Abstract: The influence of organic supplements on growth and multiplication efficiency of Phaius tankervilleae var. alba seedlings was investigated. 12 week-old seedlings were cultured on half-strength semi-solid Murashige and Skoog (MS) medium supplemented with 30 g/L sucrose, 8 g/L agar and various concentrations of coconut water (0, 50, 100, 150 and 200 mL/L) combined with potato extract (0, 25 and 50 g/L) and the pH was adjusted to 5.8 prior to autoclaving. The cultures were then kept under constant photoperiod (16 h light: 8 h dark) at 25 ± 2 °C for 12 weeks. The highest number of shoots (3.0 shoots/explant) was obtained when cultured on the medium added with 50 ml/L coconut water and 50 g/L potato extract whereas the highest number of leaves (5.9 leaves/explant) and roots (6.1 roots/explant) could receive on the medium supplemented with 150 ml/L coconut water and 50 g/L potato extract. with 150 ml/L coconut water and 50 g/L potato extract. Additionally, plantlets of P. tankervilleae var. alba were transferred to grow into seven different substrates i.e. soil, sand, coconut husk chip, soil-sand mix (1: 1), soil-coconut husk chip mix (1: 1), sand-coconut husk chip mix (1: 1) and soil-sand-coconut husk chip mix (1: 1: 1) for four weeks. The results found that acclimatized plants showed 100% of survivals when sand, coconut husk chip and sand-coconut husk chip mix are used as substrates. The number of leaves induced by sand-coconut husk chip mix was significantly higher than that planted in other substrates (P > 0.05). Meanwhile, no significant difference in new shoot formation among these substrates was observed (P < 0.05). This precursory developing protocol was likely to be applied for more large scale of plant production as well as conservation of germplasm of this orchid species.
Abstract: Salinity is one of the most widespread agricultural problems in arid and semi-arid areas that limits the plant growth and crop productivity. In this study, the salt stress effects on protein, reducing sugar, proline contents and antioxidant enzymes activities of Carum copticum L. under in vitro conditions were studied. Seeds of C. copticum were cultured in Murashige and Skoog (MS) medium containing 0, 25, 50, 100 and 150 mM NaCl and calli were cultured in MS medium containing 1 μM 2, 4-dichlorophenoxyacetic acid, 4 μM benzyl amino purine and different levels of NaCl (0, 25, 50, 100 and 150 mM). After NaCl treatment for 28 days, the proline and reducing sugar contents of shoots, roots and calli increased significantly in relation to the severity of the salt stress. The highest amount of proline and carbohydrate were observed at 150 and 100 mM NaCl, respectively. The reducing sugar accumulation in shoots was the highest as compared to roots, whereas, proline contents did not show any significant difference in roots and shoots under salt stress. The results showed significant reduction of protein contents in seedlings and calli. Based on these results, proteins extracted from the shoots, roots and calli of C. copticum treated with 150 mM NaCl showed the lowest contents. The positive relationships were observed between activity of antioxidant enzymes and the increase in stress levels. Catalase, ascorbate peroxidase and superoxide dismutase activity increased significantly under salt concentrations in comparison to the control. These results suggest that the accumulation of proline and sugars, and activation of antioxidant enzymes play adaptive roles in the adaptation of seedlings and callus of C. copticum to saline conditions.
Abstract: This study provides a regeneration protocol for Treculia africana Decne (an endangered plant) through embryo culture. Mature zygotic embryos of T. africana were excised from the seeds aseptically and cultured on varied strengths (full, half and quarter) of Murashige and Skoog (MS) basal medium supplemented. All treatments experienced 100±0.00 percent sprouting except for half and quarter strengths. Plantlets in MS full strength had the highest fresh weight, leaf area, and longest shoot length when compared to other treatments. All explants in full, half, quarter strengths and control had the same number of leaves and sprout rate. Between the treatments, there was a significant difference (P>0.05) in their effect on the length of shoot and root, number of adventitious root, leaf area, and fresh weight. Full strength had the highest mean value in all the above-mentioned parameters and differed significantly (P>0.05) from others except in shoot length, number of adventitious roots, and root length where it did not differ (P
Abstract: This research aims to investigate callus induction,
somatic embryogenesis and indirect plant regeneration of Crassula
ovata (Mill.) Druce – the famous ornamental plant. Experiment no.1:
Callus induction was obtained from leaf and stem explants on
Murashige and Skoog (MS) medium supplemented with various plant
growth regulators (PGRs). Effects of different PGRs, plant
regeneration and subsequent plantlet conversion were also assessed.
Indirect plant regeneration was achieved from the callus of stem
explants by the addition of 1.5 mg/L Kinetin (KN) alone. Best shoot
induction was achieved (6.5 shoots/per explant) after 60 days. For
successful rooting, regenerated plantlets were sub-cultured on the
same MS media supplemented with 1.5 mg/L KN alone. The rooted
plantlets were acclimatized and the survival rate was 90%.
Experiment no.2: Results revealed that 0.5 mg/L 2,4-D alone and in
combination with 1.0 mg/L 6-Benzyladenine (BA) gave 89.8% callus
from the stem explants as compared to leaf explants. Callus
proliferation and somatic embryo formation were also evaluated by
‘Double Staining Method’ and different stages of somatic
embryogenesis were revealed by scanning electron microscope. Full
Strength MS medium produced the highest number (49.6%) of
cotyledonary stage somatic embryos (SEs). Mature cotyledonary
stage SEs developed into plantlets after 12 weeks of culture. Wellrooted
plantlets were successfully acclimatized at the survival rate of
85%. Indirectly regenerated plants did not show any detectable
variation in morphological and growth characteristics when
compared with the donor plant.
Abstract: Quercetin and (+)-catechin are metabolites present in Phyllanthus niruri plant, have potential in medicinal uses as anticancer and antioxidant agents. Studies on production of quercetin and (+)-catechin from P. niruri callus culture via in vitro technique were carried out and the results were compared to the intact plant. P. niruri explants were cultured on Murashige and Skoog (MS) solidified media supplemented with several phytohormone combinations for one month. The metabolites were extracted from P. niruri callus and intact plant by using carbon dioxide supercritical fluid extraction (SFE) with ethanol as modifier and solvent extraction techniques. The extracts were analyzed by means of HPLC method. Results showed that P. niruri callus culture was successfully established. The highest content of quercetin (1.72%) was found from P. niruri callus grown in media supplemented with 0.8mg/L kinetin and 0.2mg/L 2,4-dicholophenoxyacetic acid (2,4-D), which was 1.2 fold higher than intact plant. Meanwhile, the highest amounts of (+)-catechin (0.63%) was found from P. niruri callus grown in media with addition of 0.2mg/L 1-naphthalene acetic acid (NAA) and 0.8mg/L 2,4-D. The SFE condition in this study showed better extraction efficiency when higher contents of selected metabolites were found in all SFE extracts compared to the common solvent extracts.
Abstract: Agropyron cristatum L. Gaertn. is a native grass of
semiarid region in Iran which is quit resistant to cool and drought
climate and withstand heavy grazing. This species has close
phylogenetic relationship with Triticum and Hordeum. In this
research, the effect of seven different concentrations of growth
regulator 2,4-D on callus production and somatic embryogenesis of
A. cristatum was investigated on Murashige and Skoog medium. The
results showed that the rate of callus, embryo and neomorph were
highest in 1 mg L-1 2,4-D. Callus production was increased in 1 mg
L-1 2,4-D but dramatically decreased at 5.5 and 9 mg L-1 2,4-D. The
somatic embryos were observed at 1 and 4 mg L-1 2,4-D but matured
embryos and plantlet were only occurred at 1 mg L-1 2,4-D. There
were significant differences between 1 mg L-1 2,4-D and other
treatments for producing globular and torpedo embryos, plantlet,
rooted callus and number of roots (p
Abstract: Plant tissue culture is an important in vitro technology applied for agricultural and industrial production. A sterile condition of culture medium is one of the main aspects. The alternative technique for medium sterilization to replace autoclaving was carried out. For sterilization of plant tissue culture medium without autoclaving, ten commercial pure essential oils and 5 disinfectants were tested. Each essential oil or disinfectant was added to a 20-mL Murashige and Skoog (MS) medium before medium was solidified in a 120-mL container, kept for 2 weeks before evaluating sterile conditions. Treated media, supplemented with essential oils or disinfectants, were compared to control medium, autoclaved at 121 degree Celsius for 15 min. Sterile conditions of MS medium were found 100% from betel oil or clove oil (18 mL/20 mL medium), cinnamon oil (36 mL/20 mL medium), lavender oil or holy basil oil (108 mL/20 mL medium), and lemon oil or tea tree oil or turmeric oil (252 mL/20 mL medium), compared to 100% sterile condition from autoclaved medium. For disinfectants, 2% iodine + 2.4% potassium iodide, 2% merbromine solution, 10% povidone-iodine, 6% sodium hypochlorite or 0.1% thimerosal at 36 mL/20 mL medium provided 100% sterile conditions. Furthermore, growth of new shoots from chrysanthemum node explants on treated media (fresh weight, shoot length, root length and number of node) were also reported and discussed in the comparison of those on autoclaved medium.
Abstract: Humic acids (HAs) have been shown to activate some
ion uptakes along with stimulating the lateral roots at effective
concentration of micronutrients. However, the effects of HA on ion
adsorption by plant roots are not easily explainable due to the
varieties of HAs that differ from origins. Therefore, this study was
aimed to investigate the effect of various concentrations of HA
obtained from the compost derived from mix manures and some
agricultural wastes on the growth of eggplant seedlings (Solanum
melongena L. cv. Chao Praya) in tissue cultures at low nutrient level.
Egg plant seeds were surfaced sterilized and germinated in ½
Murashige and Skoog medium (MS) without HA added or in ¼ MS
supplemented with 0, 25, 50, 75 and 100 ppm of HAs. Then, they
were cultured for 4 weeks under the controlled environment. The
results showed that seedlings grown on ¼MS supplemented with
HAs at the concentration of 25 and 50 ppm had the average plant
heights (2.49 and 2.28 cm, respectively) higher than the other
treatments. Both treatments also significantly showed the maximum
average fresh and dry weights (p
Abstract: Direct and indirect somatic embryogenesis (SE) from
petiole and leaf explants of Scaevola aemula R. Br. cv. 'Purple
Fanfare' was achieved. High frequency of somatic embryos was
obtained directly from petiole and leaf explants using an inductive
plant growth regulator signal thidiazuron (TDZ). Petiole explants
were more responsive to SE than leaves. Plants derived from somatic
embryos of petiole explants germinated more readily into plants. SE
occurred more efficiently in half-strength Murashige and Skoog
(MS) medium than in full-strength MS medium. Non-embryogenic
callus induced by 2, 4-dichlorophenoxyacetic acid was used to
investigate the feasibility of obtaining SE with TDZ as a secondary
inductive plant growth regulator (PGR) signal. Non-embryogenic
callus of S. aemula was able to convert into an “embryogenic
competent mode" with PGR signal. Protocol developed for induction
of direct and indirect somatic embryogenesis in S. aemula can
improve the large scale propagation system of the plant in future.
Abstract: Stevia rebaudiana Bertoni (natural sweetener) belongs
to Asteraceae family and can be used as substitute of artificial
sweeteners for diabetic patients. Conventionally, it is cultivated by
seeds or stem cutting, but seed viability rate is poor. A protocol for
callus induction and multiplication was developed to produce large
no. of calli in short period. Surface sterilized nodal, leaf and root
explants were cultured on Murashige and Skoog (MS) medium with
different concentrations of plant hormone like, IBA, kinetin, NAA,
2,4-D, and NAA in combination with 2,4-D. 100% callusing was
observed from leaf explants cultured on combination of NAA and
2,4-D after three weeks while with 2,4-D, only 10% callusing was
observed. Calli obtained from leaf and root explants were shiny green
while with nodal explants it was hard and brown. The present
findings deal with induction of callusing in Stevia to achieve the
rapid callus multiplication for study of steviol glycosides in callus
culture.