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Water is one of the main factors that influence the growth and development of plants. Balsa (Ochroma pyramidale) is a fast-growing tropical tree species with a great demand in the international market. The influence of soil, water deficit on morphological, physiological, and the growth of Balsa (Ochroma pyramidale) with and without irrigation was investigated. For this study, four random repetitions were carried out in a randomized complete block design. The plants were irrigated regularly with furrow irrigation. The timing of irrigation was based on the soil, water potential, according to treatment criteria. All pots were irrigated at 15, 30, 45, 60 and 75 days. Genomic DNA of O. Pyramidale was identified by ERIC-PCR and RAPD, while phylogenetic tree was determined using the UPGMA method. The results showed that leave number, shoot number, stem elongation and stem diameter were affected significantly (P 0.05) were observed, while stomata length remained high (> 20 µm) and stomata diameter was decreased during evaluation days in plants without irrigation compared to control. DNA of eight plants isolated showed different band profiles. In conclusion, the parameters evaluated can be used as a measure of water stress, growers should be careful when using O. Pyramidale in both nursery and orchards where water scarcity is present.

The use of organic fertilizers to improve the agronomic parameters of soy (Glycine max (L.) Merrill) has been considered a key production practice. During the period from December 2015 to M...

Stilbenes are naturally scarce high-added-value plant compounds with chemopreventive, pharmacological and cosmetic properties. Bioproduction strategies include engineering the metabolisms of bacterial, fungal and plant cell systems. Strikingly, one of the most effective strategies consists in the elicitation of wild grapevine cell cultures, which leads to vast stilbene resveratrol accumulation in the extracellular medium. The combination of both cell culture elicitation and metabolic engineering strategies to produce resveratrol analogs proved more efficient for the hydroxylated derivative piceatannol than for the dimethylated derivative pterostilbene, for which human hydroxylase HsCYP1B1- and grapevine O-methyltransferase VvROMT-transformed cell cultures were respectively used. Rose orcinol O-methyltransferase (OOMT) displays enzymatic properties, which makes it an appealing candidate to substitute VvROMT in the combined strategy to enhance the pterostilbene production level by engineered grapevine cells upon elicitation. Here we cloned a Rosa hybrida OOMT gene, and created a genetic construction suitable for Agrobacterium-mediated plant transformation. OOMT’s ability to catalyze the conversion of resveratrol into pterostilbene was first assessed in vitro using protein extracts of agroinfiltrated N. benthamiana leaves and transformed grapevine callus. The grapevine cell cultures transformed with RhOOMT produced about 16 mg/L culture of pterostilbene and reached an extracellular distribution of up to 34% of total production at the best, which is by far the highest production reported to date in a plant system. A bonus large resveratrol production of ca. 1500–3000 mg/L was simultaneously obtained. Our results demonstrate a viable successful metabolic engineering strategy to produce pterostilbene, a resveratrol analog with enhanced pharmacological properties. This work has been supported by grants from the Spanish Ministry of Science and Innovation (BIO2014-51861-R and BIO2017-82374-R), The Spanish Ministry of Economy and Competitiveness (PEJ-2014-A-90762/PEJ-2014-P-00289), Generalitat de Catalunya (2014SGR215) and European Funds for Regional Development (FEDER).

Se evaluo la germinacion y crecimiento de semillas de balsa aplicando siete tratamientos pre-germinativos: inmersion en HSO4 por 32 minutos, inmersion en agua a 80 °C durante tres minutos, testigo, remojo en agua a 100 °C por 15 minutos, remojo en agua de coco 12 horas, lijado de las semillas hasta que pierdan su brillo natural, calor seco 96 °C durante 5 minutos. Sembradas en siete sustratos utilizando tierra negra sola y combinada con: tamo de arroz, arcilla mas arena, humus UTEQ, zeolita, arcilla mas ceniza, humus nacaro (humus de Lombriz). Se realizo un diseno completamente al azar (DCA) en un arreglo factorial 7 (sustratos) x 7 (tratamientos pre-germinativos) con tres repeticiones y 10 unidades de observacion. A los 28 dias el factor (sustrato) que mas influyo en el porcentaje de germinacion fue tierra negra mas zeolita 7 g/k con 17,75%. El tratamiento pre-germinativo (lijado de las semillas) 20,85%. Tierra negra + humus UTEQ 0,70 g k-1 x remojo en agua de coco por 12 horas, mostro 133,11 mm de altura de plantula. Tierra negra + humus UTEQ 0,70 g/k con 3,27 mm de espesor del vastago. El mayor numero de hojas (cinco) se obtuvo en el tratamiento tierra negra + inmersion en HSO4.

Cocoa, Theobroma cacao L. is one of the main tropical industrial crops. Cocoa has a very high level of interfering substances, such as polysaccharides and phenolic compounds that could prevent the isolation of suitable protein. Efficient methods of protein extraction are a priority to successfully apply proteomic analyses. We compared and evaluated two methods (A and B) of tissue preparation for total protein extract by phenol/SDS extraction protocol. The difference in the application of the two methods was that extensively washed dry powder of pod tissue were made in Method A, whereas that crude extract were prepared Method B. Extracted proteins were examined using one-dimensional electrophoresis (1-D). Results show that each extraction method isolated a unique subset of cocoa pod proteome. Principal component analysis showed little variation in the data obtained using Method A, while that in Methods B showed no low reproducibility, thus demonstrating that Method A is a reliable for preparing cocoa pod proteins. The protocol is expected to be applicable to other recalcitrant plant tissues and to be of interest to laboratories involved in plant proteomics analyses. A combination of extraction approaches is recommended for increasing proteome coverage when using gel-based isolation techniques. El cacao, Theobroma cacao L. es uno de los principales cultivos tropicales industriales. La mazorca de cacao tiene un nivel muy alto de sustancias interferentes, tales como polisacáridos y compuestos fenólicos, que podrían impedir el aislamiento adecuado de la proteína. El uso de métodos eficientes de extracción de proteínas es una prioridad para aplicar con éxito los análisis proteómicos. Nosotros comparamos y evaluamos dos métodos preparativos (A y B) de tejidos para la extracción de proteína total mediante el protocolo de extracción con fenol/SDS. La diferencia entre los dos métodos fue extensivos lavados del polvo seco, obtenido mediante trituración con nitrógeno, de la mazorca fueron realizados en el Método A, mientras que un extracto crudo se preparó en el Método B. Extracciones proteicas fueron examinadas utilizando electroforesis monodimensional (1-D). Los resultados muestran que cada método de extracción aisló un único subconjunto del proteoma de las mazorcas de cacao. El análisis de componentes principales mostró poca variación en los datos por el Método A, mientras que el Método B fue poco reproducible, lo que demuestra que el Método A de extracción es un método fiable para la preparación de proteínas de las mazorcas de cacao. Se espera que el protocolo sea aplicable a otros tejidos de plantas recalcitrantes y podría ser de interés para los laboratorios involucrados en análisis de proteómica de plantas. Se recomienda una combinación de los enfoques de extracción para aumentar la cobertura del proteoma utilizando las técnicas de separación a base de gel. This work has been supported by grants from the Senescyt-Government of Ecuador (UTEQ-Ambiental-9-FCAmb-IFOR-2014-FOCICYT002), AMM holds a grant MAEC-AECID (2014-2015) of Spain.

Dimethyl labeling is a type of stable-isotope labeling suitable for creating isotopic variants of peptides and thus be utilized for quantitative proteomics experiments. Labeling is achieved through a reductive amination/alkylation reaction using the low-cost reagents formaldehyde and cyanoborohydride, resulting in dimethylation of free amine groups of Lys and N-termini. Availability of isotopomeric forms of these reagents allows for the generation of up to six different isotopic variants. Here we describe the application of dimethylation to create two isotopic variants, light and heavy, differing in 4 Da, to label the total tryptic digest peptides of cocoa pod extracted from healthy pods from cultivars susceptible and resistant to the fungal disease called "frosty pod" caused by Moniliophthora roreri.

Grafting is a common practice in cacao cultivation, but it has not been reported whether rootstock-scion combinations respond differently in terms of water transport, growth, or nutrient uptake under varying soil water availability conditions. The effects of water deficits on water potential (Ψf), basal diameter (db), root growth, chlorophyll and leaf concentrations of nitrogen (N) were evaluated in 16 rootstock-scion combinations that resulted from four rootstocks and four scion clones. Grafted seedlings were subjected to two water regimes: 21 days without irrigation (WD) and continuous irrigation (I). Under WD conditions, Ψf tended to be lower when using the EETP800 clone with the four rootstocks, indicating that this clone may have a higher rate of transpiration. The greater Ψf (p

Cordia alliodora (laurel) wood attractive appearance, its physical and mechanical characteristics, its abundant natural regeneration, as well as the contribution to carbon sequestration and soil protection, have made it an ideal species for reforestation. However, there is insufficient good quality planting material to meet these needs and to respond to the average annual demand. Hence, vegetative propagation has become an essential tool in breeding and it is being widely used for the conservation of genotypes in clonal banks. To this end, a methodology for laurel tree epicormic shoot induction and rooting, using plant growth regulators was established. Cytokinin concentrations used for induction of shoots from adult trees were 0, 3000, 6000, 9000 mg L-1 of BAP alone and combined with 1000, 2000, 3000 mg L-1 of IAA a Complete Randomized Design (DCA) with seven treatments and three repetitions, considering each tree as a repetition, was applied. The concentrations of auxin for rooting of shoots were 0, 1000, and 1500 mg kg-1 of NAA and IBA. A factorial arrangement DCA 3 x 3 (NAA x hormone IBA) with five repetitions and five units per repetition was used. After 105 days, the induction of epicormic shoots and epicormic buds response was evaluated 2.67 buds 16.42 cm long with 6000 mg kg-1 of BAP + 2000 mg kg-1 of IAA were obtained. At 45 days rooting percentage was 54%, the number of roots was 1.78, 2.49 length 2.59 and force with 1500 mg kg-1 of NAA + 1500 mg kg-1 of IBA. This research demonstrated that using cytokinins and auxins is effective in laurel epicormic shoots induction and rooting.

Nothofagus alpina es una de las especies forestales más importantes del Sur de Chile y Argentina, países donde se ha empezado con programas de mejoramiento genético. La micropropagación es una técnica muy útil en la clonación de árboles con fines de mejoramiento, por tanto, los objetivos planteados en esta investigación fueron: determinar la mejor concentración de fitohormonas 6-bencilaminopurina (BAP) y ácido indol butírico (AIB) y, el medio de cultivo idóneo para inducir el enraizamiento de brotes y la brotación de secciones nodales de N. alpina. Se evaluaron los medios de Murashige Skoog (MS) y Wood Plant Medium (WPM), con la mitad de los macronutrientes (enraizamiento) y los micronutrientes completos (multiplicación), suplementado con peptone (0.1%), sacarosa (2.0%), agar (0.7%) y diferentes concentraciones de auxinas. El medio MS suplementado con 2 mg L-1 de AIB resultó mejor para inducir la producción de las raíces, tanto en número como en la longitud de la raíz principal. El mayor porcentaje de enraizamiento se registró en el medio WPM suplementado con 2 mg L-1 AIB. Los tratamientos para inducir multiplicación que incluyeron citoquinina originaron callos y no enraizaron; siendo mayor la presencia de callos en el medio WPM. Se sugiere que la aplicación de citoquininas no es indispensable para la brotación, puesto que en los tratamientos testigos hubo una buena brotación; lo que hace suponer que el material vegetal utilizado tiene suficiente niveles endógenos de esta fitohormona.

El objetivo de este estudio fue determinar la erosión potencial por reconversión productiva agrícola de la subcuenca de Llay-Llay perteneciente al valle del Aconcagua, en la V Región de Valparaíso en Chile, mediante la adaptación de las unidades de respuesta a la erosión (ERU´s), según Märker et al. (2001), las mismas fueron sometidas a un proceso analítico jerárquico previo a la modelización espacial final. La validación de las ERU´s se realizó a partir de los índices morfométricos de capacidad de transporte y estabilidad de laderas. Las ERU´s destacan por su carácter cualitativo y de distribución geo-espacial homogénea, dentro del área de estudio relacionadas con factores de erosividad y erodabilidad. Para entender el proceso evolutivo de reconversión productiva de la subcuenca de Llay-Llay en los últimos treinta años se aplicó el índice normalizado de vegetación (NDVI) a partir del análisis de imágenes de satélite multiespectrales Landsat ETM, mientras que para la modelización de las variables topográficas, pendiente del suelo y exposición de laderas, así como para el cálculo de los índices morfómetricos se utilizó un modelo de elevación digital del terreno (MDE) con una apropiada resolución espacial. La categoría de ERU´s de muy alta susceptibilidad a la erosión para la subcuenca de Llay-Llay corresponde al área cubierta por el cultivo de paltos (aguacate) en laderas, sitio en donde el índice morfométrico de estabilidad de laderas se presentó alto, mientras que las áreas con menor peligro de erosión, de la clase ERU´s de baja susceptibilidad son aquellas cubiertas por matorral arborescente denso.

Targeted mass spectrometric methods such as selected/multiple reaction monitoring (SRM/MRM) have found intense application in protein detection and quantification which competes with classical immunoaffinity techniques. It provides a universal procedure to develop a fast, highly specific, sensitive, accurate, and cheap methodology for targeted detection and quantification of proteins based on the direct analysis of their surrogate peptides typically generated by tryptic digestion. This methodology can be advantageously applied in the field of plant proteomics and particularly for non-model species since immunoreagents are scarcely available. Here, we describe the issues to take into consideration in order to develop a MRM method to detect and quantify isoforms of the thylakoid-bound protein polyphenol oxidase from the non-model and database underrepresented species Eriobotrya japonica Lindl.

Targeted mass spectrometric methods such as selected/multiple reaction monitoring (SRM/MRM) have found intense application in protein detection and quantification which competes with classical immunoaffinity techniques. It provides a universal procedure to develop a fast, highly specific, sensitive, accurate, and cheap methodology for targeted detection and quantification of proteins based on the direct analysis of their surrogate peptides typically generated by tryptic digestion. This methodology can be advantageously applied in the field of plant proteomics and particularly for non-model species since immunoreagents are scarcely available. Here, we describe the issues to take into consideration in order to develop a MRM method to detect and quantify isoforms of the thylakoid-bound protein polyphenol oxidase from the non-model and database underrepresented species Eriobotrya japonica Lindl.

RNA isolation is difficult in plants that contain large amounts of polysaccharides and polyphenol compounds. To date, no commercial kit has been developed for the isolation of high-quality RNA from tissues with these characteristics, especially for fruit. The common protocols for RNA isolation are tedious and usually result in poor yields when applied to recalcitrant plant tissues. Here an efficient RNA isolation protocol based on cetyltrimethylammonium bromide (CTAB) and two successive precipitations with 10 M lithium chloride (LiCl) was developed specifically for loquat fruits but it was proved to work efficiently in other tissues of loquat and woody plants. The RNA isolated by this improved protocol was not only of high purity and integrity (A260/280 ratios ranged from 1.90 to 2.04; A260/230 ratios were >2.0), but also of high yield (up to 720 μg on average [CV 21%] total RNA per gram fresh tissue). The protocol was tested on loquat fruit (different stages of development, postharvest, ripening, bruising), leaf, root, flower, stem and bud; quince fruit and root; grapevine cells in liquid culture; and, rose petals. The RNA obtained with this method is amenable to enzymatic treatments and can be efficiently applied for research on gene characterization, expression and function. This work has been supported by Spanish Ministry of Foreign Affairs and Cooperation (A/8823/07) and (B/017931/08), the Spanish Ministry of Science and Innovation (BIO2011-29856-C02-02) and European Funds for Regional Development (FEDER).

Multiple reaction monitoring (MRM) is emerging as a promising technique for the detection and quantification of protein biomarkers in complex biological samples. Compared to Western blotting or enzyme assays, its high sensitivity, specificity, accuracy, assay speed, and sample throughput represent a clear advantage for being the approach of choice for the analysis of proteins. MRM assays are capable of detecting and quantifying proteolytic peptides differing in mass unique to particular proteins, that is, proteotypic peptides, through which different protein isoforms can be distinguished. We have focused on polyphenol oxidase (PPO), a plant conspicuous enzyme encoded by a multigenic family in loquat (Eriobotrya japonica Lindl.) and other related species. PPO is responsible for both the protection of plants from biotic stress as a feeding deterrent for herbivore insects and the enzymatic browning of fruits and vegetables. The latter makes fruit more attractive to seed dispersal agents but is also a major cause of important economic losses in agriculture and food industry. An adequate management of PPO at plant breeding level would maximize the benefits and minimize the disadvantages of this enzyme, but it would require a precise knowledge of the biological role played by each isoform in the plant. Thus, for the functional study of the PPOs, we have cloned and overexpressed fragments of three PPO isoforms from loquat to develop MRM-based methods for the quantification of each isoform. The method was developed using an ion trap instrument and validated in a QQQ instrument. It resulted in the selection of at least two peptides for each isoform that can be monitored by at least three transitions. A combination of SDS-PAGE and MRM lead to detect two out of three monitored isoforms in different gel bands corresponding to different processing stages of PPO. The method was applied to determine the amount of the PPO2 isoform in protein extracts from fruit samples using external calibrants. A.M.-M. holds a research grant from Conselleria d’Educacio, Cultura I Sport de la Generalitat Valenciana (FPA/2013/A/074), and J.M.-C. a postdoctoral grant from SENESCYT-ECUADOR (006-IECESMG5-GPLR-2012). S. S.-M. acknowledges financial support from Proteored-ISCIII. This work has been supported by grants from the Spanish Ministry of Foreign Affairs and Cooperation (A/8823/07) and (B/107931/08); University of Alicante (VIGROB-105), the Spanish Ministry of Science and Innovation (BIO2011-29856-C02-02) and European funds for Regional development (FEDER).

Tuberculosis and brucellosis remain important causes of morbidity and mortality in many countries, for the detection of both diseases requires efficient and sensitive tool for effectuate the diagnosis. This study was aimed to evaluate and compare the duplex PCR versus the nested PCR, for detection of Brucella spp. (BR) and Mycobacterium spp. (TB). A total of 100 samples of tissues from tracheo-bronchial lymph nodes, bovine lung and bacterial isolate as positive controls were used. Were evaluated ten combinations of primers which were designed to flank the segment of the 16S rRNA sequence (RB) and antigen gen MPB70 (TB), the best result for the Duplex PCR was obtained with the primers Bru-2F/Bru-2R for BR and Tub-1F/Tub-N-R for TB. The amplification of the products was 225 and 230-bp respectively. In order to compare the results of the proposed technique, all samples were initially analyzed and compared between PCR and nested PCR (Kappa, k = 0.85) and the concordance between Duplex PCR and nested PCR (k = 0.88) for the two bacteria was very good. La tuberculosis y la brucelosis siguen siendo causas importantes de morbilidad y mortalidad en muchos países, para la detección de ambas enfermedades se requieren de herramientas eficientes y sensibles para efectivizar el diagnóstico. Este trabajo fue dirigido a evaluar y comparar la técnica de Dúplex PCR frente a la PCR anidada, para la detección de Brucella spp. (BR) y Mycobacterium spp. (TB). Se utilizó un total de 100 muestras de tejidos a partir de nódulos linfáticos traqueo bronquiales, pulmón de bovinos y, aislados bacterianos de TB y BR como controles positivos. Diez combinaciones de iniciadores dirigidos a flanquear un segmento de la secuencia 16S ARNr (BR) y el gen antígeno MPB70 (TB) fueron evaluados, el mejor resultado para la Dúplex PCR se logró con los iniciadores Bru-2F/Bru-2R para BR y para TB, Tub-1F/Tub-N-R. Los productos de amplificación fueron de 225 y 230-pb respectivamente. A fin de potencializar los resultados de la técnica propuesta, todas las muestras fueron inicialmente analizadas y comparadas entre la PCR simple y la PCR anidada (Kappa, k = 0,85) y, la concordancia entre los resultados obtenidos con la PCR anidada y la Dúplex PCR (k = 0,88), para las dos bacterias fue muy buena.

En Ecuador, las plantaciones de cacao presentan bajos promedios de producción debido a la diversidad de patógenos, especialmente a la infección por Moniliophthora roreri (monilia). Se cree que existe una relación entre el ataque del hongo y el aumento de los niveles de expresión de genes codificantes de polifenol oxidasas (PPOs) como mecanismo de defensa ante patógenos y herbívoros en diferentes plantas. Para la identificación de genes que codifican para PPOs, se seleccionaron hojas de cacao Nacional, provenientes de plantas resistentes y susceptibles a monilia, ubicadas en la Finca Experimental La Represa, propiedad de la Universidad Técnica Estatal de Quevedo. Se afinó un protocolo de extracción de ARN total de alta calidad para hojas de cacao recalcitrantes. Después de su retrotranscripción a ADNc, se realizaron ensayos de amplificación por PCR con diferentes primers, diseñados a partir de secuencias conservadas de PPOs. Los productos de amplificación permitieron la identificación de un gen de 961 pb, similar a un gen que codifica para la PPO predictiva de Theobroma cacao depositada en NCBI (XP_017978715.1) La identificación de este gen, es fundamental para evaluar a futuro los niveles de expresión y cuantificación en diferentes estados de desarrollo del fruto. Dicha cuantificación permitirá proponer herramientas de control para monilia y construir las bases para el mejoramiento genético del cacao Nacional. In Ecuador, cocoa plantations have low production averages due to the diversity of pathogens, especially the infection by Moniliophthora roreri (monilia). It is believed that there is a relationship between the attack of the fungus and increased levels of expression of genes encoding polyphenol oxidases (PPOs) as a defense mechanism against pathogens and herbivores in different plants. For the identification of genes that code for PPOs, National cocoa leaves were selected, from resistant and susceptible plants to monilia, located in the Finca Experimental La Represa, property of the Universidad Técnica Estatal de Quevedo. A high-quality total RNA extraction protocol was recalculated for recalcitrant cocoa leaves. After its retrotranscription to cDNA, PCR amplification assays will be carried out with different primers, from the conserved sequences of PPOs. The amplification products allowed the identification of a 961 bp gene, similar to a gene that codes for the predictive PPO of Theobroma cacao deposited in NCBI (XP_017978715.1). The identification of this gene is essential to evaluate future levels of expression and quantification in different stages of fruit development. This reading quantification will propose control tools for monilia and build the bases for the genetic improvement of the National cacao.

Resumen La amplia distribución y abundancia natural de Tabebuia donnell-smithii (guayacán blanco) se ha visto reducida por la tala excesiva de los árboles, lo que ha ocasionado la desaparición de genotipos valiosos así como disminución de las poblaciones naturales, existiendo además carencia de alternativas de producción de plantas a gran escala. Se hace por tanto necesario orientar la investigación a establecer una técnica para la propagación vegetativa de guayacán blanco utilizando fitorreguladores. La metodología se basó en el uso de las hormonas de enraizamiento ácido naftalenacético (ANA) y ácido indolbutírico (AIB), en concentraciones de 0, 1,500 y 2,000 mg kg-1 en sustratos turba y arena. Las yemas apicales fueron colocadas en una cámara húmeda en condiciones de invernadero. Se aplicó un Diseño Completamente al Azar (DCA) en un arreglo factorial 2 sustratos x 3 dosis de hormona ANA x 3 dosis de hormona AIB, con cuatro repeticiones y cuatro unidades de observación. A los 45 días se evaluó el porcentaje de sobrevivencia y el enraizamiento, el número de raíces, la longitud de la raíz mayor, el número de brotes, la longitud de brotes, y el vigor. Los resultados no mostraron diferencias significativas entre los distintos tratamientos para todas las variables, sin embargo para el efecto simple e interacciones se observó diferencias para las variables evaluadas, siendo el mejor sustrato turba y las concentraciones óptimas de auxinas de 0 a 1,500 mg Kg-1. Se concluye que el guayacán blanco es una especie de fácil enraizamiento, ya que con y sin la aplicación de hormonas enraizadoras en el sustrato turba y arena se logró la obtención de clones, disminuyendo el tiempo de obtención de nuevas plantas.

La presente investigación se la realizó en el Laboratorio de Biotecnología de la Universidad Técnica Estatal de Quevedo, los objetivos planteados fueron establecer una metodología para la propagación vegetativa de Fernansánchez (Triplaris guayaquilensis) con el uso de hormonas de enraizamiento (ANA y AIB), y determinar las mejores concentraciones de hormonas de ANA y AIB en el enraizamiento de ramillas de esta especie. Se aplicó un diseño completamente al azar con 5 tratamientos, 3 repeticiones y diez unidades experimentales por repeticiones. A los 45 días, los mejores tratamientos para el enraizamiento del Fernansánchez con (ANA y AIB) se obtuvieron en las concentraciones 1000 mg kg-1 ANA + 1000 mg kg-1 AIB para la longitud promedio de la raíz mayor con 17.67 cm; 500 mg kg-1 ANA + 500 mg kg-1 AIB, para el número promedio de raíces y sobrevivencia con 5 raíces y 93.33% y la concentración de 1000 mg kg-1 ANA + 1000 mg kg-1 AIB fue ideal para vigor con 2.89, equivalente a 3 (vigor alto). Con esta investigación queda demostrado que las auxinas ANA y AIB inducen y mejoran la calidad de raíces sobre las estacas de Fernansánchez.

One of the major intent of metabolic engineering in cell culture systems is to increase yields of secondary metabolites. Efficient transformation methods are a priority to successfully apply metabolic engineering to cell cultures of plants that produce bioactive or therapeutic compounds, such as Vitis vinifera and Taxus x media. The aim of this study was to establish a reliable method to transform non-embryogenic cell cultures of these species. The V. vinifera cv. Gamay/cv. Monastrell cell lines and Taxus x media were used for Agrobacterium-mediated transformation using the Gateway-compatible Agrobacterium sp. binary vector system for fast reliable DNA cloning. The Taxus x media and Vitis cell lines were maintained in culture for more than 4 and 15 months, respectively, with no loss of reporter gene expression or antibiotic resistance. The introduced genes had no discernible effect on cell growth, or led to extracellular accumulation of phytoalexin trans-Resveratrol (t-R) in response to elicitation with methylated cyclodextrins (MBCD) and methyl jasmonate (MeJA) in the grapevine transgenic cell lines compared to the parental control. The method described herein provides an excellent tool to exploit exponentially growing genomic resources to enhance, optimize or diversify the production of bioactive compounds generated by grapevine and yew cell cultures, and offers a better understanding of many grapevine and yew biology areas. This work has been supported by grants from the Spanish Ministry of Science and Innovation (BIO2011-29856-C02-01, BIO2011-29856-C02-02 and BIO2014-51861-R), European Funds for Regional Development (FEDER) and Conselleria d’Educacio, Cultura i Sport de la Generalitat Valenciana (FPA/2013/A/074). J.M.C. holds a postdoctoral grant from SENESCYT-GOVERNMENT OF ECUADOR (006-IECESMG5-GPLR-2012).

La tuberculosis y la brucelosis siguen siendo causas importantes de morbilidad y mortalidad en muchos países, para la detección de ambas enfermedades se requieren de herramientas eficientes y sensibles para efectivizar el diagnóstico. Este trabajo fue diri gido a evaluar y comparar la técnica de Dúplex PCR frente a la PCR anidada, para la detección de Brucella spp. (BR) y Mycobacterium spp. (TB). Se utilizó un total de 100 muestras de tejidos a partir de nódulos linfáticos traqueo bronquiales, pulmón de bovi nos y, aislados bacterianos de TB y BR como controles positivos. Diez combinaciones de iniciadores dirigidos a flanquear un segmento de la secuencia 16S ARNr (BR) y el gen antígeno MPB70 (TB) fueron evaluados, el mejor resultado para la Dúplex PCR se logró con los iniciadores Bru - 2F/Bru - 2R para BR y para TB, Tub - 1F/Tub - N - R. Los productos de amplificación fueron de 225 y 230 - pb respectivamente. A fin de potencializar los resultados de la técnica propuesta, todas las muestras fueron inicialmente analizadas y comparadas entre la PCR simple y la PCR anidada (Kappa, k = 0,85) y, la concordancia entre los resultados obtenidos con la PCR anidada y la Dúplex PCR (k = 0,88), para las dos bacterias fue muy buena.

En el presente trabajo se estudiaron las características morfológicas de conos, semillas y plántulas de Pinus greggii Engelm. provenientes de la cosecha 2005, de una población ubicada en el municipio de Zacualpan, Veracruz, México, con la finalidad de estimar la variación porcentual atribuida a las fuentes de variación, así como el comparar los promedios de cada familia. Los resultados obtenidos mostraron que el tamaño de los conos y semillas de esta especie difieren significativamente entre familias. Inclusive, el tamaño de las semillas también fue diferente entre los conos de una misma familia. Se evaluó el porcentaje de germinación, el valor germinativo y la velocidad de germinación, identificándose a la familia 2 como la que presentó el mayor porcentaje, la que germinó mas rápido y obtuvo un mayor valor germinativo. En cuanto a plántulas, se determinaron diferencias entre familias únicamente para la altura. Los resultados encontrados en la presente investigación permitirán brindar alternativas de manejo y conservación de la especie.

This research was performed in the Biotechnology Laboratory, located on the campus's central campus of Manuel Alvares Make State Technical University Quevedo, located at Av Quito at Km 1.5 on the way Quevedo-Santo Domingo de los Tsachilas. Which was to establish an assay to standardize a protocol for in vitro propagation and development of Swietenia macrophylla King (mahogany), were isolated apical buds of mahogany which is segmented into parts 1 to 1.5 cm long. The harvested plants were 6 months old. The buds used were three weeks old, previously selected for their phenological characteristics. Which before the establishment phase in the greenhouse were sprayed with two kinds of fungicides (Agrimicin and Benlate), twice a week throughout the trial, the application of fungicides is what was done to reduce the presence of fungal pathogens in the phase emissions and thus ensure greater survival of plants in the establishment of in vitro culture. The experimental design used was in the early stages of establishment and multiplication was a completely randomized design (CRD) with 3x2 factorial arrangements with 4 replications and 5 comments and the rooting phase, DCA 4 treatments and 4 replications and 5 observations per unit experimental. Obtaining the variance and mean separation at 95% probability are performed using the statistical program MSTAC and to establish statistical differences between treatments were used Multiple Range Test of Tukey (P ≥ 0.05). In the establishment phase of testing different concentrations of calcium hypochlorite (10, 15 g) at different times (10, 15, 20 min) and were evaluated at 21 days of established explants. The best average obtained in treatment 6 (15 g of calcium hypochlorite for 20 min) with 95% survival. In the multiplication phase to increase the production of shoots explant used two types of hormones of benzylaminopurine (BAP) and indole butyric acid (IBA) at different concentrations (1, 2, 3 mg L-1 BAP and 0.5, 1 mg L-1 IBA), the shoots obtained during this stage were transferred to a medium with growth regulators auxin type to induce rooting of shoots with different concentrations (1, 1.5, 2, 2.5 mg L -1 of IBA). The treatment increased the shoot proliferation stimulated Mahogany was the concentration of hormones containing (2 mg L-1 BAP and 1 mg L-1 IBA), which yielded a level of 70% germination. Tools that the lower number of outbreaks was obtained with the concentration (3 mg L-1 BAP and 0.5 mg L-1 IBA), which reached only 40%. In regard to the rooting of shoots obtained during the multiplication phase, none of the concentrations used induced rooting. Significantly, it is recommended to test new protocols for rooting phase and thus achieve the proliferation of roots in this species.