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.
-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.
Biotransformations are chemical reactions catalyzed by cells, organs or enzymes and represent an area of biotechnology that has received considerable attention. The use of biotransformations with plant cell culture systems and fragments of plant tissue has immense potential for the production of compounds with commercial interest, especially considering the vast biochemical capability for the production of secondary metabolites from plant sources. In this context, this chapter evaluates the application of biotransformations in different plant cell culture systems, such as cell suspensions, hairy roots and cell immobilization, as well as fragments of plant tissue.
In vitro culture techniques are recognized as efficient strategies for large-scale plant production, as well as providing alternatives for plant conservation. In this study the micropropagation of Tarenaya rosea was established using petiole and foliar blade segments cultivated on MS medium with 6-benzyladenine (BA) and/or 6-furfurylaminopurine (KIN). The regeneration rate from explants was evaluated after 30-days in culture, as well as the proliferation rate from explant-derived shoots, reached after four subcultures performed at 30-days in culture. In vitro propagation occurred by both direct (DO) and indirect (IO) organogenesis. The highest regeneration rates by DO (50% to 100%) were reached on media containing only BA, while morphogenic calluses (IO) were mainly formed with BA+KIN. Explants on media with BA showed the presence of small black nodules on their surface, and histological analysis revealed the presence of trichomes with anthocyanin content. Elongation and rooting were reached on growth regulator-free MS. Acclimatization rates around 80% were achieved and the in vitro-regenerated plants were successfully maintained under field conditions. Results show significant morphogenetic potential of T. rosea from leaf explants, mainly when cultivated in the presence of 4.4 µM BA, providing a new alternative source of plant material for biotechnological and in vitro conservation studies.
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