RESUMEN

Salt stress causes several physiological and biochemical disorders and impairs plant growth. However, adequate fertilization can improve the nutritional status and may reduce significantly the harmful effects caused by salt stress. From this perspective, this study aimed to evaluate the impact of different combinations of nitrogen, phosphorus and potassium fertilization on the antioxidant activity and accumulation of organic and inorganic solutes in West Indian cherry leaves, in the second year of production. The experimental design was in randomized blocks, with treatments distributed in a 10 × 2 factorial arrangement corresponding to ten fertilization combinations (FC) of NPK (FC1: 80-100-100%, FC2:100-100-100%, FC3:120-100-100%, FC4:140-100-100%, FC5:100-80-100%, FC6:100-120-100%, FC7:100-140-100%, FC8:100-100-80%, FC9:100-100-120%, and FC10:100-100-140% of the recommendation) and two levels of electrical conductivity of irrigation water (ECw) (0.6 and 4.0 dS m-1), with three replications. The multivariate analysis showed that irrigation with water of different electrical conductivities (0.6 and 4.0 dS m-1) resulted in different responses concerning the enzyme activity, production of organic compounds, and accumulation of inorganic solutes in the leaves. Under irrigation with low salinity water, there was greater accumulation of K+, soluble carbohydrates, and proline, and lower activity of antioxidative enzymes, especially SOD and APX. Under high salinity water, greater enzyme activity and higher concentrations of Na+ and Cl- were observed. The results indicate that the response of West Indian cherry to salinity was more towards redox homeostasis than osmotic homeostasis through the accumulation of compatible solutes. Fertilization combination FC5 (100-80-100% corresponding to 200, 24 and 80 g plant-1 of NPK) modulates the enzyme activity of SOD and APX attenuating the impacts of salinity, being an efficient combination to preserve redox homeostasis in West Indian cherry plants grown under salt stress.

Asunto(s)

Fertilizantes , Potasio , Salinidad , Fertilizantes/análisis , Potasio/análisis , Potasio/metabolismo , Antioxidantes/metabolismo , Fósforo/análisis , Nitrógeno/metabolismo , Hojas de la Planta/química , Hojas de la Planta/efectos de los fármacos , Prunus avium/efectos de los fármacos , Prunus avium/enzimología , Estrés Salino

RESUMEN

The exploitation of plant genetic resources is an important and rapid strategy to release commercial cultivars. In this study, 234 sour cherry genotypes were collected from various locations of Iran and phenotypically assessed according to IPGRI and UPOV descriptors. The genotypes were grafted onto Mahaleb rootstock and were planted in Horticultural Science Research Institute (HSRI) core collection in Karaj, Iran. In this study, 22 different characteristics were measured in the sour cherry genotypes. The results showed that fruit and stone weights varied from 1.65 (G410) to 5.47 g (G125) and 0.13 (G428) to 0.59 g (G149), respectively. The fruit size index comprised average fruit length, width, and diameter, which varied from 10.57 to 19.13. The stalk length was less than 50 mm in 90.6% of the studied genotypes. Twelve of the 234 studied genotypes did not exhibit any symptoms of bacterial canker disease. Principle component analysis (PCA) and cluster analysis classified the studied genotypes into four main groups. Spearman's correlation analysis revealed that fruit size, stone shape, stone size, stalk thickness and weight, and fruit appearance correlated positively with stone and fruit weights. In contrast, fruit juice, fruit skin, and flesh color correlated negatively with the stone and fruit weights. The range of TSS varied between 12.66 (G251) and 26 (G427). Variations in pH value were between 3.66 (G236) and 5.63 (G352). In conclusion, a high level of genetic diversity was observed among the Iranian sour cherry genotypes. This diversity can be considered valuable and applicable for future breeding programs.

Asunto(s)

Prunus avium , Irán , Fitomejoramiento , Análisis por Conglomerados , Variación Genética/genética

RESUMEN

Aquaporins (AQPs) are integral transmembrane proteins well known as channels involved in the mobilization of water, small uncharged molecules and gases. In this work, the main objective was to carry out a comprehensive study of AQP encoding genes in Prunus avium (cv. Mazzard F12/1) on a genome-wide scale and describe their transcriptional behaviors in organs and in response to different abiotic stresses. A total of 28 non-redundant AQP genes were identified in Prunus spp. Genomes, which were phylogenetically grouped into five subfamilies (seven PIPs, eight NIPs, eight TIPs, three SIPs and two XIPs). Bioinformatic analyses revealed a high synteny and remarkable conservation of structural features among orthologs of different Prunus genomes. Several cis-acting regulatory elements (CREs) related to stress regulation were detected (ARE, WRE3, WUN, STRE, LTR, MBS, DRE, AT-rich and TC-rich). The above could be accounting for the expression variations associated with plant organs and, especially, each abiotic stress analyzed. Gene expressions of different PruavAQPs were shown to be preferentially associated with different stresses. PruavXIP2;1 and PruavXIP1;1 were up-regulated in roots at 6 h and 72 h of hypoxia, and in PruavXIP2;1 a slight induction of expression was also detected in leaves. Drought treatment strongly down-regulated PruavTIP4;1 but only in roots. Salt stress exhibited little or no variation in roots, except for PruavNIP4;1 and PruavNIP7;1, which showed remarkable gene repression and induction, respectively. Interestingly, PruavNIP4;1, the AQP most expressed in cherry roots subjected to cold temperatures, also showed this pattern in roots under high salinity. Similarly, PruavNIP4;2 consistently was up-regulated at 72 h of heat and drought treatments. From our evidence is possible to propose candidate genes for the development of molecular markers for selection processes in breeding programs for rootstocks and/or varieties of cherry.

Asunto(s)

Acuaporinas , Prunus avium , Prunus , Prunus avium/genética , Fitomejoramiento , Perfilación de la Expresión Génica , Prunus/genética , Estrés Fisiológico/genética , Acuaporinas/genética , Acuaporinas/metabolismo

RESUMEN

This study aimed to assess dark sweet cherry (DSC) total polyphenols (WE) and anthocyanins (ACN) against metastatic breast cancer (BC). The WE and ACN anticancer activity and underlying mechanisms were assessed in vitro using 4T1 BC cells. A pilot study using a BALB/C mouse syngeneic model bearing 4T1 tumors assessed the anti-metastatic potential of ACN in vivo. ACN inhibited cell viability with higher potency than WE and reduced reactive oxygen species (ROS) (IC50 = 58.6 µg cyanidin 3-glucoside equivalent (C3G)/mL or 122 µM). ACN induced p38 stress-related intrinsic apoptosis, leading to caspase-3 cleavage and total PARP decrease. ACN suppressed ERK1/2 and Akt/mTOR signaling pathways, which are abnormally activated in BC and promote motility and invasion. This was consistent with suppression of VCAM-1 mRNA, Scr phosphorylation and 88.6% reduction of cells migrating to wounded area. The pilot in vivo results supported the ACN-mediated suppression of angiogenesis in tumors and lungs. ACN also lowered Cenpf mRNA in lungs, associated with lung metastasis lesions and poor survival. Results demonstrated the dual Akt-ERK inhibitory role of ACN and suppression of their downstream pro-invasive targets. These results encourage a larger scale in vivo study to confirm that ACN may help to fight BC invasion and metastasis.

Asunto(s)

Prunus avium , Neoplasias de la Mama Triple Negativas , Animales , Humanos , Ratones , Antocianinas/farmacología , Antocianinas/metabolismo , Sistema de Señalización de MAP Quinasas , Ratones Endogámicos BALB C , Estrés Oxidativo , Proyectos Piloto , Proteínas Proto-Oncogénicas c-akt/metabolismo , Prunus avium/genética , ARN Mensajero/metabolismo , Transducción de Señal , Serina-Treonina Quinasas TOR/metabolismo

Asunto(s)

Actinidia , Prunus avium , Chile , Enfermedades de las Plantas , Pseudomonas syringae/genética

RESUMEN

Color acquisition is one of the most distinctive features of fruit development and ripening processes. The color red is closely related to the accumulation of polyphenolic compounds, mainly anthocyanins, during sweet cherry fruit maturity. In non-climacteric fruit species like sweet cherry, the maturity process is mainly controlled by the phytohormone abscisic acid (ABA), though other hormones may also play a role. However, the coordinated stage-specific production of polyphenolic compounds and their relation with hormone content variations have not been studied in depth in sweet cherry fruits. To further understand the accumulation dynamics of these compounds (hormones and metabolites) during fruit development, two sweet cherry cultivars ("Lapins" and "Glenred") with contrasting maturity timing phenotypes were analyzed using targeted metabolic analysis. The ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) approach revealed that phenolic acids, flavonols, and flavan-3-ols accumulated mainly until the straw-yellow stage in the early-maturing cultivar, while accumulation was mainly at the green stage in the mid-maturing cultivar, suggesting a cultivar-dependent stage-specific production of secondary metabolites. In the mid-maturing cultivar, anthocyanins were detected only from the red stage onward, whereas detection began at the pink stage in the early-maturing cultivar. ABA negatively correlated (p-value < 0.05) with the flavonols and flavan-3-ols in both cultivars. ABA and anthocyanin content increased at the same time in the early-season cultivar. In contrast, anthocyanins accumulated and the pink color initiation started several days after the ABA increase in the mid-maturing cultivar. Differential accumulation patterns of GA4, a ripening antagonizing hormone, between the cultivars could explain this difference. These results showed that both red-colored cultivars presented different accumulation dynamics of phenolic compounds and plant hormones during fruit development, suggesting underlying differences in the sweet cherry fruit color evolution.

Asunto(s)

Prunus avium , Antocianinas , Frutas , Hormonas , Espectrometría de Masas en Tándem

RESUMEN

Gibberellin (GA) negatively affects color evolution and other ripening-related processes in non-climacteric fruits. The bioactive GA, gibberellic acid (GA3), is commonly applied at the light green-to-straw yellow transition to increase firmness and delay ripening in sweet cherry (Prunus avium L.), though causing different effects depending on the variety. Recently, we reported that GA3 delayed the IAD parameter (a ripening index) in a mid-season variety, whereas GA3 did not delay IAD but reduced it at ripeness in an early-season variety. To further explore this contrasting behavior between varieties, we analyzed the transcriptomic responses to GA3 applied on two sweet cherry varieties with different maturity time phenotypes. At harvest, GA3 produced fruits with less color in both varieties. Similar to our previous report, GA3 delayed fruit color initiation and IAD only in the mid-season variety and reduced IAD at harvest only in the early-season variety. RNA-seq analysis of control- and GA3-treated fruits revealed that ripening-related categories were overrepresented in the early-season variety, including 'photosynthesis' and 'auxin response'. GA3 also changed the expression of carotenoid and abscisic acid (ABA) biosynthetic genes in this variety. In contrast, overrepresented categories in the mid-season variety were mainly related to metabolic processes. In this variety, some PP2Cs putative genes were positively regulated by GA3, which are negative regulators of ABA responses, and MYB44-like genes (putative repressors of PP2Cs expression) were downregulated. These results show that GA3 differentially modulates the transcriptome at the onset of ripening in a variety-dependent manner and suggest that GA3 impairs ripening through the modification of ripening associated gene expression only in the early-season variety; whereas in the mid-season variety, control of the ripening timing may occur through the PP2C gene expression regulation. This work contributes to the understanding of the role of GA in non-climacteric fruit ripening.

Asunto(s)

Giberelinas/metabolismo , Prunus avium/genética , Agricultura/métodos , Antocianinas/metabolismo , Secuencia de Bases/genética , Frutas/genética , Expresión Génica/genética , Perfilación de la Expresión Génica/métodos , Regulación de la Expresión Génica de las Plantas/genética , Giberelinas/farmacología , Ácidos Indolacéticos/metabolismo , Reguladores del Crecimiento de las Plantas/metabolismo , Proteínas de Plantas/genética , Prunus avium/metabolismo , Análisis de Secuencia de ARN/métodos , Factores de Transcripción/metabolismo , Transcriptoma/genética

RESUMEN

Calcium (Ca) is often supplied to crop species to prevent the occurrence of Carelated disorders. Mechanisms of Ca absorption and transport are not fully understood and the effectiveness of root and/or foliar Ca fertilization may be variable. To characterize the rate of Ca absorption and transport, trials were developed with chili pepper and sweet cherry plants, using 45CaCl2 as a tracer. The Ca treatments supplied were: (1) No 45Ca (control); (2) 45Ca soil application; (3) 45Ca supply to basal leaves, and (4) 45Ca application to apical leaves. After two months, plants were harvested for biomass and Ca content determination. The recovery of 45Ca in different plant parts was measured with a liquid scintillation counter and leaf traits were observed by scanning electronic microscopy. In general, the highest 45Ca concentrations were recovered in treated organs, while root applications led to highest 45Ca translocation rates, which varied between chili pepper and cherry plants. For chili pepper, 45Ca applied to the soil was detected mainly in roots (44 %) followed by leaves (36.6 %) stems (17.4 %) and fruits (2 %). In sweet cherry trees, soilapplied 45Ca was principally recovered in roots (45.3 %), shoots (28.5 %), leaves (14.3 %) and trunks (11.9 %). The results provide evidence of increased absorption of rootapplied Ca, as well as different degrees of Ca mobility between species. Foliar application led to major Ca increases in treated leaves, with Ca transported to other plant organs after apical leaf Ca supply chiefly in cherry trees.

Asunto(s)

Capsicum/efectos de los fármacos , Calcio/administración & dosificación , Calcio/análisis , Fertilizantes/análisis , Prunus avium/efectos de los fármacos

RESUMEN

Calcium (Ca) is often supplied to crop species to prevent the occurrence of Carelated disorders. Mechanisms of Ca absorption and transport are not fully understood and the effectiveness of root and/or foliar Ca fertilization may be variable. To characterize the rate of Ca absorption and transport, trials were developed with chili pepper and sweet cherry plants, using 45CaCl2 as a tracer. The Ca treatments supplied were: (1) No 45Ca (control); (2) 45Ca soil application; (3) 45Ca supply to basal leaves, and (4) 45Ca application to apical leaves. After two months, plants were harvested for biomass and Ca content determination. The recovery of 45Ca in different plant parts was measured with a liquid scintillation counter and leaf traits were observed by scanning electronic microscopy. In general, the highest 45Ca concentrations were recovered in treated organs, while root applications led to highest 45Ca translocation rates, which varied between chili pepper and cherry plants. For chili pepper, 45Ca applied to the soil was detected mainly in roots (44 %) followed by leaves (36.6 %) stems (17.4 %) and fruits (2 %). In sweet cherry trees, soilapplied 45Ca was principally recovered in roots (45.3 %), shoots (28.5 %), leaves (14.3 %) and trunks (11.9 %). The results provide evidence of increased absorption of rootapplied Ca, as well as different degrees of Ca mobility between species. Foliar application led to major Ca increases in treated leaves, with Ca transported to other plant organs after apical leaf Ca supply chiefly in cherry trees.(AU)

Asunto(s)

Fertilizantes/análisis , Calcio/administración & dosificación , Calcio/análisis , Prunus avium/efectos de los fármacos , Capsicum/efectos de los fármacos

RESUMEN

Abstract Sweet cherry fruit is a tasty and valuable product for consumers. In order to increase the export share of cherry, which is also very important in export, it is beneficial to grow with cherry varieties that mature at different times. The cherries offered to the market in the early period will be more attractive. In this study, morphological and biological features of pistils of early-maturing 'Cristalina', 'Early Lory', 'Prime Giant', fruit set rates and pollen germination status and some chemical applications were investigated. As a result, fruit sets of cultivars were 17.6-28.6% in two years. Significant differences were observed in pistil morphology of the cultivars and 'Cristalina' had shorter pistil (14.35-14.51 mm) and style (11.47-11.65 mm) lengths than the other cultivars. Greater deformation was observed in primary ovules of 'Early Lory' right after anthesis. There were not significant differences in pollen germination ratios of the cultivars, but boric acid treatments improved pollen germination ratios of all cultivars. Boric acid application increased pollen germination with 21%. This was followed by IAA (8%), GA3 (5%), KNO3 (4%). It was concluded based on present findings that in orchard establishment with the early cultivars, flower biology should momentously be assessed.

Asunto(s)

Flores , Óvulo Vegetal , Prunus avium , Polen

RESUMEN

In sweet cherry trees, flowering is commercially important because the flowers, after fertilization, will generate the fruits. In P. avium, the flowering induction and flower organogensis are the first developmental steps towards flower formation and they occur within specialized organs known as floral buds during the summer, nine months before blooming. During this period the number of floral buds per tree and the bud fruitfulness (number of flowers per bud) are stablished affecting the potential yield of orchards and the plant architecture. The floral bud development is sensitive to any type of stress and the hotter and drier summers will interfere with this process and are calling for new adapted cultivars. A better understanding of the underlying molecular and hormonal mechanisms would be of help, but unlike the model plant Arabidopsis, very little is known about floral induction in sweet cherry. To explore the molecular mechanism of floral bud differentiation, high-throughput RNA sequencing was used to detect differences in the gene expression of P. avium floral buds at five differentiation stages. We found 2,982 differentially expressed genes during floral bud development. We identified genes associated with floral initiation or floral organ identity that appear to be useful biomarkers of floral development and several transcription factor families (ERF, MYB, bHLH, MADS-box and NAC gene family) with novel potential roles during floral transition in this species. We analyzed in deep the MADS-box gene family and we shed light about their key role during floral bud and organs development in P. avium. Furthermore, the hormonal-related signatures in the gene regulatory networks and the dynamic changes of absicic acid, zeatin and indolacetic acid contents in buds suggest an important role for these hormones during floral bud differentiation in sweet cherry. These data provide a rich source of novel informacion for functional and evolutionary studies about floral bud development in sweet cherry and new tools for biotechnology and breeding.

Asunto(s)

Perfilación de la Expresión Génica/métodos , Proteínas de Plantas/metabolismo , Prunus avium/genética , Factores de Transcripción/metabolismo , Ácido Abscísico/metabolismo , Citocininas/metabolismo , Flores/genética , Flores/crecimiento & desarrollo , Flores/metabolismo , Regulación de la Expresión Génica de las Plantas , Biblioteca de Genes , Redes Reguladoras de Genes , Ácidos Indolacéticos/metabolismo , Filogenia , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Proteínas de Plantas/clasificación , Proteínas de Plantas/genética , Análisis de Componente Principal , Prunus avium/crecimiento & desarrollo , Prunus avium/metabolismo , RNA-Seq , Factores de Transcripción/clasificación , Factores de Transcripción/genética

RESUMEN

Sweet cherry is a valuable non-climacteric fruit with elevated phytonutrients, whose fruit quality attributes are prone to rapid deterioration after harvest, especially peel damage and water loss of stem. Here the metabolic and transcriptional response of exogenous melatonin was assessed in two commercial cultivars of sweet cherry (Santina and Royal Rainier) during cold storage. Gene expression profiling revealed that cuticle composition and water movement may underlie the effect of melatonin in delaying weight loss. An effect of melatonin on total soluble solids and lower respiration rate was observed in both cultivars. Melatonin induces overexpression of genes related to anthocyanin biosynthesis, which correlates with increased anthocyanin levels and changes in skin color (Chroma). Our results indicate that along with modulating antioxidant metabolism, melatonin improves fruit quality traits by triggering a range of metabolic and gene expression changes, which ultimately contribute to extend sweet cherry postharvest storability.

Asunto(s)

Regulación de la Expresión Génica/efectos de los fármacos , Melatonina/farmacología , Prunus avium/efectos de los fármacos , Antocianinas/metabolismo , Frutas/metabolismo , Prunus avium/metabolismo

RESUMEN

Resumen Objetivo: Evaluar mediante cuantificación de halos de inhibición el efecto antibacteriano de la cáscara y pulpa del capulí (Prunus serotina capulí) y del mortiño (Vaccinium floribundum), sobre cepas de Streptococcus mutans (ATCC 35668) a las 24 y 48 horas, comparado con arándano deshidratado y gluconato de clorhexidina al 0,12%. Materiales y métodos: Estudio experimental transversal in vitro, 15 cajas petri fueron utilizadas para sembrar 20ml de cultivo de cepas de Streptococcus mutans. En cada caja fueron colocados discos de fieltro impregnados con 20 μl de las sustancias evaluadas; mortiño y capulí, en pulpa y en cáscara, arándano deshidratado y gluconato de clorhexidina al 0,12% como control, distribuidos a una distancia equidistante. El análisis del efecto antibacteriano se realizó midiendo la zona de inhibición en un tiempo de 24 y 48 horas de incubación, los datos obtenidos se analizaron estadísticamente en el programa SPSS 22 mediante las pruebas paramétricas y de Kruskal Wallis. Resultados: No existió diferencia estadística significativa entre las variables analizadas, capulí y mortiño tanto en cáscara como en pulpa y clorhexidina empleada como control, en los dos períodos evaluados (p= > 0,05). Conclusiones: Los frutos rojos analizados tienen un efecto antibacteriano a las 24 y 48 horas, lo cual guarda relación con su capacidad antioxidante.

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Abstract Objective: To evaluate by quantification of halos of inhibition, the antibacterial effect of the shell and pulp of capulí, (Prunus serotina capuli) and mortiño (Vaccinium floribundum), on strains of Streptococcus mutans (ATCC 35668) at 24 and 48 hours, compared with dehydrated cranberry and chlorhexidine gluconate at 0,12%. Materials and methods: In vitro cross-sectional experimental study, 15 petri dishes were used to plant 20 ul of the evaluated substances were placed in each box, mortiño, and capuli, in pulp and in shell, dehydrated cranberry and 0,12% chlorhexidine gluconate as control, distributed at an equidistant distance. The analysis of the antibacterial effect was performed by measuring the zone of inhibition in a time of 24 and 48 hours of incubation, the dataobtained were statistically analyzed in the SPSS 22 program by parametric and Kruskal Wallis tests. Results: There was no significant statistical difference between the analyzed variables, capuli and mortiño, both in skin and pulp and chlorhexidine used as control, in the two evaluated periods of time (p=>0,05). Conclusions: The red fruits analyzed have an antibacterial effect 24 and 48 hours, which is related to its antioxidant capacity.

Asunto(s)

Streptococcus/efectos de los fármacos , Clorhexidina/uso terapéutico , Arándanos Azules (Planta)/inmunología , Caries Dental , Prunus avium/inmunología

RESUMEN

BACKGROUND: Sweet cherries are an excellent source of phenolic compounds, which may contribute to a healthy diet. The objective of this work was to generate dehydrated ingredients from postharvest discard of sweet cherries. RESULTS: Four dried ingredients were obtained from fresh sweet cherry discard (Lapins var.) using an osmotic dehydration pretreatment and freeze drying or air drying. The ingredients showed an important phenolic contribution (2.8-6.6 g gallic acid kg-1 of product) and preserved the natural color of the fruit to a great extent. Freeze-dried ingredients were less hygroscopic than air-dried ones, and presented with a softer texture. All the ingredients were in a supercooled state at room temperature (Tg range: -23.0 to -18.8 °C). Sugar infusion pretreatment caused a decrease in water sorption capacity and molecular mobility; it also reduced the initial rehydration rate. CONCLUSION: Relevant differences in nutritional and structural characteristics of the ingredients were observed depending on the processing method used. These ingredients could be incorporated into different processed foods, such as snacks, cereal mixtures, cereal bars, and bakery and confectionery products. Air-dried control ingredients presented better nutritional qualities and air-dried sweet cherries with sugar infusion pretreatment could be appropriate ingredients for applications where sweet flavor and slow rehydration rate are required. © 2018 Society of Chemical Industry.

Asunto(s)

Aromatizantes/análisis , Conservación de Alimentos/métodos , Frutas/química , Prunus avium/química , Color , Liofilización , Frutas/crecimiento & desarrollo , Ácido Gálico/análisis , Valor Nutritivo , Fenoles/análisis , Prunus avium/crecimiento & desarrollo

RESUMEN

Epigenetic modifications can yield information about connections between genotype, phenotype variation and environmental conditions. Bud dormancy release in temperate perennial fruit trees depends on internal and environmental signals such as cold accumulation and photoperiod. Previous investigations have noted the participation of epigenetic mechanisms in the control of this physiological process. We examined whether epigenetic modifications were modulated in MADS-box genes, potential candidates for the regulation of bud dormancy and flowering in sweet cherry (Prunus avium L.). We identified and cloned two MADS-box genes homologous to the already-characterized dormancy regulators DORMANCY-ASSOCIATED MADS-box (DAM3 and DAM5) from Prunus persica (L.) Batsch. Bisulfite sequencing of the identified genes (PavMADS1 and PavMADS2), Methylated DNA Immunoprecipitation and small RNA deep sequencing were performed to analyze the presence of DNA methylations that could be guided by non-coding RNAs in the floral buds exposed to differential chilling hours. The results obtained reveal an increase in the level of DNA methylation and abundance of matching small interference RNAs (siRNAs) in the promoter of PavMADS1 when the chilling requirement is complete. For the first intron and 5' UTR of PavMADS1, de novo DNA methylation could be associated with the increase in the abundance of 24-nt siRNA matching the promoter area. Also, in the second large intron of PavMADS1, maintenance DNA methylation in all cytosine contexts is associated with the presence of homologous siRNAs in that zone. For PavMADS2, only maintenance methylation was present in the CG context, and no matching siRNAs were detected. Silencing of PavMADS1 and PavMADS2 coincided with an increase in Flowering Locus T expression during dormancy. In conclusion, DNA methylations and siRNAs appear to be involved in the silencing of PavMADS1 during cold accumulation and dormancy release in sweet cherry.

Asunto(s)

Prunus avium/genética , Prunus avium/metabolismo , Metilación de ADN/genética , Metilación de ADN/fisiología , Epigénesis Genética/genética , Epigénesis Genética/fisiología , Flores/genética , Flores/metabolismo , Regulación de la Expresión Génica de las Plantas/genética , Regulación de la Expresión Génica de las Plantas/fisiología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo

RESUMEN

KEY MESSAGE: FT gene is expressed in leaves and buds and is involved in floral meristem determination and bud development in sweet cherry. In woody fruit perennial trees, floral determination, dormancy and bloom, depends on perception of different environmental and endogenous cues which converge to a systemic signaling gene known as FLOWERING LOCUS T (FT). In long-day flowering plants, FT is expressed in the leaves on long days. The protein travels through the phloem to the shoot apical meristem, where it induces flower determination. In perennial plants, meristem determination and flowering are separated by a dormancy period. Meristem determination takes place in summer, but flowering occurs only after a dormancy period and cold accumulation during winter. The roles of FT are not completely clear in meristem determination, dormancy release, and flowering in perennial plants. We cloned FT from sweet cherry (Prunus avium) and analyzed its expression pattern in leaves and floral buds during spring and summer. Phylogenetic analysis shows high identity of the FT cloned sequence with orthologous genes from other Rosaceae species. Our results show that FT is expressed in both leaves and floral buds and increases when the daylight reached 12 h. The peak in FT expression was coincident with floral meristem identity genes expression and morphological changes typical of floral meristem determination. The Edi-0 Arabidopsis ecotype, which requires vernalization to flower, was transformed with a construct for overexpression of PavFT. These transgenic plants showed an early-flowering phenotype without cold treatment. Our results suggest that FT is involved in floral meristem determination and bud development in sweet cherry. Moreover, we show that FT is expressed in both leaves and floral buds in this species, in contrast to annual plants.

Asunto(s)

Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/metabolismo , Prunus avium/genética , Arabidopsis/crecimiento & desarrollo , Arabidopsis/efectos de la radiación , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Frío , Flores/genética , Flores/crecimiento & desarrollo , Flores/efectos de la radiación , Expresión Génica , Meristema/genética , Meristema/crecimiento & desarrollo , Meristema/efectos de la radiación , Fenotipo , Floema/genética , Floema/crecimiento & desarrollo , Floema/efectos de la radiación , Filogenia , Hojas de la Planta/genética , Hojas de la Planta/crecimiento & desarrollo , Hojas de la Planta/efectos de la radiación , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente , Prunus avium/crecimiento & desarrollo , Prunus avium/efectos de la radiación , Reproducción , Estaciones del Año

RESUMEN

Factors regulating fine-root growth are poorly understood, particularly in fruit tree species. In this context, the effects of N addition on the temporal and spatial distribution of fine-root growth and on the fine-root turnover were assessed in irrigated sweet cherry trees. The influence of other exogenous and endogenous factors was also examined. The rhizotron technique was used to measure the length-based fine-root growth in trees fertilized at two N rates (0 and 60 kg ha(-1)), and the above-ground growth, leaf net assimilation, and air and soil variables were simultaneously monitored. N fertilization exerted a basal effect throughout the season, changing the magnitude, temporal patterns and spatial distribution of fine-root production and mortality. Specifically, N addition enhanced the total fine-root production by increasing rates and extending the production period. On average, N-fertilized trees had a length-based production that was 110-180% higher than in control trees, depending on growing season. Mortality was proportional to production, but turnover rates were inconsistently affected. Root production and mortality was homogeneously distributed in the soil profile of N-fertilized trees while control trees had 70-80% of the total fine-root production and mortality concentrated below 50 cm depth. Root mortality rates were associated with soil temperature and water content. In contrast, root production rates were primarily under endogenous control, specifically through source-sink relationships, which in turn were affected by N supply through changes in leaf photosynthetic level. Therefore, exogenous and endogenous factors interacted to control the fine-root dynamics of irrigated sweet cherry trees.

Asunto(s)

Nitrógeno/farmacología , Prunus avium/crecimiento & desarrollo , Prunus avium/metabolismo , Árboles/crecimiento & desarrollo , Árboles/metabolismo , Riego Agrícola , Chile , Relación Dosis-Respuesta a Droga , Fertilizantes/análisis , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/crecimiento & desarrollo , Hojas de la Planta/metabolismo , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/metabolismo , Prunus avium/efectos de los fármacos , Estaciones del Año , Árboles/efectos de los fármacos

RESUMEN

The present research aimed to isolate the non-polar secondary metabolites that produce the vasodilator effects induced by the dichloromethane extract of Prunus serotina (P. serotina) fruits and to determine whether the NO/cGMP and the H2S/KATP channel pathways are involved in their mechanism of action. A bioactivity-directed fractionation of the dichloromethane extract of P. serotina fruits led to the isolation of ursolic acid and uvaol as the main non-polar vasodilator compounds. These compounds showed significant relaxant effect on rat aortic rings in an endothelium- and concentration-dependent manner, which was inhibited by NG-nitro-L-arginine methyl ester (L-NAME), DL-propargylglycine (PAG) and glibenclamide (Gli). Additionally, both triterpenes increased NO and H2S production in aortic tissue. Molecular docking studies showed that ursolic acid and uvaol are able to bind to endothelial NOS and CSE with high affinity for residues that form the oligomeric interface of both enzymes. These results suggest that the vasodilator effect produced by ursolic acid and uvaol contained in P. serotina fruits, involves activation of the NO/cGMP and H2S/KATP channel pathways, possibly through direct activation of NOS and CSE.

Asunto(s)

Sulfuro de Hidrógeno/agonistas , Óxido Nítrico/agonistas , Prunus avium/química , Triterpenos/farmacología , Vasodilatación/efectos de los fármacos , Vasodilatadores/farmacología , Alquinos/antagonistas & inhibidores , Alquinos/farmacología , Animales , Aorta/citología , Aorta/efectos de los fármacos , Aorta/metabolismo , GMP Cíclico/metabolismo , Cistationina gamma-Liasa/química , Cistationina gamma-Liasa/metabolismo , Endotelio Vascular/citología , Endotelio Vascular/efectos de los fármacos , Activación Enzimática/efectos de los fármacos , Frutas/química , Gliburida/antagonistas & inhibidores , Gliburida/farmacología , Glicina/análogos & derivados , Glicina/antagonistas & inhibidores , Glicina/farmacología , Sulfuro de Hidrógeno/metabolismo , Canales KATP/agonistas , Canales KATP/metabolismo , Masculino , Simulación del Acoplamiento Molecular , NG-Nitroarginina Metil Éster/antagonistas & inhibidores , NG-Nitroarginina Metil Éster/farmacología , Óxido Nítrico/biosíntesis , Óxido Nítrico Sintasa de Tipo III/química , Óxido Nítrico Sintasa de Tipo III/metabolismo , Extractos Vegetales/química , Unión Proteica , Ratas , Triterpenos/aislamiento & purificación , Vasodilatadores/aislamiento & purificación , Ácido Ursólico

RESUMEN

The emergence of Escherichia coli isolates with multiple antibiotic resistant phenotypes is considered as a severe health concern. In the present work the antibacterial effect of following plants (Herniaria hirsuta, Prunus avium, Rubia tinctorum and Sempervivum tectorum) was examined. The bacterial model used for estimation of bacterial susceptibility is hospital multiple antibiotic resistant E. coli strain. E. coli ATCC 25922 was used for standard comparison of bacterial susceptibility. Leaves of H. hirsuta, R. tinctorum and S. tectorum as well as petioles of P. avium were collected. Ethanol and aqueous extract of each plant was prepared. Antibacterial activity was examined using the agar well diffusion method. Concentration of total phenols, flavonoids, tannins, antocyanins and saponins was determined in plant extracts. E. coli strain is resistant to four unrelated families of antibiotics. Antibacterial effect is proven for all examined plants. Ethanol extracts of H. hirsuta and P. avium have a more potent antibacterial effect than their aqueous extracts. Aqueous extracts of R. tinctorum and S. tectorum have higher antibacterial potential than theirs ethanol extracts. Examined plant extracts represent good candidates for more extensive research in view of their application in the treatment of multiple antibiotic resistant E.coli strains.

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O surgimento de Escherichia coli isoladas com vários fenótipos resistentes aos antibióticos é considerado como um grave problema de saúde. No presente trabalho o efeito antibacteriano das seguintes plantas (Herniaria hirsuta, Prunus avium, Rubia tinctorum e Sempervivum tectorum) foi analisado. O agente bacteriano modelo utilizado para estimativa de susceptibilidade bacteriana é o hospital vários resistentes a antibióticos E. coli. E. coli ATCC 25922 padrão foi utilizado para comparação de antibiogramas. Folhas de H. hirsuta, R. tinctorum e S. tectorum bem como pecíolos de P. avium foram coletados. Etanol e extrato aquoso de cada planta foi preparado. Atividade antibacteriana foi analisada através do método de difusão em ágar-bem. Total Concentração de fenóis, flavonóides, taninos e saponinas antocyanins determinou-se em extratos de plantas. E. coli estirpe é resistente às quatro famílias de antibióticos independentes. Efeito antibacteriano é comprovado para todas as plantas examinadas. Os extratos etanólicos de H. hirsuta e P. avium têm um efeito mais potente antibacteriano de seus extratos aquosos. Extratos aquosos de R. tinctorum e S. tectorum têm maior potencial antibacteriano que os extratos etanólicos. Extratos vegetais examinados representam bons candidatos para pesquisa mais ampla em vista de sua aplicação no tratamento de vários antibióticos resistentes a cepas de E. coli.

Asunto(s)

Extractos Vegetales , Escherichia coli , Medicina Tradicional , Antibacterianos , Sempervivum tectorum , Caryophyllaceae , Rubia , Prunus avium

RESUMEN

Linkage maps are valuable tools in genetic and genomic studies. For sweet cherry, linkage maps have been constructed using mainly microsatellite markers (SSRs) and, recently, using single nucleotide polymorphism markers (SNPs) from a cherry 6K SNP array. Genotyping-by-sequencing (GBS), a new methodology based on high-throughput sequencing, holds great promise for identification of high number of SNPs and construction of high density linkage maps. In this study, GBS was used to identify SNPs from an intra-specific sweet cherry cross. A total of 8,476 high quality SNPs were selected for mapping. The physical position for each SNP was determined using the peach genome, Peach v1.0, as reference, and a homogeneous distribution of markers along the eight peach scaffolds was obtained. On average, 65.6% of the SNPs were present in genic regions and 49.8% were located in exonic regions. In addition to the SNPs, a group of SSRs was also used for construction of linkage maps. Parental and consensus high density maps were constructed by genotyping 166 siblings from a 'Rainier' x 'Rivedel' (Ra x Ri) cross. Using Ra x Ri population, 462, 489 and 985 markers were mapped into eight linkage groups in 'Rainier', 'Rivedel' and the Ra x Ri map, respectively, with 80% of mapped SNPs located in genic regions. Obtained maps spanned 549.5, 582.6 and 731.3 cM for 'Rainier', 'Rivedel' and consensus maps, respectively, with an average distance of 1.2 cM between adjacent markers for both 'Rainier' and 'Rivedel' maps and of 0.7 cM for Ra x Ri map. High synteny and co-linearity was observed between obtained maps and with Peach v1.0. These new high density linkage maps provide valuable information on the sweet cherry genome, and serve as the basis for identification of QTLs and genes relevant for the breeding of the species.

Asunto(s)

Mapeo Cromosómico/métodos , Ligamiento Genético , Técnicas de Genotipaje/métodos , Repeticiones de Microsatélite/genética , Polimorfismo de Nucleótido Simple/genética , Prunus avium/genética , Análisis de Secuencia de ADN/métodos , Segregación Cromosómica/genética , Genes de Plantas , Genómica , Mapeo Físico de Cromosoma