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Screening plant growth-promoting rhizobacteria for improving growth and yield of wheat.

AIMS: Plant growth promoting rhizobacteria (PGPR) are commonly used as inoculants for improving the growth and yield of agricultural crops, however screening for the selection of effective PGPR strains is very critical. This study focuses on the screening of effective PGPR strains on the basis of their potential for in vitro auxin production and plant growth promoting activity under gnotobiotic conditions. METHODS AND RESULTS: A large number of bacteria were isolated from the rhizosphere soil of wheat plants grown at different sites. Thirty isolates showing prolific growth on agar medium were selected and evaluated for their potential to produce auxins in vitro. Colorimetric analysis showed variable amount of auxins (ranging from 1.1 to 12.1 mg l-1) produced by the rhizobacteria in vitro and amendment of the culture media with l-tryptophan (l-TRP), further stimulated auxin biosynthesis (ranging from 1.8 to 24.8 mg l-1). HPLC analysis confirmed the presence of indole acetic acid (IAA) and indole acetamide (IAM) as the major auxins in the culture filtrates of these rhizobacteria. A series of laboratory experiments conducted on two cv. of wheat under gnotobiotic (axenic) conditions demonstrated increases in root elongation (up to 17.3%), root dry weight (up to 13.5%), shoot elongation (up to 37.7%) and shoot dry weight (up to 36.3%) of inoculated wheat seedlings. Linear positive correlation (r = 0.99) between in vitro auxin production and increase in growth parameters of inoculated seeds was found. Based upon auxin biosynthesis and growth-promoting activity, four isolates were selected and designated as plant growth-promoting rhizobacteria (PGPR). Auxin biosynthesis in sterilized vs nonsterilized soil inoculated with selected PGPR was also monitored that revealed superiority of the selected PGPR over indigenous microflora. Peat-based seed inoculation with selected PGPR isolates exhibited stimulatory effects on grain yields of tested wheat cv. in pot (up to 14.7% increase over control) and field experiments (up to 27.5% increase over control); however, the response varied with cv. and PGPR strains. CONCLUSIONS: It was concluded that the strain, which produced the highest amount of auxins in nonsterilized soil, also caused maximum increase in growth and yield of both the wheat cv. SIGNIFICANCE AND IMPACT OF STUDY: This study suggested that potential for auxin biosynthesis by rhizobacteria could be used as a tool for the screening of effective PGPR strains.

Germ-Free Life↗

Genetic variation, population structure and cryptic species within the black mudfish, Neochanna diversus, an endemic galaxiid from New Zealand.

To investigate the phylogenetic relationships and geographical structure among landlocked populations of the black mudfish, Neochanna diversus, mitochondrial DNA nucleotide sequence data were sampled from seven populations from the Waikato and Northland regions of New Zealand. The complete D-loop region was sequenced from 70 individuals, with 913 bp from the tRNA-pro end used in population and phylogenetic analysis. A tandem repeat array, which ranged in size up to 200 bp, was found in most populations at the 3' end of the D-loop that was not able to be aligned for analysis. Of the seven sites sampled, two from Northland exhibited significant sequence divergence from all other sites. There was also a clear distinction among remaining Northland sites and those from the Waikato. An additional 518 bp segment of the 16S region was sequenced from all sites and compared with the other New Zealand mudfish species, N. apoda, N. burrowsius and the Tasmanian mudfish Galaxias (Neochanna) cleaveri using Galaxias maculatus as an outgroup. Both D-loop and 16S sequence data provided strong evidence for a cryptic species of mudfish present in Northland. The significant genetic structure apparent in the black mudfish appears most probably to be attributed to geological conditions during the Pliocene, where peat wetlands became apparent in the Waikato while Northland consisted of disjunct 'islands'. Conservation and management of these populations must take into account the historical processes that have shaped these patterns of genetic diversity.

Animals↗

Factors affecting the transfer of porcine parvovirus antibodies from sow to piglets.

The aim of the study was to investigate the effects of a number of environmental, behavioural and biological factors on passive immunization of piglets as assessed by transfer of porcine parvovirus (PPV) antibodies (ab) from the colostrum of PPV vaccinated mothers to the serum of the piglets. Twenty primiparous sows were housed in pens with peat, straw and branches for nest building. Half the sows were prevented from achieving feedback from a completed farrowing nest by repeated removal of the nest from 10 to 12 h after nest building had begun, whereas the other half kept their nests. Sow serum PPV-ab titres were positively related to colostrum PPV-ab titres at birth of the first piglet (BFP) (P < 0.001). Litter average piglet PPV-ab titre was positively related to both sow serum and colostrum PPV-ab titres (both P < 0.001). In addition, in the individual piglets. PPV-ab titres were reduced as time from BFP to birth and time from birth to first sucking increased and time spent sucking decreased (all P < 0.01). There were no effects of treatment, time spent in lateral recumbency by the sow, number of times the sow stood or piglet weight on day 1 on piglet serum PPV-ab titres. Preventing prolonged farrowing, while at the same time ensuring the piglets' access to the udder, is important for transfer of maternal immunity. Measurements of specific antibodies in sow serum during the periparturient period and in piglet serum at 28 days of age may provide a practical tool for evaluating transfer of maternal immunity from sow to piglets.

Animals↗

Application of nested polymerase chain reaction for the detection of as yet uncultured organisms of the class Actinobacteria in environmental samples.

Different groups of as yet uncultured, phylogenetically deeply rooting organisms of the class Actinobacteria have been shown previously by analysis of 16S rDNA clone libraries to be present in different soil and marine environments of worldwide distribution. In order to monitor specifically the occurrence of two of the actinobacterial groups among bacterial DNA extracted from different environments, a nested polymerase chain reaction (PCR) method, aiming at the sensitive detection of 16S rDNA, was used. Analysis of a clone library generated from one of these 16S rDNA-derived, group-specific PCR products of DNA from a forest soil broadened the spectrum of putative taxa within the two phylogenetic groups. Application of this PCR system demonstrated the presence of these organisms in natural environments and in a sample of dried and moistened peat. Fluctuations in the detectability of the target groups in different samples from the same site strongly suggest that the taxon diversity of a 16S rDNA clone library must be regarded as a temporal genomic snapshot taken from this environmental community.

Actinobacteria↗

Quantitative determination of secoiridoid glucosides in in vitro propagated plants of Gentiana davidii var. formosana by high performance liquid chromatography.

A simple protocol for in vitro mass propagation of Gentiana davidii var. formosana (Gentianaceae) has been developed. Multiple shoot development was achieved by culturing the stem node explants on Murashige and Skoog (MS) medium supplemented with 4.44 microM N6-benzyladenine (BA). The shoots were multiplied by subculturing on MS medium supplemented with 1.07-10.74 microM alpha-naphthaleneacetic acid (NAA) and 8.88 microM BA. Shoots were rooted on MS basal medium supplemented with various auxins. Shoots rooted on growth regulator-free medium were transferred to peat moss:vermiculite mixture and acclimatized in the growth chamber. The contents of gentiopicroside and swertiamarin, the two important secoiridoid glucosides, in different plant material were determined by high performance liquid chromatography (HPLC). The analysis revealed that the content of gentiopicroside and swertiamarin in the aerial and underground parts of G. davidii var. formosana was higher than the marketed crude drug (underground parts of G. scabra) and varied with the age of the plant.

Chromatography, High Pressure Liquid↗

[Ophthalmopathologic findings in bog victims].

More than 1800 corpses found in European bogs have been described. Most were found in northwest Germany and Denmark. The 2700-year-old corpse of a young woman found in the Borremose fen in Denmark is described. The face was crushed, though injury during cutting of the peat was ruled out. Light and scanning electron microscopy revealed vessels both in the choroid of one eye and in one ear, with intravascular corpuscles--probably erythrocytes--but no extravascular hemorrhage. This means that the face was almost certainly crushed post mortem and during the winter. The cause of death is unknown. Very few of the other bog finds in Europe are well preserved, and even fewer have been pathologically studied or radiocarbon-dated. Interpretation of their fate is a very difficult, complex problem. One reason for this is that the corpses often remained buried for several thousand years, and they are often poorly preserved. The first descriptions of it were Roman, in particular Tacitus' Germania, which appeared in 98 AD--it is thus much younger than most of the corpses so far found. Tacitus talks of hanging, drowning, and ritual sacrifices. As regards future bog finds, it is suggested that an archaeologist, an anthropologist, a forensic expert and a taxi-dermist should be called in as soon as excavation begins. International cooperation between experts could prove very fruitful.

Adult↗

Mid- to late-Holocene El Nino-Southern Oscillation dynamics reflected in the subtropical terrestrial realm.

High resolution pollen analysis of mid- to late-Holocene peat deposits from southwest Florida reveals a stepwise increase in wetland vegetation that points to an increased precipitation-driven fresh water flow during the past 5,000 years. The tight coupling between winter precipitation patterns in Florida and the strength of the El Niño-Southern Oscillation (ENSO) strongly suggests that the paleo-hydrology record reflects changes in ENSO intensity. A terrestrial subtropical record outside the Indo Pacific Warm Pool both documents ecosystem response to the known onset of modern-day ENSO periodicities, between approximately 7,000 and 5,000 years B.P., and subsequent ENSO intensification after 3,500 years B.P. The observed increases in "wetness" are sustained by a gradual rise in relative sea level that prevents a return to drier vegetation through natural succession.

Ecosystem↗

Somatic embryogenesis and plant regeneration from cotyledon explants of a timber-yielding leguminous tree, Dalbergia sissoo Roxb.

Efficient plant regeneration through somatic embryogenesis was achieved from callus cultures derived from semi-mature cotyledon explants of Dalbergia sissoo Roxb., a timber-yielding leguminous tree. Somatic embryos developed over the surface of embryogenic callus and occasionally, directly from cotyledon explants without intervening callus phase. Callus cultures were initiated from cotyledon pieces of D. sissoo on Murashige and Skoog (1962) medium supplemented with 4.52, 9.04, 13.57, and 18.09 mumol/L 2,4-dichlorophenoxyacetic acid and 0.46 mumol/L Kinetin. Maximum percentage response for callus formation was 89% on MS medium supplemented with 9.04 mumol/L 2,4-D' and 0.46 mumol/L Kn. Somatic embryogenesis was achieved after transfer of embryogenic callus clumps to 1/2-MS medium without plant growth regulators (1/2-MSO). Average numbers of somatic embryos per callus clump was 26.5 on 1/2-MSO medium after 15 weeks of culture. Addition of 0.68 mmol/L L-glutamine to 1/2-MSO medium enhanced somatic embryogenesis frequency from 55% to 66% and the number of somatic embryos per callus clump from 26.5 to 31.1. Histological studies were carried out to observe various developmental stages of somatic embryos. About 50% of somatic embryos converted into plantlets on 1/2-MSO medium containing 2% sucrose, after 20 days of culture. Transfer of somatic embryos to 1/29-MSO medium containing 10% sucrose for 15 days prior to transfer on 1/2-MS medium with 2% sucrose enhanced the conversion of somatic embryos into plantlets from 50 to 75%. The plantlets with shoots and roots were transferred to 1/2 and 1/4-liquid MS medium, each for 10 days, and then to plastic pots containing autoclaved peat moss and compost mixture (1:1). 70% of the plantiets survived after 10 weeks of transfer to pots. 120 regenerated plantlets out of 150 were successfully acclimatised. After successful acclimatisation, plants were transferred to earthen pots.

Fabaceae↗

Random amplified ribosomal DNA restriction analysis for rapid identification of thermophilic Actinomycete-like bacteria involved in hypersensitivity pneumonitis.

Hypersensitivity pneumonitis (HP) is a pulmonary disease characterised by inflammation that can be caused by, amongst other substances, a subset of 4 thermophilic mycelial bacteria: Saccharopolyspora rectivirgula, Saccharomonospora viridis, Thermoactinomyces sacchari, and Thermoactinomyces vulgaris. Air sampling analyses in highly contaminated environments are often performed to evaluate exposure to these species which are difficult and fastidious to identify by conventional techniques. The aim of this study was to use amplified ribosomal DNA restriction analysis (ARDRA) to develop a method of identification for those thermophilic organisms that would be more rapid and simple. Strains of these 4 species were obtained from the American type culture collection (ATCC) and were characterized using biochemical tests and ARDRA patterns obtained on their partial-lenght amplified 16S rDNAs. To validate this approach, ARDRA with two restriction enzymes, TaqI and HhaI, was applied to 49 thermophilic actinomycete-like strains from environmental samples (sawmills). The results obtained show that combining some cultural characteristics and biochemical tests, such as xanthine or hypoxanthine decomposition, growth in the presence of NaCl, lysozyme or novobiocin, and spore resistance over 100 degrees C provide a rough identification and selection of the genera of interest. Consequently, target species could be confirmed by digestion of partial-lenght 16S rDNA with the use of Taql and HhaI restriction enzymes that gave specific restriction patterns. ARDRA analyses on the 49 environmental actinomycete-like organisms revealed the presence of 8 Saccharopolyspora rectivirgula, 2 Saccharomonospora viridis, and 15 Thermoactinomyces vulgaris strains, the other strains had restriction patterns different than those of the species of interest. Results of the present study will be applicable to other potential HP environments such as dairy barns, peat bogs and compost plants.

Actinomycetales↗

Reduction potential of microbial, odour and ammonia emissions from a pig facility by biofilters.

The intention of this study was the determination of the potential to reduce specific microbial bioaerosol (cultivable bacteria and fungi, total cell counts of microbes, airborne endotoxins and microbial volatile organic compounds, MVOC), odour and ammonia emissions from a pig facility by biofilters. Five identical biofilter units in half technical scale were filled with different filter materials (Biochips, coconut-peat, wood-bark, pellets + bark and compost) and connected in parallel to a piggery. The results showed obvious differences between the filter materials. Numbers of airborne cultivable bacteria were decreased by ca. 70 to 95% and the total counts of bacterial cells from ca. 25 to (>) 90%. The total amount of fungal cells was reduced by at least 60%, although the percentage of cultivable moulds in the air after passing the filters was sometimes higher than before. Airborne endotoxins and MVOC were effectively reduced by all filter materials to at least 90%. Regarding odour, the average reduction was between 40 and 83%, whereas only one of the filters proved to be capable of slightly reducing the ammonia emissions. No relationships between odour/ammonia and microbial bioaerosols with regard to the reduction efficiency of the different filter materials or the total load of the emitted air could be established. A tendency could be shown, that biofilters best capable to reduce odour emitted slightly more airborne bacteria, both cultivable and total cell counts.

Air Pollutants↗

Design of laying nests in furnished cages: influence of nesting material, nest box position and seclusion.

(1) Preferences for three nesting materials and nest box positions were investigated simultaneously in two trials using a furnished cage: one with 18 individual laying hens and one with 18 groups of 5 hens. Following a habituation period in pre-test cages, every hen or group of hens was tested for 2 d: once without and once with plastic flaps at the entrance of the nest boxes. (2) Hens preferred peat and artificial turf to coated wire mesh for egg laying. (3) One nest box position was clearly preferred to both other nest boxes. The hens' choice of nest box position was influenced by the pre-test cage in which they had been habituated. (4) The presence of plastic flaps at the entrance of the nest boxes had no influence on the proportion of eggs laid on the different nesting materials or on the proportion of floor eggs. (5) Individual and group testing resulted in the same overall results despite the presence of a distinct group effect.

Animal Husbandry↗

Bodyweight and serum copper concentrations of farmed red deer stags following oral copper oxide wire administration.

A diagnosis of secondary copper deficiency was established on a deer farm with a peat soil type, on the basis of confirmed enzootic ataxia in hinds, liver and serum copper concentrations and pasture and soil element analyses. Seventy-four weaner stags were selected for a trial to investigate a growth response to copper supplementation. Thirty-seven red and red x wapiti type stags were treated with 4g copper oxide wire particles at four months of age (April). A further 8g copper oxide wire was given in June. Thirty-seven untreated animals acted as controls. Bodyweights were measured on five occasions, from April 24 to November 26. Serum copper analyses were undertaken on ten deer prior to commencement of the trial, and on seven treated and eight control deer in June, July and October. Serum copper concentrations ranged from 2.0 to 19.3 micromol/l prior to the trial. In June, July and October serum copper ranged from 0.1-6.7, 0.6-5.0, and 1.3-6.3 micromol/l respectively, in control deer. In treated deer concentrations ranged from 7.2-14.7, 5.2-10.8, and 6.9-13.7 micromol/l in June, July and October respectively. The difference between mean copper concentration at each post-treatment sampling date was highly significant, (P<0.001). At the conclusion of the trial (November 26) the treated deer averaged 3.1kg heavier than controls, but this difference was not statistically significant. In view of these results and the variation in growth response trials in other species, further investigation of the effects of copper on the growth of young deer is warranted.

Journal Article↗

Relation between selenium content and glutathione-peroxidase activity in blood of cattle.

To identify a simple method for assessing the selenium demand in cattle, the relationship between selenium content in whole blood and the concentration of the selenium containing enzyme glutathione peroxidase (EC 1.11.1.9; GSH-Px) in red blood cells was studied. On six farms with suspected low soil selenium content, blood samples were collected from groups of calves, yearlings and adults at the end of the housing period and of the grazing period. The data obtained showed a highly significant correlation between the parameters mentioned: GSH-Px (U/g Hb) = 3.261 * Se (micrograms/kg) - 40.553. In growing animals there was a decline in Se supply with age, followed by a gradual recovery in heifers. This was most pronounced on some sandy soils and on peat soil. Seasonal effects could not be demonstrated.

Age Factors↗

Photochemical behavior of parathion in the presence of humic acids from different origins.

This paper reports the effect of ultraviolet radiation on the degradation of pesticide ethyl parathion in the presence of humic acids. Ethyl parathion was completely degraded in 300 min using an artificial lamp of 7.41 x 10(-10) einstein/s. Humic acid from peat did not influence the photochemical rate (k = 8.92 x 10(-3) min). However, in the presence of aquatic humic acid, the photochemical rate was higher (11.5 x 10(-3) min). The analytical determinations show the presence of p-nitrophenol and aminophenol in the reaction medium during the photochemical experiments. The kinetic of degradation in all experiments obeyed a first-order reaction pattern.

Biodegradation, Environmental↗

Comparison of natural adsorbents for metal removal from acidic effluent.

Adsorption tests were carried out in acidic synthetic solutions (pH 2.0) using 20 g l(-1) of various natural adsorbents and 0.25 mM of 11 different metals. In decreasing order, the most efficient adsorbents tested were: oyster shells, cedar bark, vermiculite, cocoa shells and peanut shells. In contrast, weak metal adsorption was demonstrated by: red cedar wood, peat moss, pine wood, corn cobs and perlite. Metal adsorption capacities in acidic synthetic solution followed the order: Pb2+> Cr3+> Cu2+> Fe2+> Al3+> Ni2+> Cd2+ > Mn2+ > Zn2+ >> Ca2+, Mg2+. Alkaline treatment (0.75 M NaOH) increased the effectiveness of metal removal for the majority of adsorbents. In contrast, acid treatment (0.75 M H2SO4) either reduced or did not affect the adsorption capacity of the materials tested. Finally, oyster shells, red cedar wood, vermiculite, cocoa shells and peanut shells, were effective natural adsorbents for the selective recovery of lead and trivalent chromium from acidic effluent.

Adsorption↗

Methane oxidation in three Alberta soils: influence of soil parameters and methane flux rates.

Current concern over the potentially negative impacts of climate change has brought attention to anthropogenic sources of methane, a primary greenhouse gas. Two such emission sources are methane leakage at heavy oil wells and sanitary landfills. At both of these sources, substantial quantities of methane could potentially be oxidised by methanotrophic microbes living in soils. Optimisation of this phenomenon may serve as an inexpensive technique for reducing methane emissions. Soil column and batch incubation experiments were performed on a landfill loam, an agricultural loam and a sedge peat to gain a better quantitative understanding of the biological and physical processes limiting CH4 oxidation in soils that undergo the freeze-thaw cycles associated with northern climates. Moisture content emerged as a critical variable that can limit a soil's CH4 oxidation potential. For example, the oxidation rate of the agricultural soil was seen to increase by an order of magnitude after increasing its moisture content from 6% to 10% of its dry weight.

Alberta↗

Nitrogen elimination mechanisms in an organic media aerated biofilter treating pig manure.

Biofilters using organic media are known to procure efficient treatment for different types of wastewater, but the nitrogen removal pathways implied are still not well understood. In this study, a lab-scale aerated biofilter using peat and treating pig manure was operated for 180 days, in order to quantify the nitrogen transformations occurring in it. It was shown that stripping was important during the start-up, until nitrification took place. Simultaneous nitrification and denitrification, proved by N2 production, became the principal mechanism after some time. The production of N2O did not seem to come only from heterotrophic denitrification, but also from chemodenitrification and autotrophic denitrification. It has also been found that part of the influent nitrogen was retained in the system during the first 150 days, due to filtration, sorption and assimilation. During the last periods of operation, the nitrogen previously retained has been used by microorganisms, leading to an excessive N2 discharge.

Animals↗

Factors influencing stable isotope ratios in CH4 and CO2 within subenvironments of freshwater wetlands: implications for delta-signatures of emissions.

Much uncertainty still exists regarding spatial and temporal variability of stable isotope ratios (13C/12C and D/H) in different CH4-emission sources. Such variability is especially prevalent in freshwater wetlands where a range of processes can influence stable isotope compositions, resulting in variations of up to approximately 50% for delta13C-CH4 and approximately 50% for deltaD-CH4 values. Within a temperate-zone bog and marsh situated in southwestern Ontario, Canada, gas bubbles in pond sediments exhibit only minor seasonal and spatial variation in delta13C-CH4, deltaD-CH4 and delta13C-CO2 values. In pond sediments, CO2 appears to be the main source of carbon during methanogenesis either directly via CO2 reduction or indirectly through dissimilation of autotrophic acetate. In contrast, CH4 production occurs primarily via acetate fermentation at shallow depths in peat soils adjacent to ponds at each wetland. At greater depths within soils, sigmaCO2 and H2O increasingly exert an influence on delta13C- and deltaD-CH4 values. Secondary alteration processes (e.g., methanotrophy or diffusive transport) are unlikely to be responsible for depth-related changes in stable isotope values of CH4. Recent models that attempt to predict deltaD-CH4 values in freshwater environments from D/H ratios in local precipitation do not adequately account for such changes with depth. Subenvironments (i.e., soil-forming and open water areas) in wetlands should be considered separately with respect to stable isotope signatures in CH4 emission models.

Atmosphere↗