• Genetic Engineering and Biotechnology Research institute
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A simple extraction and isolation process of solasodine, a natural precursor to synthesize steroidal drugs, from fruits and leaves of Solanum laciniatum Ait. was developed. The optimum concentration of 2-propanol for the extraction of crude glycosides was 70%. The suitable hydrolysis condition of solasodine from crude glycosides was by 1 N hydrochloric acid in 2-propanol. Pure solasodine from both fruits and leaves of Solanum laciniatum Ait. was obtained without any requirement of column chromatography. The yield of pure solasodine were 0.34±0.04% and 0.44±0.16% of the dry weight of fruits and leaves, respectively. The maximum yield of 37.0% of pure 16-DPA was obtained by using tetrabutylammonium hydrogen sulfate as a phase-transfer catalyst and potassium dichromate as an oxidizing agent. The results indicated the novel economic with environmental friendly method of solasodine extraction and synthesis of 16-dehydropregnenolone acetate from solasodine by phase-transfer catalysis.
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The objective of the study was to develop an efficient method for shoot regeneration of canola (Brassica napus L.) and to compare the regeneration capacity of different explants on MS medium with several combinations of plant growth regulators. The experiments showed that the morphogenetical potential of canola depends on genotype, primary explant, hormonal structure and concentration of nutrient medium. Cotyledons possessed higher regeneration ability in comparison to hypocotyls and roots. The best regeneration capability was exhibited by the cultivar 'Quantum'. Its frequency with cotyledonary explants reached 68.8% on all used media. Addition of 3 mg/l ABA in nutrient medium considerably increased the regeneration frequency. The highest shoot regeneration (100%), however, took place when cotyledonary explants were cultivated on medium, containing 1.0 mg/l NAA, 8.0 mg/l BAP and 3.0 mg/l ABA. Precultivation of explants on callus induction medium did not affect the shoot regeneration frequency. Vitrification of regenerants was promoted by increasing the auxin NAA or cytokinin BAP, and ABA in the nutrient medium.
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In vitro control of floral transition in tomato (Lycopersicon esculentum Mill.), the model plant for autonomously flowering species has been investigated using the late flowering mutant uniflora (uf). Apices collected from truly vegetative plants were cultivated on solid media supplemented with different combinations of growth regulators and chemicals. Several chemical factors implicated in the promotion of floral transition of the uf mutant have been identified: sucrose, cytokinins and nitrogenous nutrients have all to be supplied at optimal concentrations. In contrast, gibberellic acid was found to be inhibitory. These results are discussed in relation to knowledge accumulated on the nature of the flowering signals circulating, at floral transition, in other plants, especially in photoperiodic species. This study suggests that tomato could constitute an adequate model to investigate the genetic and physiological control of floral transition and contribute in unravelling pathways which are constitutively regulating this important step of plant life cycle.
The effect of benzyladenine (BA) and thidiazuron (TDZ) in combination with α-naphthaleneacetic acid and indole-3-butyric acid (IBA) on in vitro shoot regeneration from leaf segments of several Japanese pears was investigated. A regeneration protocol was developed and regeneration was achieved from six cultivars: - 'Kosui', 'Hosui', 'Niitaka', 'Wasekaso', 'Okusankichi' and 'Whangkeumbae' pears. The highest regeneration rate was obtained in most shoots from young leaves on a medium based on MS macroelements supplemented with 0.1-1.0 mg/L TDZ and 0.1-1.0 mg/L IBA and/or NAA in each cultivar. Physiological differences with BA and TDZ treatments were compared. In the regeneration medium with BA treatments, green foci appeared on the callus surface after 8 days. Then, some adventitious buds were induced on those green foci, resulting in normal shoots. On the other hand, in the medium with TDZ, callus surface turned compact and greenish, and many adventitious buds were formed over the whole area of the callus surface. In some shoots, cultured on the medium with TDZ, there were morphologically abnormal shoots, including vitrified shoots. A-fourth strength MS medium supplemented with 0.1 mg/L NAA produced plantlets in good quality of root number and root length.
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