RESUMEN

BACKGROUND: Fermentation is an ancient technique for preserving and improving the qualities of food and beverages throughout the world. Microbial communities, not seen by the producers of fermented goods, are the actors involved in the fermentation process and are selected upon through different management processes in order to achieve a final product with culturally accepted features. This study documented the preparation of "colonche" which is a type of traditionally fermented beverages made with the fruits from several cactus species in two main producing regions of Mexico, the Altiplano and the Tehuacán Valley. We documented the selection processes of the cactus species used and the practices that could influence microbial community composition, as well as, how the producers reach the desirable sensorial attributes of the beverages. METHODS: We conducted 53 semi-structured interviews and participatory observations with colonche producers in 7 communities of the Altiplano and the Tehuacán Valley in order to characterize the practices and processes involved in the elaboration of the beverage. Opuntia and columnar cacti species used in colonche production were collected during fieldwork and identified. Selected sensorial attributes of Opuntia colonches were characterized by a ranking table and visualized by principal component analysis in order to distinguish differences of this beverage in the Altiplano localities. RESULTS: Thirteen cactus species are used for colonche production in both regions studied. In the Altiplano, the most commonly used fruit is Opuntia streptacantha because it contributes to the preferred attributes of the beverage in this region. Selection of substrates by producers depends on their preference and the availability of fruits of O. streptacantha and other species. Fermentation is mainly conducted in clay pots which is perceived to be the best type of vessel contributing to the preferred sensorial properties of colonche. The two main differences in colonche preparation between the villages are the practice of boiling the fruit juice and the use of pulque (fermented sap of Agave species) as inoculum. The most contrasting sensorial attributes selected between localities are the alcohol content and sweetness, which might be in accordance with the practices used for obtaining the final product. Colonche is produced mainly for direct consumption and secondarily used as a commercialized good to be sold for economic gains contributing to the general subsistence of households. The preparation methods are passed on by close relatives, mainly women. CONCLUSIONS: Traditional producers of colonche use several techniques in order to reach specific sensorial attributes of the final product. The production of colonche has been upheld for generations but fermentation practices are divided into two categories; (1) the use of an inoculum (either from pulque, or from colonche saved from the previous year), and (2) the use of "spontaneous" fermentation. The differing practices documented reflect the contrasts in the preferred sensorial attributes between regions. Colonche is a beverage that contributes to regional pride, cultural identity and is appreciated because of its gastronomic value. Here, we argue that there is a clear relationship of human knowledge in the management of microbiota composition in order to produce this beverage. In-depth documentation of the microbiota composition and dynamics in colonche will contribute to the preservation of this valuable biocultural heritage.

Asunto(s)

Bebidas/microbiología , Etnobotánica , Fermentación , Frutas , Opuntia , Adulto , Anciano , Etnobotánica/métodos , Femenino , Frutas/metabolismo , Humanos , Entrevistas como Asunto , Masculino , México , Persona de Mediana Edad , Opuntia/metabolismo

RESUMEN

The present study evaluated the effects of ultraviolet B (UVB) radiation and wounding stress, applied alone or combined, on the biosynthesis of phenolic compounds and ascorbic acid in the peel and pulp of red prickly pear (Opuntia ficus-indica cv. Rojo Vigor). Whole and wounded-fruit samples were treated with UVB radiation (6.4 W·m-2) for 0 and 15 min, and stored for 24 h at 16 °C. Phytochemical analyses were performed separately in the peel and pulp. The highest phenolic accumulation occurred after storage of the whole tissue treated with UVB, where the main phenolic compounds accumulated in the peel and pulp were quercetin, sinapic acid, kaempferol, rosmarinic acid, and sinapoyl malate, showing increases of 709.8%, 570.2%, 442.8%, 439.9%, and 186.2%, respectively, as compared with the control before storage. Phenylalanine ammonia-lyase (PAL) activity was increased after storage of the whole and wounded tissue treated with UVB light, and this increase in PAL activity was associated to phenolic accumulation. On the other hand, l-galactono-γ-lactone dehydrogenase (GalLDH) activity and ascorbic acid biosynthesis was enhanced due to UVB radiation, and the effect was increased when UVB was applied in the wounded tissue showing 125.1% and 94.1% higher vitamin C content after storage when compared with the control. Respiration rate was increased due to wounding stress, whereas ethylene production was increased by wounding and UVB radiation in prickly pears. Results allowed the generation of a physiological model explaining the UVB and wound-induced accumulation of phenolic compounds and ascorbic acid in prickly pears, where wounding facilitates UVB to access the underlying tissue and enhances an apparent synergistic response.

Asunto(s)

Modelos Biológicos , Opuntia/metabolismo , Fenoles/metabolismo , Proteínas de Plantas/metabolismo , Rayos Ultravioleta

RESUMEN

In plants, 14-3-3 proteins are important modulators of protein-protein interactions in response to environmental stresses. The aim of the present work was to characterize one Opuntia ficus-indica 14-3-3 and get information about its client proteins. To achieve this goal, O. ficus-indica 14-3-3 cDNA, named as Op14-3-3 µ, was amplified by 3'-RACE methodology. Op14-3-3 µ contains an Open Reading Frame of 786 bp encoding a 261 amino acids protein. Op14-3-3 µ cDNA was cloned into a bacterial expression system and recombinant protein was purified. Differential Scanning Fluorimetry, Dynamic Light Scattering, and Ion Mobility-Mass Spectrometry were used for Op14-3-3 µ protein characterization, and Affinity-Purification-Mass Spectrometry analysis approach was used to obtain information about their potential client proteins. Pyrophosphate-fructose 6-phosphate 1-phosphotransferase, ribulose bisphosphate carboxylase large subunit, and vacuolar-type H+-ATPase were identified. Interestingly chorismate mutase p-prephenate dehydratase was also identified. Op14-3-3 µ down-regulation was observed in Opuntia calluses when they were induced with Jasmonic Acid, while increased accumulation of Op14-3-3 µ protein was observed. The putative interaction of 14-3-3 µ with chorismate mutase, which have not been reported before, suggest that Op14-3-3 µ could be an important regulator of metabolites biosynthesis and responses to stress in Opuntia spp. SIGNIFICANCE: Opuntia species are important crops in arid and semiarid areas worldwide, but despite its relevance, little information about their tolerance mechanism to cope with harsh environmental conditions is reported. 14-3-3 proteins have gained attention due to its participation as protein-protein regulators and have been linked with primary metabolism and hormones responses. Here we present the characterization of the first Opuntia ficus-indica 14-3-3 (Op14-3-3) protein using affinity purification-mass spectrometry (AP-MS) strategy. Op14-3-3 has high homology with other 14-3-3 from Caryophyllales. A novel Op14-3-3 client protein has been identified; the chorismate mutase p-prephenate dehydratase, key enzyme that links the primary with secondary metabolism. The present results open new questions about the Opuntia spp. pathways mechanisms in response to environmental stress and the importance of 14-3-3 proteins in betalains biosynthesis.

Asunto(s)

Proteínas 14-3-3 , Opuntia , Proteínas de Plantas , Ácido Shikímico/metabolismo , Estrés Fisiológico , Proteínas 14-3-3/biosíntesis , Proteínas 14-3-3/química , Proteínas 14-3-3/genética , Proteínas 14-3-3/aislamiento & purificación , Sistemas de Lectura Abierta , Opuntia/química , Opuntia/genética , Opuntia/metabolismo , Proteínas de Plantas/biosíntesis , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/aislamiento & purificación , Proteínas Recombinantes

RESUMEN

BACKGROUND: Among Cactaceae, the genus Opuntia is widely known for the use of its biomass as cattle fodder and in human nutrition (e.g. species such as Opuntia ficus indica and Opuntia streptacantha). In particular, O. streptacantha (OS) produces abundant mucilage and, hence, the characterization of its properties and nutritional value is important. Accordingly, determination of the dietary fiber content of the OS mucilage and the fermentability of its hydrolysis products (oligosaccharides, OLI) is important for developing new uses of the crop as a functional food. RESULTS: The values for insoluble dietary fiber and soluble dietary fiber in the mucilage were 204.6 and 371.6 g kg-1 , respectively. After hydrolysis of OS mucilage with α-amylase, three purified fractions of OLI were evaluated (OLI-A, OLI-B and OLI-C). OLI (1% w/v) stimulated the growth of the commercial probiotic strains (Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium animalis subsp. lactis) in vitro, showing behaviors similar to those of commercial inulin. The production of short chain fatty acids (SCFAs) in the fermentation broth was also determined. The final pH of the fermentation broth as well as the identification and concentrations of SCFA depended on the type of OLI and probiotic used. CONCLUSION: The OS mucilage is an unconventional fiber source and can be used to produce non-digestible OLI as functional compounds. This knowledge will be useful for proposing new sustainable ways of processing cacti crops for food and industrial purposes. © 2018 Society of Chemical Industry.

Asunto(s)

Oligosacáridos/química , Opuntia/química , Mucílago de Planta/química , alfa-Amilasas/química , Bifidobacterium animalis/metabolismo , Biocatálisis , Fibras de la Dieta/análisis , Fibras de la Dieta/metabolismo , Ácidos Grasos Volátiles/metabolismo , Hidrólisis , Lactobacillus acidophilus/metabolismo , Modelos Biológicos , Valor Nutritivo , Oligosacáridos/metabolismo , Opuntia/metabolismo , Mucílago de Planta/metabolismo , Polisacáridos/química , Polisacáridos/metabolismo , alfa-Amilasas/metabolismo

RESUMEN

The present investigation was conducted to evaluate the effects of Escherichia coli against the ruminal microflora fermentation activities in the mitigation of CH4 and CO2 production as well as ruminal fermentation kinetics by substituting dietary corn grain with prickly pear cactus (PC) flour. Three total mixed PC rations were prepared (/kg DM): 0 g (Control), 75 g (PC75), and 150 g (PC150). Besides, E. coli was supplemented at four different levels (dose): 0, 10, 20, and 40 mg/g DM of substrates. The in vitro rumen GP, CH4, and CO2 were estimated to be affected due to various doses of E. coli up to 72 h of incubation. Asymptotic GP, fractional rate of GP, and lag time were influenced significantly (P < .05) in the presence of ration. However, E. coli doses showed minor impact on the rate of GP as well as lag time. The asymptotic CH4 production was decreased linearly (P = .005) at the ration PC150. E. coli doses reduced the asymptotic CH4 production at 10 and 20 mg/g DM. The asymptotic CO2 production was linearly (P < .001) decreased by different levels of PC. The cubic (P = .023) effect of E. coli doses as well as significant (P = .002) ration × E. coli doses impact were reported on asymptotic CO2 production. The fractional rate of GP was quadratically (P < .05) influenced by PC and E. coli doses. The rations, dose, and rations × E. coli dose interaction had no influence (P > .05) on lag time. In a nutshell, PC flour inclusion in diet has the potentiality to replace the existing conventional feedstuffs for ruminant. Most importantly, revealing the first report, PC flours along with E. coli supplementation at varied doses mitigated the ruminal biogases production. This was as consequence to the antimicrobial impacts of E. coli against ruminal microflora, and that could certainly be a promising approach in order to improve ruminant's diet constituents.

Asunto(s)

Alimentación Animal/análisis , Dióxido de Carbono/metabolismo , Escherichia coli/metabolismo , Metano/biosíntesis , Opuntia/metabolismo , Rumen/microbiología , Animales , Bovinos , Dieta , Digestión , Femenino , Fermentación/fisiología , Microbioma Gastrointestinal , Residuos Sólidos , Granos Enteros/metabolismo

RESUMEN

Dehydrins (DHNs) are intrinsically disordered proteins that play central roles in plant abiotic stress responses; however, how they work remains unclear. Herein, we report the in planta subcellular localization of Arabidopsis thaliana DHNs AtCOR47, AtERD10, and AtRAB18 through GFP translational fusions. To explore the dimerization ability of the Arabidopsis acidic DHNs AtCOR47 and AtERD10, we conducted an in planta DHN binding assay using the Bimolecular Fluorescence Complementation (BiFC) technique. Our analyses revealed homodimeric interactions for AtCOR47 and AtERD10; interestingly, heterodimeric associations also occurred with these DHNs, and these interactions were observed in the cytosol of tobacco cells. Furthermore, we evaluated whether Arabidopsis basic DHNs, such as AtRAB18, could also interact with itself and/or with AtCOR47 and AtERD10 in the BiFC system. Our data revealed homodimeric RAB18 complexes in the nucleus and cytosol, while heterodimeric associations between AtRAB18 and acidic DHNs occurred only in the cytosol. Finally, we demonstrated the presence of heterodimeric complexes among Arabidopsis AtCOR47, AtERD10, and AtRAB18 DHNs with their acidic ortholog the OpsDHN1 from Opuntia streptacantha; these heterodimeric interactions showed different subcellular distributions. Our results guide DHN research toward a new scenario where DHN/DHN oligomerization could be explored as a part of their molecular mechanism.

Asunto(s)

Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas de Unión al GTP rab/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/química , Núcleo Celular/metabolismo , Citosol/metabolismo , Dimerización , Microscopía Fluorescente , Opuntia/metabolismo , Plantas Modificadas Genéticamente/metabolismo , Unión Proteica , Nicotiana/genética , Nicotiana/metabolismo , Proteínas de Unión al GTP rab/química

RESUMEN

Opuntia species have been used for centuries as food resources and in traditional folk medicine for their nutritional properties and their benefit in chronic diseases, particularly diabetes, obesity, cardiovascular diseases, and cancer. These plants are largely distributed in America, Africa, and the Mediterranean basin. Opuntia spp. have great economic potential because they grow in arid and desert areas, and O. ficus-indica, the domesticated O. species, is used as a nutritional and pharmaceutical agent in various dietary and value-added products. Though differences in the phytochemical composition exist between wild and domesticated (O. ficus-indica) Opuntia spp., all Opuntia vegetatives (pear, roots, cladodes, seeds, and juice) exhibit beneficial properties mainly resulting from their high content in antioxidants (flavonoids, ascorbate), pigments (carotenoids, betalains), and phenolic acids. Other phytochemical components (biopeptides, soluble fibers) have been characterized and contribute to the medicinal properties of Opuntia spp. The biological properties of Opuntia spp. have been investigated on cellular and animal models and in clinical trials in humans, allowing characterization and clarification of the protective effect of Opuntia-enriched diets in chronic diseases. This review is an update on the phytochemical composition and biological properties of Opuntia spp. and their potential interest in medicine.

Asunto(s)

Enfermedades Cardiovasculares/dietoterapia , Diabetes Mellitus/dietoterapia , Dieta Mediterránea , Neoplasias/dietoterapia , Obesidad/dietoterapia , Opuntia/química , Animales , Enfermedad Crónica , Humanos , Opuntia/metabolismo

RESUMEN

The aim of this study was to evaluate the fermentation of dietary fiber from green bean (Phaseolus vulgaris) and prickly pear shell (Opuntia ficus-indica) by Lactobacillus acidophilus LA-5 and Bifidobacterium bifidum 450B growing as mono-culture and co-culture, the fermentation products, and proteins expressed during this process. The analysis of the fermentation profile showed a major growth of bacteria in the culture media of each dietary fiber supplemented with glucose, and particularly B. bifidum 450B at 48 h showed the highest growth. In the case of the co-culture, the growth was lower indicating the possible negative interaction between L. acidophilus LA-5 and B. bifidum 450B and may be due to the less amount of carbohydrates and the high content of non-soluble fiber that affected the nutrients availability for the bacterial strains. The pH changes indicated the presence of short-chain fatty acids (SCFAs), being acetate (46-100%) the main SCFA. Changes in the proteome concerned proteins that are involved in carbohydrate and other carbohydrate pathways. The characterization of the bacteria according to the growth, metabolites, and proteins expressed allows understanding the response to the change of environmental conditions and could be useful to understand L. acidophilus LA-5 and B. bifidum 450B strains' adaptation to specific applications.

Asunto(s)

Bifidobacterium bifidum/metabolismo , Fibras de la Dieta/metabolismo , Lactobacillus acidophilus/metabolismo , Opuntia/metabolismo , Phaseolus/metabolismo , Microbiología Ambiental , Ácidos Grasos Volátiles/metabolismo , Fermentación/fisiología

RESUMEN

BACKGROUND: Fat is the tissue that varies most in animals from both a quantitative and distribution perspective. It plays a fundamental biological role as energy storage during food scarcity. Renal, pelvic and internal fat are deposited first. These fats are used to identify fatty acid profiles that may be considered beneficial or unhealthy. The aim of this study is to evaluate the fatty acid profile of fat depots in Santa Inês sheep finished in confinement with spineless cactus in their diets. RESULTS: The treatments included increasing levels of spineless cactus (Opuntia fícus-indica Mill.): T1 = 0%, T2 = 30%, T3 = 50%, and T4 = 70%. The diets significantly affected the adipose depots. The orthogonal contrast between the diet with no cactus (control) and the other diets indicates that the quantity of saturated fatty acids decreased and that the levels of mono-unsaturated and polyunsaturated fatty acids increased in animals fed spineless cactus. CONCLUSION: The use of spineless cactus in the diets of Santa Inês sheep affects the lipid profile of their fat depots, reducing the quantity of saturated fatty acids and increasing the quantity of mono-unsaturated and polyunsaturated fatty acids. The fatty acid profile of the fat depots indicates that these fats can be used to formulate meat products and add economic and nutritional value to such products, which increases sheep farmers' incomes. © 2017 Society of Chemical Industry.

Asunto(s)

Alimentación Animal/análisis , Grasas/metabolismo , Ácidos Grasos/metabolismo , Opuntia/metabolismo , Ovinos/metabolismo , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Ácidos Grasos/química , Carne/análisis , Opuntia/química , Ovinos/crecimiento & desarrollo

RESUMEN

The aim of this research was to investigate the use of a natural adsorbent like nopal (Opuntia albicarpa L. Scheinvar) for removing cadmium from aqueous solutions with low concentrations of this metal. Two treatments were applied to the cladodes: a dehydration to get dehydrated nopal (DHN) and heating up to 90°C to obtain a thermally treated nopal (TN). After examining the effect of various pH values (2-7), the capacity of each biosorbent was examined in batch sorption tests at different dosages (0, 500, 1000, 1500, 2000, and 3000 mg L(-1)). The results indicated that adsorption of cadmium to biomass of DHN and TN was highly dependent on pH and biosorbent dosage. The best removal of cadmium (53.3%, corresponding to q e of 0.155 mg g(-1)) was obtained at pH 4.0 by using the TN sorbent. Infrared and Raman spectra confirmed that cadmium removal occurred via adsorption to -OH functional groups.

Asunto(s)

Cadmio/toxicidad , Contaminantes Químicos del Agua/toxicidad , Purificación del Agua , Adsorción , Biomasa , Cadmio/química , Humanos , Concentración de Iones de Hidrógeno , Cinética , Opuntia/química , Opuntia/metabolismo , Agua/química , Contaminantes Químicos del Agua/química

RESUMEN

The study was conducted with the objective to evaluate the intake, apparent digestibility of the nutrients and of ruminal fermentation characteristics of sheep fed diets based on spineless cactus, sugar cane bagasse and urea associated to diferents supplements. Five non-castrated male sheep were allotted to a 5 x 5 Latin square design, with five animals (lines) and five periods (columns). Each experimental period last 13 day. The ration without supplement was composed of 67.6% of spineless cactus, 27.7% of sugar cane bagasse, 2.7% of urea:ammonium sulphate mixture (9:1) and 2.0% of mineral mixture, in dry matter (DM) basis. The other treatments consisted of the addition of a supplement to the ration (0,4% of body weight), being: wheat meal, soybean meal, cottonseed meal or whole cottonseed. Were not observed effects of supplement association on the evaluated parameters, being: 3.29%, 60.61%, 76.37%, 6.51 and 11.85mg/100mL the means value for intake of DM (% of body weight), digestibility of DM and CP, pH and ruminal ammonium, respectively. The choice of supplement should be associated with the availability and cost.

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Objetivou-se avaliar o consumo, a digestibilidade aparente dos nutrientes e os parâmetros ruminais de ovinos alimentados com dietas à base de palma forrageira, bagaço de cana-de-açúcar e ureia associados a diferentes suplementos. Foram utilizados cinco ovinos sem padrão racial definido, machos, não-castrados, fistulados no rúmen. Os animais foram distribuídos aleatoriamente em quadrado latino 5 x 5, com cinco animais (linhas) e cinco períodos (colunas). Cada período teve duração de 13 dias. A ração sem suplemento foi composta de 67,6% de palma forrageira, 27,7% de bagaço de cana-de-açúcar in natura, 2,7% de mistura ureia:sulfato de amônio (9:1) e 2,0% de mistura mineral, em base da matéria seca (MS). Os demais tratamentos consistiram na adição de um suplemento à ração ao nível de 0,4% do peso corporal, composta por: farelo de trigo, farelo de soja, farelo de algodão ou caroço de algodão. Não foram observados efeitos da associação dos suplementos sobre os parâmetros avaliados, sendo: 3,29%, 60,61%, 76,37%, 6,51 e 11,85mg/100mL os valores médios observados para o consumo de MS em função do peso corporal, digestibilidades da MS e da PB, pH e amônia ruminal, respectivamente. A escolha do suplemento deve estar associada à disponibilidade e ao custo.

Asunto(s)

Animales , Opuntia/efectos adversos , Opuntia/metabolismo , Ovinos/crecimiento & desarrollo , Ovinos/metabolismo , Fenómenos Fisiológicos Nutricionales del Lactante , Alimentación Animal/análisis

RESUMEN

The study was conducted with the objective to evaluate the intake, apparent digestibility of the nutrients and of ruminal fermentation characteristics of sheep fed diets based on spineless cactus, sugar cane bagasse and urea associated to diferents supplements. Five non-castrated male sheep were allotted to a 5 x 5 Latin square design, with five animals (lines) and five periods (columns). Each experimental period last 13 day. The ration without supplement was composed of 67.6% of spineless cactus, 27.7% of sugar cane bagasse, 2.7% of urea:ammonium sulphate mixture (9:1) and 2.0% of mineral mixture, in dry matter (DM) basis. The other treatments consisted of the addition of a supplement to the ration (0,4% of body weight), being: wheat meal, soybean meal, cottonseed meal or whole cottonseed. Were not observed effects of supplement association on the evaluated parameters, being: 3.29%, 60.61%, 76.37%, 6.51 and 11.85mg/100mL the means value for intake of DM (% of body weight), digestibility of DM and CP, pH and ruminal ammonium, respectively. The choice of supplement should be associated with the availability and cost.(AU)

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Objetivou-se avaliar o consumo, a digestibilidade aparente dos nutrientes e os parâmetros ruminais de ovinos alimentados com dietas à base de palma forrageira, bagaço de cana-de-açúcar e ureia associados a diferentes suplementos. Foram utilizados cinco ovinos sem padrão racial definido, machos, não-castrados, fistulados no rúmen. Os animais foram distribuídos aleatoriamente em quadrado latino 5 x 5, com cinco animais (linhas) e cinco períodos (colunas). Cada período teve duração de 13 dias. A ração sem suplemento foi composta de 67,6% de palma forrageira, 27,7% de bagaço de cana-de-açúcar in natura, 2,7% de mistura ureia:sulfato de amônio (9:1) e 2,0% de mistura mineral, em base da matéria seca (MS). Os demais tratamentos consistiram na adição de um suplemento à ração ao nível de 0,4% do peso corporal, composta por: farelo de trigo, farelo de soja, farelo de algodão ou caroço de algodão. Não foram observados efeitos da associação dos suplementos sobre os parâmetros avaliados, sendo: 3,29%, 60,61%, 76,37%, 6,51 e 11,85mg/100mL os valores médios observados para o consumo de MS em função do peso corporal, digestibilidades da MS e da PB, pH e amônia ruminal, respectivamente. A escolha do suplemento deve estar associada à disponibilidade e ao custo.(AU)

Asunto(s)

Animales , Ovinos/crecimiento & desarrollo , Ovinos/metabolismo , Opuntia/efectos adversos , Opuntia/metabolismo , Fenómenos Fisiológicos Nutricionales del Lactante , Alimentación Animal/análisis

RESUMEN

Abiotic stress limits seed germination, plant growth, flowering and fruit quality, causing economic decrease. Small Heat Shock Proteins (sHSPs) are chaperons with roles in stress tolerance. Herein, we report the functional characterization of a cytosolic class CI sHSP (OpsHSP18) from Opuntia streptacantha during seed germination in Arabidopsis thaliana transgenic lines subjected to different stress and hormone treatments. The over-expression of the OpsHSP18 gene in A. thaliana increased the seed germination rate under salt (NaCl) and osmotic (glucose and mannitol) stress, and in ABA treatments, compared with WT. On the other hand, the over-expression of the OpsHSP18 gene enhanced tolerance to salt (150 mM NaCl) and osmotic (274 mM mannitol) stress in Arabidopsis seedlings treated during 14 and 21 days, respectively. These plants showed increased survival rates (52.00 and 73.33%, respectively) with respect to the WT (18.75 and 53.75%, respectively). Thus, our results show that OpsHSP18 gene might have an important role in abiotic stress tolerance, in particular in seed germination and survival rate of Arabidopsis plants under unfavorable conditions.

Asunto(s)

Arabidopsis/crecimiento & desarrollo , Germinación/efectos de los fármacos , Proteínas de Choque Térmico Pequeñas/genética , Opuntia/metabolismo , Presión Osmótica/efectos de los fármacos , Cloruro de Sodio/farmacología , Estrés Fisiológico , Ácido Abscísico/farmacología , Secuencia de Aminoácidos , Arabidopsis/efectos de los fármacos , Arabidopsis/genética , Proteínas de Choque Térmico Pequeñas/metabolismo , Datos de Secuencia Molecular , Opuntia/genética , Filogenia , Reguladores del Crecimiento de las Plantas/farmacología , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Plantones/efectos de los fármacos , Plantones/crecimiento & desarrollo , Plantones/metabolismo , Homología de Secuencia de Aminoácido

RESUMEN

In Opuntia stenopetala, flowers initiate as hermaphrodite; however, at maturity, only the stamens in male flowers and the gynoecium in female flowers become functional. At early developmental stages, growth and morphogenesis of the gynoecium in male flowers cease, forming a short style lacking stigmatic tissue at maturity. Here, an analysis of the masculinization process of this species and its relationship with auxin metabolism during gynoecium morphogenesis is presented. Histological analysis and scanning electron microscopy were performed; auxin levels were immunoanalyzed and exogenous auxin was applied to developing gynoecia. Male flower style-tissue patterning revealed morphological defects in the vascular bundles, stylar canal, and transmitting tissue. These features are similar to those observed in Arabidopsis thaliana mutant plants affected in auxin transport, metabolism, or signaling. Notably, when comparing auxin levels between male and female gynoecia from O. stenopetala at an early developmental stage, we found that they were particularly low in the male gynoecium. Consequently, exogenous auxin application on male gynoecia partially restored the defects of gynoecium development. We therefore hypothesize that, the arrest in male flower gynoecia patterning could be related to altered auxin homeostasis; alternatively, the addition of auxin could compensate for the lack of another unknown factor affecting male flower gynoecium development.

Asunto(s)

Flores/crecimiento & desarrollo , Ácidos Indolacéticos/metabolismo , Morfogénesis/fisiología , Opuntia/crecimiento & desarrollo , Opuntia/metabolismo , Arabidopsis/crecimiento & desarrollo , Arabidopsis/metabolismo , Flores/ultraestructura , Organismos Hermafroditas/citología , Organismos Hermafroditas/crecimiento & desarrollo , Caracteres Sexuales , Procesos de Determinación del Sexo

RESUMEN

Cactus pears are succulent plants of the Cactaceae family adapted to extremely arid, hot and cold environments, making them excellent models for the study of molecular mechanisms underlying abiotic stress tolerance. Herein, we report a directional cDNA library from 12-month-old cladodes of Opuntia streptacantha plants subjected to abiotic stresses. A total of 442 clones were sequenced, representing 329 cactus pear unigenes, classified into eleven functional categories. The most abundant EST (unigen 33) was characterized under abiotic stress. This cDNA of 905 bp encodes a SK(3)-type acidic dehydrin of 248 amino acids. The OpsDHN1 gene contains an intron inserted within the sequence encoding the S-motif. qRT-PCR analysis shows that the OpsDHN1 transcript is specifically accumulated in response to cold stress, and induced by abscisic acid. Over-expression of the OpsDHN1 gene in Arabidopsis thaliana leads to enhanced tolerance to freezing treatment, suggesting that OpsDHN1 participates in freezing stress responsiveness. Generation of the first EST collection for the characterization of cactus pear genes constitutes a useful platform for the understanding of molecular mechanisms of stress tolerance in Opuntia and other CAM plants.

Asunto(s)

Biblioteca de Genes , Opuntia/metabolismo , Proteínas de Plantas/metabolismo , Frío , Regulación de la Expresión Génica de las Plantas/genética , Regulación de la Expresión Génica de las Plantas/fisiología , Opuntia/genética , Opuntia/fisiología , Proteínas de Plantas/genética

RESUMEN

Proline (Pro) is one of the most accumulated osmolytes in salinity and water deficit conditions in plants. In the present study, we measured the Pro content, the activity and the expression level of delta 1-pyrroline-5-carboxylate synthetase (P5CS: gamma-glutamyl kinase, EC 2.7.2.11 and glutamate-5-semialdehyde dehydrogenase, EC 1.2.1.41), a key regulatory enzyme involved in the biosynthesis of Pro, in cactus pear (Opuntia streptacantha) subjected to 6, 9 and 11 days of salt stress. Treatment with NaCl of O. streptacantha young plants resulted in a decrease in the cladode thickness and root length, and in a significant and gradual accumulation of Pro in young cladodes, in a time- and concentration-dependent manner. P5CS activity, studied as gamma-glutamyl kinase, was reduced at all times as a consequence of salt treatment, except at the sixth day at 75 and 150mM of NaCl, where a slight increase was observed. We isolated an open reading frame (ORF) fragment of p5cs gene. The deduced amino acid sequence of the P5CS protein exhibited 90.4% of identity with the P5CS protein from Mesembryanthemum crystallinum. RT-PCR analysis revealed that the Osp5cs gene of O. streptacantha was induced by salt stress at 9 and 11 days of treatment. Furthermore, ABA-induced Osp5cs gene expression was observed in cladodes of cactus pear young plants. We observed an evident correlation between the transcript up-regulation and the Pro accumulation under salt stress; however, these results do not parallel with the changes in P5CS enzymatic activity. This Pro accumulation might function as an osmolyte for the intracellular osmotic adjustment and might be playing a critical role in protecting photosynthetic activity in O. streptacantha plants under salt stress.

Asunto(s)

Mesembryanthemum/metabolismo , Opuntia/metabolismo , Prolina/biosíntesis , Cloruro de Sodio/farmacología , Ácido Abscísico , Secuencia de Aminoácidos , Relación Dosis-Respuesta a Droga , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/fisiología , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/fisiología , Glutamato-5-Semialdehído Deshidrogenasa/biosíntesis , Glutamato-5-Semialdehído Deshidrogenasa/genética , Mesembryanthemum/genética , Datos de Secuencia Molecular , Opuntia/genética , Presión Osmótica/efectos de los fármacos , Fosfotransferasas (aceptor de Grupo Carboxilo)/biosíntesis , Fosfotransferasas (aceptor de Grupo Carboxilo)/genética , Fotosíntesis , Proteínas de Plantas/biosíntesis , Proteínas de Plantas/genética , Prolina/genética , Factores de Tiempo

RESUMEN

An experimental design considering thermal treatment of must, yeast strain, prickly pear variety and degree of ripeness was chosen to evaluate the fermentation behavior and generation of volatile compounds, during the elaboration of a distilled beverage from prickly pear. Four Mexican prickly pear varieties were characterized physically and two of them were selected for fermentation studies. The thermal treatment of the must showed the highest statistical influence on fermentation behavior and production of volatile compounds, followed by prickly pear variety, then yeast strain and finally the degree of ripeness was the least statistically significant factor. The growth rate increased when the thermal treatment was applied whereas the ethanol production rate and alcoholic efficiency were unaffected. The results also suggested that thermal treatment was effective for inhibition of microbial contamination. As regards volatile compounds production, acetic acid and methanol decreased while other volatiles increased when the thermal treatment was applied. Despite the influence of thermal treatment, prickly pear variety strongly influences the volatile profile of fermented musts.

Asunto(s)

Bebidas , Frutas/metabolismo , Opuntia/metabolismo , Levaduras/metabolismo , Acetaldehído/metabolismo , Etanol/metabolismo , Fermentación , Microbiología Industrial/métodos , Cinética , Opuntia/clasificación , Opuntia/crecimiento & desarrollo , Temperatura , Volatilización , Levaduras/crecimiento & desarrollo

RESUMEN

The genus Opuntia is a valuable forage resource in arid and semiarid lands during periods of drought and shortage of herbaceous plants. However, absolute minimum temperatures in the plains of Mendoza represent a limiting factor to cultivate several species. Opuntia ellisiana is a cold hardy species, so the goals of this study were to massively propagate is using in vitro culture techniques, and then to acclimatize plantlets obtained to field conditions. Different sterilization protocols were tested. Areoles were isolated in laminar airflow cabinet, and cultured on Murashige-Skoog medium, supplemented with sucrose and different BAP and IBA combinations. Explants were grown at 27 +/- 2 degrees C, under a 16-h photoperiod. The shoots produced were used in the rooting assay using different auxin combinations. In the most efficient growth treatment, plantlets reached 100% shooting after 35 days of culture, and a mean length of 10.2 mm after 49 days of culture. A 100% rooted plantlets was obtained on a medium containing 5 mg L-1 IBA, after 12 days of culture. Acclimatization was achieved under greenhouse conditions, showing 100% plantlet survival. This study suggests that O. ellisiana can be successfully micropropagated by areoles, and easily acclimatizated to field conditions. (AU)

Asunto(s)

RESEARCH SUPPORT, NON-U.S. GOVT , Aclimatación , Opuntia/crecimiento & desarrollo , Opuntia/metabolismo , Adaptación Fisiológica , Agricultura , Ambiente

RESUMEN

The genus Opuntia is a valuable forage resource in arid and semiarid lands during periods of drought and shortage of herbaceous plants. However, absolute minimum temperatures in the plains of Mendoza represent a limiting factor to cultivate several species. Opuntia ellisiana is a cold hardy species, so the goals of this study were to massively propagate is using in vitro culture techniques, and then to acclimatize plantlets obtained to field conditions. Different sterilization protocols were tested. Areoles were isolated in laminar airflow cabinet, and cultured on Murashige-Skoog medium, supplemented with sucrose and different BAP and IBA combinations. Explants were grown at 27 +/- 2 degrees C, under a 16-h photoperiod. The shoots produced were used in the rooting assay using different auxin combinations. In the most efficient growth treatment, plantlets reached 100% shooting after 35 days of culture, and a mean length of 10.2 mm after 49 days of culture. A 100% rooted plantlets was obtained on a medium containing 5 mg L-1 IBA, after 12 days of culture. Acclimatization was achieved under greenhouse conditions, showing 100% plantlet survival. This study suggests that O. ellisiana can be successfully micropropagated by areoles, and easily acclimatizated to field conditions.

Asunto(s)

Aclimatación , Opuntia/crecimiento & desarrollo , Opuntia/metabolismo , Adaptación Fisiológica , Agricultura , Ambiente

RESUMEN

The genus Opuntia is a valuable forage resource in arid and semiarid lands during periods of drought and shortage of herbaceous plants. However, absolute minimum temperatures in the plains of Mendoza represent a limiting factor to cultivate several species. Opuntia ellisiana is a cold hardy species, so the goals of this study were to massively propagate is using in vitro culture techniques, and then to acclimatize plantlets obtained to field conditions. Different sterilization protocols were tested. Areoles were isolated in laminar airflow cabinet, and cultured on Murashige-Skoog medium, supplemented with sucrose and different BAP and IBA combinations. Explants were grown at 27 +/- 2 degrees C, under a 16-h photoperiod. The shoots produced were used in the rooting assay using different auxin combinations. In the most efficient growth treatment, plantlets reached 100% shooting after 35 days of culture, and a mean length of 10.2 mm after 49 days of culture. A 100% rooted plantlets was obtained on a medium containing 5 mg L-1 IBA, after 12 days of culture. Acclimatization was achieved under greenhouse conditions, showing 100% plantlet survival. This study suggests that O. ellisiana can be successfully micropropagated by areoles, and easily acclimatizated to field conditions.

Asunto(s)

Aclimatación , Opuntia/crecimiento & desarrollo , Opuntia/metabolismo , Adaptación Fisiológica , Agricultura , Ambiente