Physiological development of red anthurium (Anthurium andreanum Linden) var. Tropical in three in vitro culture systems

ABSTRACT Objective: to evaluate the physiological development of Anthurium andreanum L. var. Tropical red color in three in vitro culture systems: semi-solid, partial immersion and RITA® bioreactor Design / methodology / approach: A completely randomized design was used with three treatments: semi-solid medium, partial immersion and RITA® bioreactor, with four repetitions each. Vitroplants of anthurium were selected with a size of 0.5 cm from the stem to the highest leaf, with three leaves each specimen. After 60 days of sowing, morphometric analyzes, chlorophyll content and hormone content were carried out. An analysis of variance and comparison tests of means were performed on the data obtained by Kruskal-Wallis and Tukey, respectively, using the statistical software R-STUDIO. Results: The highest shoot rate and root length was obtained in partial immersion; however, the number of leaves, shoots and root multiplication did not show differences with the RITA® bioreactors. The highest concentration of chlorophyll and indole acetic acid was observed when using RITA® bioreactors. Study limitations / implications: The results are favorable for the in vitro production of anthurium, however for commercial production the use of RITA® bioreactors is a high cost in the initial investment. Findings / conclusions: with the results obtained, it is considered that the RITA bioreactors obtained the best results for the production of anthurium, followed by the partial immersion system. Due to the liquid medium and better gas exchange, which favors the development of plants Keywords: Anthurium andreanum L.; chlorophyll; phytohormones; immersion systemsObjective: To evaluate the physiological development of red Anthurium andreanum L. var. Tropical in three in vitro culture systems: semi-solid, partial immersion and RITA® bioreactor. Design / methodology / approach: A completely randomized design with three treatments, semi-solid medium, partial immersion and RITA® bioreactor, and four repetitions each was used. Vitroplants of anthurium were selected with a size of 0.5 cm from the stem to the highest leaf, with three leaves in each specimen. Morphometric, chlorophyll content and hormone content analyses were carried out after 60 days of sowing. Analysis of variance and means comparison tests were performed on the data obtained through Kruskal-Wallis and Tukey, respectively, using the statistical software R-STUDIO. Results: The highest shoot rate and root length were obtained in partial immersion; however, the number of leaves, shoots and root multiplication did not show differences with the RITA® bioreactors. The highest concentration of chlorophylls and indole acetic acid was observed when using RITA® bioreactors. Study limitations / implications: The results are favorable for the in vitro production of anthurium, although the use of RITA® bioreactors for commercial production is a high cost in the initial investment. Findings / conclusions: With the results obtained, it is considered that the RITA® bioreactors obtained the best results for the production of anthurium, followed by the partial immersion system. This is due to the liquid medium and better gas exchange, which favors the development of plant

Obtención in vitro de plantas de Echeveria pumila cv. 'Glauca' libres de bacterias y fitoplasmas asociados a la fasciación del tallo.

Echeveria pumila cv. ‘Glauca’ es una crasulácea nativa de México de importancia ornamental, el género Echeveria destaca como un grupo que se encuentra en peligro debido a la sobreexplotación de algunas de sus especies. En algunos especímenes de Echeveria pumila por ejemplo el cv. ‘Glauca’, se han observado síntomas de fasciación, alta proliferación y reducción del área foliar; síntomas que son característicos de infecciones por bacterias y fitoplasmas. En el presente trabajo, se detectó la presencia de fitoplasmas y bacterias asociados a este síntoma en plantas de E. Pumila cv. ‘Glauca’ mediante la técnica de PCR. En el caso de bacterias utilizando iniciadores universales 27f y 1492r y para fitoplasmas los iniciadores P1 - P7 y después con una PCR anidada utilizando R16F2 y R16R2. Para poder generar material libre de estos patógenos fue necesario la multiplicación in vitro de este cultivar, el procedimiento abarcó la evaluación de medios de cultivo suplementados con fitoreguladores en diferentes concentraciones. Se evaluaron combinaciones de bencil adenina (BA) 0.2 mg•L-1 más ácido naftalén acético (ANA) 0.02 mg•L-1 y otras tres concentraciones de thidiazuron (TDZ) (0.1, 0.2 y 0.6, mg•L-1) sin la presencia de ANA. Todos los medios se suplementaron con 30 g•L-1 de sacarosa, 6 g•L-1 de agar, 100 mg•L-1 de ácido cítrico y el pH se ajustó a 5.7, antes de la esterilización con autoclave. En los explantes asintomáticos, el medio con TDZ (0.6 mg•L-1) fue el que generó el mayor número de brotes por explante (B/E) con 2.5, mientras que los explantes con síntomas en el medio con BA más ANA produjeron una media de 3.6 B/E. En los brotes con síntomas éstos se siguieron manifestando en todos los subcultivos bajo condiciones in vitro. Éstos fueron utilizados para elaborar una nueva metodología para obtener plantas libres de patógenos, basadas en la combinación de tres técnicas para superar la dificultad que presenta el manejo de los tejidos con el síntoma de fasciación. Fue posible obtener plantas libres de patógenos mediante la combinación del uso de antibióticos, la etiolación de los tejidos y la posterior extracción de los meristemos de los brotes que habían sido regenerados in vitro a partir de ápices. Los resultados fueron comprobados mediante PCR e indicaron que el 90% de los brotes generados a partir de meristemos estuvieron libres de bacterias y fitoplasmas. _______________ In vitro OBTAINING Echeveria pumila cv. 'Glauca' PLANTS FREE OF BACTERIA AND PHYTOPLASMAS ASSOCIATED FASCIATION OF STEM. ABSTRACT: Echeveria pumila cv. 'Glauca' is a native of Mexico crasulácea ornamental importance, the genus Echeveria out as a group that is at risk due to overfishing of some species. In some specimens of Echeveria pumila cv. 'Glauca' have symptoms of fasciation, proliferation and reduction in leaf area; symptoms that are characteristic in cases of infection by bacteria and phytoplasmas. In the present study, detected the presence of phytoplasmas and bacteria associated in E. pumila cv. 'Glauca' plants with this symptom by PCR. in the case of bacteria using universal primers 27f and 1492r and for phytoplasms the primers P1 - P7 and a nested PCR with R16F2 and R16R2, to generate material free of these pathogens was necessary in vitro multiplication of this species in this procedure supplemented culture media phytoregulators evaluated at different concentrations, combining benzyl adenine 0.2 mg•L-1 (BA) more naphthalene acetic acid 0.02 mg•L-1 (NAA) and three concentrations of thidiazuron (TDZ) (0.1, 0.2 and 0.6 mg•L-1) without the presence of ANA. All media were supplemented with 30 g•L-1 sucrose, 6 g•L-1 agar, 100 mg•L-1 of citric acid and the pH was adjusted to 5.7 before autoclaving. In asymptomatic explants, the medium with TDZ (0.6 mg•L-1) was the one that generated the highest number of shoots per explant (B/E) with 2.5, while the symptoms explants with medium with BA and NAA produced an average of 3.6 B/E. Besides the symptoms continued to express in all subcultures under in vitro conditions. These were used to develop a new methodology for pathogen-free plants based on the combination of three techniques to overcome the difficulty of handling tissues with symptoms of fasciation. It was possible to obtain pathogen free plants by combining antibiotics, etiolation and subsequent meristems extraction of shoot regenerated in vitro. The results were verified by PCR and indicated that 90% of shoots generated from meristems were free of bacteria and phytoplasmas.Tesis (Maestría en Ciencias, especialista en Fitopatología).- Colegio de Postgraduados, 2014.Consejo Nacional de Ciencia y Tecnología (CONACYT)