Variation in seedling growth of 11 perennial legumes in response to phosphorus supply

Phosphorus (P) deficiency is a major problem for Australian agriculture. Development of new perennial pasture legumes that acquire or use P more efficiently than the current major perennial pasture legume, lucerne (Medicago sativa L.), is urgent. A glasshouse experiment compared the response of ten perennial herbaceous legume species to a series of P supplies ranging from 0 to 384 µg g^?1 soil, with lucerne as the control. Under low-P conditions, several legumes produced more biomass than lucerne. Four species (Lotononis bainesii Baker, Kennedia prorepens F.Muell, K. prostrata R.Br, Bituminaria bituminosa (L.) C.H.Stirt) achieved maximum growth at 12 µg P g^?1 soil, while other species required 24 µg P g^?1. In most tested legumes, biomass production was reduced when P supply was ≥192 µg g^?1, due to P toxicity, while L. bainesii and K. prorepens showed reduced biomass when P was ≥24 µg g^?1 and K. prostrata at ≥48 µg P g^?1 soil. B. bituminosa and Glycine canescens F.J.Herm required less soil P to achieve 0.5 g dry mass than the other species did. Lucerne performed poorly with low P supply and our results suggest that some novel perennial legumes may perform better on low-P soils.     

Dust mediated transfer of phosphorus to alpine lake ecosystems of the Wind River Range,Wyoming, USA

Alpine lakes receive a large fraction of their nutrients from atmospheric sources and are consequently sensitive to variations in both the amount and chemistry of atmospheric deposition. In this study we explored the spatial changes in lake water chemistry and biology along a gradient of dust deposition in the Wind River Range, Wyoming. Regional differences were explored using the variation in bulk deposition, lake water, sediment, and bedrock geochemistry and catchment characteristics. Dust deposition rates in the Southwestern region averaged 3.34 g m^?2 year^?1, approximately three times higher than deposition rates in the Northwestern region (average 1.06 g m^?2 year^?1). Dust-P deposition rates ranged from 87 µg P m² day^?1 in the Northwestern region to 276 µg P m² day^?1 in the Southwestern region. Subalpine and alpine lakes in the Southwestern region had greater total phosphorus (TP) concentrations (5–13 µg L^?1) and greater sediment phosphorus (SP) concentrations (2–5 mg g^?1) than similar lakes elsewhere in the region (1–8 µg L^?1 TP, 0.5–2 mg g^?1 SP). Lake phosphorus concentrations were related to dissolved organic carbon (DOC) across vegetation gradients, but related to the percent of bare rock, catchment area to lake area, and catchment steepness across dust deposition gradients. Modern phytoplankton and zooplankton biomasses were two orders of magnitude greater in the Southwest than in the Northwest, and alpine lakes in the Southwest had a unique diatom species assemblage with relatively higher concentrations of Asterionella formosa, Pseudostaurosira pseudoconstruens, and Pseudostaurosira brevistriata. These results suggests that catchment controls on P export to lakes (i.e. DOC) are overridden in dominantly bare rock basins where poor soils cannot effectively retain dust deposited P.     

Highly thermo–halo–alkali-stable β-1,4-endoxylanase from a novel polyextremophilic strain of Bacillus halodurans

A novel bacterial isolate, capable of producing extracellular highly thermostable, halo-alkali-stable and cellulase-free xylanase, was isolated from soil and identified as Bacillus halodurans TSPV1 by polyphasic approach. The Plackett–Burman design identified wheat bran, lactose, tryptone and NaCl as the factors that significantly affect xylanase production, and thus, these were optimized by response surface methodology. The data analysis suggested that optimum levels of wheat bran (15–20 g L^?1), lactose (1.0–1.5 g L^?1), tryptone (2–2.5 g L^?1) and NaCl (7.0–8.0 g L^?1) support 6.75-fold higher xylanase production than that in the un-optimized medium. The xylanase is optimally active at 90 °C and pH 10, and stable for 4 h at 90 °C (T _1/2 60 h) over a broad range of NaCl concentrations (0–29 %). This is the first report on the isolation of polyextremophilic B. halodurans strain that produces thermo-halo-alkali-stable xylanase in submerged fermentation. This enzyme efficiently saccharifies agro residues like wheat bran and corncobs. Fifty-six percent of hemicellulose of wheat bran could be hydrolyzed by xylanase (100 U g^?1 substrate) along with cellulase (22 U FPase and 50 U CMCase g^?1). The xylanase, being thermo-alkali stable and cellulase free, can find applications in pre-bleaching of paper pulps and hydrolysis of xylan in agricultural residues.     

Zinc seed coating improves the growth,grain yield and grain biofortification of bread wheat

This study, comprising three independent experiments, was conducted to optimize the zinc (Zn) application through seed coating for improving the productivity and grain biofortification of wheat. Experiment 1 was conducted in petri plates, while experiment 2 was conducted in sand-filled pots to optimize the Zn seed coating using two sources (ZnSO₄, ZnCl₂) of Zn. In the first two experiments, seeds of two wheat cultivars Lasani-2008 and Faisalabad-2008 were coated with 0.25, 0.50, 0.75, 1.00, 1.25, 1.50, 1.75 and 2.00 g Zn kg^?1 seed using ZnSO₄ and ZnCl₂ as Zn sources. The results of experiment I revealed that seed coating with 1.25 and 1.50 g Zn kg^?1 seed using both sources of Zn improved the seedling emergence. However, seed coated with 1.25 and 1.50 g Zn kg^?1 seed using ZnSO₄ was better regarding improvement in seedling growth and seedling dry weight. The results of the second experiment indicated that seed coated with 1.25 and 1.50 g Zn kg^?1 seed using ZnSO₄ improved the seedling emergence and seedling growth of tested wheat cultivars. However, seed coating beyond 1.5 g Zn kg^?1 seed using either Zn source suppressed the seedling emergence. Third experiment was carried out in glass house in soil-filled earthen pots. Seeds of both wheat cultivars were coated with pre-optimized treatments (1.25, 1.50 g Zn kg^?1 seed) using both Zn sources. Seed coating with all treatments of ZnSO₄ and seed coating with 1.25 g Zn kg^?1 seed using ZnCl₂ improved the seedling emergence and yield-related traits of wheat cultivars. Seed coating with 1.25 g Zn kg^?1 seed also improved the chlorophyll a and b contents. Maximum straw Zn contents, before and after anthesis, were recorded from seed coated with 1.5 g Zn kg^?1 seed using either Zn source. Increase in grain yield from seed coating followed the sequence 1.25 g Zn kg^?1 seed (ZnSO₄) >1.25 g Zn kg^?1 seed (ZnCl₂) >1.5 g Zn kg^?1 seed (ZnSO₄). However, increase in grain Zn contents from seed coated was 1.5 g Zn kg^?1 seed (ZnCl₂) >1.25 and 1.5 g Zn kg^?1 seed (ZnCl₂, ZnSO₄) >1.25 g Zn kg^?1 seed (ZnSO₄). Seed coating with Zn increased the grain Zn contents from 21 to 35 %, while 33–55 % improvement in grain yield was recorded. In conclusion, wheat seeds may be coated with 1.25 g Zn kg^?1 seed using either source of Zn for improving the grain yield and grain Zn biofortification.     

Toxicity of Cd to signal grass (Brachiaria decumbens Stapf.) and Rhodes grass (Chloris gayana Kunth.)

Given that Cd accumulates within plant tissues to levels that are toxic to animals, it is necessary to understand the role of plants in highly Cd-contaminated systems and their subsequent impact on the health of animals. A solution culture experiment was conducted to elucidate the effects of increasing Cd²⁺ activity ({Cd²⁺}) on growth of Rhodes grass (Chloris gayana Kunth.) and signal grass (Brachiaria decumbens Stapf.). The shoot and root fresh mass of both Rhodes grass and signal grass was reduced by 50% at ca. 0.5 µM {Cd²⁺}. Elevated {Cd²⁺} resulted in a significant decrease in the tissue Mn concentration for both the shoots and roots, and caused a chlorosis of the veins in the shoots. Root hair growth was prolific even at high {Cd²⁺}, thus root hair growth appeared to be less sensitive to elevated Cd than was root growth per se. The critical shoot tissue concentrations (50% reduction in growth), 230 µg g^?1 for Rhodes grass and 80 µg g^?1 for signal grass, exceeded the maximum level of Cd tolerated in the diet of animals (ca. 5 µg g^?1). When assessing the risk associated with the revegetation of Cd-contaminated sites with Rhodes grass or signal grass, careful consideration must be given, therefore, to the transfer of toxic concentrations of Cd to grazing animals and through the wider food chain.     

Early plant growth and biochemical responses induced by <Emphasis Type="Italic">Azospirillum brasilense</Emphasis> Sp245 lipopolysaccharides in wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) seedlings are attenuated by procyanidin B2

This study analyzes the effects of procyanidin B2 on early wheat plant growth and plant biochemical responses promoted by lipopolysaccharides (LPS) derived from the rhizobacteria Azospirillum brasilense Sp245. Measurements of leaf, root length, fresh weight, and dry weight showed in vitro plant growth stimulation 4 days after treatment with A. brasilense as well as LPS. Superoxide anion (O₂ ^·?) and hydrogen peroxide (H₂O₂) levels increased in seedling roots treated with LPS (100 μg mL^?1). The chlorophyll content in leaf decreased while the starch content increased 24 h after treatment in seedling roots. The LPS treatment induced a high increase in total peroxidase (POX) (EC 1.11.1.7) activity and ionically bound cell wall POX content in roots, when compared to respective controls. Early plant growth and biochemical responses observed in wheat seedlings treated with LPS were inhibited by the addition of procyanidin B2 (5 μg mL^?1), a B type proanthocyanidin (PAC), plant-derived polyphenolic compound with binding properties of LPS. All results suggest first that the ionically bound cell wall POX enzymes could be a molecular target of A. brasilense LPS, and second that the recognition or association of LPS by plant cells is required to activate plant responses. This last event could play a critical role during plant growth regulation by A. brasilense LPS.     

A Review of: “North-Holland Publishing Company,Amsterdam, New York,Oxford, 1976; Softbound, 290 pages, $ 19.25.”

Several factors affecting erythritol production from glycerol by Yarrowia lipolytica Wratislavia K1 strain were examined in batch fermentations. Ammonium sulfate, monopotassium phosphate, and sodium chloride were identified as critical medium components that determine the ratio of polyols produced. The central composite rotatable experimental design was used to optimize medium composition for erythritol production. The concentrations of ammonium sulfate, monopotassium phosphate, and sodium chloride in the optimized medium were 2.25, 0.22, and 26.4 g L^?1, respectively. The C:N ratio was found as 81:1. In the optimized medium with 100 g L^?1 of glycerol the Wratislavia K1 strain produced 46.9 g L^?1 of erythritol, which corresponded to a 0.47 g g^?1 yield and a productivity of 0.85 g L^?1 hr^?1. In the fed-batch mode and medium with the total concentration of glycerol at 300 g L^?1 and C:N ratio at 81:1, 132 g L^?1 of erythritol was produced with 0.44 g g^?1 yield and a productivity of 1.01 g L^?1 hr^?1.     

Elicitor mediated enhancement of wedelolactone in cell suspension culture of <Emphasis Type="Italic">Eclipta alba</Emphasis> (L.) Hassk

The present research focused on enhancing the production of wedelolactone through cell suspension culture (CSC) in Eclipta alba (L.) Hassk. With an aim of attaining a sustainable CSC, various plant growth regulators, elicitors and agitation speed were examined. Nodal segments of in vitro propagated plantlets induced the maximum percentage (93.47?±?0.61%) of callus inoculated on Murashige and Skoog (MS) medium fortified with picloram (2 mg L^?1). The growth kinetics of CSC exhibited a sigmoid pattern with a lag phase (0–6 days), a log phase (6–18 days), a stationary phase (18–24 days) and then death phase thereafter. The highest biomass accumulation in CSC with 7.09?±?0.06 g 50 mL^?1 fresh weight, 1.52?±?0.02 g 50 mL^?1 dry cell weight, 1.34?±?0.01?×?10⁶ cell mL^?1 total cell count and 57.00?±?0.58% packed cell volume was obtained in the liquid MS medium supplemented with 1.5 mg L^?1 picloram plus 0.5 mg L^?1 kinetin at 120 rpm. High performance thin layer chromatography confirmed that yeast extract (biotic elicitor) at 150 mg L^?1 accumulated more CSC biomass with 1.22-fold increase in wedelolactone (288.97?±?1.94 µg g^?1 dry weight) content in comparison to the non-elicited CSC (237.78?±?0.04 µg g^?1 dry weight) after 120 h of incubation. Contrastingly, methyl jasmonate (abiotic elicitor) did not alter the biomass but increased the wedelolactone content (259.32?±?1.06 µg g^?1 dry weight) to an extent of 1.09-fold at 100 µM. Complete plantlet regeneration from CSC was possible on MS medium containing N⁶-benzyladenine (0.75 mg L^?1) and abscisic acid (0.5 mg L^?1). Thus, the establishment of protocol for CSC constitutes the bases for future biotechnological improvement studies in this crop.     

Soil N and C trace gas fluxes and microbial soil N turnover in a sessile oak (Quercus petraea (Matt.) Liebl.) forest in Hungary

During two intensive field campaigns in summer and autumn 2004 nitrogen (N₂O, NO/NO₂) and carbon (CO₂, CH₄) trace gas exchange between soil and the atmosphere was measured in a sessile oak (Quercus petraea (Matt.) Liebl.) forest in Hungary. The climate can be described as continental temperate. Fluxes were measured with a fully automatic measuring system allowing for high temporal resolution. Mean N₂O emission rates were 1.5 μg N m⁻² h⁻¹ in summer and 3.4 μg N m⁻² h⁻¹ in autumn, respectively. Also mean NO emission rates were higher in autumn (8.4 μg N m⁻² h⁻¹) as compared to summer (6.0 μg N m⁻² h⁻¹). However, as NO₂ deposition rates continuously exceeded NO emission rates (−9.7 μg N m⁻² h⁻¹ in summer and −18.3 μg N m⁻² h⁻¹ in autumn), the forest soil always acted as a net NO _x sink. The mean value of CO₂ fluxes showed only little seasonal differences between summer (81.1 mg C m⁻² h⁻¹) and autumn (74.2 mg C m⁻² h⁻¹) measurements, likewise CH₄uptake (summer: −52.6 μg C m⁻² h⁻¹; autumn: −56.5 μg C m⁻² h⁻¹). In addition, the microbial soil processes net/gross N mineralization, net/gross nitrification and heterotrophic soil respiration as well as inorganic soil nitrogen concentrations and N₂O/CH₄ soil air concentrations in different soil depths were determined. The respiratory quotient (ΔCO_{2 resp} ΔO_{2 resp}⁻¹) for the uppermost mineral soil, which is needed for the calculation of gross nitrification via the Barometric Process Separation (BaPS) technique, was 0.8978 ± 0.008. The mean value of gross nitrification rates showed only little seasonal differences between summer (0.99 μg N kg⁻¹ SDW d⁻¹) and autumn measurements (0.89 μg N kg⁻¹ SDW d⁻¹). Gross rates of N mineralization were highest in the organic layer (20.1–137.9 μg N kg⁻¹ SDW d⁻¹) and significantly lower in the uppermost mineral layer (1.3–2.9 μg N kg⁻¹ SDW d⁻¹). Only for the organic layer seasonality in gross N mineralization rates could be demonstrated, with highest mean values in autumn, most likely caused by fresh litter decomposition. Gross mineralization rates of the organic layer were positively correlated with N₂O emissions and negatively correlated with CH₄ uptake, whereas soil CO₂ emissions were positively correlated with heterotrophic respiration in the uppermost mineral soil layer. The most important abiotic factor influencing C and N trace gas fluxes was soil moisture, while the influence of soil temperature on trace gas exchange rates was high only in autumn.     

Salicylic acid accumulation as a result of Cu,Zn, Cd and Pb interactions in common reed (<Emphasis Type="Italic">Phragmites australis</Emphasis>) growing in natural ecosystems

Common reed (Phragmites australis (Cav.) Trin. ex Steud.) plants were harvested from four natural water ecosystems of the Bogdanka river catchment (Poznań, Poland) four times throughout the 2014 vegetative season. Over the year, average metal contents followed different decreasing trends according to the analyzed tissue: Zn > Cu ≈ Pb > Cd (rhizomes) and Zn > Pb > Cu > Cd (leaves), and mean translocation ratios (leaves/rhizomes) were found as follows: 0.93, 0.70, 0.65, 0.40 for Zn, Pb, Cd and Cu, respectively. Metal content increased gradually during the growing season, and in the case of Cu, Cd and Pb exceeded the upper limit of average concentration detected in plants from natural ecosystems. However, the content of salicylic acid did not follow the increase of metal accumulation. In rhizomes, the highest production of the metabolite was observed in May and reached 324 ng g^?1 fresh weight (FW) (mean value). Afterwards, a significant drop to 50 ng g^?1 FW was observed. Simultaneously, the highest values of total salicylic acid in P. australis leaves were observed in July and accompanied the intensive development of the aboveground biomass of the plant (11.3 µg g^?1 FW–mean value). Subsequently, its content in leaves showed a significant decrease down to 2.1 µg g^?1 FW in November. Multivariate regression analysis revealed significant interactions between analyzed metals influencing the plant response to metal-derived stress. Cu and Zn showed antagonistic properties considering their uptake and the induction of salicylic acid biosynthesis, whereas non-essential metals (Pb and Cd) acted similarly and stimulated the formation of salicylic acid glucoside.     

Accumulation of arsenic in soil and rice under wetland condition in Bangladesh

Shallow tube well (STW) water, often contaminated with arsenic (As), is used extensively in Bangladesh for irrigating rice fields in the dry season, leading to potential As accumulation in soils. In the current study the consequences of arsenic from irrigation water and direct surface (0–15 cm) soil application were studied under field conditions with wetland rice culture over 2 years. Twenty PVC cylinders (30-cm length and 30-cm diameter) were installed in field plots to evaluate the mobility and vertical distribution of soil As, As mass balance, and the resulting influences on rice yield and plant-As concentration in Boro (dry season) and transplanted (T.) Aman (wet season) rice over the 2-year growth cycle. Treatments included irrigation-water As concentrations of 0, 1 and 2 mg L^?1 (Boro season only) and soil-As concentrations of 10 and 20 mg kg^?1. Following the 2-year cropping sequence the major portion (39.3–47.6%) of the applied arsenic was retained within the rooting zone at 0–15 cm depth, with 14.7–19.5% of the total applied As at the 5–10 cm and 10–15 cm soil depths compared to 1.3–3.6% at the 35–40 cm soil depth. These results indicate the relatively low mobility of applied As and the likely continued detrimental accumulation of As within the rooting zone. Arsenic addition in either irrigation water or as soil-applied As resulted in yield reductions from 21 to 74 % in Boro rice and 8 to 80 % in T. Aman rice, the latter indicating the strong residual effect of As on subsequent crops. The As concentrations in rice grain (0.22 to 0.81 µg g^?1), straw (2.64 to 12.52 µg g^?1) and husk (1.20 to 2.48 µg g^?1) increased with increasing addition of As. These results indicate the detrimental impacts of continued long-term irrigation with As-contaminated water on agricultural sustainability, food security and food quality in Bangladesh. A critical need exists for the development of crop and water management strategies to minimize potential As hazard in wetland rice production.     

Long-term effects of boron and copper on phenolics and monoterpenes in Scots pine (Pinus sylvestris L.) needles

Backgroud and aims

Plant boron (B) status is known to affect plant secondary metabolites but most studies have been short termed and in controlled environments. Copper (Cu) effects on phenolics are better known at toxic than at low levels. Here, the chemistry of Scots pine (Pinus sylvestris L.) needles was studied 20 years after fertilisation with B and Cu in a long-term field experiment on a drained boreal peatland.

Methods

Phenolic compounds were analysed from three needle year classes using high performance liquid chromatography (HPLC) and condensed tannins with modified acid-butanol assay. Monoterpenes in the youngest needles were analysed by gas chromatography–mass spectrometry (GC–MS).

Results

Needle B concentrations were at deficient level in controls (5.7 μg g^?1), but at the optimum level (12 μg g^?1) still 20 years after fertilisation. Copper concentrations were low but not deficient (4.0 μg g^?1 in unfertilised, 4.8 μg g^?1 in fertilised). Needle ageing increased the concentrations of individual phenolics in most cases, but decreased the concentration of condensed tannins. The concentrations of several individual phenolics were reduced by B fertilisation compared to B-deficient control, significantly in the cases of (+)-catechin and a neolignan. The concentrations of eight compounds and the sum of small-molecule phenolics were higher in Cu fertilised trees. Condensed tannins and monoterpenes were not affected by the micronutrients.

Conclusions

Boron and copper additions affected mostly the same phenolic compounds, but B decreased while Cu increased their concentrations, Cu effects being clearer. The higher phenolic concentrations in B deficient trees were not likely large enough to explain leader dieback in B-deficient trees. The effects and interactions of these micronutrients need to be further studied in field conditions to establish firstly if the changes in phenolics are consistent among species, and secondly what mechanisms lead to the changes. Although small, the changes in phenolic concentrations may affect the interactions of the trees with their biotic and abiotic environment, when consistent over many years.     

Soil factors affecting selenium concentration in wheat grain and the fate and speciation of Se fertilisers applied to soil

UK crops have a low selenium (Se) status, therefore Se fertilisation of wheat (Triticum aestivum L.) at 10 field sites was investigated and the effect on the content and speciation of Se in soils determined. Soil characterisation was carried out at each field site to determine the soil factors that may influence wheat grain Se concentrations in unfertilised plots. Soil samples were taken after harvest from each treatment to determine the fate and speciation of selenate fertiliser applied to soil. Wheat grain Se concentrations could be predicted from soil Se concentration and soil extractable sulphur (S) using the following regression model: Grain Se?=?a?+?b(total soil Se)?+?c(extractable soil Se) - d(extractable soil S), with 86 % of the variance being accounted for, suggesting that these properties control Se concentrations in grain from unfertilised plots. Extractable soil Se concentrations were low (2.4 – 12.4 µg kg^?1) and predominantly consisted of selenite (up to 70 % of extractable Se) and soluble organic forms, whereas selenate was below the detection limit. Little of the added Se, in either liquid or granular form was left in the soil after crop harvest. Se fertilisation up to 20 g ha^?1 did not lead to a significant Se accumulation in the soil, suggesting losses of Se unutilised by the crop.     

Copper-Induced Modulation of Biomass Growth,Physiological Parameters,Bioactive Centellosides,and Expression of Biosynthetic Pathway Genes in an Important Medicinal Herb, <Emphasis Type="Italic">Centella asiatica</Emphasis>

Hydroponically established Centella asiatica plants were studied under different concentrations of copper (0, 0.32,1.6, and 3.2 µM) for their morphological, physiological, and biochemical characteristics. The plants grown in the medium without any exogenous supply of copper showed improved biomass accumulation with maximum fresh weight (FW?=?24.7 g plant^?1) and dry weight (DW?=?2.35 g plant^?1). The total chlorophyll content, leaf number, and the leaf area were higher (0.27 mg g^?1 DW, 20 and 100.54 cm²) in the plants grown under copper-deficient conditions. The toxic effects of increased levels of copper were evident by significant inhibition in growth and other morphological parameters. Copper treatment showed an increase in malondialdehyde (MDA) content and SOD activity. Bioactive phytochemical profiling using HPLC analysis revealed that higher levels of copper (1.6 and 3.2 µM) inhibit the accumulation of total centelloside content. The differential accumulation of centelloside content was further validated by the consistent pattern of expression of key pathway genes related to centelloside production in copper-treated C. asiatica plants. The study unveiled some of the important facts associated with the mechanism of copper tolerance in this important medicinal herb C. asiatica. The findings of the present study can be further used to provide better production of high value, in demand centellosides.     

Isolation,Selection and Characterization of Root-Associated Rock Phosphate Solubilizing Bacteria in Moroccan Wheat (Triticum aestivum L.)

Abstract

Phosphorus (P) is the most important macronutrient next to nitrogen for the growth and development of plants. But often unavailable for plants because of its high reactivity with many soil constituents. Thus, the use of phosphate solubilizing bacteria (PSBs) as biofertilizers seems to be an effective way to resolve the soluble phosphorus availability in soil. The present study was conducted to isolate and characterize rock PSB associated with the rhizosphere of wheat (Triticum aestivum L.) from fourteen different wheat-growing sites of Meknes region in Morocco. A total of one hundred ninety-eight (198) rock PSBs were isolated employing NBRIP medium amended with rock phosphate (RP), out of which five strains (A17, A81, B26, B106, and B107) were selected for their strong ability to dissolve RP and were tested in vitro for plant growth-promoting (PGP) traits including production of indole acetic acid (IAA), siderophores, hydrogen cyanide (HCN), and antifungal activity, as well as their response to the effect of extrinsic and intrinsic stress. The 16S rRNA gene sequencing and phylogenetic analysis identified these isolates belong to four genera, Pantoea, Pseudomonas, Serratia, and Enterobacter. The phosphate solubilization index (SI) of selected isolates ranged between 2.3 and 2.7, and the amount of solubilized phosphorus in the liquid medium varied from 59.1 to 90.2 µg mL^?1. HPLC analysis revealed that all the selected isolates produced multiple organic acids (oxalic, citric, gluconic succinic, and fumaric acids) from glucose under aerobic conditions. Except for the A81 strain, all selected isolates were able to produce IAA ranging between 2.9 and 21.2 µg mL^?1. The isolates A17, B26, and B107 showed the ability to produce siderophores ranging from 79.3 to 20.8% siderophore units. Only two strains (A17 and B26) were able to produce HCN. All selected isolates showed good resistance against different environmental stresses like 10–50?°C temperature, 0.5–2?g L^?1 salt concentration and 4.5–9?pH range, and against different antibiotics. The antagonistic effect showed that among the five selected strains, only two strains (B26 and A17) were able to suppress the growth of tested fungi. This study clearly indicates that our selected rock PSBs can be used as biofertilizers for grain crops after studying their interaction with the host crop and field evaluation.     

Promotion of shoot regeneration of <Emphasis Type="Italic">Swertia chirata</Emphasis> by biosynthesized silver nanoparticles and their involvement in ethylene interceptions and activation of antioxidant activity

A complete protocol for the in vitro induction of Eclipta alba tetraploids has been optimized to enhance the wedelolactone content, an anti-cancerous compound. The effects of different concentrations of colchicine (0, 0.01, 0.05, 0.1, 0.2 and 0.3%; w/v) along with treatment durations (12, 24, 36 and 48 h) were investigated on shoot tip (ST) and nodal segment (NS). The treated explants were then incubated on Murashige and Skoog (MS) medium having 1.5 mg L^?1 N⁶-benzylaminopurine and 0.5 mg L^?1 α-napthalene acetic acid for shoot regeneration and afterward root was induced on 1.0 mg L^?1 indole-3-acetic acid enriched ½MS medium. The tetraploids of E. alba were proficiently induced by the treatment of 0.1% colchicine for 24 h. The highest tetraploid induction efficiency was obtained from ST (30.56%) in comparison to the NS (22.22%). Analysis by spectrophotometry and flow cytometry showed that colchicine treated plants contained higher quantity of DNA than diploid plants. Cytological studies demonstrated doubled the chromosome number in tetraploids (2n?=?4x?=?44) than diploids (2n?=?2x?=?22). The ploidy level enhancement lead to alteration of other traits, like increased plant height, stem diameter, leaf size, stomatal size and chlorophyll content. As determined through high performance thin-layer chromatography, the ultimate achievement of this technique is the higher accumulation of wedelolactone in tetraploid plants (300.32 µg g^?1 dry weight) in evaluation to in vitro diploid (131.31 µg g^?1 dry weight) and in vivo diploid mother plants (93.26 µg g^?1 dry weight), thus improving the pharmaceutical value of E. alba.     

CITRIC ACID PRODUCTION BY Candida SPECIES GROWN ON A SOY-BASED CRUDE GLYCEROL

Citric acid was produced by five species of the yeast Candida after growth on a medium containing soy biodiesel-based crude glycerol. After growth on a medium containing 10 g L^?1 or 60 g L^?1 crude glycerol for 168 hr at 30°C, Candida parapsilosis ATCC 7330 and C. guilliermondii ATCC 9058 produced the highest citric acid levels. On 10 g L^?1 or 60 g L^?1 crude glycerol for 168 hr at 30°C, the citric acid level produced by C. parapsilosis ATCC 7330 was 1.8 g L^?1 or 11.3 g L^?1, respectively, while C. guilliermondii ATCC 9058 produced citric acid concentrations of 3.0 g L^?1 or 10.4 g L^?1, respectively. Biomass production by C. guilliermondii ATCC 9058 on 10 g L^?1 or 60 g L^?1 crude glycerol for 168 hr at 30°C was highest at 1.2 g L^?1 or 6.9 g L^?1, respectively. The citric acid yields observed for C. guilliermondii ATCC 9058 after growth on 10 g L^?1 or 60 g L^?1 crude glycerol (0.35 g g^?1 or 0.21 g g^?1, respectively) were generally higher than for the other Candida species tested. When similar crude glycerol concentrations were present in the culture medium, citric acid yields observed for some of the Candida species utilized in this study were about the same or higher compared to citric acid yields by Yarrowia lipolytica strains. Based on the findings, it appeared that C. guilliermondii ATCC 9058 was the most effective species utilized, with its citric acid production being similar to what has been observed when citric acid-producing strains of Y. lipolytica were grown on crude glycerol under batch conditions that could be of significance to biobased citric acid production.     

Bioaccumulation of Trace Mercury in Trophic Levels of Benthic, Benthopelagic, Pelagic Fish Species, and Sea Birds from Arvand River, Iran

In this study, concentration of mercury was determined in the trophic levels of benthic, benthopelagic, pelagic fish species, and river birds from Arvand River, located in the Khuzestan province in the lowlands of southwestern Iran at the head of the Persian Gulf. The order of mercury concentrations in tissues of the fish species was as follows: liver>gill>muscle and in tissues of the kingfisher species was as follows: feather>liver>kidney>muscle. Therefore, liver in fish and feather in kingfisher exhibited higher mercury concentration than the other tissues. There was a positive correlation between mercury concentrations in fish and kingfisher species with size of its food items. We expected to see higher mercury levels in tissues of female species because they are larger and can eat larger food items. The results of this study show that the highest mean mercury level were found in the kingfisher (Anas crecca), followed by benthic (Epinephelus diacanthus), benthopelagic (Chanos chanos), and pelagic fish (Strongylura strongylura). Mean value of mercury in fish species, S. strongylura were (0.61 μg g^?1 dry weight), C. chanos (0.45 μg g^?1 dry weight), E. diacanthus (0.87 μg g^?1 dry weight), and in kingfisher species A. crecca was (2.64 μg g^?1 dry weight). Significant correlation between mercury concentration in fish and kingfisher may be related to high variability of mercury in the fish.     

Handling culture medium composition for optimizing plant cell suspension culture in shake flasks

Douglas-fir is a conifer species of major economic importance worldwide, including Western Europe and New Zealand. Herein we describe some characterization and significant refinement of somatic embryogenesis in Douglas-fir, with focus on maturation. The most typical structures observed in the embryonal masses were large polyembryogenic centres (up to 800–1500 µm) with a broad meristem, creating a compact cell “package” with suspensor cells. Singulated somatic embryos composed of both a embryonal head (300–400 µm) and long, tightly arranged suspensor were also frequent. Embryo development was enhanced following embryonal mass dispersion on filter paper discs at low density (50–100 mg fresh mass). Moreover, increasing gellan gum concentration in maturation medium (up to 10 g L^?1) improved both the quantity and quality of cotyledonary somatic embryos (SEs), which were subsequently able to germinate and develop into plantlets at high frequency. Embryogenic yield was highly variable among the seven embryogenic lines tested (27–1544 SE g^?1 fresh mass). Interestingly secondary somatic embryogenesis could be induced from cotyledonary SEs of both low- and highly-productive lines with some useful practical outcomes: secondary lines from low-performance lines (30–478 SE g^?1 fresh mass) displayed significantly higher embryogenic yield (148–1343 SE g^?1 fresh mass). In our best conditions, the total protein content in cotyledonary SEs increased significantly with maturation duration (up to 150 µg mg^?1 fresh mass after 7 weeks) but remained below that of mature zygotic embryos (300 µg mg^?1). The protein pattern was similar in both somatic and zygotic embryos, with major storage proteins identified as 7S-vicilin- and legumin-like proteins.     

Evaluation of ethanol production by a new isolate of yeast during fermentation in synthetic medium and sugarcane bagasse hemicellulosic hydrolysate

A new xylose fermenting yeast was isolated from over-ripe banana by enrichment in xylose-containing medium. The phylogenetic analysis of ITS1-5.8S-ITS2 region sequences of ribosomal RNA of isolate BY2 revealed that it shows affiliation to genus Pichia and clades with Pichia caribbica. In batch fermentation, Pichia strain BY2 fermented xylose, producing 15 g l^?1 ethanol from 30 g l^?1 xylose under shaking conditions at 28°C, with ethanol yield of 0.5 g g^?1 and volumetric productivity of 0.31 g l^?1 h^?1. The optimum pH range for ethanol production from xylose by Pichia strain BY2 was 5–7. Pichia strain BY2 also produced 6.08 g l^?1 ethanol from 30 g l^?1 arabinose. Pichia strain BY2 can utilize sugarcane bagasse hemicellulose acid hydrolysate for alcohol production, efficiency of fermentation was improved by neutralization, and sequential use of activated charcoal adsorption method. Percent total sugar utilized and ethanol yield for the untreated hydrolysate was 17.14% w/v and 0.33 g g^?1, respectively, compared with 66.79% w/v and 0.45 g g^?1, respectively, for treated hemicellulose acid hydrolysate. This new yeast isolate showed ethanol yield of 0.45 g g^?1 and volumetric productivity of 0.33 g l^?1 h^?1 from sugarcane bagasse hemicellulose hydrolysate detoxified by neutralization and activated charcoal treatment, and has potential application in practical process of ethanol production from lignocellulosic hydrolysate.