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- Author or Editor: D. L. Madhavi x
Many plants can produce bioactive chemicals with medicinal or health benefits, which has stimulated a whole new research effort aimed at extracting & improving natural phytochemicals. Begonia is a rich source of biologically-active phytochemicals and an excellent donor for natural anthocyanin pigments. High levels of triterpene compounds and a host of potentially-useful flavonoids have been isolated from Begonia sp., which may account for its frequent use as a medicinal plant remedy in a diverse array of cultures worldwide. Deliberate shifting of the physical and chemical microenvironments can have a significant effect on anthocyanins and precursors produced in vitro. This realization offers the potential to thoroughly screen and study valuable phytochemicals from Begonia. Begonia genotypes from 3 species were screened to identify callus induction techniques. Contamination inherent in the vascular system of one genotype, along with spontaneous organogenesis, were found to be recurrent problems. These were partially alleviated by light and growth regulator treatments. Studies comparing callus and in vitro vegetative tissues as resources for phytochemical extraction are scheduled.
Interaction between irradiance levels (5–40 mM–m–2–s–1) and iron chelate sources (FeNa2EDTA and FeNaDTPA) on the establishment, growth, and proliferation of shoot tips of Carica papaya were tested. Reduced irradiance level (5 mM–m–2–s–1) enhanced the establishment of shoot tips regardless of the source of iron chelate tested. At higher irradiance levels (30 and 40 mM–m–2–s–1), presence of FeNaDTPA in the medium enhanced establishment of shoot tips. Continuous or alternating light/dark (16/8 h) photoperiods at high irradiance levels had no effect on the establishment or growth of the culture. At higher irradiance levels, the cultures produced smaller leaves as compared to lower irradiance levels. Low irradiance and FeNa2EDTA was preferred during the proliferation stage.
Interactions between irradiance levels (5–40 μmol·m-2·s-1) and iron chelate sources (FeEDTA and FeEDDHA) were observed for Carica papaya shoot tip cultures during both the establishment and proliferation stages of microculture. Reduced levels of irradiance (5 μmol·m-2·s-1) favored shoot tip establishment regardless of the source or level of iron. However, the highest percentage of successful explant establishment (100%), and significantly greater leaf length (1.16 cm; over double the size attained in any other treatment), resulted when a low concentration of FeEDTA alone was used at low irradiance. During the subsequent shoot proliferation stage, however, higher irradiance levels (30 and 40 μmol·m-2·s-1) were required, and FeEDTA failed to support culture growth when used as the sole iron source. The highest multiplication rates (3.6 shoots per explant) and leaf chlorophyll concentrations (0.22 mg/g fresh mass), and significantly improved shoot quality were achieved at 30 μmol·m-2·s-1 irradiance when both iron chelate formulations were combined (each at a 100 μM concentration) in the proliferation medium. Chemical names used: benzylamino purine (BA); ferric disodium ethylenediamine tetraacetate or FeNa2EDTA (FeEDTA); ferric monosodium ethylenediamine di(o-hydroxyphenylacetate), (FeNaEDDHA) or Sequestrene 138Fe (FeEDDHA); indoleacetic acid (IAA); 1-naphthaleneacetic acid (NAA).
Industrial-scale cultivation of plant cells for valuable product recovery (e.g. natural pigments, pharmaceutical compounds) can only be considered commercially-feasible when a fully-automated, predictable bioprocess is achieved. Automation of cell selection, quantification, and sorting procedures, and pinpointing of optimal microenvironmental regimes can be approached via machine vision. Macroscopic staging of Ajuga reptans callus masses (ranging between 2-6 g FW) permitted simultaneous rapid capture of top and side views. Area data used in a linear regression model yielded a reliable, non-destructive estimate of fresh mass. Suspension culture images from the same cell line were microscopically imaged at 4x (with an inverted microscope). Using color machine vision, the HSI (hue-saturation-intensity) coordinates were used to successfully separate pigmented cells and aggregates from non-pigmented cells, aggregates, and background debris. Time-course sampling of a routine suspension culture consistently allowed pigmented cells to be detected, and intensity could be correlated with the degree of pigmentation as verified using spectrophotometer analysis of parallel samples.