MULTIPLICAÇÃO E ALONGAMENTO IN VITRO DE CLONES HÍBRIDOS DE Eucalyptus globulus

Os híbridos de Eucalyptus globulus representam uma excelente alternativa para o setor de celulose e papel, em razão dos ganhos em qualidade da madeira para a fabricação de celulose. Entretanto, estes híbridos têm apresentado recalcitrância ao enraizamento adventício. Assim, a micropropagação é apontada como a técnica para o rejuvenescimento desses híbridos adultos, viabilizando a propagação clonal dos mesmos. O presente trabalho avaliou o cultivo in vitro de três clones de Eucalyptus grandis x Eucalyptus globulus e de três clones de Eucalyptus urophylla x Eucalyptus globulus, em relação à multiplicação in vitro, no meio MS suplementado com 0,5 mg L-1 de BAP e 0,01 mg L-1 de ANA, bem como o efeito das concentrações de 0,25; 0,50; 0,75 e 1,0 mg L-1 de AIB e dos meios de cultura MS e JADS no alongamento in vitro das brotações. Os clones diferiram quanto à multiplicação in vitro das brotações e apresentaram uma taxa de multiplicação média dos clones de 3,0 tufos de brotações em cada subcultivo, ao longo dos 25 subcultivos realizados. No alongamento in vitro, os clones diferiram quanto às concentrações de AIB adequadas para provocar o alongamento, bem como em relação aos meios de cultura MS e JADS. O intervalo médio entre 0,40 e 0,80 mg L-1 de AIB proporcionou o maior número e comprimento das brotações alongadas in vitro e com maior vigor.

In vitro technique for selecting onion FOR white rot disease- resistance

In vitro selection is one of the most effective and efficient techniques for plant improvement. This is due to its ability to isolate plants with the desired character(s), either by applying a selection agent on the culture media to drive the selection of somaclones with the required character(s), or by establishing particular conditions that change in the genomes of somaclones toward the required character. The objective of this study was to identify a suitable protocol for in vitro selection of Allium white rot disease ( Sclerotium cepivorum ) tolerance in commercial Egyptian onion varieties, namely Giza 20, Giza 6 and Beheri Red. Oxalic acid (OA), the phytotoxin produced by Sclerotium cepivorum, was used as the selective agent. Seeds of the three Egyptian varieties were germinated on four concentrations (0.0, 0.02, 0.2, 2 and 20 mM) of Oxalic acid. Among the tested cultivars, Beheri Red had the highest germination frequency (52%) at all concentrations tested, followed by Giza 20 (42.6%), and Giza 6 at (32%). Cotyledon explants from the varieties were cultured on toxic MSBDK medium, supplemented with 0, 3, 6 and 12 mM OA. The survival of calli on MSBDK free toxic medium was 70.7% for all tested cultivars; however, MSBDK-stressed medium, with 3 mM OA reduced the viable calli to 42.1%. The highest OA concentration (12 mM) completely inhibited calli induction from cotyledons explants. A medium supplement with 3 mM OA retarded 80% of calli growth. Among 156 tested calli of Beheri Red, only 23 calli (14.7%) survived on toxic medium for 45 days. Similarly, there was 15.6% survival for Giza 20 calli, while 40.1% of the Giza 6 calli survived. Plantlets were regenerated from surviving calli and transplanted onto ex vitro, and formed bulb after acclimatisation.

Improved production of chlorogenic acid from cell suspension cultures of Lonicera macranthoids

Purpose: To evaluate the potential of Lonicera macranthoids Hand. -Mazz. Yulei1 suspension culture system for enhanced production of the main secondary metabolite, chlorogenic acid. Methods: The callus of L. macranthoides Hand.-Mazz. “Yulei1” was suspension cultured in B5 liquid medium supplemented with different plant growth regulators. Biomass accumulation was calculated by weight method and chlorogenic acid production was measured using high performance liquid chromatography (HPLC). HPLC was carried out on C18 analytical column at 35 °C and the detection wavelength was set at 324 nm. Results: The results showed that maximum accumulation of biomass and chlorogenic acid were achieved 15 days after culture growth. The optimized conditions for biomass accumulation and chlorogenic acid production were 50 g/L of inoculum on fresh weight basis, B5 medium supplemented with plant growth regulators, 30 - 40 g/L sucrose and initial medium pH of 5.5. Maximum accumulation of chlorogenic acid and biomass were observed when the culture medium was supplemented with 2.0 mg/L6-BA. Optimal accumulation of chlorogenic acid was observed using combination of hormones 2.0 mg/L 6-Benzyladenine (BA) + 0.5 mg/L2, 4-Dichlorophenoxyacetic acid (2,4-D), while optimal accumulation of biomass was observed with 2.0 mg/L 6-BA + 2.0 mg/L2, 4-D. In addition, phenylalanine also contributed to the synthesis of chlorogenic acid at a concentration > 50 mg/L. Conclusion: Cell suspension cultures of L. macranthoides Hand.-Mazz. “Yulei1” have successfully been established. The findings provide a potential basis for large scale production of chlorogenic acid using cell suspension cultures of L. macranthoides.