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Previous results have shown that some proteins secreted in the culture medium are involved with the formation of embryogenic cells and can modify somatic embryo differentiation. Undifferentiated cell suspensions grown in the presence of 13 mM 2,4-dichlorophenoxyacetic acid (2,4-D) and obtained from embryogenic and non-embryogenic callus were used to study these events in sugarcane plants (cv. PR-62258). The cell suspension growth curves were determined and soluble proteins were extracted from embryogenic and non-embryogenic callus and culture medium from cell suspensions. In embryogenic callus we detected 1.43 times more protein than in non-embryogenic callus and the electrophoretic protein patterns show specific polypeptides for both callus types. In embryogenic callus we detected a cluster of four polypeptides in the range of 38±44 kDa and another polypeptide of 23 kDa that were not observed in non-embryogenic callus. In nonembryogenic callus there is a 35-kDa polypeptide that was not detected in embryogenic callus. In the case of extracellular proteins, the medium from embryogenic cell suspensions contained four polypeptides of 41, 38, 34 and 28 kDa that were slightly detected in the medium from non-embryogenic cell cultures; we also detected a band at 15 kDa that could not be observed in the medium from non-embryogenic cell suspensions. These results suggest that the development of embryogenic callus and cell suspensions is related to the type and amount of intracellular proteins in the callus cells and to the secreted proteins from these cells into the medium.

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Tissue culture techniques are routinely used for mass propagation and the establishment of disease free stock material. Virtually all pot type Anthuriums available in the market today are produced by tissue culture. In this chapter, we describe an efficient protocol to obtain Anthurium andreanum cv Rubrun vitro plants through micropropagation and organogenesis. Seeds from plant spadixes were germinated on MS medium supplemented with 0.5 mg/L BA. Micro-cuttings from in vitro germinated seedlings were subcultured on MS medium containing 2 mg/L BA and 0.5 mg/L NAA. Four-week-old in vitro plants obtained from microcuttings, showed callus proliferation at the stem base. The development of shoots and plantlets was observed from callus tissue. We also describe a detailed method for the histological analysis of callus tissue and a vitro plants acclimatization protocol.

To establish an efficient regeneration system for Anthurium andreanum cv Rubrun, seeds from plant spadixes were germinated on a medium supplemented with 2.2 µ M BA. After 2 weeks, 74% of the seeds germinated and four weeks later, micro-cuttings from these plantlets were subcultured on a medium containing 4.4 µ M BA and 0.05 µ M NAA. On average, 3.6 shoots per explant were obtained. Four weeks old in vitro plants from germinated seeds and the plantlets obtained from micro-cuttings, showed callus proliferation *Corresponding author at the stem base. These tissues were subcultured on a medium supplemented with 8.9 µ M BA and 2.7 µ M NAA. After 6 weeks of culture, about 43.8 plantlets per square cm of callus were obtained. Anatomical studies showed the organogenic nature of these calli. Anthurium andreanum plants regenerated by organogenesis were transferred to pots and a rate of 80% of plant acclimatization was obtained.The Anthurium genus comprises about 1500 tropical

RESUMENExisten numerosos factores que afectan la micropropagación y la microtuberización en papa; entre ellos, las hormonas vegetales y el fotoperiodo. Para estudiar el efecto de estos dos factores en las variedades 'Arbolona negra' (AN) y 'Granola' (G), se cultivaron microesquejes de cada variedad en medio MS líquido con o sin giberelinas (GA) y con 25 g/L de sacarosa e incubados bajo condiciones de luz blanca continua. Para inducir la microtuberización, las vitroplántulas obtenidas fueron sub-cultivadas en medio MS suplementado con 50 g/L sacarosa, tres concentraciones de BA (0, 1 y 5 mg/L) e incubadas bajo diferentes regímenes lumínicos. El pre-tratamiento con GA favoreció el alargamiento del vástago en AN pero no en G. Ambas variedades produjeron el mayor número de microtubérculos en medio MS suplementado con 5 mg/L de BA, bajo condiciones fotoperiódicas, sin la adición previa de GA. El cultivo in vitro de microesquejes de papa en medios de cultivo suplementados con BA y sacarosa, y la incubación bajo condiciones de días cortos permite obtener microtubérculos de papa en condiciones in vitro, en un tiempo más corto que el que podría esperarse en condiciones tradicionales de cultivo.Palabras clave: microtuberización, benciladenina, giberelinas, fotoperiodo. ABSTRACTMicropropagation and microtuberization in potato plants are both affected by plant hormones and photoperiod. To study the effect of these factors on potato cultivars 'Arbolona negra' (AN) and 'Granola' (G), microcuttings of each cultivar were cultured on MS liquid medium supplemented with sucrose 25 g/L, with or without gibberellins (GA). These microcuttings were incubated under continuous light. In order to induce microtuberization, the obtained vitro- plantlets were subcultured on MS medium supplemented with sucrose 50 g/L, three concentrations of BA (0, 1 and 5 mg/L) and incubated under different light patterns. GA pre-treatment induced shoots elongation on AN but not on G cultivar. The highest microtubers production for both cultivars was achieved on MS medium supplemented with BA 5 mg/L under short day light condition without GA pretreatment. The in vitro culture of microcuttings on culture media supplemented with BA and sucrose, incubated under short day conditions, allowed microtubers production in a shorter time lapse.

<p><strong>Título en ingles</strong>: Efecto de Dicamba y de ácido 2,4 diclorofenoxiacético sobre la embriogénesis somática en caña de azúcar</p><p>El cultivo <em>in vitro</em> de la caña de azúcar ha sido establecido en muchas variedades comerciales con el propósito de producir material libre de enfermedades microbianas, conservar germoplasma, detectar resistencia a enfermedades y plagas, etc. En este sentido, el objetivo de este trabajo fue analizar la efectividad de las auxinas sintéticas ácido 2,4-diclorofenoxiacético (2,4D) y ácido 3,6-dicloro-2-metoxibenzoico (Dicamba), en la inducción del proceso de embriogénesis somática y la regeneración de vitroplántulas de distintas variedades de caña de azúcar (C26670, RB855546, V99245, V756, V781, V0050, CC8592, CC8475). Para esto se cultivaron discos de hojas en fase de macollamiento, de 1 cm de diámetro y 2 mm de grosor, en medio Murashige-Skoog, 1962 (MS) suplementado con 50 ml.l-1 agua de coco, 30 g.l-1 sacarosa y dos tratamientos diferentes: 3 mg.l-1 2,4-D ó 6.63 mg.l-1 Dicamba, ambos en completa oscuridad a 25ºC, durante 1 mes. Los callos obtenidos se colocaron en medio de regeneración, conteniendo ½ sales MS, 200 ml.l-1 agua de coco y 60 g.L-1 sacarosa, incubándose bajo luz continua, 25ºC, por 2 meses. El mayor porcentaje de callo embriogénico se obtuvo en medios suplementados con Dicamba un promedio de 70,83 % de callo embriogénico por variedad ; mientras que en los medios con 2,4D se obtuvo 62,08 % de callo embriogénico por variedad. Se obtuvo un promedio de 89,00 % de plantas regeneradas a partir de los callos obtenidos en medios con Dicamba y 66,12 % de plantas a partir de callos obtenidos en medios con 2,4D. Con el uso de Dicamba se estableció un sistema eficiente de embriogénesis somática para estas variedades de caña de azúcar.</p>