In vitro culture is an important aid for ex situ conservation of rare, endemic or threatened plants. In this work, we establish an efficient method for the seed germination, seedling development, and axillary shoot propagation of Centaurea zeybekii Wagenitz. The seeds, collected from a wild population, were surface sterilised and cultured on various in vitro germination media. The effects of photoperiod and temperature on seed germination were also investigated. Germinations were obtained after 6 weeks in culture and the radicle emergence was evaluated as a main indicator. A high frequency of germination was obtained on distilled water supplemented with vitamines and 1 mg/L GA3. Although the seed germination frequencies were not affected by photoperiod, the highest germination frequency was obtained at 24 ± 2 • C. A high frequency of axillary shoot proliferation was produced on MS medium supplemented with 1 mg/L BA. Then, the axillary shoots were separated and transferred to MS medium with or without plant growth regulators for rooting. Rhizogenezis was promoted after 6 weeks only in MS and 1 /2 MS media containing 0.5 mg/L IBA. The rooting process was very slow and the percentage of shoot rooting was also very low (15%). The present study not only enables reinforcement of wild plant populations using ex situ growth of individuals, but it also helps to large number of aseptic seedling to use it in clonaly micropropagation studies.
Background: Hypericum adenotrichum contain many biologically active compounds, some of which, especially hypericins (hypericin and pseudohypericin), have antidepressant, antimicrobial, antiviral, and antitumor properties. In this paper, we report the effects of osmotic stress on the production of hypericins in H. adenotrichum under in vitro conditions. Osmotic stress is an abiotic elicitor that can alter the physiological and biochemical properties of plants, as well as decrease or increase the concentrations of secondary metabolites in plant tissues. Material and Methods: Sucrose and polyethylene glycol (PEG) were used to cause osmotic stress. Seedlings of H. adenotrichum were grown on a modified MS medium containing sucrose (15, 30, 45, and 60 g/L) or PEG (2.5, 10, and 15 g/L) for 15 and 30 days. Then, H. adenotrichum seedlings were extracted with methanol. These extracts were analysed by HPLC to investigate the changes in hypericins levels.Results: Under osmotic stress conditions, the concentrations of hypericins changed in seedlings of H. adenotrichum. Treatment with 10 g/L PEG for 15 days increased production of hypericin (2.1-fold) and pseudohypericin (2.3-fold), but PEG treatment for 30 days affected less hypericins levels when compared to PEG treatment for 15 days. The amount that the hypericins increased was minimal and proportional with the amount of sucrose up to treatment with 45 g/L sucrose, and then the hypericins decreased at 60 g/L of sucrose treatment for 15 days. In sucrose treatment, the highest hypericins levels were observed in the control seedlings at 30 days treatment period of sucrose. Conclusions: These results can be evaluated in experimental botany and in the technology of Hypericum species cultivation for pharmaceutical applications. Yamaner O, Erdag B (2013) Effects of sucrose and polyethylene glycol on hypericins content in Hypericum adenotrichum. Eurasia J Biosci 7: 100-110. http://dx.
An in vitro micropropagation method by somatic embryogenesis was developed for Gladiolus anatolicus (Boiss.) Stapf using leaves of in vitro shoots obtained from lateral buds. Lateral buds removed from sterilized fresh corms were placed on Murashige and Skoog (MS) medium supplemented with various concentrations of N6-benzyladenine (BA) for shoot culture establishment. The highest number of shoot per lateral bud explant was on MS medium supplemented with 2 mg L(-1) BA (11.00 +/- 0.38). To induce somatic embryogenesis, leaves of in vitro shoots obtained from lateral buds were used as explant. Calli were obtained from middle and basal region of leaf explant cultured on MS basal medium supplemented with different concentrations of alpha-naphthaleneacetic acid: N6-benzyladenine (NAA:BA) ratio and without growth regulators. The highest rate of callus formation was obtained from basal part of leaves cultured on MS medium containing 5 mg L(-1) NAA in darkness (80 +/- 0.41%). Creamy-white and friable calli produced numerous somatic embryos on MS basal medium supplemented with 0.1 mg L(-1) BA within 4 weeks in light (On avarage 30 structures per callus). Well-developed somatic embryos were germinated on MS medium supplemented with 0.1 mg L(-1) BA and reduced sucrose concentration (20 g L(-1)). On this medium 40% of the somatic embryos developed into plantlets. Cormlet formation was observed on MS basal medium (30 g L(-1) sucrose) containing same concentration of BA.
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