Purpose The purpose of this paper is to validate the antimicrobial activity (both antibacterial and antifungal) of in vivo and in vitro ethanolic anthocyanin extracts of Clitoria ternatea L. (vivid blue flower butterfly-pea) and Dioscorea alata L. (purple yam) against selected bacteria (Bacillus subtilis, Staphylococcus aureus and Escherichia coli) and fungi (Fusarium sp., Aspergillus niger and Trichoderma sp.). Design/methodology/approach The freeze-dried samples (0.2 g) from in vivo vivid blue flowers of C. ternatea L. were extracted using 10 mL ethanol (produced ethanolic red extraction) and 10 mL distilled water (produced aqueous blue extraction) separately. Two-month-old in vitro callus samples (0.2 g) were only extracted using 10 mL ethanol. The anthocyanin extractions were separated with the addition (several times) of ethyl acetate and distilled water (1:2:3) to remove stilbenoids, chlorophyll, less polar flavonoids and other non-polar compounds. Furthermore, the antimicrobial properties were determined using agar diffusion technique. Three bacteria (B. subtilis, S. aureus and E. coli) and fungi (F. sp., A. niger and T. sp.) were streaked on bacteria agar and dextrose agar, respectively, using “hockey stick”. Then, the sterile paper discs (6 mm diameter) were pipetted with 20 µL of 1,010 CFU/mL chloramphenicol (as control for antibacterial) and carbendazim (as control for antifungal) in vivo and in vitro extracts. The plates were incubated at room temperature for 48 h, and the inhibition zones were measured. Findings Based on the results, both in vivo and in vitro ethanolic extracts from vivid blue flowers of C. ternatea L. showed the best antibacterial activity against the same bacteria (B. subtilis), 11 and 10 mm inhibition zones, respectively. However, different antifungal activity was detected in in vitro ethanolic callus extract (12 mm), which was against T. sp., contrary to in vivo ethanolic extract (10 mm), which was against F. sp.; antibacterial activity of D. alata L. was seen against the same bacteria (E. coli) with the highest inhibition zone for in vivo extract (8.8 mm), followed by in vitro extract (7.8 mm). Research limitations/implications Anthocyanins are responsible for the water soluble and vacuolar, pink, red, purple and blue pigments present in coloured plant pigments. These pigments (pink, red, purple and blue) are of important agronomic value in many crops and ornamental plants. However, anthocyanins are not stable and are easy to degrade and fade whenever exposed to light. Social implications Plant extracts containing bioactive agents with antimicrobial properties have been found to be useful in treating bacterial and fungal infections, as well as showed multiple antibiotic resistance. Originality/value Both in vivo and in vitro extracts from vivid blue flower petals (C. ternatea L.) and purple yam (D. alata L.) have important applications as natural antimicrobial (antibacterial and antifungal) agents in the coating industry, instead of natural pharmaceutical products.
The aromatic and medicinal properties of Ocimum basilicum L. (sweet basil) are related to the unique essential oil chemistry in different cultivars. This study describes efficient micropropagation and in vitro flowering protocols from shoot tips and reveals information on seed germination capability, glandular trichomes ultrastructure, and essential oil content and composition at different plant developmental stages from micropropagated O. basilicum 'Sweet Thai', chemotype methyl chavicol. Shoot tips from 2-mo-old aseptic seedlings were induced to proliferate shoots on Murashige and Skoog (MS) medium supplemented with 6-benzyl-aminopurine (BAP) and gibberellic acid (GA 3) either alone or in combination with α-naphthaleneacetic acid (NAA). Maximum shoot formation was achieved in MS medium supplemented with 1.0 mg L −1 BAP. The micropropagated plants were successfully acclimatized ex vitro with an 80% survival rate. All of the micropropagated plants flowered in vitro on MS medium supplemented with 1.0 mg L −1 GA 3. Relative to the mother plant, in vitro plants flowered at a younger stage of plant development but showed a lack of seed formation, fewer fully filled peltate glandular trichomes, lower essential oil content, and higher methyl chavicol content. Ex vitro plants flowered at an intermediate stage of plant development and formed seed with nearly the same seed germinability, essential oil content, and methyl chavicol content as the mother plant.
In vitro direct regeneration of Nelumbo nucifera Gaertn. was successfully achieved from immature explants (yellow plumule) cultured on a solid MS media supplemented with combinations of 0.5 mg/L BAP and 1.5 mg/L NAA which resulted in 16.00 ± 0.30 number of shoots per explant and exhibited a new characteristic of layered multiple shoots, while normal roots formed on the solid MS basal media. The double-layered media gave the highest number of shoots per explant with a ratio of 2 : 1 (liquid to solid) with a mean number of 16.67 ± 0.23 shoots per explant with the formation of primary and secondary roots from immature explants. In the study involving light distance, the tallest shoot (16.67 ± 0.23 mm) obtained from the immature explants was at a light distance of 200 mm from the source of inflorescent light (1000 lux). The plantlets were successfully acclimatized in clay loam soil after 8 months being maintained under in vitro conditions.
Abstract-Encapsulated embryogenic callus of Clitoria ternatea L. were successfully created from leaf explants within 3 weeks after germination on Murashige and Skoog (MS) media. The seeds were initially washed with tap water and teepol, then the seeds were sterilised with 99% (v/v) sodium hypochlorite solution for 1 minute and rinsed with distilled water three times. In a laminar flow cabinet, the seeds were dipped in 70% (v/v) ethanol for 1 minute and blotted with steriled tissue. The 3 mm 2 leaf explants were encapsulated with 3% alginate (w/v) which were suplemented with various concentrations (0.5-2.5 mg l -1 ) and combinations of NAA, BAP and adenine. The optimum concentration for the formation of encapsulation matrix was 3.0% sodium alginate (NaC 6 H 7 O 6 ). Encapsulated beads were soaked in 100 mM calcium chloride dehydrate (CaCl 2 .2H 2 O) solution for 30 minutes. No suitable beads were formed with low concentration (1-2%) of sodium alginate. Within 10 minutes soaking in calcium chloride dehydrate, clear and bead formation with no definite shape was observed. While, within 20 minutes in calcium chloride dehydrate, clear beads, solid and round in shape was observed, however, inside the bead was still in liquid condition. In the present study, the rate of germination of synthetic seeds were slightly decreased from 100% to 77% after 60 days of storage at 4°C. Embryogenic tissue from leaf explants of Clitoria ternatea was distinguished by double staining method with bright red of acetocarmine. This technology is an alternative and supplementary method for regeneration, mass propagation and conservation of this medicinal, attractive ornamental and also forage crop for future uses and exploitation.Index Terms-BAP, encapsulated embryogenic callus, In vitro regeneration, NAA. I. INTRODUCTIONClitoria ternatea L. is a climber plant that usually grows in various garden hedges. Clitoria ternatea L. or its common name including butterfly-pea, blue-pea, cordofan-pea and Asian pigeon wings, belongs to the family Fabaceae. The United State Development Agency (USDA) intends to conserve Clitoria ternatea L. along with other 16 leguminous Manuscript received May 19, 2015; revised July 15, 2015 species with potentially useful phytochemicals [1]. It is a strongly persistent, a herbaceous perennial legume with stems fine twining, sparsely pubescent, suberect at base, 0.5-3 m long [2]. In addition, Clitoria ternatea L. is a tropical twining herb, growing wild and also in gardens, bearing white or ink blue flowers resemble a conch-shell. It is widely planted as an ornamental on fencerows. Originally it is selected as a cover crop, but now used for short and medium-term pastures and as green manure, cover crops and protein bank.Clitoria ternatea contains flavonoids such as quarcetin, kaemferol, robinin and Clitorin. It also contains starch, tannin, resin and anthocyanins [3] and several glycosides including malvidin-3-β-glycoside, deiphinidin-3-β-glycoside [4]. In Malaysia, petals of Clitoria ternatea L. or commonly known as "B...
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