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This paper describes a protocol for the efficient vegetative propagation of

Alternative methods for in vitro shoot culture of Cleome rosea, a Brazilian herbaceous species with ornamental value and medicinal potential, were evaluated. A protocol for rapid in vitro multiplication of roots, a valuable source of medicinal compounds, was also developed. Stem explants were cultured in liquid media (continuous immersion and paper bridge), while root explants were cultivated in continuous immersion and on solidified media. The highest numbers of shoots, 20 ± 4.6 shoots/ explant, were obtained from stem explants incubated in a continuous immersion system in a liquid medium supplemented with 2.2 lM BA. Root explants cultivated in liquid media produced only hyperhydrous adventitious shoots. However, these explants generated 5.8 ± 0.8 shoots/ explant by indirect organogenesis when cultivated on solidified medium supplemented with 2.2 lM BA. In addition, root multiplication was achieved in liquid medium in the presence of a-naphthaleneacetic acid. Adventitious shoots developed on newly formed roots when inoculated on solidified medium supplemented with 2.2 lM BA. Shoot microcuttings developed roots when transferred onto solidified MS medium without growth regulators. Rooted microcuttings were efficiently acclimatized when transferred ex vitro.

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The effects of different levels of Murashige and Skoog (MS) basal medium, 2,4-dichlorophenoxyacetic acid (2,4-D), and sucrose on anthocyanin production and biomass accumulation of cell suspension cultures of Cleome rosea were investigated. Cultures were established in liquid MS medium containing 30 g l -1 sucrose and supplemented with 0.90 lM 2,4-D. Proliferating cell suspension cultures achieved the highest growth capacity, a fourfold increase in biomass accumulation, following subculture at the exponential growth phase, 14-18 days of culture. Moreover, the presence of 2,4-D was essential for anthocyanin production and biomass accumulation. On the other hand, increasing levels of sucrose above 30 g l -1 resulted in a drastic reduction in biomass accumulation. Anthocyanin production was highest in cell suspension cultures grown on halfstrength MS medium (1/2 MS), 30 g l -1 sucrose, and 0.45 lM 2,4-D. These cell suspension cultures were mainly composed of small aggregates of spherical cells with similar morphology observed in anthocyanin-producing and non-producing cultures. Moreover, microscopic analysis of anthocyanin-producing cultures showed the presence of mixtures of non-pigmented, low-pigmented, and high-pigmented cells.

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-The genus Cleome is widely distributed in drier areas of the tropics and subtropics. Cleome dendroides and C. rosea are Brazilian native species that occur mainly in Atlantic Forest and sandy coastal plains, respectively ecosystems negatively affected by human impacts. Cleome spinosa is frequently found in urban areas. Many Cleome species have been used in traditional medicine, as C. spinosa. In the present work, was investigated C. dendroides, C. rosea and C. spinosa germinative behavior under in vivo conditions, as well as was established suitable conditions to in vitro germination and seedling development. The in vivo germination was performed evaluating the influence of temperature, substrate and light. It was observed that only C. spinosa seeds presents physiological dormancy, which was overcome by using alternate temperatures. The substrate influenced significantly the germination of C. rosea and the seeds of C. dendroides showed the highest germination percentages in the different conditions evaluated. The post-seminal development stages under in vivo and in vitro conditions were defined. It was observed that the development was faster under in vitro than in vivo conditions. An effective methodology for in vitro germination, enabling the providing of material to experiment on plant tissue culture was established to C. dendroides and C. spinosa.