Influencing in vitro clonal propagation of Chonemorpha fragrans(moon) Alston by culture media strength,plant growth regulators,carbon source and photo periodic incubation

Chonemorpha fragrans is an endangered medicinal woody climber,regarded among alternative plant sources of camptothecin.Camptothecin is a monoterpene indole anti-cancer alkaloid with annual trade value of over three billion U.S.dollars in the recent,and is used in the production of its analog drugs approved for the chemotherapy of cancer of varied types.Effects of plant growth regulators,culture media strength and photoperi-odic duration on the micropropagation ef ciency of C.fragrans from nodal segment explants were studied on Murashige and Skoog(MS)medium amended with Thidiazuron(TDZ),Benzylaminopurine(BAP)or Kinetin(Kin).Thidiazuron was more ef cient over BAP and Kin when half basal MS medium was used over full or quarter strength.Results of carbon source experiment showed sucrose as the most effective over glucose,fructose,and maltose in the clonal production.Studies on the photope-riodic incubation duration showed 12 h as the best light period and sub or supra-optimal resulted in the production of abnormal and albino micro shoots.Experimental results on the evaluation of physiological,biochemical parameters showed the role of pigment molecules and antioxidant systems in the production of albino micro shoots.     

Evaluation the effect of subchronic feeding of transgenic cotton line (CKC1) on the faecal microbiota of albino rabbits

Recent studies have demonstrated a strong relationship between the intestinal microbiota and the host health. As such, consumers are increasingly becoming more concerned about the potential effect of certain foods/feeds, particularly of transgenic origin on the gut microbiota. Although the European Food Safety Authority has recommended in their guidelines, to study the effect of transgenic food/feed on host-microbiota, yet, few studies have focused on the evaluation of such effects mainly due to culturing difficulties. Therefore, this study was intended to evaluate the potential adverse effects of transgenic diet consumption on some specific gut microflora (Lactobacillus group, Bifidobacterium genus, Escherichia coli subgroup and Enterococcus genus) of rabbits. A total of forty-eight rabbits were randomly assigned into four groups and fed a diet containing a variable proportion of transgenic cottonseeds at 0, 20, 30 and 40% inclusion level, respectively. Changes in the specific or total faecal bacterial population were monitored at five different experimental stages (i.e. 0, 45, 90, 135 and 180 days) using both the traditional plate count method (TM) and quantitative real-time PCR (qPCR). No significant differences (p > .05) were observed concerning numbers of specific bacteria or total bacteria between the control and experimental groups, though qPCR showed numerically higher values in terms of 16S rRNA gene copies as compared to the values obtained from TM. However, such numerical differences were biologically insignificant (p > .05). Similarly, no significant variations were noticed in the calculated B/E (log₁₀ copies of Bifidobacterium per g faces/log10 copies of E. coli genome per g faeces) ratios in all the groups. All the ratios were in the range of 1.24 to 1.30 throughout the experiment, indicating a good balance of intestinal microflora and greater resistance to intestinal disorders. It is therefore concluded that feeding transgenic cottonseeds could not adversely affect the gut microflora of rabbits during a long-term study.     

Efficiency of rice bran for the removal of selected organics from water: kinetic and thermodynamic investigations

The sorption efficiency of indigenous rice (Oryza sativa) bran for the removal of organics, that is, benzene, toluene, ethylbenzene, and cumene (BTEC), from aqueous solutions has been studied. The sorption of BTEC by rice bran is observed over a wide pH range of 1-10, indicating its high applicability to remove these organics from various industrial effluents. Rice bran effectively adsorbs BTEC of 10 microg mL(-1) sorbate concentration from water at temperatures of 283-323 +/- 2 K. The effect of pH, agitation time between solid and liquid phases, sorbent dose, its particle size, and temperature on the sorption of BTEC onto rice bran has been studied. The pore area and average pore diameter of rice bran by BET method are found to be 19 +/- 0.7 m(2) g(-1) and 52.8 +/- 1.3 nm. The rice bran exhibits appreciable sorption of the order of 85 +/- 3.5, 91 +/- 1.8, 94 +/- 1.4, and 96 +/- 1.2% for 10 microg mL(-1) concentration of benzene, toluene, ethylbenzene, and cumene, respectively, in 60 min of agitation time using 0.1 g of rice bran at pH 6 and 303 K. The sorption data follow Freundlich, Langmuir, and Dubinin-Radushkevich (D-R) models. Sorption capacities have been computed for BTEC by Freundlich (32 +/- 3, 61 +/- 14, 123 +/- 28, and 142 +/- 37 m mol g(-1)), Langmuir (6.6 +/- 0.1, 7.5 +/- 0.13, 9.5 +/- 0.22, and 9.4 +/- 0.18 m mol g(-1)), and D-R isotherms (11 +/- 0.5, 16 +/- 1.3, 30 +/- 2.2, and 33 +/- 2.5 m mol g(-1)), respectively. The Lagergren equation is employed for the kinetics of the sorption of BTEC onto rice bran and first-order rate constants (0.03 +/- 0.002, 0.04 +/- 0.003, 0.04 +/- 0.003, and 0.05 +/- 0.004 min(-1)) have been computed for BTEC at their concentration of 100 mug mL(-1) at 303 K. Studies on the variation of sorption with temperatures (283-323 K) at 100 mug mL(-1) sorbate concentration gave thermodynamic constants DeltaH (kJ mol(-1)), DeltaG (kJ mol(-1)), and DeltaS (J mol(-1) K(-1)). The results indicate that the sorption of organics onto rice bran is exothermic and spontaneous in nature under the optimized experimental conditions selected. This sorbent has been used successfully to accumulate and then to determine benzene, toluene, and ethylbenzene in wastewater sample.     

Biological control of red rot in sugarcane by native pyoluteorin‐producing Pseudomonas putida strain NH‐50 under field conditions and its potential modes of action

BACKGROUND: Rhizobacteria have a good potential to suppress soilborne diseases, but their efficacy against sugarcane pests is rarely reported. Bacterial strains isolated from the rhizosphere of sugarcane were evaluated for their potential to suppress red rot disease on two susceptible varieties, Co‐1148 and SPF‐234, under field conditions. The strains were also characterised for the production of secondary metabolites associated with their antagonistic activity. RESULTS: One out of four strains, the Pseudomonas putida strain NH‐50 (EU627168), reduced disease severity by 44–60% in different field trials. This potent antagonistic strain produced pyoluteorin antibiotic, as confirmed by high‐performance liquid chromatography (HPLC). The PltB gene involved in pyoluteorin synthesis was amplified from the P. putida strain NH‐50 and sequenced. The extracellular metabolites and volatile and diffusible antibiotics secreted by the tested strains inhibited mycelial growth of Glomerella tucumensis (Speg.) Arx & E Mull in vitro by 7–55%. CONCLUSION: The pyoluteorin‐producing bacteria P. putida strain NH‐50 significantly reduced disease severity on both sugarcane varieties, irrespective of fungal inoculation, i.e. either inoculated through stem or through soil. This strain also possesses other plant growth characteristics and can be used as a biopesticide for sugarcane crop. Copyright © 2011 Society of Chemical Industry     

Suppression of root-knot disease by Pseudomonas fluorescens CHA0 in tomato: importance of bacterial secondary metabolite, 2,4-diacetylpholoroglucinol

The antimicrobial metabolites 2,4-diacetylphloroglucinol (2,4-DAPG) and pyoluteorin contribute to the ability of Pseudomonas fluorescens strain CHA0 to control plant diseases caused by soil-borne pathogens. P. fluorescens strain CHA0 and its derivatives CHA89 (antibiotics-deficient) and CHA0/pME3424 (antibiotics overproducing) were investigated as potential biocontrol agents against Meloidogyne javanica the root-knot nematode. Exposure of root-knot nematode to culture filtrates of P. fluorescens under in vitro conditions significantly reduced egg hatch and caused substantial mortality of M. javanica juveniles. Nutrient broth yeast extract (NBY) medium amended with 2% (w/v) glucose or 1 mM EDTA markedly repressed hatch inhibition activity of the strain CHA0 but not that of CHA0/pME3424 or CHA89. On the other hand, NBY medium amended with glucose significantly enhanced nematicidal activity of the strain CHA0/pME3424. Neither glucose nor EDTA had an influence on the nematicidal activity of the strains CHA0 and CHA89. Under in vitro conditions, antibiotic overproducing strain CHA0/pME3424 and CHA0 expressed phl‘-’lacZ reporter gene but strain CHA89 did not. Expression of the reporter gene reflects actual production of DAPG. In general, CHA0/pME3424 expressed reporter gene to a greater extent compared to its wild type counterpart CHA0. Regardless of the bacterial strains, reporter gene expression was markedly enhanced when NBY medium was amended with glucose but EDTA had no such effect. A positive correlation between the degree of juvenile mortality and extent of phl‘-’lacZ reporter gene expression was also observed in vitro. Strain CHA0 produced zones of 4-6 mm on MM medium containing gelatin while strain CHA0/pME3424 and CHA89 did not. When MM medium containing gelatin was amended with 2% glucose of 1 mM EDTA size of haloes produced by the strain CHA0 reduced to 2 mm. Under glasshouse conditions aqueous cell suspension of the strains CHA0 or CHA0/pME3424 at various inoculum levels (10⁷, 10⁸ or 10⁹ cfu ml⁻¹) significantly reduced root-knot development. CHA89 caused significant reduction in galling when applied at 10⁹ cfu ml⁻¹. To better understand the mechanism of nematode suppression, split root bioassay was performed. Split-root experiments, that guarantee a spatial separation of inducing agent and a challenging pathogen, showed that soil treatment of one half of the root system with cell suspension of CHA0 or CHA0/pME3424 resulted in a significant systemic induced resistance leading to reduction of M. javanica infection of tomato roots in the non-baterized nematode treated half. The results clearly suggest that the antibiotic 2,4-DAPG from P. fluorescens CHA0 act as the inducing agents of systemic resistance in tomato roots. Populations of CHA0 and its derivatives declined progressively by 10-fold between first and fourth harvests (0-21 days after inoculation). However, bacterial populations increased at final harvest (28 days after application).     

Management of human health risk in the context of kitchen gardens polluted by lead and cadmium near a lead recycling company

Purpose

At the global scale, gardening activities are often performed in urban areas with a historical background of pollution. In this study, a participatory program was developed with citizens concerned by gardening activities near a 50-year-old regulated lead recycling company, with the aim of co-constructing the tools for the assessment and management of potential sanitary risks induced by historic pollution with persistent (eco) toxic metals: lead and cadmium.

Materials and methods

Soils and vegetables (lettuce, leek, celery, carrot, chard, pumpkin, and celeriac) samples were collected from four kitchen gardens neighboring a 50-year-old secondary lead smelter. Both total and in vitro human bioaccessible metal concentrations in the cultivated plants were measured in relation to soil characteristics.

Results and discussion

The results showed that the soils of these gardens were slightly contaminated by metals (Pb, 77 to 236 mg kg^?1; and Cd, 0.5 to 1 mg kg^?1) in comparison with the natural geologic background. However, significant pollution of vegetables can occur especially with lead (Pb up to 9.8 mg kg^?1 in lettuce) and certainly as a result of direct foliar transfer. The washing of plants before consumption is therefore recommended in the context of atmospheric fallout of ultrafine particles enriched with metals.

Conclusions

Metal bioaccessibility measure integrates the influence of metal type, plant type, and soil physico-chemical properties. Based on the results, it is proposed that human bioaccessible fraction of metals may also be currently taken into account as well as total metal quantities and bioaccumulation factors in risk assessment studies performed in gardens. Overall, this study has led to reflections and functional recommendations aimed at reducing human exposure and to finally developing sustainable gardening practices.

     

Suppression of Meloidogyne javanica by Pseudomonas aeruginosa IE-6S in tomato: the influence of NaCl, oxygen and iron levels

Understanding the environmental factors that influence the suppression of disease-suppressive strains of Pseudomonas fluorescens is an essential step toward improving the level and reliability of their biocontrol activity. A 0.8 M NaCl concentration was optimal for in vitro survival and growth of IE-6S⁺ while, nematicidal activity by IE-6S⁺ was maximal when the bacterium was exposed to 0.4 M NaCl. The bacterium was highly sensitive to high (1.6 M) NaCl concentration. Culture filtrate of the bacterium resulting from the medium supplemented with 0.2 or 0.4 M NaCl showed the presence of secondary metabolite, hydrogen cyanide (HCN). Soil amendment with IE-6S⁺ alone or in conjunction with up to 0.8 M NaCl enhanced bacterial efficacy towards Meloidogyne javanica, the root-knot nematode. Soil amendment with NaCl up to 0.8 M also resulted in enhanced bacterial rhizosphere colonization and growth of tomato seedlings. Protein content of the shoot was reduced when soil was amended with 1.6 M NaCl. Inner root establishment of the bacterium was greatly affected in the soils treated with 1.6 M NaCl. Under in vitro conditions, IE-6S⁺ showed enhanced growth when kept at ambient oxygen conditions while the growth of bacterium affected when incubated at low oxygen conditions. Culture filtrate of the bacterium resulting from low oxygen level caused greater mortality of M. javanica juveniles in vitro compared with the filtrates obtained from ambient oxygen conditions. Culture filtrate from low oxygen conditions also showed the presence of hydrogen cyanide while those from ambient oxygen condition did not. Under glasshouse conditions, regardless of bacterial application, nematode penetration rate was greater when the pots were watered from the top; nematode penetration was lowered in bacterized pots compared with non-bacterized controls. IE-6S⁺ applied in the pots either watered from the top or bottom had no significant impact on growth of tomato but protein contents of the leaves increased after treatment with the bacterium. Rhizosphere and inner root colonization of the bacterium increased when the pots were watered from the top. Under in vitro conditions, with an increased iron concentration in the form of FeEDDHA, growth of IE-6S⁺ and its nematicidal activity increased. Culture filtrate of IE-6S⁺ obtained from liquid King's B medium supplemented with 0.5 or 1.0 mM FeEDDHA showed the presence of HCN. Under glasshouse conditions, soil treated with FeEDDHA alone did not reduce nematode penetration rates but did reduce greatly when applied in conjunction with IE-6S⁺. FeEDDHA applied at 0.5 mg/kg of soil in combination with IE-6S⁺ significantly enhanced plant growth and leaf protein contents. FeEDDHA at 1 mg/kg of soil increased bacterial populations both in the rhizosphere and inner root tissues of tomato.     

Mineral bioavailability in grains of Pakistani bread wheat declines from old to current cultivars

Estimating variation in grain mineral concentration and bioavailability in relation to grain yield and the year of cultivar release is important for breeding wheat with increased content of bioavailable minerals. The grain yield and yield components, grain phytate concentration, and concentration and bioavailability of minerals (zinc Zn, iron Fe and calcium Ca) in wheat grains were estimated in 40 wheat cultivars released in Punjab (Pakistan) during the last five decades. Mean grain Zn and Ca concentrations in current-cultivars were significantly lower (≥14%) than in obsolete cultivars released during the Green Revolution (1965–1976). Much of this variation was related to increased grain weight in current-cultivars. There was a positive correlation among minerals (r = 0.39 or higher, n = 40) and minerals with phytate in wheat grains (r = 0.38 or higher, n = 40). The tested cultivars varied widely in grain yield and grain phytate-to-mineral molar ratios (phytate:mineral). Compared to obsolete cultivars, the current-cultivars had a higher phytate:mineral ratio in grains, indicating poor bioavailability of minerals to humans. The study revealed a non-significant relationship between grain yield and phytate:mineral ratios in grains. Therefore, breeding for lower phytate:mineral ratios in wheat grains can ensure increased mineral bioavailability without significant reduction in the yield potential. Future breeding should be focused on developing new genotypes suitable for mineral biofortification and with increased mineral bioavailability in grains.