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Over the last 10 years, global raspberry production has increased by 47.89%, based mainly on the red raspberry species (Rubus idaeus). However, the black raspberry (Rubus occidentalis), although less consumed, is resistant to one of the most important diseases for the crop, the late leaf rust caused by Acculeastrum americanum fungus. In this context, genetic resistance is the most sustainable way to control the disease, mainly because there are no registered fungicides for late leaf rust in Brazil. Therefore, the aim was to understand the genetic architecture that controls resistance to late leaf rust in raspberries. For that, we used an interspecific multiparental population using the species mentioned above as parents, 2 different statistical approaches to associate the phenotypes with markers [GWAS (genome-wide association studies) and copula graphical models], and 2 phenotyping methodologies from the first to the 17th day after inoculation (high-throughput phenotyping with a multispectral camera and traditional phenotyping by disease severity scores). Our findings indicate that a locus of higher effect, at position 13.3 Mb on chromosome 5, possibly controls late leaf rust resistance, as both GWAS and the network suggested the same marker. Of the 12 genes flanking its region, 4 were possible receptors, 3 were likely defense executors, 1 gene was likely part of signaling cascades, and 4 were classified as nondefense related. Although the network and GWAS indicated the same higher effect genomic region, the network identified other different candidate regions, potentially complementing the genetic control comprehension.
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Basidiomycota , Resistencia a la Enfermedad , Estudio de Asociación del Genoma Completo , Fenotipo , Enfermedades de las Plantas , Rubus , Resistencia a la Enfermedad/genética , Rubus/microbiología , Rubus/genética , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/genética , Sitios de Carácter Cuantitativo , Hojas de la Planta/microbiología , Hojas de la Planta/genética , Polimorfismo de Nucleótido Simple , Mapeo CromosómicoRESUMEN
ABSTRACTThis study evaluated the effects of Rubus sp. extract on behavioral and neurochemical parameters in female mice submitted to experimental model of depression induced by lipopolysaccharide (LPS). The results indicated that Rubus sp. extract protected against depressive-like behavior induced by LPS. Moreover, the administration of Rubus sp. extract was effective in preventing the increase in reactive species and nitrites levels, as well as the decrease in catalase activity induced by LPS in the cerebral cortex. In the serum, the Rubus sp. extract was effective in preventing the decrease in catalase activity induced by LPS. Treatment with Rubus sp. extract attenuated the increase in acetylcholinesterase activity induced by LPS in the cerebral cortex. Finally, blackberry extract also downregulated IL-1ß levels in cerebral cortex. In conclusion, our findings demonstrated that treatment with Rubus sp. exerted antidepressant, antioxidant, anticholinesterase and anti-inflammatory effects in a model of depressive - like behavior induced by LPS in female mice. This highlights Rubus sp. as a potential therapeutic agent for individuals with major depressive disorder.
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The research objective was to experimentally optimize the fluidized bed agglomeration process of an agglomerated blackberry powder mixture (ABPM) using the response surface methodology. As a raw material, a powdered mixture of blackberry from Castile (Rubus glaucus Benth) obtained by spray drying (SD) was used. In the evaluation of the agglomeration process, the response surface methodology was applied using a central design with a face-centered composition (α = 1), considering the independent variables: fluidisation air inlet temperature (T) (50-70 °C), the binder solution atomization air pressure (P) (1-2 bar) and process time (t) (20-35 min); and the dependent variable: moisture content (Xw), solubility (S), wettability (We), apparent density (ρa), total phenols (TP), radical scavenging (ABTS·+ and DPPH· methods), anthocyanins (Ant) (cyanidin-3-glucoside (C3G)), ellagic acid (EA) and vitamin C (Vit. C). In general, the ABPM exhibited higher porosity and particle size, which generated changes in S, We and ρa, and a better rehydration capacity of the ABPM. The optimal process conditions (T = 70 °C, P = 1.7 bar and t = 21.7 min) defined the most favourable attributes of the ABPM (Xw = 9.7 ± 0.1%, S = 74.9 ± 4.9%, We = 13.7 ± 3.6 min, ρa = 0.312 ± 0.009 g/mL, TP = 4084.6 ± 30.6 mg AGE/100g dry base (db), ABTS·+ = 4511.4 ± 124.5 mg TE/100 g db, DPPH· = 4182.7 ± 66.4 mg TE/100 g db, Ant = 213.6 ± 15.9 mg C3G/100 g db, EA = 1878.2 ± 45.9 mg/100 g db and Vit. C = 29.8 ± 7.4 mg/100 g db. The agglomeration process improved the instantaneous properties and the flow behaviour of the ABPM. Additionally, it offers significant nutritional value with potential use as an instant drink and raw material for the food industry.
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The raspberry (Rubus idaeus L.) fruit is characterized by its richness in functional molecules and high nutritional value, but the high rate of fruit softening limits its quality during postharvest. Raspberry drupelets have a particular ripening regulation, depending partially on the effect of ethylene produced from the receptacle. However, the possible role of abscisic acid (ABA) in the modulation of quality parameters during the ripening of raspberry is unclear. This study characterized the fruit quality-associated parameters and hormonal contents during fruit development in two seasons. The quality parameters showed typical changes during ripening: a drastic loss of firmness, increase in soluble solids content, loss of acidity, and turning to a red color from the large green stage to fully ripe fruit in both seasons. A significant increase in the ABA content was observed during the ripening of drupelets and receptacles, with the higher content in the receptacle of ripe and overripe stages compared to the large green stage. Moreover, identification of ABA biosynthesis-(9-cis-epoxycarotenoid dioxygenase/NCED) and ABA receptor-related genes (PYRs-like receptors) showed three genes encoding RiNCEDs and nine genes for RiPYLs. The expression level of these genes increased from the large green stage to the full-ripe stage, specifically characterized by a higher expression of RiNCED1 in the receptacle tissue. This study reports a consistent concomitant increase in the ABA content and the expression of RiNCED1, RiPYL1, and RiPYL8 during the ripening of the raspberry fruit, thus supporting the role for ABA signaling in drupelets.
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Plants with medicinal potential may also produce adverse effects in humans. This seems to be the case for the species Rubus rosifolius, where preliminary studies demonstrated genotoxic effects attributed to extracts obtained from leaves and stems of this plant using on HepG2/C3A human hepatoma cells as a model. Considering the beneficial properties of this plant as an antidiarrheal, analgesic, antimicrobial, and antihypertensive and its effects in the treatment of gastrointestinal diseases, the present study was developed with the aim of determining the cytotoxic and genotoxic potential of extracts of leaves and stems of R. rosifolius in primary without metabolic competence in human peripheral blood mononuclear cells (PBMC). Cell viability analyses at concentrations of between 0.01 and 100 µg/ml of both extracts did not markedly affect cell viability. In contrast, assessment of the genotoxic potential using the comet assay demonstrated significant damage to DNA within PBMC from a concentration of 10 µg/ml in the stem extract, and a clastogenic/aneugenic response without cytokinesis-block proliferation index (CBPI) alterations at concentrations of 10, 20, or 100 µg/ml for both extracts. Under our experimental conditions, the data obtained demonstrated genotoxic and mutagenic effects attributed to extracts from leaves and stems of R. rosifolius in cells in the absence of hepatic metabolism.
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Leucocitos Mononucleares , Rubus , Humanos , Extractos Vegetales/toxicidad , Pruebas de Micronúcleos , Ensayo Cometa , Daño del ADN , Mutágenos , Hojas de la PlantaRESUMEN
Berry consumption is increasing worldwide due to their high content of bioactive compounds. However, such fruits have a very short shelf life. To avoid this drawback and to offer an effective alternative for its consumption at any time of the year, an agglomerated berry powder mix (APB) was developed. The aim of this work was to evaluate the stability of APB during a 6-months-period storage at 3 temperatures. The stability of APB was determined by moisture, aw, antioxidant activity, total phenolics, total anthocyanins, vitamin C, color, phenolic profiles, and MTT assay. APB showed differences in antioxidant activity between 0 and 6 months. It experimented non-enzymatic browning, which was more remarkable at 35 °C. APB at time 0 exhibited growth inhibitory effects against HT-29 human cancer cells. Most properties were significantly modified by storage temperature and time, which induces a significant decreasing of bioactive compounds.
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Late leaf rust is a fungal disease in raspberries caused by Aculeastrum americanum (Farl.) M. Scholler U. Braun (syn. Thekopsora americana (Farl.) Aime McTaggart) leading to early defoliation and yield losses. Red raspberries (Rubus idaeus L.) are susceptible to this pathogen, although this susceptibility varies among cultivars. In contrast, black raspberries were previously reported to be more resistant (Rubus occidentalis L.) and immune (Rubus niveus Thunb.) to this pathogen, raising their importance in plant breeding programs. However, what features make them respond differently to the same pathogen? In this study, we characterize for the first time the pre- and post-formed structural and biochemical defense mechanisms of R. idaeus cv. Autumn Bliss, R. occidentalis and R. niveus. Ultrastructural and histopathological analyses were used to uncover the interactions between these raspberries and A. americanum. The ultrastructural results indicate that the pathogen germinates on both leaf surfaces but can only form appressoria on the stomata. Although the three raspberry species were infected and colonized by A. americanum, a clear difference in susceptibility was observed between them. A compact mesophyll, pre- and post-formed phenolic compounds, and post-formed pectic compounds were the main plant defense mechanisms against fungal colonization. These findings provide new information about raspberries' defense mechanisms in response to A. americanum and elucidate the interactions occurring in these pathosystems.
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Background: Today, cardiovascular, oncological, and neurodegenerative diseases are the main causes of death in the world, according to official World Health Organization (WHO) statistics. Antioxidants are used to treat and prevent these diseases. In order to develop optimal technology for obtaining drugs based on plant extracts with antioxidant action, it is necessary to determine the total antioxidant capacity of raspberry shoots. Objectives: The study aimed to determine the total antioxidant capacity of red raspberry shoots, study the content of biologically active substances (BAS), and the antioxidant activity of red raspberry shoot extracts obtained during subsequent exhaustive extraction. Methods: The number of phenolic compounds, catechins, flavonoids, and hydroxycinnamic acids was determined by a spectrophotometric analysis method, whereas organic acids were determined by the alkalimetric method in red raspberry shoot extracts; the antioxidant activity of obtained extracts was evaluated by potentiometric method. Results: The total antioxidant capacity of red raspberry shoots was 164.12 mmol-equiv./m dry weight, the sum of the total content of phenolic compounds was 24.40 mg gallic acid (GA)/mL, catechins 21.36 mg epigallocatechin-3-O-gallate (EGCG)/mL, flavonoids 0.77 mg rutin (R)/mL, hydroxycinnamic acids derivatives 2.56 mg chlorogenic acid (ChA)/mL and organic acids 1.88 mg citric acid (CA)/mL in red raspberry shoot extracts obtained during subsequent exhaustive extraction. The analysis showed that there is a very high positive correlation between antioxidant activity and total phenolic compounds, catechin, flavonoid, hydroxycinnamic acids derivatives, and organic acids content in red raspberry shoot extracts. Conclusions: Total red raspberry shoots' antioxidant capacity has been determined. The study results can be used to develop optimal technology for obtaining drugs based on the extract of red raspberry shoots, which has an antioxidant effect
Contexto: Hoy en día, las enfermedades cardiovasculares, oncológicas y neurodegenerativas son las principales causas de muerte en el mundo según estadísticas oficiales de la Organización Mundial de la Salud OMS. Los antioxidantes se utilizan para tratar y prevenir estas enfermedades. Para desarrollar una tecnología óptima para la obtención de fármacos a base de extractos de plantas con acción antioxidante, es necesario determinar la capacidad antioxidante total de los brotes de frambuesa.Objetivos: El estudio tuvo como objetivo determinar la capacidad antioxidante total de los brotes de frambuesa roja, estudiar el contenido de sustancias biológicamente activas (SBA) y la actividad antioxidante de los extractos de brotes de frambuesa roja obtenidos mediante extracción exhaustiva. Métodos: La cantidad de compuestos fenólicos, catequinas, flavonoides y ácidos hidroxicinámicos se determinó por método de análisis espectrofotométrico, mientras que los ácidos orgánicos por método alcalimétrico en extractos de brotes de frambuesa roja; La actividad antioxidante de los extractos obtenidos se evaluó por método potenciométrico. Resultados: La capacidad antioxidante total de los brotes de frambuesa roja fue de 164.12 mmol-equiv./m de peso seco, la suma del contenido total de compuestos fenólicos fue de 24.40 mg gálico ácido (GA)/mL, catequinas 21.36 mg epigalocatequina-3-O-galato (EGCG)/mL, flavonoides 0.77 mg rutina (R)/mL, derivados de ácidos hidroxicinámicos 2.56 mg clorogénico ácido (ChA)/mL y ácidos orgánicos 1.88 mg cítrico ácido (CA)/mL en extractos de brotes de frambuesa roja obtenidos durante extracción exhaustiva. La correlación analizada mostró que existe una correlación positiva entre la actividad antioxidante y el contenido de compuestos fenólicos totales, catequinas, flavonoides, derivados de ácidos hidroxicinámicos y ácidos orgánicos en extractos de brotes de frambuesa roja. Conclusiones: Gracias a nuestros resultados se ha determinado la capacidad antioxidante total de los brotes de frambuesa roja. Los resultados del estudio se pueden utilizar para desarrollar una tecnología óptima para la obtención de fármacos basados en el extracto de brotes de frambuesa roja, que tiene un efecto antioxidante
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Humanos , Antioxidantes , Fenoles , Extracción Seriada , Ácidos Orgánicos , Correlación de DatosRESUMEN
Rubus rosifolius, popularly known as "red mulberry", is a common medicinal plant in southern Brazil and is used as an antidiarrheal, analgesic, antimicrobial and antihypertensive, and to treat stomach diseases. The aim of this study was to analyze the R. rosifolius stem extract (RrSE) for possible in vitro cytotoxic and genotoxic effects, using the comet assay and the micronucleus test to assess genotoxicity, and flow cytometry to assess the impact on the cell cycle and apoptosis in HepG2/C3A cells, in addition to evaluating the expression of genes linked to the induction of DNA damage, cell cycle, apoptosis and metabolism of xenobiotics. The MTT assay observed no cytotoxic effects at concentrations between 0.01 and 100 µg/mL of the extract. However, genotoxic effects occurred in treatments with the extract from a 1 µg/mL concentration. Flow cytometry analysis revealed a significant increase in cells in the G2/M phase after treatment with 10 µg/mL, a decrease in cells in the G0/G1 phase in the treatment with 100 µg/mL, and a significant increase in total apoptotic cells. In the gene expression analysis, an increase in the CYP1A2 xenobiotics metabolizing gene expression was observed. Despite the promising pharmacological effects of R. rosifolius, the results revealed that the RrSE has genotoxic effect and induces apoptosis in HepG2/C3A cells, indicating danger in using this plant extract by humans.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , Rubus , Humanos , Apoptosis , Daño del ADN , Extractos Vegetales/toxicidad , Extractos Vegetales/análisis , Células Hep G2 , Línea CelularRESUMEN
Blackberry fruits are appreciated as a source of nutrients and compounds related to benefit human health. However, they are highly perishable and very susceptible to decay factors. Current methods to improve and maintain blackberry quality are limited in use because of the fruit's fragile physical properties. Regarding these properties, it has been reported that the activities of certain enzymes are linked to senescence and fruit softening processes. This study was aimed to assess the effect of salicylic acid (SA) and chitosan (COS) as preharvest treatments on the physiology related to improving fruit conservation and preserving the marketability index of blackberry fruit. The preharvest treatments were foliar sprayed on blackberry plants at different concentrations. The activities of enzymes superoxide dismutase (SOD), catalase (CAT), phenylalanine ammonia-lyase (PAL), and polygalacturonase (PG) were measured. Total soluble solids (TSS), titratable acidity (TA), TSS/TA ratio, and marketability index (MI) were analyzed after 144 h of storage. The application of 3 mM of SA and 0.25% of COS treatments preserved the MI of blackberries by reducing leakage, red drupelet reversion (RDR), and mycelium presence in the fruit. SA application increased SOD, CAT, and PAL activities. Our results also showed that SA and COS preharvest treatments modified the activity of the cell wall degrading enzyme PG, which might play a role in improving the shelf life and resistance to decay factors of blackberry fruit without any significant effects on physicochemical properties like TSS, TA, and the TSS/TA ratio.
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Fusarium wilt of blackberry (FWB) is an emerging disease caused by a Fusarium oxysporum species complex. More than 3,000 ha of blackberry (Rubus spp.) crops have been lost in Mexico since 2011. The objectives of this research were: to evaluate the sensitivity of pathogenic F. oxysporum isolates recovered from symptomatic blackberry plants to fungicides with different modes of action; to assess the potential of these fungicides and plant resistance inducers against FWB in the greenhouse; and to determine the effects of commercial biofungicides and two indigenous strains of Trichoderma spp. on the incidence of FWB. The EC50 values of the fungicides prochloraz, thiabendazole, azoxystrobin, thiophanate-methyl, difenoconazole, triflumizole, and potassium phosphite for six pathogenic F. oxysporum isolates were determined. In a separate experiment, the fungicides acibenzolar-s-methyl (ASM), potassium phosphite, and commercial biofungicides, as well as two soil microbial inoculants and two indigenous Trichoderma strains, were tested for protection against wilt development in blackberry plants in the greenhouse. Prochloraz showed an average sensitivity for EC50 of 0.01 µg ml-1 for the tested F. oxysporum isolates, followed by difenoconazole and thiabendazole. Prochloraz and ASM proved to be the most effective treatments in the greenhouse. In contrast, potassium phosphite was ineffective in both the in vitro and in vivo experiments. The soil bioinoculants MicroSoil, Baktillis, T. koningiopsis, and T. asperellum significantly reduced the incidence of disease in the greenhouse. These results provide evidence for the potential of the various tools as useful components of integrated FWB management in the field.
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Fungicidas Industriales , Fusarium , Rubus , Trichoderma , Fungicidas Industriales/farmacología , Enfermedades de las Plantas/prevención & control , Suelo , TiabendazolRESUMEN
Caracterização da ação local do Rubus coreanus através da funcionalização da superfície dos implantes a serem instalados. Realizado em 2 etapas. Na primeira etapa, foi realizada a caracterização da superfície de implantes funcionalizadas pelos fármacos, a partir de teste físico químico e testes biológicos em culturas de células. Para a cultura de células foram utilizadas células mesenquimais indiferenciadas, isoladas de fêmures de ratas. Foram avaliadas a atividade e a diferenciação celular. Posteriormente, foi realizada a segunda etapa onde foram executados os experimentos in vivo. Para tanto, foram utilizadas 80 ratas Wistar adultas jovens, fêmeas, sendo 32 ratas para o primeiro período de eutanásia (14 dias), e 48 ratas para o segundo período de eutanásia (28 dias). Os grupos experimentais são: SHAM CONV, SHAM DMSO, SHAM RC 200, SHAM RC 400, OVX CONV, OVX DMSO, OVX RC 200 e OVX RC 400, sendo que os animais OVX foram submetidos a ovariectomia bilateral, enquanto que os animais SHAM foram submetidos à cirurgia fictícia de ovariectomia bilateral. Passados 30 dias, todos animais foram submetidos à cirurgia de instalação dos implantes nas metáfises tibiais. Os implantes tiveram as superfícies tratadas com DMSO ou RC 200 ou RC 400, e houve o controle negativo realizado através dos implantes convencionais. A eutanásia dos animais do primeiro período de eutanásia aconteceu aos 14 dias após a instalação dos implantes e as tíbias foram destinadas para a análise biomecânica (contra-torque). Para os animais do segundo período de eutanásia, aos 28 dias após a instalação dos implantes foi realizada a eutanásia dos animais, e as tíbias foram destinadas para as seguintes análises: biomecânica (contra-torque), enquanto do lado contralateral foi realizada a microtomografia computadorizada para avaliação morfométrica do osso formado ao redor dos implantes instalados. Os dados quantitativos foram submetidos ao teste de homocedasticidade para a seleção do teste estatístico apropriado (paramétrico ou não paramétrico), com nível de significância de 5%. Através das análises biomecânica e microtomográfica foi possível verificar resultados semelhantes entre os grupos experimentais. Contudo, o rubus coreanus não foi capaz de atuar de forma positiva no processo de reparo ósseo perimplantar(AU)
Characterization of the local action of Rubus coreanus through the functionalization of the surface of the implants to be installed. Carried out in 2 steps. In the first stage, the characterization of the surface of implants functionalized by drugs was carried out, based on physical-chemical tests and biological tests on cell cultures. For cell culture, undifferentiated mesenchymal cells, isolated from femurs of rats, were used. Cell activity and differentiation were evaluated. Subsequently, the second stage was carried out, where the in vivo experiments were performed. For that, 80 young adult female Wistar rats were used, 32 rats for the first period of euthanasia (14 days), and 48 rats for the second period of euthanasia (28 days). The experimental groups are: SHAM CONV, SHAM DMSO, SHAM RC 200, SHAM RC 400, OVX CONV, OVX DMSO, OVX RC 200 and OVX RC 400, with the OVX animals undergoing bilateral ovariectomy, while the SHAM animals underwent submitted to sham surgery of bilateral ovariectomy. After 30 days, all animals underwent surgery to install the implants in the tibial metaphysis. The implants had their surfaces treated with DMSO or RC 200 or RC 400, and there was a negative control performed using conventional implants. The animals from the first period of euthanasia were euthanized 14 days after implant placement and the tibiae were destined for biomechanical analysis (counter-torque). For animals in the second period of euthanasia, 28 days after implant placement, the animals were euthanized, and the tibiae were destined for the following analyses: biomechanics (counter-torque), while on the contralateral side, computerized microtomography was performed for morphometric evaluation of the bone formed around the installed implants. Quantitative data were submitted to the homoscedasticity test to select the appropriate statistical test (parametric or non-parametric), with a significance level of 5%. Through biomechanical and microtomographic analyzes it was possible to verify similar results between the experimental groups. However, rubus coreanus was not able to act positively in the peri-implant bone repair process(AU)
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Animales , Ratas , Osteoporosis , Regeneración Ósea , Implantes Dentales , Oseointegración , Osteogénesis , Implantación Dental Endoósea , EstrógenosRESUMEN
Raspberries are important sources of bioactive compounds, whose synthesis is influenced by the fertilization system and the maturity stage. This study evaluated the effect of organic and conventional fertilization systems on raspberries at three maturity stages, pink, ripe, and overripe. Physicochemical characteristics, bioactive compounds (phenolic profile, vitamin C), antioxidant capacity (DPPH, FRAP, TEAC, and ORAC), phenolic-associated enzyme, phenylalanine ammonia lyase (PAL), and antioxidant enzymes (SOD, CAT, GPx, and APX) were evaluated. The physicochemical determination of the fruit did not reveal differences between fertilization systems. Regarding bioactive compounds, higher content of anthocyanins was found in organic raspberries at all maturity stages. Organic fertilization increased the content of ellagic acid and gallic acid at all stages of maturity. Higher content of caffeic, hydroxybenzoic, protocatechuic acid, and vitamin C was observed in organic raspberry at the overripe stage. Raspberries grown with organic fertilization exhibited higher values of antioxidant capacity by the DPPH, FRAP, and TEAC methods at all maturity stages. Raspberries under organic fertilization showed significantly greater activity of CAT, SOD, APX, GPX, and PAL. The present study suggests that organic fertilization induces oxidative stress causing an increase in antioxidant defense mechanisms, enhancing bioactive compound production, and improving antioxidant capacity in raspberries.
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Fusarium oxysporum f. sp. mori, the causal agent of Fusarium wilt of blackberry, was first reported in California and Mexico in 2016. A limited survey of the population revealed this pathogen to be one of the most diverse formae speciales of F. oxysporum. We explored the possibility that strains of F. oxysporum pathogenic to commercial blackberry could also be recovered from wild blackberry (Rubus spp.) in California. For this purpose, wild Rubus species in blackberry nurseries, fruit production fields, and nearby areas were collected between 2017 and 2019. Thirty-four isolates of F. oxysporum were recovered from asymptomatic Rubus armeniacus and Rubus ursinus plants. Based on sequence of the translation elongation factor 1-α, somatic compatibility, and pathogenicity to blackberry, 16 isolates were confirmed as F. oxysporum f. sp. mori. These isolates were associated with three somatic compatibility groups, one of which was first identified in this study. Recovery of the pathogen confirmed that wild blackberry plants can act as a reservoir of inoculum of F. oxysporum f. sp. mori and that it can move from wild blackberry plants to commercial cultivars or vice versa.
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Fusarium , Rubus , California , México , Enfermedades de las PlantasRESUMEN
Trichoderma is used as a biocontrol agent against different plant pathogens in different crops. In Costa Rica, Trichoderma isolates from blackberry fruits (Rubus adenotrichos Schltdl.) have shown antagonism in laboratory and field trials against Botrytis cinerea. Quantifying fungal antagonistic activity directly on target organs or target tissues is of interest to estimate the performance of biocontrol agents. However, this is difficult due to the lack of visual manifestations of fungal structures. As part of a larger study to quantify antagonistic activity by quantitative PCR, we detail here a method to isolate and purify each fungus and then inoculate or co-inoculate them onto surface-sterilized blackberry fruits. ⢠A procedure to co-inoculate the surfaces of blackberry fruits with monosporic fungal suspensions for molecular analyses is described. ⢠The protocol described herein was implemented for subsequent qPCR analysis.
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Andean blackberries (Rubus glaucus Benth) are fruits rich in phytocomponents with high antioxidant activity. In this work, the changes in the total polyphenol content (TPC), the total flavonoid content (TFC), and the total anthocyanin content (TAC) of four blackberry varieties at three maturity stages (E1-25%, E2-50%, and E3-100%) were measured. The antioxidant activity (AA) was evaluated using the 2,2'azinobis-(3-ethylbenzthiazolin 6-sulphonic acid (ABTS) and ferric reducing antioxidant power (FRAP) methods. TPC and TFC content decreased with the increase in the maturity stage. The blackberry Brazos cultivar presented TPC values of 51.26, 38.16, and 31.59 mg of gallic acid equivalents (GAE)/g dry weight (DW) at E1, E2, and E3, respectively. The TAC and soluble solids increased with the increase in the maturity stage of the fruits. The Andimora variety at E3 presented a high TPC content, and the Colombiana variety presented a high TFC content. The blackberry Colombiana cultivar presented TAC values of 1.40, 2.95, and 12.26 mg cy-3-glu/100g DW at E1, E2, and E3, respectively. The blackberry Colombiana cultivar presented a high AA value at 1278.63 µmol TE/g DW according to the ABTS method and 1284.55 µmol TE/g DW according to the FRAP method. The TPC and TFC showed a high correlation with the AA according to the ABTS and the FRAP methods. The Pearson correlation between the TFC and AA/ABTS has a value of r = 0.92. The TFC and AA/FRAP present a value of r = 0.94.
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Raspberry (Rubus sp.) is a berries fruit with an ongoing agricultural and commercial interest due to its high contents of flavonoids and nutrients beneficial for human health. The growing demand for raspberries is facing great challenges associated mainly with the dispersal of diseases, which produces a decrease in productivity and fruit quality. A broad range of genomic resources is available for other Rosaceae species; however, genomic resources for species of the Rubus genus are still limited. Here, we characterize the transcriptome of the Rubus idaeus (Var. Amira) in order to 1) provide clues in the transcriptional changes of R. idaeus against tomato ringspot virus (ToRSV); and 2) generate genomic resources for this economically important species. We generate more than 200 million sequencing reads from two mRNA samples of raspberry, infected and not infected by ToRSV, using Illumina technology. After de novo assembly, we obtained 68,853 predicted protein-coding sequences of which 71.3% and 61.3% were annotated using Gene Ontology and Pfam databases, respectively. Moreover, we find 2,340 genes with differential expression between raspberries infected and not infected by ToRSV. Analysis of these genes shows functional enrichments of the oxidation-reduction process, cell wall biogenesis, terpene synthase activity, and lyase activity. These genes could be involved in the raspberry immune response through the interaction of different metabolic pathways; however, this statement needs further investigations. Up-regulation of genes encoding terpene synthases, multicopper oxidases, laccases, and beta-glucosidases might suggest that these enzymes appear to be the predominant transcriptome immune response of R. idaeus against ToRSV. Furthermore, we identify thousands of molecular markers (i.e., SSRs and SNPs), increasing considerably the genomic resources currently available for raspberries. This study is the first report on investigating the transcriptional changes of R. idaeus against ToRSV.
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Oral glucose tolerance test on male Sprague-Dawley rats was done to determine the hypoglycemic effect of the n-hexane, ethyl acetate and methanol fruit extracts (50 mg/kg BW; oral administration) of two varieties of Rubus rosifolius. Metformin was used as a positive control (15 mg/kg BW; intravenous administration). The n-hexane extract was most potent and was investigated phytochemically to yield compound 1, a mixture of triacylglycerols. In its oxidized state compound 1 produced a significant hypoglycemic effect which was more effective than metformin for the first 30 min of the assay (p = .03) and not significantly different for up to 120 min. Mechanisms through which the oxidized triacylglycerol species could form were explored and presented. This is the first account of the hypoglycemic activity of R. rosifolius and it is also the first account of this activity being credited to compounds other than polyphenols and terpenes in Rubus plants. PRACTICAL APPLICATIONS: A significant percentage of the world's population is affected by diabetes and diabetes-related illnesses. One of the most popular antidiabetic drug on the market is metformin which is used to lower blood glucose concentrations. The findings of this study indicate that an oxidized mixture of triacylglycerols is more fast-acting than metformin for the first 30 min of an oral glucose tolerance test. This effect was also not significantly different from that of the popular drug up to 120 min. These results demonstrate the hypoglycemic activity of an oxidized R. rosifolius fruit extract and indicate its potential use in applications such as functional food product development and drug discovery. Therapeutic applications can also include the plant extract as a potential primary treatment or as adjunct therapy to conventional medications.
Asunto(s)
Hipoglucemiantes , Rubus , Animales , Frutas , Hipoglucemiantes/farmacología , Masculino , Extractos Vegetales/farmacología , Ratas , Ratas Sprague-DawleyRESUMEN
The aim of this study was to design a Trichoderma atroviride-specific qPCR oligo set, evaluate its specificity, and standardize a methodology that quantifies antagonism against Botrytis cinerea in blackberry fruits (Rubus adenotrichos Schltdl.). Primers and probe were designed based on the nuclear translation elongation factor 1-alpha (tef1-α) of T. atroviride. A commercial IGS-based oligo set was used to quantify B. cinerea. The specificity of the designed oligo set, along with ITS-based oligo sets, was assessed using other Trichoderma species and B. cinerea. Multiplex qPCR assays were performed using DNA from B. cinerea, T. atroviride, and blackberries inoculated with these fungi. Assays with the tef1-α oligo set showed high sensitivity and reproducibility. In inoculated fruits, T. atroviride and B. cinerea were quantified simultaneously, including in symptomless tissues. This work standardized a qPCR methodology that specifically targets a T. atroviride isolate. This newly-designed qPCR oligo set could be useful in future biological control programs.
RESUMEN
The blackberry is a fragile fruit with a high degree of decomposition, which limits its shelf life. The effect of an edible coating (EC) based on cassava starch, whey protein, beeswax, chitosan, glycerol, stearic acid, and glacial acetic acid on the shelf life of fruit stored at 4 °C was evaluated. The physical, chemical, physical, microbiological, and sensorial quality was evaluated, comparing with a fresh control fruit. The EC had a positive effect on the physicochemical and sensorial properties (mainly in texture, flavor, and aromas), due to the reduction of physiological processes, whereas the color changes are mainly due to anthocyanin losses. After 10 days of storage, weight losses were 39.6% lower and firmness was 81.4% higher; while chitosan reduced the mold and yeast count. The EC increased the useful life of the Andean blackberries by 100%.