced from nitrate and nitrite, at least partially by NR, in nectaries and nectar. Hypoxia-related processes are induced in nectaries during secretion, including lactic acid and ethanolic fermentation. Finally, treatments that alter nitrate supply affect levels of hypoxic metabolites, nectar volume and nectar sugar composition. The induction of N metabolism in C. pepo nectaries thus plays an important role in the synthesis and secretion of nectar sugar.
Assuntos
Alanina/biossíntese , Cucurbita/metabolismo , Óxido Nítrico/metabolismo , Néctar de Plantas/metabolismo , Alanina Transaminase/genética , Alanina Transaminase/metabolismo , Aminoácidos/metabolismo , Cucurbita/efeitos dos fármacos , Cucurbita/fisiologia , Flores/metabolismo , Regulação da Expressão Gênica de Plantas , Hipóxia , Nitrato Redutase/genética , Nitrato Redutase/metabolismo , Nitratos/metabolismo , Nitratos/farmacologia , Nitrogênio/metabolismo , Néctar de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estresse Fisiológico/fisiologia , Açúcares/metabolismoRESUMO
Nectar is a primary reward mediating plant-animal mutualisms to improve plant fitness and reproductive success. Four distinct trichomatic nectaries develop in cotton (Gossypium hirsutum), one floral and three extrafloral, and the nectars they secrete serve different purposes. Floral nectar attracts bees for promoting pollination, while extrafloral nectar attracts predatory insects as a means of indirect protection from herbivores. Cotton therefore provides an ideal system for contrasting mechanisms of nectar production and nectar composition between different nectary types. Here, we report the transcriptome and ultrastructure of the four cotton nectary types throughout development and compare these with the metabolomes of secreted nectars. Integration of these datasets supports specialization among nectary types to fulfill their ecological niche, while conserving parallel coordination of the merocrine-based and eccrine-based models of nectar biosynthesis. Nectary ultrastructures indicate an abundance of rough endoplasmic reticulum positioned parallel to the cell walls and a profusion of vesicles fusing to the plasma membranes, supporting the merocrine model of nectar biosynthesis. The eccrine-based model of nectar biosynthesis is supported by global transcriptomics data, which indicate a progression from starch biosynthesis to starch degradation and sucrose biosynthesis and secretion. Moreover, our nectary global transcriptomics data provide evidence for novel metabolic processes supporting de novo biosynthesis of amino acids secreted in trace quantities in nectars. Collectively, these data demonstrate the conservation of nectar-producing models among trichomatic and extrafloral nectaries.
Assuntos
Produtos Agrícolas/metabolismo , Flores/metabolismo , Gossypium/metabolismo , Néctar de Plantas/biossíntese , Tricomas/metabolismo , Vias BiossintéticasRESUMO
Floral nectar is a sugary solution produced by plants to entice pollinator visitation. A general mechanism for nectar secretion has been established from genetic studies in Arabidopsis (Arabidopsis thaliana); however, supporting metabolic and biochemical evidence for this model is scarce in other plant species. We used squash (Cucurbita pepo) to test whether the genetic model of nectar secretion in Arabidopsis is supported at the metabolic level in other species. As such, we analyzed the expression and activity of key enzymes involved in carbohydrate metabolism in squash nectaries throughout floral maturation and the associated starch and soluble sugars, as well as nectar volume and sugar under different growth conditions. Here we show that the steps that are important for nectar secretion in Arabidopsis, including nectary starch degradation, Suc synthesis, and Suc export, are supported by metabolic and biochemical data in C. pepo Additionally, our findings suggest that sugars imported from the phloem during nectar secretion, without prior storage as starch, are important for generating C. pepo nectar. Finally, we predict that trehalose and trehalose 6-P play important regulatory roles in nectary starch degradation and nectar secretion. These data improve our understanding of how nectar is produced in an agronomically relevant species with the potential for use as a model to help us gain insight into the biochemistry and metabolism of nectar secretion in flowering plants.
Assuntos
Metabolismo dos Carboidratos/fisiologia , Cucurbita/metabolismo , Flores/metabolismo , Néctar de Plantas/metabolismo , Polinização/fisiologia , Transdução de Sinais/fisiologiaRESUMO
Angiosperms developed floral nectaries that reward pollinating insects. Although nectar function and composition have been characterized, the mechanism of nectar secretion has remained unclear. Here we identify SWEET9 as a nectary-specific sugar transporter in three eudicot species: Arabidopsis thaliana, Brassica rapa (extrastaminal nectaries) and Nicotiana attenuata (gynoecial nectaries). We show that SWEET9 is essential for nectar production and can function as an efflux transporter. We also show that sucrose phosphate synthase genes, encoding key enzymes for sucrose biosynthesis, are highly expressed in nectaries and that their expression is also essential for nectar secretion. Together these data are consistent with a model in which sucrose is synthesized in the nectary parenchyma and subsequently secreted into the extracellular space via SWEET9, where sucrose is hydrolysed by an apoplasmic invertase to produce a mixture of sucrose, glucose and fructose. The recruitment of SWEET9 for sucrose export may have been a key innovation, and could have coincided with the evolution of core eudicots and contributed to the evolution of nectar secretion to reward pollinators.
Assuntos
Arabidopsis/metabolismo , Glucosiltransferases/metabolismo , Néctar de Plantas/metabolismo , Proteínas de Plantas/metabolismo , Sacarose/metabolismo , Alquil e Aril Transferases/metabolismo , Animais , Arabidopsis/citologia , Arabidopsis/enzimologia , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Brassica rapa/anatomia & histologia , Brassica rapa/enzimologia , Brassica rapa/metabolismo , Metabolismo dos Carboidratos , Espaço Extracelular/metabolismo , Flores/fisiologia , Glucosiltransferases/genética , Células HEK293 , Humanos , Proteínas de Membrana Transportadoras/metabolismo , Oócitos , Néctar de Plantas/biossíntese , Polinização , Transporte Proteico , Homologia de Sequência , Amido/metabolismo , Nicotiana/anatomia & histologia , Nicotiana/enzimologia , Nicotiana/metabolismo , Xenopus , beta-Frutofuranosidase/metabolismoRESUMO
Nectar is one of the key rewards mediating plant-mutualist interactions. In addition to sugars, nectars often contain many other compounds with important biological functions, including proteins. This study was undertaken to assess the proteinaceous content of Brassica rapa nectar. SDS-PAGE analysis of raw B. rapa nectar revealed the presence of ~10 proteins, with a major band at ~10 kDa. This major band was found to contain a non-specific lipid transfer protein encoded by B. rapa locus Bra028980 and subsequently termed BrLTP2.1. Sequence analysis of BrLTP2.1 predicted the presence of a signal peptide required for secretion from the cell, eight cysteines, and a mature molecular mass of 7.3 kDa. Constitutively expressed BrLTP2.1-GFP in Arabidopsis displayed accumulation patterns consistent with secretion from nectary cells. BrLTP2.1 was also found to have relatively high sequence similarity to non-specific lipid-transfer proteins with known functions in plant defense, including Arabidopsis DIR1. Heterologously expressed and purified BrLTP2.1 was extremely heat stable and bound strongly to saturated free fatty acids, but not methyl jasmonate. Recombinant BrLTP2.1 also had direct antimicrobial activity against an extensive range of plant pathogens, being particularly effective against necrotrophic fungi. Taken together, these results suggest that BrLTP2.1 may function to prevent microbial growth in nectars.
Assuntos
Antifúngicos/química , Brassica rapa/genética , Proteínas de Transporte/genética , Néctar de Plantas/química , Proteínas de Plantas/genética , Sequência de Aminoácidos , Brassica rapa/metabolismo , Proteínas de Transporte/metabolismo , Filogenia , Proteínas de Plantas/metabolismoRESUMO
BACKGROUND: Pennycress [Thlaspi arvense L (Brassicaceae)] is being domesticated as a renewable biodiesel feedstock that also provides crucial ecosystems services, including as a nutritional resource for pollinators. However, its flowers produce significantly less nectar than other crop relatives in the Brassicaceae. This study was undertaken to understand the basic biology of the pennycress nectary as an initial step toward the possibility of enhancing nectar output from its flowers. RESULTS: Pennycress flowers contain four equivalent nectaries located extrastaminally at the base of the insertion sites of short and long stamens. Like other Brassicaceae, the nectaries have open stomates on their surface, which likely serve as the sites of nectar secretion. The nectaries produce four distinct nectar droplets that accumulate in concave structures at the base of each of the four petals. To understand the molecular biology of the pennycress nectary, RNA was isolated from 'immature' (pre-secretory) and 'mature' (secretory) nectaries and subjected to RNA-seq. Approximately 184 M paired-end reads (368 M total reads) were de novo assembled into a total of 16,074 independent contigs, which mapped to 12,335 unique genes in the pennycress genome. Nearly 3700 genes were found to be differentially expressed between immature and mature nectaries and subjected to gene ontology and metabolic pathway analyses. Lastly, in silico analyses identified 158 pennycress orthologs to Arabidopsis genes with known enriched expression in nectaries. These nectary-enriched expression patterns were verified for select pennycress loci by semi-quantitative RT-PCR. CONCLUSIONS: Pennycress nectaries are unique relative to those of other agriculturally important Brassicaceae, as they contain four equivalent nectaries that present their nectar in specialized cup-shaped structures at the base of the petals. In spite of these morphological differences, the genes underlying the regulation and production of nectar appear to be largely conserved between pennycress and Arabidopsis thaliana. These results provide a starting point for using forward and reverse genetics approaches to enhance nectar synthesis and secretion in pennycress.
Assuntos
Néctar de Plantas/genética , Thlaspi/genética , Flores/anatomia & histologia , Flores/genética , Flores/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genes de Plantas/genética , Filogenia , Néctar de Plantas/metabolismo , RNA de Plantas/genética , RNA de Plantas/metabolismo , Thlaspi/metabolismoRESUMO
The PIN family of proteins is best known for its involvement in polar auxin transport and tropic responses. PIN6 (At1g77110) is one of the remaining PIN family members in Arabidopsis thaliana to which a biological function has not yet been ascribed. Here we report that PIN6 is a nectary-enriched gene whose expression level is positively correlated with total nectar production in Arabidopsis, and whose function is required for the proper development of short stamens. PIN6 accumulates in internal membranes consistent with the ER, and multiple lines of evidence demonstrate that PIN6 is required for auxin-dependent responses in nectaries. Wild-type plants expressing auxin-responsive DR5:GFP or DR5:GUS reporters displayed intense signal in lateral nectaries, but pin6 lateral nectaries showed little or no signal for these reporters. Further, exogenous auxin treatment increased nectar production more than tenfold in wild-type plants, but nectar production was not increased in pin6 mutants when treated with auxin. Conversely, the auxin transport inhibitor N-1-naphthylphthalamic acid (NPA) reduced nectar production in wild-type plants by more than twofold, but had no significant effect on pin6 lines. Interestingly, a MYB57 transcription factor mutant, myb57-2, closely phenocopied the loss-of-function mutant pin6-2. However, PIN6 expression was not dependent on MYB57, and RNA-seq analyses of pin6-2 and myb57-2 mutant nectaries showed little overlap in terms of differentially expressed genes. Cumulatively, these results demonstrate that PIN6 is required for proper auxin response and nectary function in Arabidopsis. These results also identify auxin as an important factor in the regulation of nectar production, and implicate short stamens in the maturation of lateral nectaries.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Ácidos Indolacéticos/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Arabidopsis/efeitos dos fármacos , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Transporte Biológico , Flores/efeitos dos fármacos , Flores/genética , Flores/crescimento & desenvolvimento , Flores/metabolismo , Genes Reporter , Homeostase , Ácidos Indolacéticos/farmacologia , Proteínas de Membrana Transportadoras/genética , Mutagênese Insercional , Fenótipo , Reguladores de Crescimento de Plantas/farmacologia , Néctar de Plantas/metabolismo , Regiões Promotoras Genéticas , Fatores de Transcrição/genéticaRESUMO
Nectar is a sugary, aqueous solution that plants offer as a reward to animal mutualists for visitation. Since nectars are so nutrient-rich, they often harbor significant microbial communities, which can be pathogenic, benign, or even sometimes beneficial to plant fitness. Through recent advances, it is now clear that these microbes alter nectar chemistry, which in turn influences mutualist behavior (e.g. pollinator visitation). To counteract unwanted microbial growth, nectars often contain antimicrobial compounds, especially in the form of proteins, specialized (secondary) metabolites, and metals. This review covers our current understanding of nectar antimicrobials, as well as their interplay with both microbes and insect visitors.
Assuntos
Anti-Infecciosos/metabolismo , Flores/fisiologia , Insetos/fisiologia , Néctar de Plantas/metabolismo , Proteínas de Plantas/fisiologia , Animais , Comportamento Animal , Metais , Polinização , Metabolismo SecundárioRESUMO
BACKGROUND: In microarray gene expression profiling experiments, differentially expressed genes (DEGs) are detected from among tens of thousands of genes on an array using statistical tests. It is important to control the number of false positives or errors that are present in the resultant DEG list. To date, more than 20 different multiple test methods have been reported that compute overall Type I error rates in microarray experiments. However, these methods share the following dilemma: they have low power in cases where only a small number of DEGs exist among a large number of total genes on the array. RESULTS: This study contrasts parallel multiplicity of objectively related tests against the traditional simultaneousness of subjectively related tests and proposes a new assessment called the Error Discovery Rate (EDR) for evaluating multiple test comparisons in microarray experiments. Parallel multiple tests use only the negative genes that parallel the positive genes to control the error rate; while simultaneous multiple tests use the total unchanged gene number for error estimates. Here, we demonstrate that the EDR method exhibits improved performance over other methods in specificity and sensitivity in testing expression data sets with sequence digital expression confirmation, in examining simulation data, as well as for three experimental data sets that vary in the proportion of DEGs. The EDR method overcomes a common problem of previous multiple test procedures, namely that the Type I error rate detection power is low when the total gene number used is large but the DEG number is small. CONCLUSIONS: Microarrays are extensively used to address many research questions. However, there is potential to improve the sensitivity and specificity of microarray data analysis by developing improved multiple test comparisons. This study proposes a new view of multiplicity in microarray experiments and the EDR provides an alternative multiple test method for Type I error control in microarray experiments.
Assuntos
Perfilação da Expressão Gênica/métodos , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Simulação por Computador , Modelos Genéticos , Sensibilidade e EspecificidadeRESUMO
To date, no genes have been reported to directly affect the de novo production of floral nectar. In an effort to identify genes involved in nectar production, the Affymetrix((R)) ATH1 GeneChip was previously used to examine global gene expression profiles in Arabidopsis thaliana nectaries. One of the genes displaying highly enriched expression in nectaries was CELL WALL INVERTASE 4 (AtCWINV4, At2g36190), which encodes an enzyme that putatively catalyses the hydrolysis of sucrose into glucose and fructose. RT-PCR was used to confirm the nectary-enriched expression of AtCWINV4, as well as an orthologue from Brassica rapa. To probe biological function, two independent Arabidopsis cwinv4 T-DNA mutants were isolated. Unlike wild-type plants, cwinv4 lines did not produce nectar. While overall nectary morphology appeared to be normal, cwinv4 flowers accumulated higher than normal levels of starch in the receptacle, but not within the nectaries themselves. Conversely, wild-type, but not cwinv4, nectarial stomata stained intensely for starch. Cell wall extracts prepared from mutant flowers displayed greatly reduced invertase activity when compared with wild-type plants, and cwinv4 flowers also accumulated significantly lower levels of total soluble sugar. Cumulatively, these results implicate CWINV4 as an absolutely required factor for nectar production in the Brassicaceae, specifically by maintaining constant sink status within nectaries, thus allowing them to accumulate the sugars necessary for nectar production. In addition, CWINV4 is probably responsible for the hexose-rich composition observed for many Brassicaceae nectars.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimologia , Néctar de Plantas/metabolismo , beta-Frutofuranosidase/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Brassicaceae/enzimologia , Brassicaceae/genética , Brassicaceae/metabolismo , Flores/enzimologia , Flores/genética , Flores/metabolismo , Mutação , Amido/metabolismo , beta-Frutofuranosidase/genéticaRESUMO
BACKGROUND: Many flowering plants attract pollinators by offering a reward of floral nectar. Remarkably, the molecular events involved in the development of nectaries, the organs that produce nectar, as well as the synthesis and secretion of nectar itself, are poorly understood. Indeed, to date, no genes have been shown to directly affect the de novo production or quality of floral nectar. To address this gap in knowledge, the ATH1 Affymetrix GeneChip array was used to systematically investigate the Arabidopsis nectary transcriptome to identify genes and pathways potentially involved in nectar production. RESULTS: In this study, we identified a large number of genes differentially expressed between secretory lateral nectaries and non-secretory median nectary tissues, as well as between mature lateral nectaries (post-anthesis) and immature lateral nectaries (pre-anthesis). Expression within nectaries was also compared to thirteen non-nectary reference tissues, from which 270 genes were identified as being significantly upregulated in nectaries. The expression patterns of 14 nectary-enriched genes were also confirmed via RT PCR. Upon looking into functional groups of upregulated genes, pathways involved in gene regulation, carbohydrate metabolism, and lipid metabolism were particularly enriched in nectaries versus reference tissues. CONCLUSION: A large number of genes preferentially expressed in nectaries, as well as between nectary types and developmental stages, were identified. Several hypotheses relating to mechanisms of nectar production and regulation thereof are proposed, and provide a starting point for reverse genetics approaches to determine molecular mechanisms underlying nectar synthesis and secretion.
Assuntos
Arabidopsis/genética , Flores/genética , Perfilação da Expressão Gênica , Arabidopsis/metabolismo , Análise por Conglomerados , Flores/metabolismo , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Análise de Sequência com Séries de Oligonucleotídeos , Regiões Promotoras Genéticas , RNA de Plantas/genéticaRESUMO
Nectaries and nectar have received much research attention for well over 200 years due to their central roles in plant-pollinator interactions. Despite this, only a few genes have demonstrated impacts on nectary development, and none have been reported to mediate de novo nectar production. This scarcity of information is largely due to the lack of a model that combines sizeable nectaries, and high levels of nectar production, along with suitable genomics resources. For example, even though Arabidopsis thaliana has been useful for developmental studies, it has been largely overlooked as a model for studying nectary function due to the small size of its flowers. However, Arabidopsis nectaries, along with those of related species, are quite operational and can be used to discern molecular mechanisms of nectary form and function. A current understanding of the machinery underlying nectary function in plants is briefly presented, with emphasis placed on the prospects of using Arabidopsis as a model for studying these processes.
Assuntos
Arabidopsis/metabolismo , Néctar de Plantas/metabolismo , Arabidopsis/química , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Flores/química , Flores/genética , Flores/crescimento & desenvolvimento , Flores/metabolismo , Modelos Biológicos , Néctar de Plantas/química , Néctar de Plantas/genéticaRESUMO
Nectar is the main reward that flowers offer to pollinators to entice repeated visitation. Cucurbita pepo (squash) is an excellent model for studying nectar biology, as it has large nectaries that produce large volumes of nectar relative to most other species. Squash is also monoecious, having both female and male flowers on the same plant, which allows comparative analyses of nectary function in one individual. Here, we report the nectary transcriptomes from both female and male nectaries at four stages of floral maturation. Analysis of these transcriptomes and subsequent confirmatory experiments revealed a metabolic progression in nectaries leading from starch synthesis to starch degradation and to sucrose biosynthesis. These results are consistent with previously published models of nectar secretion and also suggest how a sucrose-rich nectar can be synthesized and secreted in the absence of active transport across the plasma membrane. Nontargeted metabolomic analyses of nectars also confidently identified 40 metabolites in both female and male nectars, with some displaying preferential accumulation in nectar of either male or female flowers. Cumulatively, this study identified gene targets for reverse genetics approaches to study nectary function, as well as previously unreported nectar metabolites that may function in plant-biotic interactions.
RESUMO
Over 75% of crop species produce nectar and are dependent on pollinators to achieve maximum seed set, yet little is known about the mechanisms regulating nectar secretion. The phytohormone jasmonic acid (JA) is recognized to be involved in several plant processes including development and defense. JA was also recently shown to positively influence nectar secretion in both floral and extrafloral nectaries. For example, endogenous JA levels peak in flowers just prior to nectar secretion, but the details of how JA regulates nectar secretion have yet to be elucidated. We have found that the octadecanoid pathway does indeed play a role in the production and regulation of floral nectar in Arabidopsis. Null alleles for several JA biosynthesis and response genes had significantly reduced amounts of nectar, as well as altered expression of genes known to be involved in nectar production. We additionally identified crosstalk between the JA and auxin response pathways in nectaries. For example, the nectar-less JA synthesis mutant aos-2 showed no auxin response in nectaries, but both nectar production and the auxin response were restored upon exogenous JA and auxin treatment. Conversely, coi1-1, a JA-Ile-insensitive receptor mutant, displayed no auxin response in nectaries under any circumstance, even in older flowers that produced nectar. Surprisingly, opr3-1, a mutant for 12-oxophytodienoate reductase 3 [an enzyme further down the JA biosynthetic pathway that reduces 12-oxo phytodienoic acid (OPDA)], produced no nectar in newly opened flowers, but did secrete nectar in older flowers. Furthermore, a similar phenotype was observed in coi1-1. Cumulatively, these observations strongly suggest an indispensable role for an octadecanoic acid- and auxin-dependent, but JA- and COI1-dispensible, pathway in regulating nectar production in Arabidopsis.
RESUMO
Nectar is a floral reward that sustains mutualisms with pollinators, which in turn, improves fruit set. While it is known that nectar is a chemically complex solution, extensive identification and quantification of this complexity has been lacking. Cucurbita maxima cv. Big Max, like many cucurbits, is monoecious with separate male and female flowers. Attraction of bees to the flowers through the reward of nectar is essential for reproductive success in this economically valuable crop. In this study, the sex-dependent variation in composition of male and female nectar and the nectaries were defined using a combination of GC-MS based metabolomics and LC-MS/MS based proteomics. Metabolomics analysis of nectar detected 88 metabolites, of which 40 were positively identified, and includes sugars, sugar alcohols, aromatics, diols, organic acids, and amino acids. There are differences in 29 metabolites between male and female nectar. The nectar proteome consists of 45 proteins, of which 70% overlap between nectar types. Only two proteins are unique to female nectar, and 10 are specific to male nectar. The nectary proteome data, accessible at ProteomeXchange with identifier PXD009810, contained 339 identifiable proteins, 71% of which were descriptively annotatable by homology to Plantae. The abundance of 45 proteins differs significantly between male and female nectaries, as determined by iTRAQ labeling. This rich dataset significantly expands the known complexity of nectar composition, supports the hypothesis of H+-driven nectar solute export, and provides genetic and chemical targets to understand plant-pollinator interactions.
RESUMO
Floral nectar and other reward facilitate crop pollination, and in so doing, increase the amount and breadth of food available for humans. Though abundance and diversity of pollinators (particularly bees) have declined over the past several decades, a concomitant increase in reliance on pollinators presents a challenge to food production. Development of crop varieties with specific nectar or nectar-related traits to attract and retain pollinating insects is an appealing strategy to help address needs of agriculture and pollinators for several reasons. First, many crops have specific traits which have been identified to enhance crop-pollinator interactions. Also, an improved understanding of mechanisms that govern nectar-related traits suggest simplified phenotyping and breeding are possible. Finally, the use of nectar-related traits to enhance crop pollination should complement other measures promoting pollinators and will not limit options for crop production or require any changes by growers (other than planting varieties that are more attractive or rewarding to pollinators). In this article, we review the rationale for improving crop-pollinator interactions, the effects of specific plant traits on pollinator species, and use cultivated sunflowers as a case study. Recent research in sunflower has (i) associated variation in bee visitation with specific floral traits, (ii) quantified benefits of pollinators to hybrid yields, and (iii) used genetic resources in sunflower and other plants to find markers associated with key floral traits. Forthcoming work to increase pollinator rewards should enable sunflower to act as a model for using nectar-related traits to enhance crop-pollinator interactions.