Micropropagation of Kaempferia angustifolia Roscoe via Direct Regeneration

This study has established an efficient and reproducible protocol to micropropagation Kaempferia angustifolia Roscoe via direct regeneration. The use of young shoots as explants showed the best results compared to rhizome shoots, where the young shoots showed a low percentage of contamination of 10% - 30% (agar 6 g/L) and 45% - 55% (agar 3 g/L), respectively, compared to the use of rhizome shoots, where the contamination rate exceeded 80%. For shoot initiation, the combination of BAP (6 Benzylaminopurine) and NAA (1-Naphthaleneacetic acid) showed higher results for the percentage of initial shoots and number of micro shoots/explants compared to BAP with Kin (Kinetin). The highest concentration of BAP (5 mg/L) combined with the lowest concentration of NAA (0.5 mg/L) resulted in 90% of initial shoots and a number of shoots/explants of 5.8. The highest number of shoots for micropropagation was in treatment with 30 g/L sucrose that was segmented with 3 mg/L BAP + 0.5 mg/L NAA. For the number of roots, the highest number of roots was 11.8 recorded at sucrose (45) with only BAP (1 mg/L) used as the plant growth regulator, while the longest length of roots was 7 - 8 cm, recorded both at sucrose with the combination of BAP and NAA.

Direct Shoot Regeneration from Callus of <i>Melicope lunu-ankenda</i>

Melicope lunu-ankenda is commonly used in traditional medicine. The conventional propagation method for this species is inefficient due to low propagation rate and its lengthy period to maturity. In addition, insufficient planting materials often pose a problem for the plantation sector. The tissue culture technique is best alternative to overcome the problems. The callus induction and direct shoot regeneration protocols for M. lunu-ankenda were established. Callus was successfully initiated from leaves explants cultured in MS medium added with 2,4-D at concentrations 0.5 to 5.0 mg/L singly or in combination with NAA at concentrations 1.0 to 10 mg/L. Shoot was regenerated from callus in phytohormone-free medium, BAP at concentrations 0.5 - 5.0 mg/L singly or in combination of BAP with NAA or 2,4-D at concentration 0.5 and 1.0 mg/L, respectively. BAP at 1.0 mg/L induced the highest shoot regeneration rate (80%) and number of plantlet per calli. The established methods might be used for production of phytochemicals and plantlets in large scale.

Influence of Agitation Rate on the Growth of MD2 Pineapple Protocorm-Like Bodies and Shoots in Liquid-Shake Culture

The present study was conducted to investigate the effect of agitation rate on the increase in fresh weight of MD2 pineapple protocorm-like bodies (PLBs) and shoots cultured in liquid medium. PLBs were cultured in 250 ml Erlenmeyer flasks (7 g per flask) containing MS medium and plant growth regulators (1.5 mg/L 6-Benzylaminopurine, BAP and 0.2 mg/L 1-Naphthaleneacetic acid, NAA). The orbital shaker was set at speeds of 50, 80, 100, 120, and 150 rpm. After 40 days, the cultures shaken at 80 rpm showed the highest fresh weight and the highest number of shoots at 76 g and 41 shoots, respectively. A comparative study of agitation found that 80 rpm was the best speed which enhanced both PLB and shoot formation. The findings in the present study would be helpful in setting up large-scale in vitro mass propagation of MD2 pineapple.

<i>In Vitro</i>Micropropagation of a Valuable Medicinal Plant, <i>Plectranthus amboinicus</i>

The effect of the plant growth regulators benzyl amino purine (BAP), 1-naphthaleneacetic acid (NAA) and kinetin (KIN) on in vitro shoot induction and proliferation of Plectranthus amboinicus was examined. Explants obtained from lateral shoots and apical shoots of P. amboinicus were inoculated on Murashige and Skoog (MS) culture medium supplemented with different concentrations of BAP, NAA and KIN. When the effect of each growth regulator was considered singly, the highest rate of shoot induction (80% of explants producing shoots) and highest number of shoots produced (2.4 shoots per explant) were obtained from lateral shoot explants cultured on MS media supplemented with 3.0 mg/L BAP within 6 - 7 weeks. Better results were obtained using MS medium supplemented with 1 mg/L BAP + 5 mg/L NAA. Shoot proliferation rose to 85%, while 5.7 shoots per explants were recorded. Among the different media tested for rooting, MS medium supplemented with 1.0 mg/L IBA was the most effective for root induction. The quality of the roots obtained was better than that obtained using MS media supplemented with NAA or IAA.

Optimization of Extraction Conditions for Total Phenolics and Total Flavonoids from <i>Kaempferia parviflora</i>Rhizomes

Kaempferia parviflora plants derived from in vitro culture were grown in the glasshouse. A comparison of the yield of total phenolics and total flavonoids under varying extraction conditions from rhizomes harvested from plants of different ages was undertaken. The results showed that phenolic and flavonoid contents in the rhizomes were highest 8 months after planting. Another study found that 2 g rhizomes extracted in 50 ml of water at 90°C for 120 minutes gave the best yield of phenolics and flavonoids. Under these conditions, an average of 210 mg GAE/g dry weight of total phenolics and 81 μg QCE/g dry weight of total flavonoids were obtained.

Optimizing Extraction of Phenolics and Flavonoids from <i>Solanum ferox</i>Fruit

Various phenolic and flavonoid compounds that are secondary plant metabolites are known to contribute to physiological wellbeing. Extraction efficiency of such compounds from plant sources is dependent on the extraction solvent type and composition, and its pH. In this study, different extraction variables were examined: heating time (20 to 180 min), temperature (60°C, 75°C and 90°C) and pH (2.5, 3.0, 4.0, 5.0, 6.0 and 7.0). Hot water was used in the extraction of dry samples. For phenolics, the most efficient extraction was by using water at 60°C for 180 min, whereby 5.95 mg GA equivalent/dry extract was achieved. The most efficient extraction of flavonoids was achieved with water at 60°C for 150 min, whereby 43 μg Quercetin equivalent/dry extract was obtained. Adjusting the solvent to pH 2.5 increased the yield to 45.3 μg Quercetin equivalent/dry extract.

Callus Induction of Young Leaf Coconut cv. MATAG with Combination of 2,4-Dichlorophenoxyacetic Acid (2,4-D), α-Naphthalene Acetic Acid (NAA) and Benzyl Amino Purin (BAP)

This research was to study in vitro callus induction in Coconut cv MATAG from young leaf explants. Young leaf segments from mature coconut were cultured on Y3 medium supplemented with different concentrations of 2,4-D and a combination of NAA and BAP. Each of these plant growth regulators (PGR) gives different responses toward callus formation, the percentage of explants producing callus, the percentage of callus proliferation, and the morphology of callus. A series of different concentrations were used for 2,4-D (1, 5, 10, 20, 40, 60, 100 mg/L), NAA (1, 3, 5 mg/L) and BAP (1, 3, 5 mg/L) respectively. The range of days of callus formation using 2,4-D treatments is 7 - 12 months, while the 2,4-D combined with NAA is recorded at 2 - 5 months. Despite the variety of different months between these plant growth regulators for callus formation, the percentages of explants producing callus and callus proliferation are different. The highest percentage of explants producing callus (2.9%) was observed at 2,4-D (40 mg/mL), followed by 2.7% at 2,4-D (10.0 mg/mL) with NAA (1 mg/mL). At a concentration of 100 mg/mL of 2,4-D, the highest percentage of callus proliferation was found, as well.