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1.
Plant Cell ; 35(8): 2799-2820, 2023 08 02.
Artigo em Inglês | MEDLINE | ID: mdl-37132634

RESUMO

Actinomorphic flowers usually orient vertically (relative to the horizon) and possess symmetric nectar guides, while zygomorphic flowers often face horizontally and have asymmetric nectar guides, indicating that floral symmetry, floral orientation, and nectar guide patterning are correlated. The origin of floral zygomorphy is dependent on the dorsoventrally asymmetric expression of CYCLOIDEA (CYC)-like genes. However, how horizontal orientation and asymmetric nectar guides are achieved remains poorly understood. Here, we selected Chirita pumila (Gesneriaceae) as a model plant to explore the molecular bases for these traits. By analyzing gene expression patterns, protein-DNA and protein-protein interactions, and encoded protein functions, we identified multiple roles and functional divergence of 2 CYC-like genes, i.e. CpCYC1 and CpCYC2, in controlling floral symmetry, floral orientation, and nectar guide patterning. CpCYC1 positively regulates its own expression, whereas CpCYC2 does not regulate itself. In addition, CpCYC2 upregulates CpCYC1, while CpCYC1 downregulates CpCYC2. This asymmetric auto-regulation and cross-regulation mechanism might explain the high expression levels of only 1 of these genes. We show that CpCYC1 and CpCYC2 determine asymmetric nectar guide formation, likely by directly repressing the flavonoid synthesis-related gene CpF3'5'H. We further suggest that CYC-like genes play multiple conserved roles in Gesneriaceae. These findings shed light on the repeated origins of zygomorphic flowers in angiosperms.


Assuntos
Magnoliopsida , Néctar de Plantas , Néctar de Plantas/genética , Filogenia , Magnoliopsida/genética , Flores/genética , Genes de Plantas/genética
2.
Proc Natl Acad Sci U S A ; 117(36): 22552-22560, 2020 09 08.
Artigo em Inglês | MEDLINE | ID: mdl-32848061

RESUMO

The evolution of novel features, such as eyes or wings, that allow organisms to exploit their environment in new ways can lead to increased diversification rates. Therefore, understanding the genetic and developmental mechanisms involved in the origin of these key innovations has long been of interest to evolutionary biologists. In flowering plants, floral nectar spurs are a prime example of a key innovation, with the independent evolution of spurs associated with increased diversification rates in multiple angiosperm lineages due to their ability to promote reproductive isolation via pollinator specialization. As none of the traditional plant model taxa have nectar spurs, little is known about the genetic and developmental basis of this trait. Nectar spurs are a defining feature of the columbine genus Aquilegia (Ranunculaceae), a lineage that has experienced a relatively recent and rapid radiation. We use a combination of genetic mapping, gene expression analyses, and functional assays to identify a gene crucial for nectar spur development, POPOVICH (POP), which encodes a C2H2 zinc-finger transcription factor. POP plays a central role in regulating cell proliferation in the Aquilegia petal during the early phase (phase I) of spur development and also appears to be necessary for the subsequent development of nectaries. The identification of POP opens up numerous avenues for continued scientific exploration, including further elucidating of the genetic pathway of which it is a part, determining its role in the initial evolution of the Aquilegia nectar spur, and examining its potential role in the subsequent evolution of diverse spur morphologies across the genus.


Assuntos
Aquilegia/fisiologia , Néctar de Plantas/metabolismo , Proteínas de Plantas , Fatores de Transcrição , Dedos de Zinco/genética , Aquilegia/genética , Flores/genética , Flores/metabolismo , Néctar de Plantas/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
3.
Planta ; 250(5): 1703-1715, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31414205

RESUMO

MAIN CONCLUSION: The tobacco nectar proteome mainly consists of pathogenesis-related proteins with two glycoproteins. Expression of nectarins was non-synchronous, and not nectary specific. After secretion, tobacco nectar changed from sucrose rich to hexose rich. Floral nectar proteins (nectarins) play important roles in inhibiting microbial growth in nectar, and probably also tailoring nectar chemistry before or after secretion; however, very few plant species have had their nectar proteomes thoroughly investigated. Nectarins from Nicotiana tabacum (NT) were separated using two-dimensional gel electrophoresis and then analysed using mass spectrometry. Seven nectarins were identified: acidic endochitinase, ß-xylosidase, α-galactosidase, α-amylase, G-type lectin S-receptor-like serine/threonine-protein kinase, pathogenesis-related protein 5, and early nodulin-like protein 2. An eighth nectarin, a glycoprotein with unknown function, was identified following isolation from NT nectar using a Qproteome total glycoprotein kit, separation by SDS-PAGE, and identification by mass spectrometry. Expression of all identified nectarins, plus four invertase genes, was analysed by qRT PCR; none of these genes had nectary-specific expression, and none had synchronous expression. The total content of sucrose, hexoses, proteins, phenolics, and hydrogen peroxide were determined at different time intervals in secreted nectar, both within the nectar tube (in vivo) and following extraction from it during incubation at 30 °C for up to 40 h in plastic tubes (in vitro). After secretion, the ratio of hexose to sucrose substantially increased for in vivo nectar, but no sugar composition changes were detected in vitro. This implies that sucrose hydrolysis in vivo might be done by fixed apoplastic invertase. Both protein and hydrogen peroxide levels declined in vitro but not in vivo, implying that some factors other than nectarins act to maintain their levels in the flower, after secretion.


Assuntos
Nicotiana/enzimologia , Néctar de Plantas/metabolismo , Proteoma , Proteômica , Eletroforese em Gel Bidimensional , Flores/genética , Flores/metabolismo , Glicoproteínas/genética , Glicoproteínas/metabolismo , Espectrometria de Massas , Proteínas de Membrana/metabolismo , Néctar de Plantas/genética , Proteínas de Plantas/metabolismo , Nicotiana/genética
4.
Ann Bot ; 123(2): 247-261, 2019 01 23.
Artigo em Inglês | MEDLINE | ID: mdl-30032269

RESUMO

Background: Floral nectar is an important determinant of plant-pollinator interactions and an integral component of pollination syndromes, suggesting it is under pollinator-mediated selection. However, compared to floral display traits, we know little about the evolutionary ecology of nectar. Combining a literature review with a meta-analysis approach, we summarize the evidence for heritable variation in nectar traits and link this variation to pollinator response and plant fitness. We further review associations between nectar traits and floral signals and discuss them in the context of honest signalling and targets of selection. Scope: Although nectar is strongly influenced by environmental factors, heritable variation in nectar production rate has been documented in several populations (mean h2 = 0.31). Almost nothing is known about heritability of other nectar traits, such as sugar and amino acid concentrations. Only a handful of studies have quantified selection on nectar traits, and few find statistically significant selection. Pollinator responses to nectar traits indicate they may drive selection, but studies tying pollinator preferences to plant fitness are lacking. So far, only one study conclusively identified pollinators as selective agents on a nectar trait, and the role of microbes, herbivores, nectar robbers and abiotic factors in nectar evolution is largely hypothetical. Finally, there is a trend for positive correlations among floral cues and nectar traits, indicating honest signalling of rewards. Conclusions: Important progress can be made by studies that quantify current selection on nectar in natural populations, as well as experimental approaches that identify the target traits and selective agents involved. Signal-reward associations suggest that correlational selection may shape evolution of nectar traits, and studies exploring these more complex forms of natural selection are needed. Many questions about nectar evolution remain unanswered, making this a field ripe for future research.


Assuntos
Evolução Biológica , Néctar de Plantas/genética , Seleção Genética , Animais , Flores/fisiologia , Variação Genética , Néctar de Plantas/análise , Característica Quantitativa Herdável
5.
Mol Ecol ; 27(8): 2067-2076, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29446179

RESUMO

Both dispersal limitation and environmental sorting can affect genetic variation in populations, but their contribution remains unclear, particularly in microbes. We sought to determine the contribution of geographic distance (as a proxy for dispersal limitation) and phenotypic traits (as a proxy for environmental sorting), including morphology, metabolic ability and interspecific competitiveness, to the genotypic diversity in a nectar yeast species, Metschnikowia reukaufii. To measure genotypic diversity, we sequenced the genomes of 102 strains of M. reukaufii isolated from the floral nectar of hummingbird-pollinated shrub, Mimulus aurantiacus, along a 200-km coastline in California. Intraspecific genetic variation showed no detectable relationship with geographic distance, but could be grouped into three distinct lineages that correlated with metabolic ability and interspecific competitiveness. Despite ample evidence for strong competitive interactions within and among nectar yeasts, a full spectrum of the genotypic and phenotypic diversity observed across the 200-km coastline was represented even at a scale as small as 200 m. Further, more competitive strains were not necessarily more abundant. These results suggest that dispersal limitation and environmental sorting might not fully explain intraspecific diversity in this microbe and highlight the need to also consider other ecological factors such as trade-offs, source-sink dynamics and niche modification.


Assuntos
Flores/microbiologia , Variação Genética/genética , Metschnikowia/genética , Néctar de Plantas/genética , Animais , Aves/fisiologia , California , Flores/genética , Genômica , Genótipo , Metschnikowia/isolamento & purificação , Metschnikowia/patogenicidade , Fenótipo , Polinização/genética
6.
Ann Bot ; 121(3): 501-512, 2018 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-29300875

RESUMO

Background and Aims: Floral nectar can be variable in composition, influencing pollinator behaviour and the composition of honey derived from it. The non-peroxide antibacterial activity of manuka (Leptospermum scoparium, Myrtaceae) honey results from the chemical conversion of the triose sugar dihydroxyacetone (DHA), after DHA accumulates for an unknown reason in the nectar. This study examined variation in nectar DHA, glucose, fructose and sucrose content with floral stage of development, between manuka genotypes with differing flower morphology, and in response to water stress. Methods: Six manuka genotypes were grown without nectar-feeding insects. Stages of flower development were defined, nectar was harvested and its composition was compared between stages and genotypes, and with floral morphology. Water stress was imposed and its effect on nectar composition was examined. Key Results: Nectar was present from soon after flower opening until the end of petal abscission, with the quantity of accumulated nectar sugars rising, then stabilizing or falling, indicating nectar secretion followed by reabsorption in some genotypes. The quantity of DHA, the ratio of DHA to other nectar sugars and the fructose to glucose ratio also varied with stage of development, indicating differences in rates of production and reabsorption between nectar components. Nectar composition and yield per flower also differed between genotypes, although neither was positively related to nectary area or stomatal density. Drying soil had no effect on nectar composition or yield, but variation in nectar yield was correlated with temperature prior to nectar sampling. Conclusions: Manuka nectar yield and composition are strongly influenced by plant genotype, flower age and the environment. There were clear stoichiometric relationships between glucose, fructose and sucrose per flower, but DHA per flower was only weakly correlated with the amount of other sugars, suggesting that accumulation of the triose sugar is indirectly coupled to secretion of the larger sugars by the nectary parenchyma.


Assuntos
Flores/crescimento & desenvolvimento , Leptospermum/genética , Néctar de Plantas/genética , Desidratação , Di-Hidroxiacetona/análise , Flores/genética , Flores/ultraestrutura , Estudos de Associação Genética , Leptospermum/anatomia & histologia , Leptospermum/crescimento & desenvolvimento , Microscopia Eletrônica de Varredura , Néctar de Plantas/química
7.
BMC Plant Biol ; 17(1): 201, 2017 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-29137608

RESUMO

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/metabolismo
8.
Sci Rep ; 7: 45838, 2017 04 04.
Artigo em Inglês | MEDLINE | ID: mdl-28374829

RESUMO

Amongst gynodioecious plant breeding systems, there can exist intermediate morphs with a reduction in their male function (i.e. reduced number of functional anthers). Along with this sexual trimorphism, plants can also show floral colour polymorphism. Such intricate mixtures of phenotypes within a species may have complex effects on floral rewards. Floral rewards are known to vary between sexually dimorphic species and to a lesser extent between colour morphs. However, the interactive effect of sexual trimorphism and colour polymorphism is unexplored. We measured nectar's sugar content in the sexually trimorphic Geranium sylvaticum, a gynodioecious plant with a light/dark floral polymorphism. We found that nectar reward differed across genders and colour morphs. Results were not however consistent within the three genders; dark female and hermaphrodite flowers had higher sugar content than light morphs, whereas intermediate flowers did not. As expected, females and hermaphrodites had different nectar reward, with intermediate morphs being midway between the other genders. In intermediates, the sugar content was not related to the number of functional stamens. We show for the first time the existence of sex-specific differences between flower gender and colour morphs in nectar rewards. Our results demonstrate the importance of considering multiple and conflicting selection pressures to explain rewards.


Assuntos
Flores/genética , Geranium/genética , Melhoramento Vegetal , Polinização/genética , Feminino , Geranium/fisiologia , Humanos , Fenótipo , Pigmentação/genética , Néctar de Plantas/genética , Polinização/fisiologia , Comportamento Sexual/fisiologia
9.
Science ; 355(6320): 75-78, 2017 01 06.
Artigo em Inglês | MEDLINE | ID: mdl-28059766

RESUMO

Plants pollinated by hummingbirds or bats produce dilute nectars even though these animals prefer more concentrated sugar solutions. This mismatch is an unsolved evolutionary paradox. Here we show that lower quality, or more dilute, nectars evolve when the strength of preferring larger quantities or higher qualities of nectar diminishes as magnitudes of the physical stimuli increase. In a virtual evolution experiment conducted in the tropical rainforest, bats visited computer-automated flowers with simulated genomes that evolved relatively dilute nectars. Simulations replicated this evolution only when value functions, which relate the physical stimuli to subjective sensations, were nonlinear. Selection also depended on the supply/demand ratio; bats selected for more dilute nectar when competition for food was higher. We predict such a pattern to generally occur when decision-makers consider multiple value dimensions simultaneously, and increases of psychological value are not fully proportional to increases in physical magnitude.


Assuntos
Evolução Biológica , Quirópteros/psicologia , Cognição , Flores/química , Néctar de Plantas/química , Polinização , Alelos , Animais , Simulação por Computador , Tomada de Decisões , Flores/genética , Modelos Psicológicos , Néctar de Plantas/genética , Sacarose/análise
10.
Plant Biol (Stuttg) ; 19(2): 101-107, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27770594

RESUMO

The evolution of monomorphisms from heterostylous ancestors has been related to the presence of homostyly and the loss of self-incompatibility, allowing the occurrence of selfing, which could be advantageous under pollinator limitation. However, flowers of some monomorphic species show herkogamy, attraction and rewarding traits that presumably favour cross-pollination and/or a mixed mating system. This study evaluated the contributions of pollinators, breeding system and floral traits to the reproduction of Turnera velutina, a herkogamous monomorphic species. Floral visitors and frequency of visits were recorded, controlled hand cross-pollinations were conducted under greenhouse and natural conditions, and individual variation in floral traits was characterised to determine their contribution to seed production. Apis mellifera was the most frequent floral visitor. Flowers presented approach herkogamy, high variation in nectar features, and a positive correlation of floral length with nectar volume and sugar concentration. Seed production did not differ between manual self- and cross-pollinations, controls or open cross-pollinations, but autonomous self-pollination produced, on average, 82.74% fewer seeds than the other forms, irrespective of the level of herkogamy. Differences in seed production among autonomous self-pollination and other treatments showed that T. velutina flowers depend on insect pollination for reproduction, and that approach herkogamy drastically reduced seed production in the absence of pollen vectors. The lack of differences in seed production from manual cross- and self-pollinations suggests the possible presence of a mixed mating system in the studied population. Overall, this species was possibly derived from a distylous ancestor but appears fully capable of outcrossing despite being monomorphic.


Assuntos
Polinização , Turnera/fisiologia , Animais , Abelhas/fisiologia , Evolução Biológica , Cruzamento , Cruzamentos Genéticos , Flores/genética , Flores/fisiologia , Insetos/fisiologia , Fenótipo , Néctar de Plantas/genética , Néctar de Plantas/fisiologia , Pólen/genética , Pólen/fisiologia , Poliploidia , Reprodução , Sementes/genética , Sementes/fisiologia , Autofertilização , Autoincompatibilidade em Angiospermas , Turnera/genética
11.
Ann Bot ; 119(3): 395-408, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28025289

RESUMO

BACKGROUND AND AIMS: The detailed relationships in Geraniales in their current circumscription have only recently been clarified. The disparate floral morphologies and especially the nectaries of the corresponding group have consequently not previously been studied in a phylogenetic context. METHODS: The present study investigates floral and especially nectary morphology and structure for representatives of 12 of the 13 currently accepted genera in the five families of the Geraniales. Flowers were studied using light microscopy and scanning electron microscopy. KEY RESULTS: The data demonstrate the derivation of even the most disparate floral morphologies from a basic pentamerous and pentacyclic organization, with an obdiplostemonous androecium and receptacular nectaries associated with the antesepalous stamens. Divergent morphologies are explained by modifications of merosity (tetramerous flowers), symmetry (several transitions to zygomorphic flowers) and elaboration of the nectaries into variously shaped outgrowths and appendages, especially in Francoaceae. The divergent development of nectar glands ultimately leads to either a reduction in their number (to one in some Geraniaceae and Melianthaceae) or their total loss (some Vivianiaceae). CONCLUSIONS: Floral morphology of the Geraniales shows a high degree of similarity, despite the variation in overall floral appearance and nectary morphology. A hypothesis on the transformation of the nectaries within the Geraniales is presented.


Assuntos
Flores/anatomia & histologia , Geraniaceae/genética , Flores/ultraestrutura , Geraniaceae/anatomia & histologia , Geraniaceae/ultraestrutura , Microscopia Eletrônica de Varredura , Néctar de Plantas/genética
12.
Planta ; 244(4): 805-18, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27189006

RESUMO

MAIN CONCLUSION: Class II and III chitinases belonging to different glycoside hydrolase families were major nectarins in Rhododendron irroratum floral nectar which showed significant chitinolytic activity. Previous studies have demonstrated antimicrobial activity in plant floral nectar, but the molecular basis for the mechanism is still poorly understood. Two chitinases, class II (Rhchi2) and III (Rhchi3), were characterized from alkaline Rhododendron irroratum nectar by both SDS-PAGE and mass spectrometry. Rhchi2 (27 kDa) and Rhchi3 (29 kDa) are glycoside hydrolases (family 19 and 18) with theoretical pI of 8.19 and 7.04. The expression patterns of Rhchi2 and Rhchi3 were analyzed by semi-quantitative RT-PCR. Rhchi2 is expressed in flowers (corolla nectar pouches) and leaves while Rhchi3 is expressed in flowers. Chitinase in concentrated protein and fresh nectar samples was visualised by SDS-PAGE and chitinolytic activity in fresh nectar was determined spectrophotometrically via chitin-azure. Full length gene sequences were cloned with Tail-PCR and RACE. The amino acid sequence deduced from the coding region for these proteins showed high identity with known chitinases and predicted to be located in extracellular space. Fresh R. irroratum floral nectar showed significant chitinolytic activity. Our results demonstrate that class III chitinase (GH 18 family) also exists in floral nectar. The functional relationship between class II and III chitinases and the role of these pathogenesis-related proteins in antimicrobial activity in nectar is suggested.


Assuntos
Quitinases/genética , Néctar de Plantas/genética , Proteínas de Plantas/genética , Rhododendron/genética , Álcalis , Sequência de Aminoácidos , Quitinases/metabolismo , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Flores/enzimologia , Flores/genética , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas , Concentração de Íons de Hidrogênio , Espectrometria de Massas , Folhas de Planta/enzimologia , Folhas de Planta/genética , Néctar de Plantas/química , Néctar de Plantas/metabolismo , Proteínas de Plantas/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rhododendron/enzimologia , Homologia de Sequência de Aminoácidos
14.
J Genet ; 92(3): 445-59, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24371166

RESUMO

Gossypium tomentosum, a wild tetraploid cotton species with AD genomes, possesses genes conferring strong fibers and high heat tolerance. To effectively transfer these genes into Gossypium hirsutum, an entire microsatellite (simple sequence repeat, SSR)-based genetic map was constructed using the interspecific cross of G. hirsutum x G. tomentosum (HT). We detected 1800 loci from 1347 pairs of polymorphic primers. Of these, 1204 loci were grouped into 35 linkage groups at LOD ≥ 4. The map covers 3320.8 cM, with a mean density of 2.76 cM per locus. We detected 420 common loci (186 in the At subgenome and 234 in Dt) between the HT map and the map of TM-1 (G. hirsutum) and Hai 7124 (G. barbadense; HB map). The linkage groups were assigned chromosome numbers based on location of common loci and the HB map as reference. A comparison of common markers revealed that no significant chromosomal rearrangement exist between G. tomentosum and G. barbadense. Interestingly, however, we detected numerous (33.7%) segregation loci deviating from 3:1 ratio (P < 0.05) in HT, mostly clustering on eight chromosomes in the Dt subgenome, with some on three chromosomes in At. Two morphological traits, leaf hairiness and leaf nectarilessness were mapped on chromosomes 6 (A6) and 26 (D12), respectively. The SSR-based map constructed in this study will be useful for further genetic studies on cotton breeding, including mapping loci controlling quantitative traits associated with fiber quality, stress tolerance and developing chromosome segment specific introgression lines from G. tomentosum into G. hirsutum using marker-assisted selection.


Assuntos
Genes de Plantas , Gossypium/genética , Repetições de Microssatélites , Néctar de Plantas/genética , Mapeamento Cromossômico , Cromossomos de Plantas/genética , Estudos de Associação Genética , Ligação Genética , Marcadores Genéticos , Gossypium/anatomia & histologia , Fenótipo , Folhas de Planta/anatomia & histologia , Folhas de Planta/genética , Polimorfismo Genético
15.
BMC Plant Biol ; 13: 101, 2013 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-23848992

RESUMO

BACKGROUND: Floral nectar (FN) contains not only energy-rich compounds to attract pollinators, but also defense chemicals and several proteins. However, proteomic analysis of FN has been hampered by the lack of publically available sequence information from nectar-producing plants. Here we used next-generation sequencing and advanced proteomics to profile FN proteins in the opportunistic outcrossing wild tobacco, Nicotiana attenuata. RESULTS: We constructed a transcriptome database of N. attenuata and characterized its nectar proteome using LC-MS/MS. The FN proteins of N. attenuata included nectarins, sugar-cleaving enzymes (glucosidase, galactosidase, and xylosidase), RNases, pathogen-related proteins, and lipid transfer proteins. Natural variation in FN proteins of eleven N. attenuata accessions revealed a negative relationship between the accumulation of two abundant proteins, nectarin1b and nectarin5. In addition, microarray analysis of nectary tissues revealed that protein accumulation in FN is not simply correlated with the accumulation of transcripts encoding FN proteins and identified a group of genes that were specifically expressed in the nectary. CONCLUSIONS: Natural variation of identified FN proteins in the ecological model plant N. attenuata suggests that nectar chemistry may have a complex function in plant-pollinator-microbe interactions.


Assuntos
Flores/metabolismo , Variação Genética , Nicotiana/metabolismo , Néctar de Plantas/metabolismo , Proteínas de Plantas/metabolismo , Eletroforese em Gel Bidimensional , Flores/química , Flores/genética , Flores/crescimento & desenvolvimento , Espectrometria de Massas , Filogenia , Néctar de Plantas/química , Néctar de Plantas/genética , Proteínas de Plantas/química , Proteínas de Plantas/genética , Nicotiana/classificação , Nicotiana/genética , Nicotiana/crescimento & desenvolvimento
16.
Planta ; 238(1): 77-89, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23568404

RESUMO

Plant desiccation-related proteins (DRPs) were first identified as pcC13-62 from the resurrection plant Craterostigma plantagineum and it has been suggested they are involved in plant desiccation tolerance. We identified and characterized a plant DRP, which we called MS-desi, in the floral nectar of a subtropical bean species, Mucuna sempervirens (MS). MS-desi is a major nectar protein (nectarin) of the bean plant and expresses exclusively in the stylopodium, where the nectary is located. The full-length MS-desi gene encodes for a protein of 306 amino acids with a molecular mass of 33,248 Da, and possesses a ferritin-like domain and a signal peptide of 30 amino acids. Structural and phylogenetic analysis demonstrated MS-desi has high similarity to members of the plant DRPs, including pcC 13-62 protein. MS-desi has a similar hydropathy profile to that of pcC13-62 with a grand average of hydropathy index of 0.130 for MS-desi and 0.106 for pcC13-62 protein, which is very different from those of dehydrins and late embryogenesis abundant proteins. The protein's secondary structures, both predicted from the amino acid sequence and directly analysed by far UV circular dichroism, showed that MS-desi is mainly composed of alpha helices and is relatively temperature dependent. The structure change is reversible within a wide range of temperatures. Purified MS-desi and raw MS floral nectar showed dose-dependent citrate synthase inhibition activity, but insensitivity to lactate dehydrogenase, suggesting that, unlike dehydrins, it does not act as a chaperone. The overall results constitute, to our knowledge, the first study on a desiccation-related protein in plant floral nectar.


Assuntos
Mucuna/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Dicroísmo Circular , Citrato (si)-Sintase/metabolismo , Clonagem Molecular , Regulação da Expressão Gênica de Plantas , Dados de Sequência Molecular , Mucuna/genética , Filogenia , Néctar de Plantas/genética , Estabilidade Proteica , Estrutura Secundária de Proteína , Estrutura Terciária de Proteína , Homologia de Sequência de Aminoácidos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
17.
PLoS Genet ; 8(2): e1002506, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22346763

RESUMO

For self-pollinating plants to reproduce, male and female organ development must be coordinated as flowers mature. The Arabidopsis transcription factors AUXIN RESPONSE FACTOR 6 (ARF6) and ARF8 regulate this complex process by promoting petal expansion, stamen filament elongation, anther dehiscence, and gynoecium maturation, thereby ensuring that pollen released from the anthers is deposited on the stigma of a receptive gynoecium. ARF6 and ARF8 induce jasmonate production, which in turn triggers expression of MYB21 and MYB24, encoding R2R3 MYB transcription factors that promote petal and stamen growth. To understand the dynamics of this flower maturation regulatory network, we have characterized morphological, chemical, and global gene expression phenotypes of arf, myb, and jasmonate pathway mutant flowers. We found that MYB21 and MYB24 promoted not only petal and stamen development but also gynoecium growth. As well as regulating reproductive competence, both the ARF and MYB factors promoted nectary development or function and volatile sesquiterpene production, which may attract insect pollinators and/or repel pathogens. Mutants lacking jasmonate synthesis or response had decreased MYB21 expression and stamen and petal growth at the stage when flowers normally open, but had increased MYB21 expression in petals of older flowers, resulting in renewed and persistent petal expansion at later stages. Both auxin response and jasmonate synthesis promoted positive feedbacks that may ensure rapid petal and stamen growth as flowers open. MYB21 also fed back negatively on expression of jasmonate biosynthesis pathway genes to decrease flower jasmonate level, which correlated with termination of growth after flowers have opened. These dynamic feedbacks may promote timely, coordinated, and transient growth of flower organs.


Assuntos
Arabidopsis/crescimento & desenvolvimento , Arabidopsis/genética , Flores/crescimento & desenvolvimento , Flores/genética , Redes Reguladoras de Genes/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Ciclopentanos/metabolismo , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Regulação da Expressão Gênica de Plantas , Mutação , Oxilipinas/metabolismo , Fenótipo , Néctar de Plantas/genética , Plantas Geneticamente Modificadas/genética , Pólen/genética , Pólen/crescimento & desenvolvimento , Sesquiterpenos/metabolismo , Transdução de Sinais , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
18.
J Exp Bot ; 61(11): 2951-65, 2010 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20460362

RESUMO

Plants requiring an insect pollinator often produce nectar as a reward for the pollinator's visitations. This rich secretion needs mechanisms to inhibit microbial growth. In Nicotiana spp. nectar, anti-microbial activity is due to the production of hydrogen peroxide. In a close relative, Petunia hybrida, limited production of hydrogen peroxide was found; yet petunia nectar still has anti-bacterial properties, suggesting that a different mechanism may exist for this inhibition. The nectar proteins of petunia plants were compared with those of ornamental tobacco and significant differences were found in protein profiles and function between these two closely related species. Among those proteins, RNase activities unique to petunia nectar were identified. The genes corresponding to four RNase T2 proteins from Petunia hybrida that show unique expression patterns in different plant tissues were cloned. Two of these enzymes, RNase Phy3 and RNase Phy4 are unique among the T2 family and contain characteristics similar to both S- and S-like RNases. Analysis of amino acid patterns suggest that these proteins are an intermediate between S- and S-like RNases, and support the hypothesis that S-RNases evolved from defence RNases expressed in floral parts. This is the first report of RNase activities in nectar.


Assuntos
Petunia/enzimologia , Néctar de Plantas/metabolismo , Proteínas de Plantas/metabolismo , Ribonucleases/metabolismo , Sequência de Aminoácidos , Dados de Sequência Molecular , Petunia/química , Petunia/classificação , Petunia/genética , Filogenia , Néctar de Plantas/química , Néctar de Plantas/genética , Proteínas de Plantas/química , Proteínas de Plantas/genética , Ribonucleases/química , Ribonucleases/genética , Alinhamento de Sequência
19.
Sex Plant Reprod ; 22(4): 235-46, 2009 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-20033445

RESUMO

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/genética
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