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1.
Chembiochem ; 24(18): e202300234, 2023 09 15.
Artículo en Inglés | MEDLINE | ID: mdl-37249120

RESUMEN

Cocaine and hyoscyamine are two tropane alkaloids (TA) from Erythroxylaceae and Solanaceae, respectively. These famous compounds possess anticholinergic properties that can be used to treat neuromuscular disorders. While the hyoscyamine biosynthetic pathway has been fully elucidated allowing its de novo synthesis in yeast, the cocaine pathway remained only partially elucidated. Recently, the Huang research group has completed the cocaine biosynthetic route by characterizing its two missing enzymes. This allowed the whole pathway to be transferring into Nicotiana benthamiana to achieve cocaine production. Here, besides highlighting the impact of this discovery, we discuss how TA biosynthesis evolved via the recruitment of two distinct and convergent pathways in Erythroxylaceae and Solanaceae. Finally, while enriching our knowledge on TA biosynthesis, this diversification of the molecular actors involved in cocaine and hyoscyamine biosynthesis opens perspectives in metabolic engineering by exploring enzyme biochemical plasticity that can ease and shorten TA pathway reconstitution in heterologous organisms.


Asunto(s)
Cocaína , Hiosciamina , Solanaceae , Cocaína/metabolismo , Tropanos/química , Tropanos/metabolismo , Solanaceae/metabolismo , Antagonistas Colinérgicos/metabolismo
2.
Molecules ; 27(15)2022 Jul 25.
Artículo en Inglés | MEDLINE | ID: mdl-35897915

RESUMEN

COVID-19, caused by the coronavirus SARS-CoV-2, emerged in late December 2019 in Wuhan, China. As of 8 April 2022, the virus has caused a global pandemic, resulting in 494,587,638 infections leading to 6,170,283 deaths around the world. Although several vaccines have received emergency authorization from USA and UK drug authorities and two more in Russia and China, it is too early to comment on the prolonged effectiveness of the vaccines, their availability, and affordability for the developing countries of the world, and the daunting task to vaccinate 7 billion people of the world with two doses of the vaccine with additional booster doses. As a result, it is still worthwhile to search for drugs and several promising leads have been found, mainly through in silico studies. In this study, we have examined the binding energies of several alkaloids and anthocyanin derivatives from the Solanaceae family, a family which contains common consumable vegetables and fruit items such as eggplant, pepper, and tomatoes. Our study demonstrates that Solanaceae family alkaloids such as incanumine and solaradixine, as well as anthocyanins and anthocyanidins, have very high predicted binding energies for the 3C-like protease of SARS-CoV-2 (also known as Mpro). Since Mpro is vital for SARS-CoV-2 replication, the compounds merit potential for further antiviral research towards the objective of obtaining affordable drugs.


Asunto(s)
Alcaloides , Tratamiento Farmacológico de COVID-19 , Solanaceae , Alcaloides/farmacología , Antocianinas , Antivirales/química , Proteasas 3C de Coronavirus , Cisteína Endopeptidasas/química , Humanos , Simulación del Acoplamiento Molecular , Simulación de Dinámica Molecular , Péptido Hidrolasas/metabolismo , Fitoquímicos/farmacología , Inhibidores de Proteasas/química , SARS-CoV-2 , Solanaceae/metabolismo , Verduras/metabolismo , Proteínas no Estructurales Virales/metabolismo
3.
Commun Biol ; 4(1): 671, 2021 06 03.
Artículo en Inglés | MEDLINE | ID: mdl-34083720

RESUMEN

Wolfberry Lycium, an economically important genus of the Solanaceae family, contains approximately 80 species and shows a fragmented distribution pattern among the Northern and Southern Hemispheres. Although several herbaceous species of Solanaceae have been subjected to genome sequencing, thus far, no genome sequences of woody representatives have been available. Here, we sequenced the genomes of 13 perennial woody species of Lycium, with a focus on Lycium barbarum. Integration with other genomes provides clear evidence supporting a whole-genome triplication (WGT) event shared by all hitherto sequenced solanaceous plants, which occurred shortly after the divergence of Solanaceae and Convolvulaceae. We identified new gene families and gene family expansions and contractions that first appeared in Solanaceae. Based on the identification of self-incompatibility related-gene families, we inferred that hybridization hotspots are enriched for genes that might be functioning in gametophytic self-incompatibility pathways in wolfberry. Extremely low expression of LOCULE NUBER (LC) and COLORLESS NON-RIPENING (CNR) orthologous genes during Lycium fruit development and ripening processes suggests functional diversification of these two genes between Lycium and tomato. The existence of additional flowering locus C-like MADS-box genes might correlate with the perennial flowering cycle of Lycium. Differential gene expression involved in the lignin biosynthetic pathway between Lycium and tomato likely illustrates woody and herbaceous differentiation. We also provide evidence that Lycium migrated from Africa into Asia, and subsequently from Asia into North America. Our results provide functional insights into Solanaceae origins, evolution and diversification.


Asunto(s)
Cromosomas de las Plantas/genética , Genoma de Planta/genética , Lycium/genética , Solanaceae/genética , Secuenciación Completa del Genoma/métodos , África , Asia , Evolución Molecular , Frutas/genética , Frutas/metabolismo , Regulación de la Expresión Génica de las Plantas , Geografía , Lycium/clasificación , Lycium/metabolismo , América del Norte , Filogenia , Poliploidía , Polisacáridos/metabolismo , Solanaceae/clasificación , Solanaceae/metabolismo , Especificidad de la Especie
4.
Genes (Basel) ; 11(9)2020 09 19.
Artículo en Inglés | MEDLINE | ID: mdl-32961813

RESUMEN

Floral organ size, especially the size of the corolla, plays an important role in plant reproduction by facilitating pollination efficiency. Previous studies have outlined a hypothesized organ size pathway. However, the expression and function of many of the genes in the pathway have only been investigated in model diploid species; therefore, it is unknown how these genes interact in polyploid species. Although correlations between ploidy and cell size have been shown in many systems, it is unclear whether there is a difference in cell size between naturally occurring and synthetic polyploids. To address these questions comparing floral organ size and cell size across ploidy, we use natural and synthetic polyploids of Nicotiana tabacum (Solanaceae) as well as their known diploid progenitors. We employ a comparative transcriptomics approach to perform analyses of differential gene expression, focusing on candidate genes that may be involved in floral organ size, both across developmental stages and across accessions. We see differential expression of several known floral organ candidate genes including ARF2, BIG BROTHER, and GASA/GAST1. Results from linear models show that ploidy, cell width, and cell number positively influence corolla tube circumference; however, the effect of cell width varies by ploidy, and diploids have a significantly steeper slope than both natural and synthetic polyploids. These results demonstrate that polyploids have wider cells and that polyploidy significantly increases corolla tube circumference.


Asunto(s)
Flores/metabolismo , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/metabolismo , Poliploidía , Solanaceae/metabolismo , Evolución Biológica , Flores/anatomía & histología , Flores/genética , Flores/crecimiento & desarrollo , Fenotipo , Proteínas de Plantas/genética , Polinización , Solanaceae/genética , Solanaceae/crecimiento & desarrollo
5.
BMC Plant Biol ; 20(1): 266, 2020 Jun 09.
Artículo en Inglés | MEDLINE | ID: mdl-32517797

RESUMEN

BACKGROUND: Cellular membranes are dynamic structures, continuously adjusting their composition, allowing plants to respond to developmental signals, stresses, and changing environments. To facilitate transmembrane transport of substrates, plant membranes are embedded with both active and passive transporters. Aquaporins (AQPs) constitute a major family of membrane spanning channel proteins that selectively facilitate the passive bidirectional passage of substrates across biological membranes at an astonishing 108 molecules per second. AQPs are the most diversified in the plant kingdom, comprising of five major subfamilies that differ in temporal and spatial gene expression, subcellular protein localisation, substrate specificity, and post-translational regulatory mechanisms; collectively providing a dynamic transportation network spanning the entire plant. Plant AQPs can transport a range of solutes essential for numerous plant processes including, water relations, growth and development, stress responses, root nutrient uptake, and photosynthesis. The ability to manipulate AQPs towards improving plant productivity, is reliant on expanding our insight into the diversity and functional roles of AQPs. RESULTS: We characterised the AQP family from Nicotiana tabacum (NtAQPs; tobacco), a popular model system capable of scaling from the laboratory to the field. Tobacco is closely related to major economic crops (e.g. tomato, potato, eggplant and peppers) and itself has new commercial applications. Tobacco harbours 76 AQPs making it the second largest characterised AQP family. These fall into five distinct subfamilies, for which we characterised phylogenetic relationships, gene structures, protein sequences, selectivity filter compositions, sub-cellular localisation, and tissue-specific expression. We also identified the AQPs from tobacco's parental genomes (N. sylvestris and N. tomentosiformis), allowing us to characterise the evolutionary history of the NtAQP family. Assigning orthology to tomato and potato AQPs allowed for cross-species comparisons of conservation in protein structures, gene expression, and potential physiological roles. CONCLUSIONS: This study provides a comprehensive characterisation of the tobacco AQP family, and strengthens the current knowledge of AQP biology. The refined gene/protein models, tissue-specific expression analysis, and cross-species comparisons, provide valuable insight into the evolutionary history and likely physiological roles of NtAQPs and their Solanaceae orthologs. Collectively, these results will support future functional studies and help transfer basic research to applied agriculture.


Asunto(s)
Acuaporinas/genética , Nicotiana/genética , Proteínas de Plantas/genética , Solanaceae/genética , Aminoácidos/metabolismo , Perfilación de la Expresión Génica , Genes de Plantas/genética , Estudio de Asociación del Genoma Completo , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Filogenia , Análisis de Secuencia de ADN , Solanaceae/metabolismo , Solanum tuberosum/genética , Solanum tuberosum/metabolismo , Nicotiana/metabolismo
6.
Plant Signal Behav ; 15(2): 1707348, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-31884882

RESUMEN

Solanaceous plants produce sesquiterpenoid phytoalexins to defend themselves against a variety of pathogens. These toxic compounds are not only harmful to the pathogen but also to the plant, and thus need to be detoxified by the plant after the threat has been eliminated. We report that the detoxification of rishitin, the major phytoalexin in potato tubers and tomato fruits, is mediated by a cytochrome P450 CYP76 family enzyme via the hydroxylation of the isopropenyl group resulting in the formation of 13-hydroxyrishitin, also known as rishitin-M1. We further observed hydroxylation of the potato phytoalexins solavetivone, lubimin and oxylubimin by the same enzyme. Constitutive expression of CYP76 in Nicotiana benthamiana also led to a reduction of the non-potato phytoalexins capsidiol and its derivative capsidiol 3-acetate. We therefore annotated this enzyme as sesquiterpenoid phytoalexins hydroxylase, SPH. This broad range of substrates indicates that SPH functions as a general phytoalexin detoxification enzyme in Solanaceae, and is therefore relevant for a better understanding of plant-pathogen interaction in solanaceous plants, which comprise many economically important crops, such as potato, tomato, eggplant and pepper.


Asunto(s)
Sistema Enzimático del Citocromo P-450/metabolismo , Furanos/metabolismo , Sesquiterpenos/metabolismo , Terpenos/metabolismo , Solanaceae/metabolismo , Solanum tuberosum/metabolismo , Fitoalexinas
7.
Genes (Basel) ; 10(8)2019 07 25.
Artículo en Inglés | MEDLINE | ID: mdl-31349565

RESUMEN

Solanaceae have played an important role in elucidating how flower color is specified by the flavonoid biosynthesis pathway (FBP), which produces anthocyanins and other secondary metabolites. With well-established reverse genetics tools and rich genomic resources, Solanaceae provide a robust framework to examine the diversification of this well-studied pathway over short evolutionary timescales and to evaluate the predictability of genetic perturbation on pathway flux. Genomes of eight Solanaceae species, nine related asterids, and four rosids were mined to evaluate variation in copy number of the suite of FBP enzymes involved in anthocyanin biosynthesis. Comparison of annotation sources indicated that the NCBI annotation pipeline generated more and longer FBP annotations on average than genome-specific annotation pipelines. The pattern of diversification of each enzyme among asterids was assessed by phylogenetic analysis, showing that the CHS superfamily encompasses a large paralogous family of ancient and recent duplicates, whereas other FBP enzymes have diversified via recent duplications in particular lineages. Heterologous expression of a pansy F3'5'H gene in tobacco changed flower color from pink to dark purple, demonstrating that anthocyanin production can be predictably modified using reverse genetics. These results suggest that the Solanaceae FBP could be an ideal system to model genotype-to-phenotype interactions for secondary metabolism.


Asunto(s)
Antocianinas/biosíntesis , Genoma de Planta , Metabolismo Secundario , Solanaceae/metabolismo , Antocianinas/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Solanaceae/genética
8.
ACS Chem Biol ; 14(7): 1546-1555, 2019 07 19.
Artículo en Inglés | MEDLINE | ID: mdl-31246411

RESUMEN

Plant pathogenic bacteria possess sophisticated mechanisms to detect the presence of host plants by sensing host-derived compounds. Ralstonia solanacearum, the causative agent of bacterial wilt on solanaceous plants, employs quorum sensing to control the production of the secondary metabolite ralfuranones/ralstonins, which have been suggested to be involved in virulence. Here, we report that d-galactose and d-glucose, plant sugars, activate the production of ralfuranones/ralstonins in R. solanacearum. As a result, two new derivatives, ralfuranone M (1) and ralstonin C (2), were found in the culture extracts, and their structures were elucidated by spectroscopic and chemical methods. Ralstonin C (2) is a cyclic lipopeptide containing a unique fatty acid, (2S,3S,Z)-3-amino-2-hydroxyicos-13-enoic acid, whereas ralfuranone M (1) has a common aryl-furanone structure with other ralfuranones. d-Galactose and d-glucose activated the expression of the biosynthetic ralfuranone/ralstonin genes and in part became the biosynthetic source of ralfuranones/ralstonins. Ralfuranones and ralstonins were detected from the xylem fluid of the infected tomato plants, and their production-deficient mutants exhibited reduced virulence on tomato and tobacco plants. Taken together, these results suggest that activation of ralfuranone/ralstonin production by host sugars functions in R. solanacearum virulence.


Asunto(s)
Galactosa/metabolismo , Glucosa/metabolismo , Lactonas/metabolismo , Enfermedades de las Plantas/microbiología , Ralstonia solanacearum/fisiología , Solanaceae/microbiología , Interacciones Huésped-Patógeno , Solanum lycopersicum/metabolismo , Solanum lycopersicum/microbiología , Percepción de Quorum , Ralstonia solanacearum/patogenicidad , Solanaceae/metabolismo , Nicotiana/metabolismo , Nicotiana/microbiología
9.
Plant J ; 99(4): 686-702, 2019 08.
Artículo en Inglés | MEDLINE | ID: mdl-31009131

RESUMEN

The genetic mechanisms underlying fruit development have been identified in Arabidopsis and have been comparatively studied in tomato as a representative of fleshy fruits. However, comparative expression and functional analyses on the bHLH genes downstream the genetic network, ALCATRAZ (ALC) and SPATULA (SPT), which are involved in the formation of the dehiscence zone in Arabidopsis, have not been functionally studied in the Solanaceae. Here, we perform detailed expression and functional studies of ALC/SPT homologs in Nicotiana obtusifolia with capsules, and in Capsicum annuum and Solanum lycopersicum with berries. In Solanaceae, ALC and SPT genes are expressed in leaves, and all floral organs, especially in petal margins, stamens and carpels; however, their expression changes during fruit maturation according to the fruit type. Functional analyses show that downregulation of ALC/SPT genes does not have an effect on gynoecium patterning; however, they have acquired opposite roles in petal expansion and have been co-opted in leaf pigmentation in Solanaceae. In addition, ALC/SPT genes repress lignification in time and space during fruit development in Solanaceae. Altogether, some roles of ALC and SPT genes are different between Brassicaceae and Solanaceae; while the paralogs have undergone some subfunctionalization in the former they are mostly redundant in the latter.


Asunto(s)
Proteínas de Plantas/metabolismo , Solanaceae/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Brassicaceae/genética , Brassicaceae/metabolismo , Capsicum/genética , Capsicum/metabolismo , Regulación de la Expresión Génica de las Plantas , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Proteínas de Plantas/genética , Solanaceae/genética
10.
Biochemistry (Mosc) ; 83(6): 727-737, 2018 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-30195329

RESUMEN

Galectins are involved in various biological processes, e.g. cell-cell and cell-matrix adhesion and the transmission of cellular signals. Despite the diversity of functions, little is known about the nature of their physiological cognate ligands on the cell surface and the localization of galectins in the glycocalyx, although this information is important for understanding the functional activity of galectins. In this work, localization of endogenous and exogenously loaded galectins in the glycocalyx was studied. The following main conclusions are drawn: 1) galectins are not evenly distributed within the glycocalyx, they are accumulated in patches. Patching is not the result of a cross-linking of cellular glycans by galectins. Instead, patch-wise localization is the consequence of irregular distribution of glycans forming the glycocalyx; 2) galectins are accumulated in the inner zone of the glycocalyx rather than at its outer face or directly in vicinity of the cell membrane; 3) patches are not associated with cell rafts.


Asunto(s)
Galectinas/metabolismo , Glicocálix/metabolismo , Línea Celular Tumoral , Células HT29 , Humanos , Células Jurkat , Lectinas/metabolismo , Microscopía Confocal , Solanaceae/metabolismo
11.
Ecotoxicol Environ Saf ; 155: 109-116, 2018 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-29510305

RESUMEN

To determine whether the living hyperaccumulator plants and their straws have the same effects on the growth and heavy metal accumulation of common plants, two pot experiments (intercropping experiment and straw mulch experiment) were conducted to study the effects of living hyperaccumulator plants (Solanum photeinocarpum, Tagetes erecta, Galinsoga parviflora and Bidens pilosa) and their straws on the growth and cadmium (Cd) accumulation of common plant Cyphomandra betacea seedlings. Intercropping with T. erecta or B. pilosa promoted the growth of C. betacea seedlings compared with the monoculture, while intercropping with S. photeinocarpum or G. parviflora inhibited that. Intercropping with S. photeinocarpum decreased the Cd contents in the roots and shoots of C. betacea seedlings compared with the monoculture, but intercropping with the other plants did not. In the straw mulch experiment, the straw of S. photeinocarpum or T. erecta promoted the growth of C. betacea seedlings compared with the control, while the straw of G. parviflora or B. pilosa did not. The straw of S. photeinocarpum or T. erecta decreased the Cd contents in the shoots of C. betacea seedlings, and the straw of G. parviflora or B. pilosa increased the shoot Cd contents. Thus, intercropping with S. photeinocarpum and applying S. photeinocarpum or T. erecta straw can reduce the Cd uptake of C. betacea.


Asunto(s)
Asteraceae/metabolismo , Cadmio/toxicidad , Contaminantes del Suelo/toxicidad , Solanaceae/efectos de los fármacos , Biodegradación Ambiental , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/metabolismo , Brotes de la Planta/efectos de los fármacos , Brotes de la Planta/crecimiento & desarrollo , Brotes de la Planta/metabolismo , Plantones/efectos de los fármacos , Plantones/crecimiento & desarrollo , Plantones/metabolismo , Solanaceae/crecimiento & desarrollo , Solanaceae/metabolismo
12.
Chem Biodivers ; 14(8)2017 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-28581196

RESUMEN

Twelve Salpichroa taxa have been phytochemically analyzed. From the aerial parts of S. scandens, four known salpichrolides A, C, I, S, and an unreported withanolide named salpichrolide V (1), were isolated. In S. dependens, S. gayi, S. glandulosa subsp. glandulosa, S. glandulosa subps. weddellii, S. leucantha, S. micrantha, S. microloba, S. proboscidea, S. ramosissima, S. tristis var. tristis, and S. weberbauerii, no withanolides were found. The chemical content of ca. 85% of the Salpichroa taxa is in agreement with molecular studies, which suggest that Salpichroa and Jaborosa, a genus considered morphologically close to Salpichroa, are distant in the systematic of the Solanoideae subfamily. Moreover, the in vitro cytotoxic activity of a set of natural salpichrolides and derivatives was examined against two prostate carcinoma cell lines (PC3 and LNCaP) and two human breast cancer cell lines (MCF-7 and T47D). Several compounds showed moderate activity (IC50  = 64.91 - 29.97 µm).


Asunto(s)
Fitoquímicos/química , Fitoquímicos/farmacología , Solanaceae/química , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Cromatografía en Capa Delgada , Ensayos de Selección de Medicamentos Antitumorales , Femenino , Humanos , Células MCF-7 , Masculino , Fitoquímicos/aislamiento & purificación , Neoplasias de la Próstata/metabolismo , Neoplasias de la Próstata/patología , Solanaceae/metabolismo
13.
Int J Mol Sci ; 17(10)2016 Oct 02.
Artículo en Inglés | MEDLINE | ID: mdl-27706100

RESUMEN

This review provides a current summary of plant resistance inducers (PRIs) that have been successfully used in the Solanaceae plant family to protect against pathogens by activating the plant's own defence. Solanaceous species include many important crops such as potato and tomato. We also present findings regarding the molecular processes after application of PRIs, even if the number of such studies still remains limited in this plant family. In general, there is a lack of patterns regarding the efficiency of induced resistance (IR) both between and within solanaceous species. In many cases, a hypersensitivity-like reaction needs to form in order for the PRI to be efficient. "-Omics" studies have already given insight in the complexity of responses, and can explain some of the differences seen in efficacy of PRIs between and within species as well as towards different pathogens. Finally, examples of field applications of PRIs for solanaceous crops are presented and discussed. We predict that PRIs will play a role in future plant protection strategies in Solanaceae crops if they are combined with other means of disease control in different spatial and temporal combinations.


Asunto(s)
Solanaceae/metabolismo , Aminobutiratos/metabolismo , Aminobutiratos/farmacología , Bacterias/efectos de los fármacos , Productos Agrícolas , Ciclopentanos/metabolismo , Ciclopentanos/farmacología , Etilenos/metabolismo , Etilenos/farmacología , Hongos/efectos de los fármacos , Oxilipinas/metabolismo , Oxilipinas/farmacología , Especies Reactivas de Oxígeno/metabolismo , Solanaceae/genética , Solanaceae/microbiología
14.
J Exp Bot ; 67(16): 4801-12, 2016 08.
Artículo en Inglés | MEDLINE | ID: mdl-27222514

RESUMEN

Cyclotides are plant-derived cyclic peptides that have a head-to-tail cyclic backbone and three conserved disulphide bonds that form a cyclic cystine knot motif. They occur in plants from the Violaceae, Rubiaceae, Cucurbitaceae, Fabaceae, and Solanaceae families, typically with 10-100 cyclotides in a given plant species, in a wide range of tissues, including flowers, leaves, stems, and roots. Some cyclotides are expressed in large amounts (up to 1g kg(-1) wet plant weight) and their natural function appears to be to protect plants from pests or pathogens. This article provides a brief overview of their discovery, distribution in plants, and applications. In particular, their exceptional stability has led to their use as peptide-based scaffolds in drug design applications. They also have potential as natural 'ecofriendly' insecticides, and as protein engineering frameworks.


Asunto(s)
Ciclotidas/genética , Magnoliopsida/genética , Proteínas de Plantas/genética , Cucurbitaceae/genética , Cucurbitaceae/metabolismo , Ciclotidas/metabolismo , Fabaceae/genética , Fabaceae/metabolismo , Magnoliopsida/metabolismo , Proteínas de Plantas/metabolismo , Rubiaceae/genética , Rubiaceae/metabolismo , Solanaceae/genética , Solanaceae/metabolismo , Violaceae/genética , Violaceae/metabolismo
15.
Phytochemistry ; 127: 12-22, 2016 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-26988730

RESUMEN

Brugmansia arborea is a woody plant species that produces tropane alkaloids (TAs). The gene encoding tropine-forming reductase or tropinone reductase I (BaTRI) in this plant species was functionally characterised. The full-length cDNA of BaTRI encoded a 272-amino-acid polypeptide that was highly similar to tropinone reductase I from TAs-producing herbal plant species. The purified 29kDa recombinant BaTRI exhibited maximum reduction activity at pH 6.8-8.0 when tropinone was used as substrate; it also exhibited maximum oxidation activity at pH 9.6 when tropine was used as substrate. The Km, Vmax and Kcat values of BaTRI for tropinone were 2.65mM, 88.3nkatmg(-1) and 2.93S(-1), respectively, at pH 6.4; the Km, Vmax and Kcat values of TRI from Datura stramonium (DsTRI) for tropinone were respectively 4.18mM, 81.20nkatmg(-1) and 2.40S(-1) at pH 6.4. At pH 6.4, 6.8 and 7.0, BaTRI had a significantly higher activity than DsTRI. Analogues of tropinone, 4-methylcyclohexanone and 3-quinuclidinone hydrochloride, were also used to investigate the enzymatic kinetics of BaTRI. The Km, Vmax and Kcat values of BaTRI for tropine were 0.56mM, 171.62nkat.mg(-1) and 5.69S(-1), respectively, at pH 9.6; the Km, Vmax and Kcat values of DsTRI for tropine were 0.34mM, 111.90nkatmg(-1) and 3.30S(-1), respectively, at pH 9.6. The tissue profiles of BaTRI differed from those in TAs-producing herbal plant species. BaTRI was expressed in all examined organs but was most abundant in secondary roots. Finally, tropane alkaloids, including hyoscyamine, anisodamine and scopolamine, were detected in various organs of B. arborea by HPLC. Interestingly, scopolamine constituted most of the tropane alkaloids content in B. arborea, which suggests that B. arborea is a scopolamine-rich plant species. The scopolamine content was much higher in the leaves and stems than in other organs. The gene expression and TAs accumulation suggest that the biosynthesis of hyoscyamine, especially scopolamine, occurred not only in the roots but also in the aerial parts of B. arborea.


Asunto(s)
Oxidorreductasas de Alcohol/metabolismo , Medicamentos Herbarios Chinos/aislamiento & purificación , Solanaceae , Tropanos/metabolismo , Secuencia de Aminoácidos , Cromatografía Líquida de Alta Presión , Medicamentos Herbarios Chinos/química , Concentración de Iones de Hidrógeno , Datos de Secuencia Molecular , Estructura Molecular , Filogenia , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de Proteína , Solanaceae/genética , Solanaceae/metabolismo , Tropanos/química
16.
Plant J ; 85(4): 561-77, 2016 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-26749139

RESUMEN

Exploring the diversity of plant secondary metabolism requires efficient methods to obtain sufficient structural insights to discriminate previously known from unknown metabolites. De novo structure elucidation and confirmation of known metabolites (dereplication) remain a major bottleneck for mass spectrometry-based metabolomic workflows, and few systematic dereplication strategies have been developed for the analysis of entire compound classes across plant families, partly due to the complexity of plant metabolic profiles that complicates cross-species comparisons. 17-hydroxygeranyllinalool diterpene glycosides (HGL-DTGs) are abundant defensive secondary metabolites whose malonyl and glycosyl decorations are induced by jasmonate signaling in the ecological model plant Nicotiana attenuata. The multiple labile glycosidic bonds of HGL-DTGs result in extensive in-source fragmentation (IS-CID) during ionization. To reconstruct these IS-CID clusters from profiling data and identify precursor ions, we applied a deconvolution algorithm and created an MS/MS library from positive-ion spectra of purified HGL-DTGs. From this library, 251 non-redundant fragments were annotated, and a workflow to characterize leaf, flower and fruit extracts of 35 solanaceous species was established. These analyses predicted 105 novel HGL-DTGs that were restricted to Nicotiana, Capsicum and Lycium species. Interestingly, malonylation is a highly conserved step in HGL-DTG metabolism, but is differentially affected by jasmonate signaling among Nicotiana species. This MS-based workflow is readily applicable for cross-species re-identification/annotation of other compound classes with sufficient fragmentation knowledge, and therefore has the potential to support hypotheses regarding secondary metabolism diversification.


Asunto(s)
Diterpenos/química , Glicósidos/química , Metabolómica/métodos , Solanaceae/química , Espectrometría de Masas en Tándem/métodos , Capsicum/química , Capsicum/metabolismo , Ciclopentanos/metabolismo , Diterpenos/clasificación , Diterpenos/aislamiento & purificación , Glicósidos/clasificación , Glicósidos/aislamiento & purificación , Lycium/química , Lycium/metabolismo , Oxilipinas/metabolismo , Reguladores del Crecimiento de las Plantas/metabolismo , Hojas de la Planta/química , Hojas de la Planta/metabolismo , Transducción de Señal , Solanaceae/metabolismo , Especificidad de la Especie , Nicotiana/química , Nicotiana/metabolismo
17.
Acta Biol Hung ; 66(3): 304-15, 2015 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-26344026

RESUMEN

The family Solanaceae includes several melliferous plants, which tend to produce copious amounts of nectar. Floral nectar is a chemically complex aqueous solution, dominated by sugars, but minor components such as amino acids, proteins, flavonoids and alkaloids are present as well. This study aimed at analysing the protein and alkaloid profile of the nectar in seven solanaceous species. Proteins were examined with SDS-PAGE and alkaloids were analyzed with HPLC. The investigation of protein profile revealed significant differences in nectar-protein patterns not only between different plant genera, but also between the three Nicotiana species investigated. SDS-PAGE suggested the presence of several Nectarin proteins with antimicrobial activity in Nicotiana species. The nectar of all tobacco species contained the alkaloid nicotine, N. tabacum having the highest nicotine content. The nectar of Brugmansia suaveolens, Datura stramonium, Hyoscyamus niger and Lycium barbarum contained scopolamine, the highest content of which was measured in B. suaveolens. The alkaloid concentrations in the nectars of most solanaceous species investigated can cause deterrence in honeybees, and the nectar of N. rustica and N. tabacum can be considered toxic for honeybees.


Asunto(s)
Néctar de las Plantas/química , Proteínas de Plantas/análisis , Solanaceae/química , Alcaloides Solanáceos/análisis , Animales , Abejas , Néctar de las Plantas/metabolismo , Proteínas de Plantas/metabolismo , Solanaceae/metabolismo , Alcaloides Solanáceos/metabolismo
18.
New Phytol ; 208(1): 210-23, 2015 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-26009937

RESUMEN

Plant nucleotide-binding, leucine-rich repeat (NB-LRR) proteins confer immunity to pathogens possessing the corresponding avirulence proteins. Activation of NB-LRR proteins is often associated with induction of the hypersensitive response (HR), a form of programmed cell death. NRC1 (NB-LRR Required for HR-Associated Cell Death-1) is a tomato (Solanum lycopersicum) NB-LRR protein that participates in the signalling cascade leading to resistance to the pathogens Cladosporium fulvum and Verticillium dahliae. To identify mutations in NRC1 that cause increased signalling activity, we generated a random library of NRC1 variants mutated in their nucleotide-binding domain and screened them for the ability to induce an elicitor-independent HR in Nicotiana tabacum. Screening of 1920 clones retrieved 11 gain-of-function mutants, with 10 of them caused by a single amino acid substitution. All substitutions are located in or very close to highly conserved motifs within the nucleotide-binding domain, suggesting modulation of the signalling activity of NRC1. Three-dimensional modelling of the nucleotide-binding domain of NRC1 revealed that the targeted residues are centred around the bound nucleotide. Our mutational approach has generated a wide set of novel gain-of-function mutations in NRC1 and provides insight into how the activity of this NB-LRR is regulated.


Asunto(s)
Resistencia a la Enfermedad/genética , Mutación , Enfermedades de las Plantas/microbiología , Proteínas de Plantas/genética , Dominios y Motivos de Interacción de Proteínas/genética , Proteínas/genética , Solanaceae/genética , Secuencia de Aminoácidos , Proteínas Bacterianas/metabolismo , Sitios de Unión , Muerte Celular , Cladosporium/metabolismo , Cladosporium/patogenicidad , Genes de Plantas , Leucina/metabolismo , Proteínas Repetidas Ricas en Leucina , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Solanum lycopersicum/microbiología , Estructura Molecular , Mutagénesis , Nucleótidos/metabolismo , Proteínas de Plantas/metabolismo , Proteínas/metabolismo , Transducción de Señal , Solanaceae/metabolismo , Solanaceae/microbiología , Nicotiana/genética , Nicotiana/microbiología , Verticillium/metabolismo , Verticillium/patogenicidad
19.
J Exp Bot ; 66(17): 5315-25, 2015 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-26002971

RESUMEN

Plant elicitor peptides (Peps) are potent inducers of pattern-triggered immunity and amplify the immune response against diverse pathogens. Peps have been discovered and studied extensively in Arabidopsis and only recently orthologues in maize were also identified and characterized in more detail.Here, the presence of PROPEPs, the Pep precursors, and PEPRs, the Pep receptors, was investigated within the plant kingdom. PROPEPs and PEPRs were identified in most sequenced species of the angiosperms. The conservation and compatibility of the Pep-PEPR-system was analysed by using plants of two distantly related dicot families, Brassicaceae and Solanaceae, and a representative family of monocot plants, the Poaceae. All three plant families contain important crop plants, including maize, rice, tomato, potato, and canola. Peps were not recognized by species outside of their plant family of origin, apparently because of a divergence of the Pep sequences. Three family-specific Pep motifs were defined and the integration of such a motif into the Pep sequence of an unrelated Pep enabled its perception. Transient transformation of Nicotiana benthamiana with the coding sequences of the AtPEPR1 and ZmPEPR1a led to the recognition of Pep peptides of Brassicaceae or Poaceae origin, respectively, and to the proper activation of downstream signalling. It was concluded that signalling machinery downstream of the PEPRs is highly conserved whereas the leucine-rich repeat domains of the PEPRs co-evolved with the Peps, leading to distinct motifs and, with it, interfamily incompatibility.


Asunto(s)
Evolución Biológica , Brassicaceae/genética , Péptidos/genética , Proteínas de Plantas/genética , Poaceae/genética , Transducción de Señal , Solanaceae/genética , Brassicaceae/metabolismo , Evolución Molecular , Péptidos/metabolismo , Inmunidad de la Planta , Proteínas de Plantas/metabolismo , Poaceae/metabolismo , Solanaceae/metabolismo
20.
Nucleic Acids Res ; 43(Database issue): D1036-41, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25428362

RESUMEN

The Sol Genomics Network (SGN, http://solgenomics.net) is a web portal with genomic and phenotypic data, and analysis tools for the Solanaceae family and close relatives. SGN hosts whole genome data for an increasing number of Solanaceae family members including tomato, potato, pepper, eggplant, tobacco and Nicotiana benthamiana. The database also stores loci and phenotype data, which researchers can upload and edit with user-friendly web interfaces. Tools such as BLAST, GBrowse and JBrowse for browsing genomes, expression and map data viewers, a locus community annotation system and a QTL analysis tools are available. A new tool was recently implemented to improve Virus-Induced Gene Silencing (VIGS) constructs called the SGN VIGS tool. With the growing genomic and phenotypic data in the database, SGN is now advancing to develop new web-based breeding tools and implement the code and database structure for other species or clade-specific databases.


Asunto(s)
Bases de Datos de Ácidos Nucleicos , Genoma de Planta , Solanaceae/genética , Cruzamiento , Cruzamientos Genéticos , Genómica , Genotipo , Internet , Fenotipo , Solanaceae/metabolismo
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