Establishment Of Tissue Culture And Evaluation Of Biological Activities Of Jarum Tujuh Bilah (Pereskia Bleo Kunth)

Pereskia bleo is a leafy cactus which belongs to Cactaceae family and it is commonly used among the traditional medicine practitioners to prevent or treat cancer by consuming the leaves. As there is lack of information available on this plant, this research was carried out to determine the biological activities in natural plants and tissue culture materials. In order to produce standardize plant material for secondary metabolite production, tissue culture systems for Pereskia bleo was initiated and established. In vitro plantlets and tissues of Pereskia bleo were micropropagated in order to provide enough uniform explants for further tissue culture work and for extraction. The highest number of sterile explants (65.6%) was observed using fungicide for 20 minutes followed by 20% Chlorox® for 10 minutes. The growth characteristics on different types of basal media containing plant growth regulators were analyzed for maximum production of biomass: plantlets, calli and cell suspension. The highest number of multiple shoots (5.3 ± 0.5) was formed in MS basal medium supplemented with 2.22 μM BAP using shoot tips explants. Using node explants cultured onto MS supplemented with 8.88 μM BAP gave the highest number of multiple shoots (6.7 ± 0.58). Cell suspension growth was highest in MS basal media supplemented with 2.26 μM 2,4-D. Hairy root experiments revealed that both leaf and stem explants of Pereskia bleo were susceptible to all Agrobacterium rhizogenes (TR 105, LBA 9402, 8196, ATCC 15834) tested. Addition of acetosringone in the Agrobacterium rhizogenes culture increased the transformation frequencies of hairy root in Pereskia bleo. Results from brine shrimp lethality test showed no toxicity of the Pereskia bleo extracts occurred. The petroleum ether, chloroform and methanol leaf extracts showed strong in vitro antiproliferative activities (IC50 : 6.5 - 25.4 μg/ml) for MCF-7 (human hormone-dependent breast cancer cell line) and (IC50 : 19.3 - 26.8 μg/ml) for MDA-MB-231 (human hormone non-dependent breast cancer cell line). The IC50 value against HL-60 (human leukemia cell line) between 6.91 and 9.98 μg/ml for chloroform, petroleum ether and methanol extracts. Non-cytotoxic activity towards the non-tumour 3T3 mouse fibroblast indicated that the extracts exhibited selective mode of inhibition between tumor and non-tumor cells. The findings of antioxidative and antiproliferative activities of the plant extracts in vitro suggested that this species does contain antioxidant and cytotoxic compounds. The results obtained support the use of this species in traditional medicine for the prevention and treatment of cancer

Bio-potential of fermented fruits waste solutions on in vitro seed germination and regeneration of Lycium barbarum and Aquilaria malaccensis Lamk.

There are many synthetic growth media for plant tissue culture available in the market such as Murashige and Skoog (MS) Medium, Woody Plant Medium (WPM), Schenk and Hildebrandt (SH) Medium and Gamborg’s B-5 Medium. The aim of this study was to substitute the synthetic media used in the plant tissue culture by organic additives which are pineapple, banana, papaya, calamansi lime, kaffir lime and key lime peels. Two formulated fermented fruits waste solutions composed of these organic additives were prepared in different concentrations (Formula A- calamansi lime, kaffir lime, and key lime peel; Formula B -banana, pineapple, and papaya peels) to study their effects on in vitro seed germination and regeneration of Lycium barbarum and Aquilaria malaccensis Lamk. Statistical results showed that they were significantly different in interaction effects (p<0.05) in promoting the plant growth in the formulated media as compared to control medium determined by ANOVA test. Application of this formulated fermented fruits waste solutions should be considered since it is found to be responsive in in vitro seed germination and regeneration of L. barbarum and A. malaccensis Lamk and will potentially minimize the operational cost

Effects of organic substrates on growth and yield of ginger cultivated using soilless culture

The effects of organic soilless substrates on growth and yield of ginger were studied. In soilless production system, many types of growing media or substrates such as rockwool, perlite, vermiculite and peat have been used to grow many kinds of crops. Alternative substrates that are cheaper and locally available such as coconut fibres and burnt paddy husks should be used as alternative media. The main objective of the study was to determine the most suitable organic growth substrate for cultivation of ginger using fertigation technique. The study was conducted under the side-netted rain shelter equipped with an irrigation system to supply fertiliser solution at a regulated time schedule. Five combinations of growth substrates were evaluated: 100% coir dust; 100% burnt paddy husks; 70% coir dust + 30% burnt paddy husks; 30% coir dust + 70% burnt paddy husks; and 50% coir dust + 50% burnt paddy husks. The ginger plants were selected randomly and the rhizomes were harvested 3 – 9 months after sowing. Plants grown in 100% coir dust gave the best growth performance and yield compared to the other treatments. They produced the highest shoot height (123 ± 23 cm), shoot fresh weight (1,340 ± 235 g) and rhizome yield (5,480 ± 325 g per plant). The lowest rhizome yield (2,570 ± 135 g) was obtained from plants planted in 30% coir dust + 70% burnt paddy husks. Hence, it can be concluded that the ginger plants cultivated in 100% coir dust substrate using fertigation technique gave the best plant growth and yields

Effects of N-methyl-N-nitrosourea on seed germination of Stevia rebaudiana

Stevia rebaudiana is a unique plant that contains non-caloric natural sweetener and has gained much interest among Malaysians. In this study, the effect of different concentrations of N‑methyl-N-nitrosourea (MNU) was assessed in inducing mutation in Stevia seeds to produce genetic variations, which is valuable for crop improvement. Stevia seeds were soaked in six concentrations of MNU (0.0, 0.13, 0.25, 0.38, 0.50, and 1.00 mM) for four different durations (15, 30, 45, and 60 min) at room temperature. As a result, application of MNU reduced the germination percentage and germination rate of Stevia seeds as compared to the control group. Prolonged exposure to the highest concentration of MNU recorded the lowest percentage of germination (2.5 ± 1.4%) and the lowest germination rate (0.21 ± 0.16). Tricots were observed among seedlings treated with 0.13, 0.38 and 1.0 mM of MNU for 30 min. Presence of seedlings with albino colour proved the mutagenic effect of MNU on Stevia genome. Based on the percentage of seedlings with chlorophyll mutation, the most effective and efficient mutagenic treatment to induce mutation was 60 min in 0.25 mM of MNU

Effects of N-Methyl-N-Nitrosourea on seed germination of Stevia rebaudiana

Stevia rebaudiana is a unique plant that contains non-caloric natural sweetener and has gained much interest among Malaysians. In this study, the effect of different concentrations of N‑methyl-N-nitrosourea (MNU) was assessed in inducing mutation in Stevia seeds to produce genetic variations, which is valuable for crop improvement. Stevia seeds were soaked in six concentrations of MNU (0.0, 0.13, 0.25, 0.38, 0.50, and 1.00 mM) for four different durations (15, 30, 45, and 60 min) at room temperature. As a result, application of MNU reduced the germination percentage and germination rate of Stevia seeds as compared to the control group. Prolonged exposure to the highest concentration of MNU recorded the lowest percentage of germination (2.5 ± 1.4%) and the lowest germination rate (0.21 ± 0.16). Tricots were observed among seedlings treated with 0.13, 0.38 and 1.0 mM of MNU for 30 min. Presence of seedlings with albino colour proved the mutagenic effect of MNU on Stevia genome. Based on the percentage of seedlings with chlorophyll mutation, the most effective and efficient mutagenic treatment to induce mutation was 60 min in 0.25 mM of MNU

Surface sterilization method for reducing contamination of

Surface sterilization is a vital step in preparation of healthy and viable explants in tissue culture. Most surface contaminants can be eliminated by surface sterilization with a suitable sterilizing agent. The study aimed to present an effective disinfection method for Clinacanthus nutans shoot regeneration using nodal segments. A total of four different sterilization approaches were conducted by treating nodal explants with various concentrations of sterilizing agent. Sterilizing agents used were Rhizophora apiculata Pyroligneous acid (PA), sodium hypochlorite (Clorox) thiophanate-methyl (fungicide), and Mercuric chloride (HgCl2). Nodal explant then was cultured on plant growth regulator-free Murashige and Skoog (MS) basal medium. This study sterilizing agents revealed that PA showed strong bactericidal activity. However, it led to a high number of fungal contaminations. The pyroligneous acid did not exhibit a strong potential as a disinfectant for C. nutans nodal explant. Overall, HgCl2 exhibits the best reduction in fungal contamination and gives a significant result with thiophanate-methyl fungicide. Surface sterilization with mercuric chloride (0.2%) for 1 hour was the optimum concentration and duration, which resulted in the highest percentage of nodal explant survival and viability. All viable nodal segments developed into shoots. It had been concluded that the best surface sterilization agent was HgCl2

The effects of different strength of MS media in solid and liquid media on in vitro growth of Typhonium flagelliforme

Objective: To determine the effects of different strength of Murashige and Skoog (MS) media (full, ½ and ¼) in solid and liquid media on in vitro growth of Typhonium flagelliforme (T. flagelliforme), whereby an optimum media composition can be provided for mass propagation of T. flagelliforme. Methods: Rhizome bud of T. flagelliforme was obtained from the axenic in vitro established T. flagelliforme plantlets in Plant Tissue Culture Laboratory, Universiti Teknologi MARA, Shah Alam. Rhizome bud was used as explant and cultured onto shoot proliferation medium under different strength of MS media (full, ½, ¼) in solid and liquid culture media. Results: After 6 weeks of culture, the number of shoot, number of leaf, number of root, height of shoot, fresh weight, dry weight and chlorophyll content of T. flagelliforme were analyzed. A comparison was made between liquid and solid culture media. The results revealed that the liquid culture media were more effective for all the growth parameters (shoot height, shoot number, leaf number, root number, fresh weight, dry weight, chlorophyll a and chlorophyll b content) compared to solid culture media. Apart from that, this study revealed the positive relationship between strength of MS media and type of culture media (solid and liquid media) to the growth of T. flagelliforme. Growth of T. flagelliforme was improved when MS strength was increased in liquid media. In contrast, growth of T. flagelliforme was improved when MS strength was decreased in solid media. Conclusions: Through this study, an optimum media composition for mass propagation of T. flagelliforme had been established by observing effects of MS media strength and type of culture media (solid and liquid media) on the growth of T. flagelliforme