Kinetin, BAP and NAA increased glucotropaeolin content in dry weight of nasturtium hairy roots but they inhibited biomass growth. Salicylates most strongly (by about 100-200% above control) and DL-?-aminobutyric acid and methyl jasmonate to a lesser degree increased glucotropaeolin yield. They also slightly increased myrosinase activity. Trans-cinnamic acid was a much stronger inducer of myrosinase activity than glucotropaeolin biosynthesis: it strongly inhibited biomass growth. L-1-amino-2-phenylethylphosphonic acid, inhibitor of L-phenylalanine ammonia-lyase (PAL), induced glucotropaeolin production and enhanced the effect of salicylates and jasmonate on glucotropaeoplin yield but it did not affect myrosinase activity.
The influence of a xenobiotic, a substrate and elicitors on glutathione S-transferase, peroxidase activities and glutathione concentration was studied in reed cannarygrass. The induction of both enzymes? activities and glutathione concentrations by 4-chlorophenol (xenobiotic), benzyl isothiocyanate (substrate), salicylic acid and 3-aminobutyric acid (substrate) was shown. Salicylic acid treatment increased glutathione level, but did not decrease GSH/GSSG ratio. The ability of reed cannarygrass to adsorb 4-chlorophenol in the hydroponic culture was demonstrated. The obtained results suggest that the reed cannarygrass has significant potential to conjugate xenobiotics with glutathione and it may be useful for phytoremediation.
The conditions of endogenous hydrolysis of glucotropaeolin to bioactive benzyl isothiocyanate for T. majus hairy root cultures giving high yields of glucotropaeolin and myrosinase were optimized. After in vitro glucotropaeolin hydrolysis at pH 5.5, optimal for myrosinase activity, 54% of glucotropaeolin were converted to benzyl isothiocyanate, 22% to benzyl cyanide, and 1.9% to benzyl thiocyanate. During endogenous glucotropaeolin hydrolysis, maximal benzyl isothiocyanate yield, 87% of tissue glucotropaeolin, was detected at pH 7.5, in the 60th min of incubation. In the presence of 100 M ascorbic acid, the rate of hydrolysis increased and in the 60th min 99% of glucotropaeolin was converted to benzyl isothiocyanate. The highest benzyl isothiocyanate yield, 0.746 mmol/10 g fresh weight, was obtained during endogenous hydrolysis at pH 7.5, in the presence of 100 M ascorbic acid from the hairy roots with glucotropaeolin content, enhanced by precursors (phenylalanine + cystein) and inhibitor of phenylalanine ammonia-lyase activity (l-1-amino-2-phenylethylphosphonic acid).
The Oxalis triangularis transformed plants were regenerated from hairy roots induced by A. rhizogenes LBA 9402. In vitro shoot proliferation of the transformed roots-regenerated plants was 65% higher by than that of the non-transformed ones. In vivo the hairy root syndrome was observed for the transformed roots-regenerated plants. Anthocyanin content in the leaves of non-transformed plants was about 40% higher than in the transformed roots-regenerated ones. In the non-transformed plants, GSH (1,5 mM) enhanced anthocyanin production by 30% at the most, while in the transformed roots-regenerated plants the maximal increase in its production was found on the 2nd and 9th days after the treatment, it reached 230-320% and 750-600% of control, respectively.
In this study, the transformed root culture of Centaurium erythraea was established, as a result of infection of leaves culture with agropine strain of Agrobacterium rhizogenes (LBA 9402). Frequency of root formation depended on the kind of explants and the presence of acetosyringone in the bacteria medium. The transformation was confirmed according to the analysis of opines and RAPD-PCR. As the result of transformation 30 clones of hairy roots were obtained. These clones have grown in liquid Woody Plant (WPM) medium without growth regulators. The curve of growth indicates that fresh weight of roots increased almost 17 times , and dry weight increased 12 times during 55 days. Hairy roots of C. erythraea were able to produce secoiridoid glycosides (sweroside, gentiopicroside, swertiamarine).
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