Avaliação da produção e concentração viral em pimentão infectado por tospovírus com o uso de piraclostrobina + metiram

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influence of Osmotic Dehydration on Ascorbic Acid Loss in Pickled Dry Peppers (Capsicum chinense)

The objective of this work was (1) to develop a dehydrated pepper with 45% humidity, determining the drying curves for pepper, with and without osmotic pre-treatment and (2) to evaluate the influence of both drying and osmotic treatment on the content ascorbic acid (vitamin C) in fresh pepper and pepper with 45% humidity. The experiments were carried out using the peppers cut in half, with and without osmotic pre-treatment, followed by drying in an oven at 70 degrees C. The results showed that the osmotic pretreatment did not influence the retention of ascorbic acid during the drying of pepper. The sensory analysis regarding the color, flavor, and texture attributes revealed that there was no difference in the acceptability.

Digeribilità e bioaccessibilità di composti bioattivi in diverse cultivar di peperoni dolci

Un discreto numero di molecole biologicamente attive contenute nei cibi vegetali si suppone esercitino un ruolo preventivo e favorevole su molteplici funzioni dell’organismo, con meccanismi d’azione spesso legati alla modulazione diretta e indiretta dello stress ossidativo. Acido ascorbico, tocoferoli, carotenoidi, polifenoli, posseggono attività antiossidante e giocano un ruolo positivo nella conservazione dello stato di salute. L’elevato contenuto di essi nei peperoni dolci (Capsicum Annuum L.) ha incrementato l’interesse nei confronti di questi vegetali da parte del settore agronomico e dell’industria alimentare. È tuttavia noto che la concentrazione di composti bioattivi può essere molto diversa anche tra cultivar della stessa specie vegetale ed è pertanto importante evidenziare il contenuto quali-quantitativo delle varie molecole nelle diverse cultivar di peperoni dolci, in modo da evidenziare le più ricche di tali componenti. Occorre però tenere conto anche della biodisponibilità e bioaccessibilità dei diversi componenti funzionali. Infatti il possibile effetto positivo di tali molecole non dipende solo dal loro contenuto nell’alimento ma soprattutto dalla quantità che viene rilasciata dalla matrice alimentare durante il processo digestivo, e che quindi risulta essere potenzialmente biodisponibile e attivo nell’organismo. Scopo della ricerca presentata è stato valutare e confrontare la digeribilità e la bioaccessibilità di alcuni composti bioattivi antiossidanti in peperoni dolci rossi e gialli appartenenti a due diverse cultivar, Lamuyo e Corno di Toro. Il contenuto fenolico totale e di vitamina C, l’attività antiossidante totale sono stati determinati nei campioni di peperone digeriti in vitro, e comparati ai prodotti freschi, evidenziando differenze significative in termini di bioaccessibilità in particolare tra i peperoni rossi delle due cultivar. Sebbene il processo di digestione in vitro sia una simulazione parziale ed incompleta di quanto accade in vivo, la valutazione di un alimento dopo averlo sottoposto a tale processo rappresenta un importante progresso nello studio delle proprietà e del valore nutrizionale degli alimenti.

Functional analysis of the amine substrate specificity domain of pepper tyramine and serotonin N-hydroxycinnamoyltransferases.

Pepper (Capsicum annuum) serotonin N-hydroxycinnamoyltransferase (SHT) catalyzes the synthesis of N-hydroxycinnamic acid amides of serotonin, including feruloylserotonin and p-coumaroylserotonin. To elucidate the domain or the key amino acid that determines the amine substrate specificity, we isolated a tyramine N-hydroxycinnamoyltransferase (THT) gene from pepper. Purified recombinant THT protein catalyzed the synthesis of N-hydroxycinnamic acid amides of tyramine, including feruloyltyramine and p-coumaroyltyramine, but did not accept serotonin as a substrate. Both the SHT and THT mRNAs were found to be expressed constitutively in all pepper organs. Pepper SHT and THT, which have primary sequences that are 78% identical, were used as models to investigate the structural determinants responsible for their distinct substrate specificities and other enzymatic properties. A series of chimeric genes was constructed by reciprocal exchange of DNA segments between the SHT and THT cDNAs. Functional characterization of the recombinant chimeric proteins revealed that the amino acid residues 129 to 165 of SHT and the corresponding residues 125 to 160 in THT are critical structural determinants for amine substrate specificity. Several amino acids are strongly implicated in the determination of amine substrate specificity, in which glycine-158 is involved in catalysis and amine substrate binding and tyrosine-149 plays a pivotal role in controlling amine substrate specificity between serotonin and tyramine in SHT. Furthermore, the indisputable role of tyrosine is corroborated by the THT-F145Y mutant that uses serotonin as the acyl acceptor. The results from the chimeras and the kinetic measurements will direct the creation of additional novel N-hydroxycinnamoyltransferases from the various N-hydroxycinnamoyltransferases found in nature.

Medallones de carne vacuna : reducción del contenido de sal por incorporación de oleorresinas de especias

Para evaluar la posible disminución de la adición de cloruro de sodio, mediante el agregado de oleorresinas de especias, se elaboró una masa en la que se empleó 85 % de carne vacuna y 15 % de grasa. Se fraccionó en tres lotes, adicionándoles: 1000 mg/kg de oleorresina de Origanum x Majoricum, 750 mg/kg de la de Capsicum annum o de la de Acantholippia seriphioides, respectivamente. Luego se dividieron en cinco porciones iguales y se agregó sal, hasta alcanzar contenidos de 0.00 %; 0.25 %; 0.50 %; 0.75 % y 1.00 %. Se homogeneizaron y se elaboraron medallones de 100 g, que se cocinaron en horno hasta alcanzar una temperatura interna de 72 °C. Se realizó una evaluación sensorial con 15 jueces semientrenados, solicitándoles que asignaran puntajes, mediante escalas estructuradas de siete puntos e indicaran cuál/es rechazarían. Los puntajes asignados por el panel se sometieron a un análisis exploratorio de datos, a las pruebas de Page y de comparaciones múltiples. Los medallones adicionados con los tres tipos de oleorresinas para las dosis de sal ensayadas presentaron diferencias (= 0.05): las dosis de 0 y 1% fueron las menos aceptadas y la de 0.25 % fue la más aceptada. El 50 % de los jueces rechazó el medallón con 0 % de sal, para los tres tipos de oleorresinas. En las condiciones ensayadas, la incorporación de oleorresinas en dosis de 1000 mg/kg para orégano o de 750 mg/kg para pimiento y tomillo mendocino, permite formular medallones de carne vacuna, con bajo contenido de sal y alta aceptación.

Oidiopsis taurica : nuevo patógeno de pimiento (Capsicum annum L.) y ají (Capsicum frutescens L.) en Mendoza (Argentina)

En febrero de 1998 se observó en cultivos de pimiento y ají de Mendoza (Argentina) muerte anticipada de plantas con severos daños en la producción. La magnitud del problema motivó una prospección en los principales oasis de la provincia, a fin de describir la sintomatología, conocer su dispersión, estimar los daños causados y recolectar material para realizar estudios en laboratorio tendientes a confirmar su etiología. Se constató que la afección se encontraba distribuida en todos los oasis del centro y norte. La sintomatología en hojas se caracterizó por la presencia en la cara adaxial de manchas amarillentas, de forma circular de 15 a 20 mm de diámetro, con puntuaciones necróticas, con tendencia a presentarse en anillos concéntricos. En la cara abaxial, generalmente las lesiones eran acompañadas de un micelio blanco tenue. Posteriormente siguió una severa defoliación. La producción disminuyó notablemente y además en pimiento los frutos fueron afectados por escaldaduras de sol y perdieron su valor comercial. A partir de aislamientos de pimiento y ají, mantenidos en plantas de brincos (Impatiens balsamina L.), se inocularon plantas sanas y se logró reproducir la enfermedad en condiciones de invernáculo.

Control de la salinización del suelo mediante sistemas de drenes en producciones intensivas de cultivos bajo cubierta

Los suelos en los cuales se desarrolla la actividad hortícola en el Gran La Plata presentan como limitante natural restricciones en el drenaje debido a la presencia de elevada cantidad de arcillas desde superficie (expansivas en alta proporción). Estas características edáficas, junto a la calidad del agua de riego, y al excesivo aporte de enmiendas y fertilizantes, conducen a la ocurrencia de procesos degradativos tales como, salinización, alcalinización, anegamiento, mayor incidencia de plagas, y efectos sobre el cultivo. Se tuvo como objetivo estudiar la eficiencia de drenes subsuperficiales en el control de la salinidad-alcalinidad de un suelo Hapludert típico con cultivos protegidos, que presentaba en promedio pH 8,25; CE 4,45 dS.m-1; RAS 22, aunque con amplia variabilidad. Se comparan dos tratamientos con drenes (T1 10cm y T2 15cm de diámetro), un tratamiento con yeso (T3) y un testigo. Se evaluó el suelo y el cultivo de pimiento "Capsicum annuum" durante 3 años (2009-2011). Los drenes subsuperficiales condujeron a una homogenización de todo el sitio de estudio y evitaron la ocurrencia de pulsos salinos característicos de estos sistemas productivos, sin ocasionar cambios significativos en la RAS, la cual se mantuvo en los valores dados por el agua de riego. El diámetro del dren no influyó de una manera significativa. El yeso presentó mejoras temporales en la salinidad-alcalinidad, pero no condujo a una mejora permanente del sitio. El desarrollo fenológico y rendimiento del cultivo en los tratamientos fue mejor que en el testigo en los años evaluados. Los tratamientos con drenes se diferenciaron significativamente del yeso en 2009 (P 0,01). El distanciamiento entre drenes (5m), resultó suficiente como para mejorar todo el invernáculo (460m2).

Long-term foliar persistence and efficacy of spinosad against beet armyworm under greenhouse conditions.

BACKGROUND: The immediate lethality caused by spinosad has been widely studied on Spodoptera exigua (H ¿ ubner). However, long-term effects can also provide valuable information on insecticide toxic action. Here, the persistence of spinosad on Capsicum annuum L. foliage and the lethal and sublethal effects of greenhouse-aged foliar residues of this insecticide on third instars of S. exigua are reported. RESULTS: Foliage was collected at 0, 3, 5, 10, 20, 30, 40 and 50 days after application, and spinosad residues were measured. Residues decreased over time according to first-order kinetics. The average rate constant and half-life of disappearance were 4.44×10?3 and156 daysand5.80×10?3 and120 days for60and120 mg L?1 respectively. Larval mortalitygradually decreased, corresponding to the residues, but was still appreciable (35 and 65% for 60 and 120 mg L?1 respectively) when the larvae were fed with foliage collected 50 days after treatment. Subsequently, pupal development was reduced and varied between 20 and 60% and between 21 and 41% for 60 and 120 mg L?1, respectively, in all ages of leaf residues that were bioassayed. At all time points, the consumption rate by the larvae was reduced between 62 and 84% for both concentrations that were bioassayed. CONCLUSION: It is concluded that, under the present greenhouse conditions, the degradation of spinosad was slower than that reported by other authors in the field, and, because of that, its residues could cause lethal and sublethal effects to S. exigua larvae.

Red Bell Pepper Chromoplasts Exhibit in Vitro Import Competency and Membrane Targeting of Passenger Proteins from the Thylakoidal Sec and ΔpH Pathways but Not the Chloroplast Signal Recognition Particle Pathway1

Chloroplast to chromoplast development involves new synthesis and plastid localization of nuclear-encoded proteins, as well as changes in the organization of internal plastid membrane compartments. We have demonstrated that isolated red bell pepper (Capsicum annuum) chromoplasts contain the 75-kD component of the chloroplast outer envelope translocon (Toc75) and are capable of importing chloroplast precursors in an ATP-dependent fashion, indicating a functional general import apparatus. The isolated chromoplasts were able to further localize the 33- and 17-kD subunits of the photosystem II O2-evolution complex (OE33 and OE17, respectively), lumen-targeted precursors that utilize the thylakoidal Sec and ΔpH pathways, respectively, to the lumen of an internal membrane compartment. Chromoplasts contained the thylakoid Sec component protein, cpSecA, at levels comparable to chloroplasts. Routing of OE17 to the lumen was abolished by ionophores, suggesting that routing is dependent on a transmembrane ΔpH. The chloroplast signal recognition particle pathway precursor major photosystem II light-harvesting chlorophyll a/b protein failed to associate with chromoplast membranes and instead accumulated in the stroma following import. The Pftf (plastid fusion/translocation factor), a chromoplast protein, integrated into the internal membranes of chromoplasts during in vitro assays, and immunoblot analysis indicated that endogenous plastid fusion/translocation factor was also an integral membrane protein of chromoplasts. These data demonstrate that the internal membranes of chromoplasts are functional with respect to protein translocation on the thylakoid Sec and ΔpH pathways.

Molecular Characterization of an Arabidopsis Gene Encoding Hydroperoxide Lyase, a Cytochrome P-450 That Is Wound Inducible1

Hydroperoxide lyase (HPL) cleaves lipid hydroperoxides to produce volatile flavor molecules and also potential signal molecules. We have characterized a gene from Arabidopsis that is homologous to a recently cloned HPL from green pepper (Capsicum annuum). The deduced protein sequence indicates that this gene encodes a cytochrome P-450 with a structure similar to that of allene oxide synthase. The gene was cloned into an expression vector and expressed in Escherichia coli to demonstrate HPL activity. Significant HPL activity was evident when 13S-hydroperoxy-9(Z),11(E),15(Z)-octadecatrienoic acid was used as the substrate, whereas activity with 13S-hydroperoxy-9(Z),11(E)-octadecadienoic acid was approximately 10-fold lower. Analysis of headspace volatiles by gas chromatography-mass spectrometry, after addition of the substrate to E. coli extracts expressing the protein, confirmed enzyme-activity data, since cis-3-hexenal was produced by the enzymatic activity of the encoded protein, whereas hexanal production was limited. Molecular characterization of this gene indicates that it is expressed at high levels in floral tissue and is wound inducible but, unlike allene oxide synthase, it is not induced by treatment with methyl jasmonate.

Dedicated Roles of Plastid Transketolases during the Early Onset of Isoprenoid Biogenesis in Pepper Fruits1

Isopentenyl diphosphate (IPP), which is produced from mevalonic acid or other nonmevalonic substrates, is the universal precursor of isoprenoids in nature. Despite the presence of several isoprenoid compounds in plastids, enzymes of the mevalonate pathway leading to IPP formation have never been isolated or identified to our knowledge. We now describe the characterization of two pepper (Capsicum annuum L.) cDNAs, CapTKT1 and CapTKT2, that encode transketolases having distinct and dedicated specificities. CapTKT1 is primarily involved in plastidial pentose phosphate and glycolytic cycle integration, whereas CapTKT2 initiates the synthesis of isoprenoids in plastids via the nonmevalonic acid pathway. From pyruvate and glyceraldehyde-3-phosphate, CapTKT2 catalyzes the formation of 1-deoxy-xylulose-5-phosphate, the IPP precursor. CapTKT1 is almost constitutively expressed during the chloroplast-to-chromoplast transition, whereas CapTKT2 is overexpressed during this period, probably to furnish the IPP necessary for increased carotenoid biosynthesis. Because deoxy-xylulose phosphate is shared by the plastid pathways of isoprenoid, thiamine (vitamin B1), and pyridoxine (vitamin B6) biosynthesis, our results may explain why albino phenotypes usually occur in thiamine-deficient plants.

Identification of a plastid protein involved in vesicle fusion and/or membrane protein translocation.

Structural evidence has accumulated suggesting that fusion and/or translocation factors are involved in plastid membrane biogenesis. To test this hypothesis, we have developed an in vitro system in which the extent of fusion and/or translocation is monitored by the conversion of the xanthophyll epoxide (antheraxanthin) into the red ketocarotenoid (capsanthin). Only chromoplast membrane vesicles from red pepper fruits (Capsicum annuum) contain the required enzyme. Vesicles prepared from the mutant yellow cultivar are devoid of this enzyme and accumulate antheraxanthin. The fusion and/or translocation activity is characterized by complementation due to the synthesis of capsanthin and the parallel decrease of antheraxanthin when the two types of vesicles are incubated together in the presence of plastid stroma. We show that the extent of conversion is dependent upon an ATP-requiring protein that is sensitive to N-ethylmaleimide. Further purification and immunological analysis have revealed that the active factor, designated plastid fusion and/or translocation factor (Pftf), resides in a protein of 72 kDa. cDNA cloning revealed that mature Pftf has significant homology to yeast and animal (NSF) or bacterial (Ftsh) proteins involved in vesicle fusion or membrane protein translocation.

Ocurrencia de mosca blanca (Bemisia tabaci Genn.) (hemíptera: Aleyrodidae) e incidencia de virosis en chiltoma (Capsicum annum L. ) con dos densidades de siembra, en condición semiprotegida

El objetivo de este estudio fue generar información sobre la ocurrencia poblacional de mosca blanca Bemisia tabaci Genn., e incidencia de virosis a través del tiempo en el cultivo de chiltoma Capsicum annuum L., con dos densidades de siembra en condición semiprotegida, en El Centro Experimental El Plantel. Se muestreó la población de mosca blanca desde los 30 hasta los 93 ddt y la incidencia de virosis desde los 34 hasta los 109 ddt; la rentabilidad del cultivo se evaluó por tratamiento realizando tres cortes cada 20 días después de los 70 ddt. Se detectó un incremento gradual de adultos y ninfas de mosca blanca e incidencia de virosis en el cultivo, con poblaciones más bajas durante la etapa de cimiento del cultivo de chiltoma, teniendo promedios más altos en el tratamiento de surcos sencillos, alcanzando su nivel crítico durante la etapa de floración y al finalizar la cosecha. El modelo epidémico apropiado para describir la curva de progreso de virosis resulto ser el monomolecular. Los parámetros de progreso de virosis, tasa y el Área Bajo la Curva de Progreso de virosis en los tratamientos de surcos sencillos y dobles no presentaron significancia estadísticas. La incidencia de virosis con la ocurrencia poblacional de mosca blanca mostró correlación positiva. En condición semiprotegida es conveniente sembrar con surcos dobles, ya que, la ocurrencia de mosca blanca e incidencia de virosis es menor, además resulta más rentable al momento de la cosecha. También el tratamiento de surcos sencillos fue dominado por el tratamiento de surcos dobles, el cual presentó una alta tasa de retorno marginal.

Calidad fisiológica de semillas de berenjena, morrón y tomate tratadas con tiametoxam, un producto bioactivador

Considerando la escasa información referente al efecto del bioactivador tiametoxam y los potenciales beneficios que este compuesto puede proporcionar en producción de semillas, el objetivo de este estudio fue evaluar la influencia del producto en el desempeño fisiológico de semillas de berenjena, morrón y tomate. Semillas de tres lotes de cada una de las especies fueron tratadas con seis dosis crecientes de tiametoxam (0; 0,2; 0,4; 0,6; 0,8 y 1,0 mL del producto por cada mil semillas) y sometidas a germinación, primer conteo de germinación, test de frío, test de envejecimiento acelerado, longitud total de las plántulas y emergencia de plántulas en invernadero. El bioactivador tuvo efecto positivo sobre los parámetros analizados, estimulando el desarrollo fisiológico de las semillas, con intensidades variables de acuerdo con el lote. En berenjena, el aumento medio porcentual de la germinación en el primer conteo tuvo un incremento de aproximadamente 11 %, en morrón de 13 % y para los lotes de tomate aproximadamente de 17 %. La dosis de 0,6 mL cada mil semillas resultó ser la más efectiva para aumentar el vigor de las semillas de las tres especies.

Transcriptome Analysis of Capsicum Chlorosis Virus-Induced Hypersensitive Resistance Response in Bell Capsicum

Background Capsicum chlorosis virus (CaCV) is an emerging pathogen of capsicum, tomato and peanut crops in Australia and South-East Asia. Commercial capsicum cultivars with CaCV resistance are not yet available, but CaCV resistance identified in Capsicum chinense is being introgressed into commercial Bell capsicum. However, our knowledge of the molecular mechanisms leading to the resistance response to CaCV infection is limited. Therefore, transcriptome and expression profiling data provide an important resource to better understand CaCV resistance mechanisms. Methodology/Principal Findings We assembled capsicum transcriptomes and analysed gene expression using Illumina HiSeq platform combined with a tag-based digital gene expression system. Total RNA extracted from CaCV/mock inoculated CaCV resistant (R) and susceptible (S) capsicum at the time point when R line showed a strong hypersensitive response to CaCV infection was used in transcriptome assembly. Gene expression profiles of R and S capsicum in CaCV- and buffer-inoculated conditions were compared. None of the genes were differentially expressed (DE) between R and S cultivars when mock-inoculated, while 2484 genes were DE when inoculated with CaCV. Functional classification revealed that the most highly up-regulated DE genes in R capsicum included pathogenesis-related genes, cell death-associated genes, genes associated with hormone-mediated signalling pathways and genes encoding enzymes involved in synthesis of defense-related secondary metabolites. We selected 15 genes to confirm DE expression levels by real-time quantitative PCR. Conclusion/Significance DE transcript profiling data provided comprehensive gene expression information to gain an understanding of the underlying CaCV resistance mechanisms. Further, we identified candidate CaCV resistance genes in the CaCV-resistant C. annuum x C. chinense breeding line. This knowledge will be useful in future for fine mapping of the CaCV resistance locus and potential genetic engineering of resistance into CaCV-susceptible crops.