Callus and azadirachtin related limonoids production through in vitro culture of neem (Azadirachta indica A. Juss)

A protocol was established for the induction of callus and suspension cultures for azadirachtin production from neem explants. Different concentrations and combinations of plant growth regulators (2,4-D, NAA, IAA and BAP) were supplemented in MS medium. Immature flowers, nodular stem sections, leaves immature embryos and mature seeds were used as explants. The highest callus development (78%) was observed when immature flowers were inoculated on MS basal medium with addition of 1.0 mg/l 2,4-D, 1.0 mg/l BAP, 0.2 mg/l NAA and 3% sucrose. The azadirachtin containing liminoids were determined from calli obtained from different explants. Effect of sucrose, glucose, NH4NO3, KNO3 and urea on cell suspension cultures and azadirachtin contents were also investigated. The dry cell weight and azadirachtin contents increased to 373.1 and 359.2 mg/ 50 ml when 0.25 and 0.5 g/l NH4NO3 was added in MS liquid media and supplemented with 1.0 mg/l 2,4-D, 0.2 mg/l BAP and 3% sucrose

Effect of immersion systems on chlorophyll contents in micro-propagating banana

Banana is a nutritionally as well as economically important plant for which (Basrai variety) an efficient micro-propagation protocol was developed by using micro-stem cutting, as an explant. The maximum numbers of plantlets with higher chlorophyll and lower carotenoid contents were observed, which developed through routinely used tissue culture system (10.0 ìM BA; 8.0 ìM IAA; 3.0 g/L phytagel) for organogenesis, permanent immersion system (10.0 ìM BA; 1.0 g/L phytagel) for shoot induction andtemporary immersion system (10.0 ìM BA; 2.0 g/L phytagel) for shoot multiplication. The developed plantlets were cultured on 0.5 MS medium with IBA (0.01 mg/L). Rooted plantlets were successfully transferred to field after initial acclimatization

Optimization of cultural conditions for protease production by Bacillus subtilis EFRL 01

Molasses was used as a sole carbon source for the protease production from Bacillus subtilis EFRL 01 in batch wise submerged condition. The bacterial culture was grown on mineral medium and maximum production was noted after 8 h of incubation. The effect of different variable such as carbon sources (0.5 and 1.0%), nitrogen sources (0.75), sodium chloride, potassium chloride, zinc chloride (0.5 - 3.0%), pH (3 - 12) and temperature (25 - 55°C) on the protease production was checked. The maximum enzyme production was noted when B. subtilis EFRL 01 was grown on mineral medium containing 1.0% molasses, 0.75% peptone and 2.0% sodium chloride when incubated at 45°C for 8 h with initial pH 8.5. The enzyme produced by B. subtilis is pH stable and thermostable, which can be utilized in local detergent and leather industry.Key words: Bacillus subtilis EFRL 01, molasses, protease

Growth responses of NaCl stressed rice (Oryza sativa L.) plants germinated from seed in aseptic nutrient cultures supplemented with proline

Negative impact of salinity on plant germination is significant because of abundance of Na+ in culture medium, which causes growth inhibition. Effect of salinity (NaCl) in the presence of proline was assessed in rice (Oryza sativa L.) variety Khushbo-95 at seedling stage. Seeds were cultured on MS0 (MS basal medium), MS1 (MS0 + 100 mM NaCl) and MS2 (MS1 + 5 mM proline) for 20 days. Seedlings and its biomass decreased in saline culture. Similarly, total protein and sugar contents also decreased, while reducing sugars and proline contents increased. These parameters were observed to be slightly adverse in cultures supplemented with proline (MS2) and NaCl (MS2). Among cultures, leaf demography (cell size) was affected significantly; this may be the reflection of accumulation of proline, Na+ and Cl- and exclusion of K+ in developed rice seedlings.Key words: Oryza sativa L., seedling biomass, epidermal cells, proline content

Characteristics of micro-propagated banana (Musa spp.) cultures stressed with NaCl and polyethylene glycol

The effect of NaCl and PEG was assessed on plant micro-propagation rate in banana (Musa spp.) cv., Basrai. Well micro-propagated plantlets were cultured on four different stresses of NaCl and PEG-4000 including control level: MS2b (MS0 + 3.0 mg l-1 BAP), MS2c (MS0 + 100 mol m-3 NaCl), MS2d (MS0 + 5% PEG) and MS2e (MS0 + 100 mol m-3 NaCl + 5 % PEG) for 6-weeks. Efficiency of plant micro-propagation was reduced significantly among the stressed cultures. Similarly, photosynthetic pigments like chl a was decreased non-significantly but chl b, chl ab were decreased significantly. Total carotenoids were increased in the saline as well as PEG stressed cultures. Cell size of epidermis and aerenchyma was increased (p < 0.05), while parenchyma decreased. Proline and glycinebetain contents were increased (p < 0.05) in each stressed culture but were high in MS2 than in MS3 and MS4 cultures. Meanwhile, proteins, sugars, phenolics and nitrates were observed to be in the reversed (p < 0.05) phenomena. In conclusion, NaCl treatment was observed to be most toxic than the PEG or PEG with NaCl on the banana micro-propagation.Key words: Musa spp., micro-propagation, NaCl (sodium chloride), PEG (polyethylene glycol), chlorophyll contents, proline, reducing sugars

Blechnum Orientale Linn - a fern with potential as antioxidant, anticancer and antibacterial agent

<p>Abstract</p> <p>Background</p> <p><it>Blechnum orientale </it>Linn. (<it>Blechnaceae</it>) is used ethnomedicinally for the treatment of various skin diseases, stomach pain, urinary bladder complaints and sterilization of women. The aim of the study was to evaluate antioxidant, anticancer and antibacterial activity of five solvent fractions obtained from the methanol extract of the leaves of <it>Blechnum orientale </it>Linn.</p> <p>Methods</p> <p>Five solvent fractions were obtained from the methanol extract of <it>B. orientale</it> through successive partitioning with petroleum ether, chloroform, ethyl acetate, butanol and water. Total phenolic content was assessed using Folin-Ciocalteu's method. The antioxidant activity was determined by measuring the scavenging activity of DPPH radicals. Cytotoxic activity was tested against four cancer cell lines and a non-malignant cell using MTT assay. Antibacterial activity was assessed using the disc diffusion and broth microdilution assays. Standard phytochemical screening tests for saponins, tannins, terpenoids, flavonoids and alkaloids were also conducted.</p> <p>Results</p> <p>The ethyl acetate, butanol and water fractions possessed strong radical scavenging activity (IC₅₀8.6-13.0 μg/ml) and cytotoxic activity towards human colon cancer cell HT-29 (IC₅₀27.5-42.8 μg/ml). The three extracts were also effective against all Gram-positive bacteria tested: <it>Bacillus cereus, Micrococcus luteus</it>, methicillin-susceptible <it>Staphylococcus aureus </it>(MSSA), methicillin-resistant <it>Staphylococcus aureus </it>(MRSA) and <it>Stapylococcus epidermidis</it>(minimum inhibitory concentration MIC 15.6-250 μg/ml; minimum bactericidal concentration MBC 15.6-250 μg/ml). Phytochemical analysis revealed the presence of flavonoids, terpenoids and tannins. Ethyl acetate and butanol fractions showed highest total phenolic content (675-804 mg gallic acid equivalent/g).</p> <p>Conclusions</p> <p>The results indicate that this fern is a potential candidate to be used as an antioxidant agent, for colon cancer therapy and for treatment of MRSA infections and other MSSA/Gram-positive bacterial infectious diseases.</p