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1.
Phytochemistry ; 185: 112668, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33743499

RESUMO

We review glucosinolate (GSL) diversity and analyze phylogeny in the crucifer tribe Cardamineae as well as selected species from Brassicaceae (tribe Brassiceae) and Resedaceae. Some GSLs occur widely, while there is a scattered distribution of many less common GSLs, tentatively sorted into three classes: ancient, intermediate and more recently evolved. The number of conclusively identified GSLs in the tribe (53 GSLs) constitute 60% of all GSLs known with certainty from any plant (89 GSLs) and apparently unique GSLs in the tribe constitute 10 of those GSLs conclusively identified (19%). Intraspecific, qualitative GSL polymorphism is known from at least four species in the tribe. The most ancient GSL biosynthesis in Brassicales probably involved biosynthesis from Phe, Val, Leu, Ile and possibly Trp, and hydroxylation at the ß-position. From a broad comparison of families in Brassicales and tribes in Brassicaceae, we estimate that a common ancestor of the tribe Cardamineae and the family Brassicaceae exhibited GSL biosynthesis from Phe, Val, Ile, Leu, possibly Tyr, Trp and homoPhe (ancient GSLs), as well as homologs of Met and possibly homoIle (intermediate age GSLs). From the comparison of phylogeny and GSL diversity, we also suggest that hydroxylation and subsequent methylation of indole GSLs and usual modifications of Met-derived GSLs (formation of sulfinyls, sulfonyls and alkenyls) occur due to conserved biochemical mechanisms and was present in a common ancestor of the family. Apparent loss of homologs of Met as biosynthetic precursors was deduced in the entire genus Barbarea and was frequent in Cardamine (e.g. C. pratensis, C. diphylla, C. concatenata, possibly C. amara). The loss was often associated with appearance of significant levels of unique or rare GSLs as well as recapitulation of ancient types of GSLs. Biosynthetic traits interpreted as de novo evolution included hydroxylation at rare positions, acylation at the thioglucose and use of dihomoIle and possibly homoIle as biosynthetic precursors. Biochemical aspects of the deduced evolution are discussed and testable hypotheses proposed. Biosyntheses from Val, Leu, Ile, Phe, Trp, homoPhe and homologs of Met are increasingly well understood, while GSL biosynthesis from mono- and dihomoIle is poorly understood. Overall, interpretation of known diversity suggests that evolution of GSL biosynthesis often seems to recapitulate ancient biosynthesis. In contrast, unprecedented GSL biosynthetic innovation seems to be rare.


Assuntos
Barbarea , Brassicaceae , Acilação , Brassicaceae/genética , Glucosinolatos , Filogenia
2.
Phytochemistry ; 185: 112658, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33744557

RESUMO

A library of ion trap MS2 spectra and HPLC retention times reported here allowed distinction in plants of at least 70 known glucosinolates (GSLs) and some additional proposed GSLs. We determined GSL profiles of selected members of the tribe Cardamineae (Brassicaceae) as well as Reseda (Resedaceae) used as outgroup in evolutionary studies. We included several accessions of each species and a range of organs, and paid attention to minor peaks and GSLs not detected. In this way, we obtained GSL profiles of Barbarea australis, Barbarea grayi, Planodes virginica selected for its apparent intermediacy between Barbarea and the remaining tribe and family, and Rorippa sylvestris and Nasturtium officinale, for which the presence of acyl derivatives of GSLs was previously untested. We also screened Armoracia rusticana, with a remarkably diverse GSL profile, the emerging model species Cardamine hirsuta, for which we discovered a GSL polymorphism, and Reseda luteola and Reseda odorata. The potential for aliphatic GSL biosynthesis in Barbarea vulgaris was of interest, and we subjected P-type and G-type B. vulgaris to several induction regimes in an attempt to induce aliphatic GSL. However, aliphatic GSLs were not detected in any of the B. vulgaris types. We characterized the investigated chemotypes phylogenetically, based on nuclear rDNA internal transcribed spacer (ITS) sequences, in order to understand their relation to the species B. vulgaris in general, and found them to be representative of the species as it occurs in Europe, as far as documented in available ITS-sequence repositories. In short, we provide GSL profiles of a wide variety of tribe Cardamineae plants and conclude aliphatic GSLs to be absent or below our limit of detection in two major evolutionary lines of B. vulgaris. Concerning analytical chemistry, we conclude that availability of authentic reference compounds or reference materials is critical for reliable GSL analysis and characterize two publicly available reference materials: seeds of P. virginica and N. officinale.


Assuntos
Barbarea , Brassicaceae , Resedaceae , Barbarea/genética , Brassicaceae/genética , Cromatografia Líquida de Alta Pressão , Europa (Continente) , Glucosinolatos , Filogenia , Espectrometria de Massas em Tandem
3.
Nutrients ; 12(10)2020 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-33036498

RESUMO

Lactic acid bacteria (LAB) "fermentates" confer a beneficial effect on intestinal function. However, the ability of new fermentations to improve LAB broth activity in preventing pathogen-induced intestinal inflammation and barrier dysfunction has not yet been studied. The objective of this study was to determine if broths of LAB fermented with Eruca sativa or Barbarea verna seed extracts prevent gut barrier dysfunction and interleukin-8 (CXCL8) release in vitro in human intestinal Caco-2 cells infected with enterohemorrhagic Escherichia coli (EHEC) O157:H7. LAB broths were assayed for their effects on EHEC growth and on Caco-2 viability; thereafter, their biological properties were analysed in a co-culture system consisting of EHEC and Caco-2 cells. Caco-2 cells infected with EHEC significantly increased CXCL8 release, and decreased Trans-Epithelial Electrical Resistance (TEER), a barrier-integrity marker. Notably, when Caco-2 cells were treated with LAB broth enriched with E. sativa seed extract and thereafter infected, both CXCL8 expression and epithelial dysfunction reduced compared to in untreated cells. These results underline the beneficial effect of broths from LAB fermented with E. sativa seed extracts in gut barrier and inflammation after EHEC infection and reveal that these LAB broths can be used as functional bioactive compounds to regulate intestinal function.


Assuntos
Brassicaceae/química , Escherichia coli O157/efeitos dos fármacos , Escherichia coli O157/crescimento & desenvolvimento , Fermentação , Gastroenterite/prevenção & controle , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiologia , Lactobacillus acidophilus , Extratos Vegetais/farmacologia , Probióticos/farmacologia , Sementes/química , Antibacterianos , Barbarea/química , Células CACO-2 , Sobrevivência Celular/efeitos dos fármacos , Técnicas de Cocultura , Farmacorresistência Bacteriana , Impedância Elétrica , Infecções por Escherichia coli , Escherichia coli O157/patogenicidade , Gastroenterite/microbiologia , Humanos , Interleucina-8/metabolismo , Mucosa Intestinal/fisiologia , Fitoterapia , Extratos Vegetais/isolamento & purificação
4.
Nutrients ; 12(4)2020 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-32290311

RESUMO

Iron deficiency is a global epidemic affecting a third of the world's population. Current efforts are focused on investigating sustainable ways to improve the bioavailability of iron in plant-based diets. Incorporating microgreens into the diet of at-risk groups in populations could be a useful tool in the management and prevention of iron deficiency. This study analysed and compared the mineral content and bioavailability of iron from microgreen and mature vegetables. The mineral content of rocket, broccoli and fenugreek microgreens and their mature counterparts was determined using microwave digestion and ICP-OES. Iron solubility and bioavailability from the vegetables were determined by a simulated gastrointestinal in vitro digestion and subsequent measurement of ferritin in Caco-2 cells as a surrogate marker of iron uptake. Iron contents of mature fenugreek and rocket were significantly higher than those of the microgreens. Mature fenugreek and broccoli showed significantly (p < 0.001) higher bioaccessibility and low-molecular-weight iron than found in the microgreens. Moreover, iron uptake by Caco-2 cells was significantly higher only from fenugreek microgreens than the mature vegetable. While all vegetables except broccoli enhanced FeSO4 uptake, the response to ferric ammonium citrate (FAC) was inhibitory apart from the mature rocket. Ascorbic acid significantly enhanced iron uptake from mature fenugreek and rocket. Microgreen fenugreek may be bred for a higher content of enhancers of iron availability as a strategy to improve iron nutrition in the populace.


Assuntos
Barbarea/química , Disponibilidade Biológica , Brassica/química , Dieta , Análise de Alimentos , Trato Gastrointestinal/metabolismo , Ferro/análise , Ferro/metabolismo , Fenômenos Fisiológicos da Nutrição/imunologia , Trigonella/química , Células CACO-2 , Humanos , Técnicas In Vitro , Absorção Intestinal , Solubilidade
5.
BMC Genomics ; 20(1): 371, 2019 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-31088355

RESUMO

BACKGROUND: Barbarea vulgaris is a wild cruciferous plant and include two distinct types: the G- and P-types named after their glabrous and pubescent leaves, respectively. The types differ significantly in resistance to a range of insects and diseases as well as glucosinolates and other chemical defenses. A high-density linkage map was needed for further progress to be made in the molecular research of this plant. RESULTS: We performed restriction site-associated DNA sequencing (RAD-Seq) on an F2 population generated from G- and P-type B. vulgaris. A total of 1545 SNP markers were mapped and ordered in eight linkage groups, which represents the highest density linkage map to date for the crucifer tribe Cardamineae. A total of 722 previously published genome contigs (50.2 Mb, 30% of the total length) can be anchored to this high density genetic map, an improvement compared to a previously published map (431 anchored contigs, 38.7 Mb, 23% of the assembly genome). Most of these (572 contigs, 31.2 Mb) were newly anchored to the map, representing a significant improvement. On the basis of the present high-density genetic map, 37 QTL were detected for eleven traits, each QTL explaining 2.9-71.3% of the phenotype variation. QTL of glucosinolates, leaf size and color traits were in most cases overlapping, possibly implying a functional connection. CONCLUSIONS: This high-density linkage map and the QTL obtained in this study will be useful for further understanding of the genetic of the B. vulgaris and molecular basis of these traits, many of which are shared in the related crop watercress.


Assuntos
Barbarea/genética , Mapeamento Cromossômico/métodos , Locos de Características Quantitativas , Análise de Sequência de DNA/métodos , Barbarea/fisiologia , DNA de Plantas/genética , Ligação Genética , Fenótipo , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Polimorfismo de Nucleotídeo Único
6.
New Phytol ; 222(3): 1599-1609, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30661245

RESUMO

Plants continuously evolve new defense compounds. One class of such compounds is triterpenoid saponins. A few species in the Barbarea genus produce saponins as the only ones in the large crucifer family. However, the molecular mechanism behind saponin biosynthesis and their role in plant defense remains unclear. We used pathway reconstitution in planta, enzymatic production of saponins in vitro, insect feeding assays, and bioinformatics to identify a missing gene involved in saponin biosynthesis and saponin-based herbivore defense. A tandem repeat of eight CYP72A cytochromes P450 colocalise with a quantitative trait locus (QTL) for saponin accumulation and flea beetle resistance in Barbarea vulgaris. We found that CYP72A552 oxidises oleanolic acid at position C-23 to hederagenin. In vitro-produced hederagenin monoglucosides reduced larval feeding by up to 90% and caused 75% larval mortality of the major crucifer pest diamondback moth and the tobacco hornworm. Sequence analysis indicated that CYP72A552 evolved through gene duplication and has been under strong selection pressure. In conclusion, CYP72A552 has evolved to catalyse the formation of hederagenin-based saponins that mediate plant defense against herbivores. Our study highlights the evolution of chemical novelties by gene duplication and selection for enzyme innovations, and the importance of chemical modification in plant defense evolution.


Assuntos
Barbarea/imunologia , Barbarea/parasitologia , Sistema Enzimático do Citocromo P-450/metabolismo , Herbivoria/fisiologia , Ácido Oleanólico/análogos & derivados , Saponinas/biossíntese , Animais , Barbarea/enzimologia , Barbarea/genética , Sistema Enzimático do Citocromo P-450/genética , Duplicação Gênica , Genoma de Planta , Herbivoria/efeitos dos fármacos , Insetos/fisiologia , Mariposas/fisiologia , Ácido Oleanólico/biossíntese , Ácido Oleanólico/química , Ácido Oleanólico/farmacologia , Oxirredução , Filogenia , Locos de Características Quantitativas/genética , Saponinas/química , Saponinas/farmacologia
7.
J Chem Ecol ; 44(12): 1190-1205, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30218254

RESUMO

We investigated the influences of two structurally similar glucosinolates, phenethylglucosinolate (gluconasturtiin, NAS) and its (S)-2-hydroxyl derivative glucobarbarin (BAR), as well as their hydrolysis products on larvae of the generalist Mamestra brassicae (Lepidoptera: Noctuidae). Previous results suggested a higher defensive activity of BAR than NAS based on resistance toward M. brassicae larvae of natural plant genotypes of Barbarea vulgaris R. Br. (Brassicaceae) dominated by BAR. In the present study, the hypothesis of a higher defensive activity of BAR than NAS was tested by comparing two Barbarea species similarly dominated either by BAR or by NAS and by testing effects of isolated BAR and NAS on larval survival and feeding preferences. Larvae reared on leaf disks of B. verna (Mill.) Asch. had a lower survival than those reared on B. vulgaris P- and G-chemotypes. Leaves of B. verna were dominated by NAS, whereas B. vulgaris chemotypes were dominated by BAR or its epimer. In addition, B. verna leaves showed a threefold higher activity of the glucosinolate-activating myrosinase enzymes. The main product of NAS from breakdown by endogenous enzymes including myrosinases ("autolysis") in B. verna leaves was phenethyl isothiocyanate, while the main products of BAR in autolyzed B. vulgaris leaves were a cyclized isothiocyanate product, namely an oxazolidine-2-thione, and a downstream metabolite, an oxazolidin-2-one. The glucosinolates BAR and NAS were isolated and offered to larvae on disks of cabbage. Both glucosinolates exerted similar negative effects on larval survival but effects of NAS tended to be more detrimental. Low concentrations of BAR, but not of NAS, stimulated larval feeding, whereas high BAR concentrations acted deterrent. NAS only tended to be deterrent at the highest concentration, but the difference was not significant. Recoveries of NAS and BAR on cabbage leaf disks were similar, and when hydrolyzed by mechanical leaf damage, the same isothiocyanate-type products as in Barbarea plants were formed with further conversion of BAR to cyclic products, (R)-5-phenyloxazolidine-2-thione [(R)-barbarin] and (R)-5-phenyloxazolidin-2-one [(R)-resedine]. We conclude that a previously proposed generally higher defensive activity of BAR than NAS to M. brassicae larvae could not be confirmed. Indeed, the higher resistance of NAS-containing B. verna plants may be due to a combined effect of rather high concentrations of NAS and a relatively high myrosinase activity or other plant traits not investigated yet.


Assuntos
Antibiose , Barbarea/química , Glucosinolatos/metabolismo , Glicosídeo Hidrolases/metabolismo , Herbivoria , Mariposas/fisiologia , Animais , Glucosinolatos/análise , Glicosídeo Hidrolases/análise , Larva/crescimento & desenvolvimento , Larva/fisiologia , Mariposas/crescimento & desenvolvimento , Especificidade da Espécie
8.
Plant Mol Biol ; 97(1-2): 37-55, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29603041

RESUMO

KEY MESSAGE: This study identifies six UGT73Cs all able to glucosylate sapogenins at positions 3 and/or 28 which demonstrates that B. vulgaris has a much richer arsenal of UGTs involved in saponin biosynthesis than initially anticipated. The wild cruciferous plant Barbarea vulgaris is resistant to some insects due to accumulation of two monodesmosidic triterpenoid saponins, oleanolic acid 3-O-ß-cellobioside and hederagenin 3-O-ß-cellobioside. Insect resistance depends on the structure of the sapogenin aglycone and the glycosylation pattern. The B. vulgaris saponin profile is complex with at least 49 saponin-like metabolites, derived from eight sapogenins and including up to five monosaccharide units. Two B. vulgaris UDP-glycosyltransferases, UGT73C11 and UGT73C13, O-glucosylate sapogenins at positions 3 and 28, forming mainly 3-O-ß-D-glucosides. The aim of this study was to identify UGTs responsible for the diverse saponin oligoglycoside moieties observed in B. vulgaris. Twenty UGT genes from the insect resistant genotype were selected and heterologously expressed in Nicotiana benthamiana and/or Escherichia coli. The extracts were screened for their ability to glycosylate sapogenins (oleanolic acid, hederagenin), the hormone 24-epibrassinolide and sapogenin monoglucosides (hederagenin and oleanolic acid 3-O-ß-D-glucosides). Six UGTs from the UGT73C subfamily were able to glucosylate both sapogenins and both monoglucosides at positions 3 and/or 28. Some UGTs formed bisdesmosidic saponins efficiently. At least four UGT73C genes were localized in a tandem array with UGT73C11 and possibly UGT73C13. This organization most likely reflects duplication events followed by sub- and neofunctionalization. Indeed, signs of positive selection on several amino acid sites were identified and modelled to be localized on the UGT protein surface. This tandem array is proposed to initiate higher order bisdesmosidic glycosylation of B. vulgaris saponins, leading to the recently discovered saponin structural diversity, however, not directly to known cellobiosidic saponins.


Assuntos
Barbarea/enzimologia , Glicosiltransferases/isolamento & purificação , Sapogeninas/metabolismo , Saponinas/biossíntese , Barbarea/genética , Barbarea/metabolismo , Brassinosteroides/metabolismo , Escherichia coli/genética , Genes de Plantas , Glicosídeos/metabolismo , Glicosilação , Glicosiltransferases/química , Glicosiltransferases/genética , Glicosiltransferases/metabolismo , Modelos Moleculares , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/metabolismo , Saponinas/química , Saponinas/isolamento & purificação , Esteroides Heterocíclicos/metabolismo , Sequências de Repetição em Tandem , Tabaco/genética , Transcriptoma
9.
Sci Rep ; 7: 40728, 2017 01 17.
Artigo em Inglês | MEDLINE | ID: mdl-28094805

RESUMO

The genus Barbarea has emerged as a model for evolution and ecology of plant defense compounds, due to its unusual glucosinolate profile and production of saponins, unique to the Brassicaceae. One species, B. vulgaris, includes two 'types', G-type and P-type that differ in trichome density, and their glucosinolate and saponin profiles. A key difference is the stereochemistry of hydroxylation of their common phenethylglucosinolate backbone, leading to epimeric glucobarbarins. Here we report a draft genome sequence of the G-type, and re-sequencing of the P-type for comparison. This enables us to identify candidate genes underlying glucosinolate diversity, trichome density, and study the genetics of biochemical variation for glucosinolate and saponins. B. vulgaris is resistant to the diamondback moth, and may be exploited for "dead-end" trap cropping where glucosinolates stimulate oviposition and saponins deter larvae to the extent that they die. The B. vulgaris genome will promote the study of mechanisms in ecological biochemistry to benefit crop resistance breeding.


Assuntos
Barbarea/genética , Genoma de Planta , Genômica , Barbarea/química , Barbarea/classificação , Barbarea/metabolismo , Biologia Computacional/métodos , Resistência à Doença/genética , Variação Genética , Genômica/métodos , Glucosinolatos/metabolismo , Metaboloma , Metabolômica/métodos , Anotação de Sequência Molecular , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Locos de Características Quantitativas , Característica Quantitativa Herdável , Sequenciamento Completo do Genoma
10.
Molecules ; 21(12)2016 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-27886152

RESUMO

Recently the number of studies investigating triterpenoid saponins has drastically increased due to their diverse and potentially attractive biological activities. Currently the literature contains chemical structures of few hundreds of triterpenoid saponins of plant and animal origin. Triterpenoid saponins consist of a triterpene aglycone with one or more sugar moieties attached to it. However, due to similar physico-chemical properties, isolation and identification of a large diversity of triterpenoid saponins remain challenging. This study demonstrates a methodology to screen saponins using hyphenated analytical platforms, GC-MS, LC-MS/MS, and LC-SPE-NMR/MS, in the example of two different phenotypes of the model plant Barbarea vulgaris (winter cress), glabrous (G) and pubescent (P) type that are known to differ by their insect resistance. The proposed methodology allows for detailed comparison of saponin profiles from intact plant extracts as well as saponin aglycone profiles from hydrolysed samples. Continuously measured 1D proton NMR data during LC separation along with mass spectrometry data revealed significant differences, including contents of saponins, types of aglycones and numbers of sugar moieties attached to the aglycone. A total of 49 peaks were tentatively identified as saponins from both plants; they are derived from eight types of aglycones and with 2-5 sugar moieties. Identification of two previously known insect-deterrent saponins, hederagenin cellobioside and oleanolic acid cellobioside, demonstrated the applicability of the methodology for relatively rapid screening of bioactive compounds.


Assuntos
Barbarea/química , Cromatografia Líquida/métodos , Cromatografia Gasosa-Espectrometria de Massas/métodos , Ressonância Magnética Nuclear Biomolecular/métodos , Saponinas/química , Triterpenos/química , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/química , Folhas de Planta/química
11.
Phytochemistry ; 132: 33-56, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27743600

RESUMO

As a basis for future investigations of evolutionary trajectories and biosynthetic mechanisms underlying variations in glucosinolate structures, we screened members of the crucifer tribe Cardamineae by HPLC-MS/MS, isolated and identified glucosinolates by NMR, searched the literature for previous data for the tribe, and collected HPLC-MS/MS data for nearly all glucosinolates known from the tribe as well as some related structures (70 in total). This is a considerable proportion of the approximately 142 currently documented natural glucosinolates. Calibration with authentic references allowed distinction (or elucidation) of isomers in many cases, such as distinction of ß-hydroxyls, methylthios, methylsulfinyls and methylsulfonyls. A mechanism for fragmentation of secondary ß-hydroxyls in MS was elucidated, and two novel glucosinolates were discovered: 2-hydroxy-3-methylpentylglucosinolate in roots of Cardamine pratensis and 2-hydroxy-8-(methylsulfinyl)octylglucosinolate in seeds of Rorippa amphibia. A large number of glucosinolates (ca. 54 with high structural certainty and a further 28 or more suggested from tandem MS), representing a wide structural variation, is documented from the tribe. This included glucosinolates apparently derived from Met, Phe, Trp, Val/Leu, Ile and higher homologues. Normal side chain elongation and side chain decoration by oxidation or methylation was observed, as well as rare abnormal side chain decoration (hydroxylation of aliphatics at the δ rather than ß-position). Some species had diverse profiles, e.g. R. amphibia and C. pratensis (19 and 16 individual glucosinolates, respectively), comparable to total diversity in literature reports of Armoracia rusticana (17?), Barbarea vulgaris (20-24), and Rorippa indica (>20?). The ancestor or the tribe would appear to have used Trp, Met, and homoPhe as glucosinolate precursor amino acids, and to exhibit oxidation of thio to sulfinyl, formation of alkenyls, ß-hydroxylation of aliphatic chains and hydroxylation and methylation of indole glucosinolates. Two hotspots of apparent biochemical innovation and loss were identified: C. pratensis and the genus Barbarea. Diversity in other species mainly included structures also known from other crucifers. In addition to a role of gene duplication, two contrasting genetic/biochemical mechanisms for evolution of such combined diversity and redundancy are discussed: (i) involvement of widespread genes with expression varying during evolution, and (ii) mutational changes in substrate specificities of CYP79F and GS-OH enzymes.


Assuntos
Brassicaceae/química , Glucosinolatos/análise , Filogenia , Barbarea/química , Cromatografia Líquida de Alta Pressão , Duplicação Gênica , Glucosinolatos/química , Humanos , Estrutura Molecular , Sementes/química
12.
Ann Bot ; 117(4): 667-79, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26975314

RESUMO

BACKGROUND AND AIMS: Senescence is the process of losing fitness when growing old, and is shaped by the trade-off between maintenance and reproduction that makes reproduction more unsure and maintenance more costly with age. In repeatedly reproducing plants, reductions in growth and fertility are signs of senescence. Disturbance, however, provides an opportunity to reset the ageing clock and consequently potentially ameliorate senescence. METHODS: To test the effects of disturbance on traits closely related to fitness and thus to senescence, a long-term garden experiment was established with two short-lived perennial congeners,Barbarea vulgaris and Barbarea stricta, that differ in their ability to resprout after injury. In the experiment, five damage treatments were applied to plants in four different phenophases. KEY RESULTS: It was found that damage to the plant body significantly prolonged life span in B. vulgaris but decreased whole-life seed production in both species. High concentration of seed production in one growing season characterized short life spans. Both more severe damage and a more advanced phenological phase at the time of damage caused reproduction to be spread over more than one growing season and equalized per-season seed production. In terms of seed quality, average weight of a single seed decreased and seed germination rate increased with age regardless of damage. CONCLUSIONS: Although disturbance is able to reset the ageing clock of plants, it is so harmful to plant fitness that resprouting serves, at best, only to alleviate slightly the signs of senescence. Thus, in terms of whole-life seed production, injured plants were not more successful than uninjured ones in the two studied species. Indeed, in these species, injury only slightly postponed or decelerated senescence and did not cause effective rejuvenation.


Assuntos
Barbarea/crescimento & desenvolvimento , Germinação , Raízes de Plantas/crescimento & desenvolvimento , Tamanho Corporal , Modelos Biológicos , Modelos Estatísticos , Reprodução , Sementes/crescimento & desenvolvimento , Especificidade da Espécie , Fatores de Tempo
13.
J Therm Biol ; 56: 109-12, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26857984

RESUMO

Endotherms foraging at temperatures outside of their thermal neutral zone (TNZ) pay an increased energetic cost. We asked if thermally-induced changes in foraging costs influence quitting harvest rate (QHR) of mice. We predicted that mice foraging during the winter would have a higher QHR in more costly colder conditions. We conducted our study with wild caught Peromyscus leucopus in an enclosure located in West Terre Haute, Indiana. We assayed changes in QHR using the forager's giving up density (GUD), which is the amount of uneaten seeds reaming in a tray after foraging activity. Each night from January 12th to March 13th, we assigned 4 trays as "cold trays" (at ambient temperature), and 4 trays as "hot trays" (trays with a ceramic heat element that increased the temperatures of feeding trays ca. 10-15°C). GUDs (and therfore QHRs) increased as a function of decreasing ambient temperature. Furthermore there was an interaction between tray temperature and ambient temperature; namely, on cool nights mice had lower GUDs in the "hot trays", but on warm nights mice had lower GUDs in the "cold trays". The TNZ for P. leucopus actively foraging during winter may be closer to the environmental average temperature than typically measured in the laboratory. Overall, these results support the idea that QHR is related to an animal's foraging in thermally challenged conditions. We present a unique way of measuring an animal's TNZ in the field using behavioral indicators.


Assuntos
Comportamento Alimentar/fisiologia , Peromyscus/fisiologia , Estações do Ano , Temperatura , Animais , Barbarea , Ambiente Controlado , Camundongos
14.
Food Microbiol ; 55: 7-15, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26742611

RESUMO

The aim of the present study was to assess the expression of key virulence genes, during growth of a Listeria monocytogenes isolate in liquid medium, on melon and rocket at different temperatures and time. For that purpose, BHI broth, rocket and melon were inoculated at 7.0-7.5 log CFU mL(-1) or g(-1)and stored at 4, 10 and 30 °C. Sampling took place upon inoculation and after 0.5, 6 and 24 h of incubation. The RNA was stabilized and the expression of hly, plcA, plcB, sigB, inlA, inlB, inlC, inlJ, lmo2672 and lmo2470 was assessed by RT-qPCR. The results obtained were summarized into two observations; the first one referring to the interactive effect of incubation temperature and type of substrate and the second one to the effect of time on gene expression. Regarding the latter, nearly all genes were regulated upon inoculation and exhibited differential expression in the subsequent sampling times indicating the existence of additional regulatory mechanisms yet to be explored.


Assuntos
Proteínas de Bactérias/genética , Barbarea/microbiologia , Cucurbitaceae/microbiologia , Meios de Cultura/análise , Frutas/microbiologia , Regulação Bacteriana da Expressão Gênica , Listeria monocytogenes/genética , Fatores de Virulência/genética , Proteínas de Bactérias/metabolismo , Listeria monocytogenes/crescimento & desenvolvimento , Listeria monocytogenes/metabolismo , Temperatura , Fatores de Virulência/metabolismo
15.
Plant J ; 84(3): 478-90, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26333142

RESUMO

The ability to evolve novel metabolites has been instrumental for the defence of plants against antagonists. A few species in the Barbarea genus are the only crucifers known to produce saponins, some of which make plants resistant to specialist herbivores, like Plutella xylostella, the diamondback moth. Genetic mapping in Barbarea vulgaris revealed that genes for saponin biosynthesis are not clustered but are located in different linkage groups. Using co-location with quantitative trait loci (QTLs) for resistance, transcriptome and genome sequences, we identified two 2,3-oxidosqualene cyclases that form the major triterpenoid backbones. LUP2 mainly produces lupeol, and is preferentially expressed in insect-susceptible B. vulgaris plants, whereas LUP5 produces ß-amyrin and α-amyrin, and is preferentially expressed in resistant plants; ß-amyrin is the backbone for the resistance-conferring saponins in Barbarea. Two loci for cytochromes P450, predicted to add functional groups to the saponin backbone, were identified: CYP72As co-localized with insect resistance, whereas CYP716As did not. When B. vulgaris sapogenin biosynthesis genes were transiently expressed by CPMV-HT technology in Nicotiana benthamiana, high levels of hydroxylated and carboxylated triterpenoid structures accumulated, including oleanolic acid, which is a precursor of the major resistance-conferring saponins. When the B. vulgaris gene for sapogenin 3-O-glucosylation was co-expressed, the insect deterrent 3-O-oleanolic acid monoglucoside accumulated, as well as triterpene structures with up to six hexoses, demonstrating that N. benthamiana further decorates the monoglucosides. We argue that saponin biosynthesis in the Barbarea genus evolved by a neofunctionalized glucosyl transferase, whereas the difference between resistant and susceptible B. vulgaris chemotypes evolved by different expression of oxidosqualene cyclases (OSCs).


Assuntos
Barbarea/genética , Barbarea/metabolismo , Saponinas/metabolismo , Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Regulação da Expressão Gênica de Plantas , Genoma de Planta , Herbivoria , Transferases Intramoleculares/genética , Transferases Intramoleculares/metabolismo , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/metabolismo , Triterpenos Pentacíclicos/metabolismo , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Locos de Características Quantitativas , Sapogeninas/metabolismo , Saponinas/genética , Tabaco/genética , Triterpenos/metabolismo
16.
Phytochemistry ; 118: 131-8, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26318326

RESUMO

The metabolites produced in leaves of the crucifers winter cress (Barbarea vulgaris) and upland cress (Barbarea verna) abiotically elicited were investigated and their chemical structures were elucidated by analyses of spectroscopic data and confirmed by syntheses. Nasturlexins C and D and their sulfoxides are cruciferous phytoalexins displaying antifungal activity against the crucifer pathogens Alternaria brassicicola, Leptosphaeria maculans and Sclerotinia sclerotiorum. The biosynthesis of these metabolites is proposed based on pathways of cruciferous indolyl phytoalexins. This work indicates that B. vulgaris and B. verna have great potential as sources of defense pathways transferable to agriculturally important crops within the Brassica species.


Assuntos
Barbarea/química , Sesquiterpenos/química , Sesquiterpenos/isolamento & purificação , Sulfóxidos/isolamento & purificação , Alternaria/metabolismo , Antifúngicos/química , Ascomicetos/metabolismo , Brassica/metabolismo , Ressonância Magnética Nuclear Biomolecular , Folhas de Planta/química , Sesquiterpenos/farmacologia , Sulfóxidos/química , Sulfóxidos/farmacologia
17.
BMC Genomics ; 16: 486, 2015 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-26126637

RESUMO

BACKGROUND: Barbarea vulgaris contains two genotypes: the glabrous type (G-type), which confers resistance to the diamondback moth (DBM) and other insect pests, and the pubescent type (P-type), which is susceptible to the DBM. Herein, the transcriptomes of P-type B. vulgaris before and after DBM infestation were subjected to Illumina (Solexa) pyrosequencing and comparative analysis. RESULTS: 5.0 gigabase pairs of clean nucleotides were generated. Non-redundant unigenes (33,721) were assembled and 94.1 % of them were annotated. Compared with our previous G-type transcriptome, the expression patterns of many insect responsive genes, including those related to secondary metabolism, phytohormones and transcription factors, which were significantly induced by DBM in G-type plants, were less sensitive to DBM infestation in P-type plants. The genes of the triterpenoid saponin pathway were identified in both G- and P-type plants. The upstream genes of the pathway showed similar expression patterns between the two genotypes. However, gene expression for two downstream enzymes, the glucosyl transferase (UGT73C11) and an oxidosqualene cyclase (OSC), were significantly upregulated in the P-type compared with the G-type plant. The homologous genes from P- and G-type plants were detected by BLAST unigenes with a cutoff level E-value < e(-10). 12,980 gene families containing 26,793 P-type and 36,944 G-type unigenes were shared by the two types of B. vulgaris. 38,397 single nucleotide polymorphisms (SNPs) were found in 9,452 orthologous genes between the P- and G-type plants. We also detected 5,105 simple sequence repeats (SSRs) in the B. vulgaris transcriptome, comprising mono-nucleotide-repeats (2,477; 48.5 %) and triple-nucleotide-repeats (1,590; 31.1 %). Of these, 1,657 SSRs displayed polymorphisms between the P- and G-type. Consequently, 913 SSR primer pairs were designed with a resolution of more than two nucleotides. We randomly chose 30 SSRs to detect the genetic diversity of 32 Barbarea germplasms. The distance tree showed that these accessions were clearly divided into groups, with the G-type grouping with available Western and Central European B. vulgaris accessions in contrast to the P-type accession, B. stricta and B. verna. CONCLUSIONS: These data represent useful information for pest-resistance gene mining and for the investigation of the molecular basis of plant-pest interactions.


Assuntos
Barbarea/classificação , Barbarea/genética , Perfilação da Expressão Gênica/métodos , Mariposas/parasitologia , Proteínas de Plantas/genética , Animais , Barbarea/parasitologia , Resistência à Doença , Regulação da Expressão Gênica de Plantas , Variação Genética , Sequenciamento de Nucleotídeos em Larga Escala , Repetições de Microssatélites , Filogenia , Análise de Sequência de RNA
18.
Oecologia ; 179(2): 353-61, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26001606

RESUMO

It is well known that many parasitic wasps use herbivore-induced plant odours (HIPVs) to locate their inconspicuous host insects, and are often able to distinguish between slight differences in plant odour composition. However, few studies have examined parasitoid foraging behaviour under (semi-)field conditions. In nature, food plants of parasitoid hosts are often embedded in non-host-plant assemblages that confer both structural and chemical complexity. By releasing both naïve and experienced Cotesia glomerata females in outdoor tents, we studied how natural vegetation surrounding Pieris brassicae-infested Sinapis arvensis and Barbarea vulgaris plants influences their foraging efficiency as well as their ability to specifically orient towards the HIPVs of the host plant species on which they previously had a positive oviposition experience. Natural background vegetation reduced the host-encounter rate of naïve C. glomerata females by 47 %. While associative learning of host plant HIPVs 1 day prior to foraging caused a 28 % increase in the overall foraging efficiency of C. glomerata, it did not reduce the negative influence of natural background vegetation. At the same time, however, females foraging in natural vegetation attacked more host patches on host-plant species on which they previously had a positive oviposition experience. We conclude that, even though the presence of natural vegetation reduces the foraging efficiency of C. glomerata, it does not prevent experienced female wasps from specifically orienting towards the host-plant species from which they had learned the HIPVs.


Assuntos
Borboletas/parasitologia , Ecossistema , Herbivoria , Vespas/fisiologia , Animais , Barbarea/química , Borboletas/fisiologia , Comportamento Alimentar , Feminino , Interações Hospedeiro-Parasita , Larva/parasitologia , Larva/fisiologia , Odorantes , Oviposição , Sinapis/química
19.
Oecologia ; 178(4): 1181-92, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25783488

RESUMO

The preference-performance hypothesis posits that the host plant range of plant-feeding insects is ultimately limited by larval costs associated with feeding on multiple resources, and that female egg-laying preferences evolve in response to these costs. The trade-off of either using few host plant species and being a strong competitor on them due to effective utilization or using a wide host plant range but being a poor competitor is further predicted to result in host plant specialization. This follows under the hypothesis that both females and offspring are ultimately favoured by utilizing only the most suitable host(s). We develop an experimental approach to identify such trade-offs, i.e. larval costs associated with being a host generalist, and apply a suite of experiments to two sympatric and syntopic populations of the closely related butterflies Pieris napi and Pieris rapae. These butterflies show variation in their level of host specialization, which allowed comparisons between more and less specialized species and between families within species. Our results show that, first, the link between female host preference and offspring performance was not significantly stronger in the specialist compared to the generalist species. Second, the offspring of the host plant specialist did not outperform the offspring of the generalist on the former's most preferred host plant species. Finally, the more generalized species, or families within species, did not show higher survival or consistently higher growth rates than the specialists on the less preferred plants. Thus, the preference and performance traits appear to evolve as largely separated units.


Assuntos
Barbarea , Evolução Biológica , Borboletas/fisiologia , Herbivoria , Oviposição , Animais , Feminino , Especificidade de Hospedeiro , Larva/fisiologia , Plantas
20.
Oecologia ; 177(2): 441-52, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25380645

RESUMO

It is well known that pathogens and arthropod herbivores attacking the same host plant may affect each other. Little is known, however, about their combined impact on plant fitness, which may differ from simple additive expectations. In a 2-year common garden field experiment, we tested whether the pathogen Albugo sp. (white blister rust) and the herbivorous flea beetle Phyllotreta nemorum affected each other's performance on two resistance types (G-type and P-type) of the crucifer Barbarea vulgaris ssp. arcuata, and whether biomass, reproduction and survival of the plants were affected by interactive impacts of the antagonists. Most of the insect-resistant G-plants were severely affected by white rust, which reduced biomass and reproductive potential compared to the controls. However, when also exposed to flea beetles, biomass loss was mitigated in G-plants, even though apparent disease symptoms were not reduced. Most of the insect-susceptible P-plants were resistant to white rust; however, the number of flea beetle mines tended to increase in plants also exposed to Albugo, and biomass at the last harvest was slightly lower in the combined treatment. Thus, interactive impacts of the herbivore and pathogen differed between the two resistance types, with an antagonistic combined impact in G-plants, which lasted surprisingly long, and a slight synergistic impact in P-plants.


Assuntos
Barbarea/microbiologia , Besouros/microbiologia , Herbivoria , Oomicetos/fisiologia , Animais , Barbarea/genética , Resistência à Doença , Doenças das Plantas/microbiologia
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