In Vitro Mass Propagation of Cymbopogon citratus Stapf., a Medicinal Gramineae.

Cymbopogon citratus (D.C.) Stapf. is a medicinal plant source of lemon grass oils with multiple uses in the pharmaceutical and food industry. Conventional propagation in semisolid culture medium has become a fast tool for mass propagation of lemon grass, but the production cost must be lower. A solution could be the application of in vitro propagation methods based on liquid culture advantages and automation. This chapter provides two efficient protocols for in vitro propagation via organogenesis and somatic embryogenesis of this medicinal plant. Firstly, we report the production of shoots using a temporary immersion system (TIS). Secondly, a protocol for somatic embryogenesis using semisolid culture for callus formation and multiplication, and liquid culture in a rotatory shaker and conventional bioreactors for the maintenance of embryogenic culture, is described. Well-developed plants can be achieved from both protocols. Here we provide a fast and efficient technology for mass propagation of this medicinal plant taking the advantage of liquid culture and automation.

Influencia de la 6-Bencilaminopurina y el agente gelificante en la reducción de la hiperhidricidad en Tectona grandis L / Influence of 6-Benzyladenine and gelling agent on the reduction of hyperhydricity in Tectona grandis L

The influence of different factors on shoot proliferation and the occurrence of hyperhydricity in teak (Tectona grandis L.) have been studied. Four concentrations of BA (2.22, 4.44, 6.66 and 8.88 µM) and a control treatment with 0 BA were examined. Aiming at reducing the costs during commercial propagation by using gelrite in stead of agar, the use of both gelling agent in the proliferation and hyperhydricity was tested. In order to evaluate if hyperhydricity can be reduced by increasing the gelrite concentration in the culture medium, three concentrations (2.0, 2.5 and 3.0 g L-1) were tested in combination with 4.44 µM BA. The proliferation and occurrence of hyperhydricity during 21 successive subcultures were evaluated. The highest proliferation was achieved in the treatments with 6.66 or 8.88 µM BA. They yielded 5.22 and 5.56 shoots/explant, respectively. But also, the highest percent of hyperhydric shoots was achieved in this treatment. Gelrite resulted in a higher proliferation, but also an almost two times higher hyperhydricity as compared to agar-solidified media. Satisfactory reduction in hyperhydricity (18%) was achieved with 3.0 g L-1 gelrite. However, the successive subcultures onto proliferation in this treatment favored hyperhydricity compromising shoot quality and it´s competence to proliferate. in vitro teak plants were ex vitro rooted and then transferred to greenhouse conditions for acclimatization; ten weeks after transfer they were ready for field plantation.

Se estudió la influencia de diferentes factores en la proliferación y la ocurrencia de la hiperhidricidad in teca (Tectona grandis L.). Se probaron cuatro concentraciones de BA (2,22; 4,44; 6,66 and 8,88 µM) y un control sin BA. Con el objetivo de reducir los costos durante la propagación comercial se experimentó sustituir el agar por el gelrite, para lo cual se estudió en efecto de ambos gelificantes en la proliferación y la hiperhidricidad de los brotes. Se estudiaron, tres concentraciones de gelrite (2,0; 2,5 and 3,0 g L-1) combinadas con 4,44 µM BA, con el objetivo de evaluar si la hiperhidricidad podía ser reducida incrementando la concentración de gelrite. Se evaluó la proliferación de brotes y la ocurrencia de la hiperhidricidad durante 21 subcultivos. Se logró una alta proliferación de brotes en los tratamientos con 6,66 y 8,88 µM BA (5,22 y 5,56 brotes), pero el porcentaje de brotes hiperhídricos también se incrementó. El gelrite resultó en una alta proliferación de brotes, pero con mayor incidencia de la hiperhidricidad que el medio gelificado con agar. Se obtuvo una reducción satisfactoria de la hiperhidricidad (18%), cuando la concentración de gelrite se incrementó hasta 3,0 g L-1. No obstante, la multiplicación de los brotes en este tratamiento más allá del 11no subcultivo favoreció la hiperhidricidad, lo que afectó la calidad de los brotes y su competencia para la proliferación. Las plantas fueron enraizadas ex vitro, transferidas a condiciones de invernadero para su aclimatización y diez semanas después de la transferencia estaban listas para la plantación en campo.

Proteomic profiling of Tectona grandis L. leaf.

Tectona grandis L. (teak) is one of the premier hardwood timbers in the world, ranking at present in the top five tropical hardwood species in terms of worldwide plantation area. Characterization of the proteins present in teak leaves will provide a basis for the development of new tools aimed at assisting tree selection, the monitoring of plant propagation, and the certification of clonal and phenotypic identities. In this paper, we describe the extraction, separation, and identification of leaf proteins from T. grandis using a TCA/acetone protocol, 2DE, and MALDI-TOF. After TCA/acetone protein extraction of leaves, 998 well-resolved spots were detected in Coomassie-stained gels within the 10-114 kDa relative molecular mass (Mr) range at a pH ranging from 3 to 11. A total of 120 spots were digested and subjected to MS. Of these, 100 nonredundant protein species were successfully identified. Functional classification of the identified proteins revealed that proteins involved in photosynthesis, protein translation, and energy production were the most abundant. This work is the first high-throughput attempt to study the T. grandis leaf proteome and represents a stepping stone for further differential expression proteomic studies related to growth, development, biomass production, and culture-associated physiological responses.