Nodulation capacity in legume forest species (Fabaceae) according to their phylogeny and morphological characteristics

Gran parte del éxito de las plantas leguminosas (Familia Fabaceae) en los diferentes ecosistemas de Costa Rica podría deberse a su capacidad de establecer relaciones simbióticas con microorganismos de la familia Rhizobiaceae y al nitrógeno fijado que estas bacterias les pueden proveer. En este trabajo, evaluamos la presencia de nódulos, tejido donde se realiza la fijación de nitrógeno, en 24 especies forestales de leguminosas pertenecientes a cuatro subfamilias de Fabaceae y se anotaron varias características morfológicas de las raíces. Se detectó la presencia de nódulos en 14 de las 24 especies (58.3 %), sin embargo, hubo grandes diferencias entre las subfamilias. En las dos subfamilias consideradas las más basales (Detarioideae y Cercidoideae) no se detectaron nódulos. En la subfamilia Caesalpinioideae, se reportaron nódulos en el 58.3% de las especies, restringidas mayoritariamente al clado Mimosoideae. En la familia Papilionoideae, considerada la más derivada, se observó la presencia de nódulos en el 87.5 % de las especies evaluadas. La mayoría de especies con nódulos presentaron una coloración clara en sus raíces y las especies sin nódulos una coloración más oscura, con algunas excepciones. Estas tendencias están relacionadas con la historia filogenética de las subfamilias y los orígenes de la nodulación. Sin embargo, también podrían responder a diferentes mecanismos de infección e incluso podrían relacionarse con características fisiológicas de las plantas, como la producción de compuestos secundarios inhibitorios de la nodulación. De esta forma, este estudio establece un primer paso para entender mejor las interacciones entre los microorganismos fijadores de nitrógeno con sus hospederos.A large part of the success of legume plants (Fabaceae) in the different ecosystems of Costa Rica could be due to their ability to establish symbiotic relationships with microorganisms of the Rhizobiaceae family and to the fixed nitrogen that these bacteria can provide them. In this work, we evaluated the presence of nodules, the tissue where nitrogen fixation is carried out, in 24 forest species of legumes belonging to four subfamilies of Fabaceae, and several morphological characteristics of the roots were determined. The presence of nodules was detected in 14 of the 24 species (58.3 %); however, there were large differences between the subfamilies. In the two subfamilies considered the most basal (Detarioideae and Cercidoideae), no nodules were detected. In the Caesalpinioideae subfamily, nodules were reported in 58.3 % of the species, mostly restricted to the clade Mimosoideae. In the Papilionoideae family, considered the most derived, the presence of nodules was observed in 87.5 % of the evaluated species. Most species with nodules had a light coloration on their roots, and species without nodules had a darker coloration, with some exceptions. These trends are related to the phylogenetic history of the subfamilies and the origins of nodulation. However, they could also respond to different infection mechanisms and could even be related to physiological characteristics of plants, such as the production of secondary inhibitory compounds of the nodulation. In this way, this study constitutes. the first step to understand better the interactions between nitrogen-fixing microorganisms with their hosts.Universidad de Costa Rica/[111-B9-204]/UCR/Costa RicaUCR::Vicerrectoría de Docencia::Ciencias Básicas::Facultad de Ciencias::Escuela de Biologí

Endogenous GUS-like activity in Capsicum chinense Jacq.

The gene uidA, codes for β-glucuronidase, which is one of the reporters more frequently utilized in transgenic plants. However, this can only be use if the selected organism does not present endogenous GUS-like activity. In tissues of C. chinense we found a GUS-like activity showing different levels of intensity. Histochemical screening showed that endogenous GUS-like activity decreased, or reduced significantly, in almost all tissues with exception of stament, when phosphate buffer was adjusted to pH 8. Subsequently, C. chinense zygotic embryo explants were transient transformed with Agrobacterium tumefaciens LBA4404 (pCAMBIA2301) and plantlets regenerated were histochemically stained in phosphate buffer pH 8. Observations of incubated tissues of C. chinense regenerants showed blue staining, suggesting expression of uidA. Incubated tissues of non-transformed regenerants did not show blue staining in phosphate buffer pH 8. The results show that for transformation experiments of C. chinense with uidA gene, pH 8 is recommended for histochemical staining

Effect of Gamma Irradiation and Selection with Fungus Filtrate (Rhizoctonia solani Kuhn) on the in Vitro Culture of Common Bean (Phaseolus vulgaris)

The present investigation was undertaken to study the effect of gamma irradiation (dose from 10 to 100 Gy) and in vitro selection with fungus filtrate as selecting agent (concentration from 20% to 100%) on the susceptibility of the common bean to Rhizoctonia solani. The best results were found with a dose of 20 Gy or a concentration of 20% of fungus filtrate applied separately. These conditions were used to evaluate the combined effect of both approaches in a second experiment. The combined effect of irradiation and then selection adversely affected growth (height and roots) and survival of the in vitro plants. It may not be necessary to combine the variation generated by irradiation with the selection technique. For future assays we propose the application of: 1) gamma radiation, thereby inducing not only mutants with pathogen resistance, but also with other agronomic traits of interest. Later in the subculture MV4 potential fungus-resistant mutants will be evaluated in the field; or 2) selection pressure using fungus filtrate during three subcultures, which may be sufficient to induce the variation necessary to obtain in vitro plants resistant to fungus.International Atomic Energy Agency/[COS5028]/IAEA/AustriaUniversidad de Costa Rica/[111-A8-210]/UCR/Costa RicaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biología Celular y Molecular (CIBCM)UCR::Vicerrectoría de Docencia::Ciencias Básicas::Facultad de Ciencias::Escuela de BiologíaUCR::Vicerrectoría de Docencia::Ciencias Sociales::Facultad de Ciencias Económicas::Escuela de Estadístic

Establecimiento de suspensiones celulares de dos especies Jatropha (Euphorbiaceae) de Costa Rica

J. curcas has been studied in different countries and some interesting agronomic, pharmacological and industrial properties have been reported. More recently, it has been considered an important alternative source for biofuel production. The objective of this study was to establish a long-term method for the maintenance of calli and cell suspension cultures of the local species J. curcas and J. gossypifolia, in order to allow future studies for novel compounds with pharmaceutical or industrial applications. For this, friable calli were successfully induced from hypocotyl segments of J. curcas and J. gossypifolia that were cultured in semisolid MS media supplemented with 1.5mg/L, and 0.5mg/L of 2,4-D, respectively. Cell suspension cultures of J. curcas were established using 1g of 35 and 60-day calli, in 50mL of liquid MS media supplied with 1.5mg/L of 2,4-D; sucrose and maltose were additionally evaluated as carbon sources. After 35 days, cell suspension cultures initiated with 35-day calli, showed greater cell growth with a maximum biomass of 194.9g/L fresh weight, 6.59g/L dry weight and 17.3% packed volume. The exponential phase ended at day 35 for cultures initiated with 35-day calli, and at day 21 for cultures initiated with 60-day calli. Higher biomass production was obtained with sucrose. Cell cultures were established with 35-day calli in MS media with the same 2,4-D concentration used for calli induction and 30g/L sucrose. This medium was considered optimum for the maintenance and growth of cell suspensions for both species, with sub-cultures every 20 days. The biotechnological potential for the production of bioactive compounds in these species for pharmacological, agricultural and industrial applications is being evaluated.J. curcas es un importante recurso alternativo de biocombustible. Por otro lado, propiedades de interés agronómico, farmacológico e industrial han sido reportadas para esta especie. El objetivo de este estudio fue el establecimiento y mantenimiento a largo plazo de callos y cultivos celulares en suspensión de J. curcas y J. gossypifolia, con el objetivo de permitir futuros estudios para nuevos compuestos con aplicaciones farmaceúticas e industriales. Los callos friables fueron exitosamente inducidos a partir de segmentos de hipocótilos J. curcas and J. gossypifolia cultivados en medio MS semisólido suplementado con 1.5mg/L y 0.5mg/L of 2,4-D, respectivamente. Los cultivos celulares en suspensión de J. curcas fueron establecidos utilizando 1g de callos de 35 y 60 días de edad en 50mL de medio MS líquido adicionado con 1.5mg/L de 2,4-D. Después de 35 días, los cultivos en suspensión celular iniciados con callos de 35 días, mostraron mayor crecimiento celular con una biomasa máxima de 194.9g/L de peso fresco y 6.59g/L de peso seco y 17.3% de volumen empacado. La fase exponencial finalizó al día 35 en los cultivos iniciados con callos de 35 días, y al día 21 en los cultivos iniciados con callos de 60 días. Dos fuentes de carbono fueron evaluadas: sacarosa y maltosa. La producción de mayor biomasa fue obtenida con sacarosa. Los cultivos celulares se establecieron con callos de 35 días cultivados en medio MS con la misma concentración de 2,4-D utilizada para la inducción de callos y 30g/L de sacarosa. Este medio fue considerado el óptimo para el mantenimiento y crecimiento de suspensiones celulares en ambas especies con subcultivos cada 20 días. El potencial biotecnológico para la producción de compuestos bioactivos en estas especies, para aplicaciones farmacológicas, agrícolas e industriales está siendo evaluado.Universidad de Costa Rica/[111-A9-097]/UCR/Costa RicaUCR::Vicerrectoría de Docencia::Ciencias Básicas::Facultad de Ciencias::Escuela de BiologíaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Agroalimentarias::Centro de Investigaciones Agronómicas (CIA

Establishment of cell suspension cultures of two Costa Rican Jatropha species (Euphorbiaceae)

J. curcas has been studied in different countries and some interesting agronomic, pharmacological and industrial properties have been reported. More recently, it has been considered an important alternative source for biofuel production. The objective of this study was to establish a long-term method for the maintenance of calli and cell suspension cultures of the local species J. curcas and J. gossypifolia, in order to allow future studies for novel compounds with pharmaceutical or industrial applications. For this, friable calli were successfully induced from hypocotyl segments of J. curcas and J. gossypifolia that were cultured in semisolid MS media supplemented with 1.5mg/L, and 0.5mg/L of 2,4-D, respectively. Cell suspension cultures of J. curcas were established using 1g of 35 and 60-day calli, in 50mL of liquid MS media supplied with 1.5mg/L of 2,4-D; sucrose and maltose were additionally evaluated as carbon sources. After 35 days, cell suspension cultures initiated with 35-day calli, showed greater cell growth with a maximum biomass of 194.9g/L fresh weight, 6.59g/L dry weight and 17.3% packed volume. The exponential phase ended at day 35 for cultures initiated with 35-day calli, and at day 21 for cultures initiated with 60-day calli. Higher biomass production was obtained with sucrose. Cell cultures were established with 35-day calli in MS media with the same 2,4-D concentration used for calli induction and 30g/L sucrose. This medium was considered optimum for the maintenance and growth of cell suspensions for both species, with sub-cultures every 20 days. The biotechnological potential for the production of bioactive compounds in these species for pharmacological, agricultural and industrial applications is being evaluated