Effect of inoculation with wild type Azospirillum brasilense and A. irakense strains on development and nitrogen uptake of spring wheat and grain maize

Under the controlled conditions of the greenhouse and by varying some biotic and abiotic factors, we tried to identify some of the factors critical to obtain successful Azospirillum inoculation. Spring wheat and grain maize were inoculated with different concentrations of the wild type strains A. brasilense Sp245 and A. irakense KBC1, and grown in a substrate with varying concentrations of organic matter (OM) and N fertiliser. The inoculum concentration was one of the factors that influenced most the outcome of an inoculation experiment on wheat, with lower inoculum concentrations (10⁵-10⁶ cfu plant^-1) stimulating root development and plant dry weight and higher inoculum concentrations (10⁷-10⁸ cfu plant^-1) having no effect or sometimes even inhibiting root development. The effect of inoculation was most pronounced at low to intermediate N fertilisation levels, while the OM content of the substrate had no effect. Inoculation was found to affect early plant and root development, plant and root dry weight, grain yield and the N-uptake efficiency of plants. However, inoculation did not change the N concentration in plants or grains. In addition, a difference in the ability of both strains to stimulate plant growth and N uptake of wheat and maize was observed, with A. brasilense Sp245 having most effect on spring wheat and A. irakense KBC1 being more effective on grain maize. The significance of the obtained results for agriculture is discussed.     

A spontaneous chlorate-resistant mutant of Azospirillum brasilense Sp245 displays defects in nitrate reduction and plant root colonization

Azospirillum, a soil bacterium capable of colonizing plant roots, can reduce NO₃^-. In this work, a spontaneous chlorate-resistant mutant of Azospirillum brasilense Sp245, named Sp245chl1, was phenotypically characterized. The mutant is defective in both assimilatory and periplasmic dissimilatory nitrate reductase activity. Using the gusA reporter gene methodology, Sp245chl1 was found to be significantly affected in its ability to colonize roots of wheat and rice seedlings.     

Nitrogen fixation in biological soil crusts from southeast Utah,USA

Biological soil crusts can be the dominant source of N for arid land ecosystems. We measured potential N fixation rates biweekly for 2 years, using three types of soil crusts: (1) crusts whose directly counted cells were >98% Microcoleus vaginatus (light crusts); (2) crusts dominated by M. vaginatus, but with 20% or more of the directly counted cells represented by Nostoc commune and Scytonema myochrous (dark crusts); and (3) the soil lichen Collema sp. At all observation times, Collema had higher nitrogenase activity (NA) than dark crusts, which had higher NA than light crusts, indicating that species composition is critical when estimating N inputs. In addition, all three types of crusts generally responded in a similar fashion to climate conditions. Without precipitation within a week of collection, no NA was recorded, regardless of other conditions being favorable. Low (<1°C) and high (>26°C) temperatures precluded NA, even if soils were moist. If rain or snow melt had occurred 3 or less days before collection, NA levels were highly correlated with daily average temperatures of the previous 3 days (r²=0.93 for Collema crusts; r²=0.86 for dark crusts and r²=0.83 for light crusts) for temperatures between 1°C and 26°C. If a precipitation event followed a long dry period, NA levels were lower than if collection followed a time when soils were wet for extended periods (e.g., winter). Using a combination of data from a recording weather datalogger, time-domain reflectometry, manual dry-down curves, and N fixation rates at different temperatures, annual N input from the different crust types was estimated. Annual N input from dark crusts found at relatively undisturbed sites was estimated at 9 kg ha^-1 year^-1. With 20% cover of the N-fixing soil lichen Collema, inputs are estimated at 13 kg ha^-1 year^-1. N input from light crusts, generally indicating soil surface disturbance, was estimated at 1.4 kg ha^-1 year^-1. The rates in light crusts are expected to be highly variable, as disturbance history will determine cyanobacterial biomass and therefore N fixation rates.     

The role of potassium in sustaining yields in a long-term rice-wheat experiment in the Indo-Gangetic Plains of Nepal

A long-term soil fertility experiment (1988-1999) at the Regional Agricultural Research Station, Bhairhawa, Nepal, was analysed to determine: (1) how long the yields of rice (Oryza sativa L.) and wheat (Triticum aestivum L.) can be sustained without K but with N and N+P (NP) applied with or without farmyard manure (FYM) and green manure, and (2) the impact of K application on yields. Starting from the 1995 wheat season, the experiment was modified to accommodate K at 0, 42, and 84 kg ha^-1 in plots receiving NP to study the response of rice and wheat to K. Both rice and wheat responded to K application but the response of wheat was substantially higher, indicating that the availability of native K may have been lower in wheat. Rice yields were lower in treatments without P than with P, and yields declined significantly (0.11-0.20 Mg ha^-1 year^-1) in all the treatments except in NP and NP+FYM. Wheat yield was more adversely affected than rice yield when P and K were not applied. In addition, wheat yields were low (average 0.5-2.1 Mg ha^-1 in various treatments). Wheat yields declined (0.08-0.12 Mg ha^-1 year^-1) in all but FYM treatments indicating the role of FYM in sustaining yields. The interaction of K deficiency with Helminthosporium leaf blight (spot blotch and tan spot) is also suggested as one of the factors limiting wheat yields. The estimated K balance in soil was highly negative. Results suggest that farmers should apply adequate amount of K for higher and sustainable rice and wheat yields.     

Alginate microbeads as inoculant carriers for plant growth-promoting bacteria

A method of inoculating wet and dry seeds with plant growth-promoting bacteria (PGPB) using alginate microbeads as a substrate and Azospirillum brasilense as the model PGPB was developed. The microbeads were produced by low pressure spraying of an alginate solution mixed with liquid bacterial culture suspended in a very rich medium through a small nozzle resulting in small-diameter droplets. These droplets, when sprayed into a slowly stirred solution of CaCl₂, immediately hardened into microbeads at diameters ranging between 100 and 200 µm. Although the process killed part of the entrapped bacteria, the remaining bacteria residing in the microbeads were sufficient [>10¹¹ colony-forming units (CFU) g^-1 inoculant] for seed inoculation. Further, it was found that the bacterial population in the inoculant could be enhanced by secondary multiplication in the same medium for an additional 16 h. It was found that the microbeads could be used either wet or dry. Dry inoculant was produced using dry air at 38°C, creating a powdery substance loaded with >10⁹ CFU g^-1 beads. Alternatively, dry microbeads were produced using a standard freeze-drying procedure. This dry preparation was easily attached to dry seed surfaces with the addition of 1% alcohol-diluted lecithin or with 0.5% synthetic paper adhesive (Resistol). The bacteria were slowly released from the microbeads in amounts ranging from 10⁴ to 10⁶ CFU g^-1 depending on the type (wet or dry, with or without skim milk) and the time of incubation (the longer the incubation period, the smaller the amount of bacteria released with time). The wet and dry inoculants enhanced the development of wheat and tomato seedlings growing in unfertile soil, and biodegraded within 15 days in moist soil.     

Denitrification, N2O and CO2 fluxes in rice-wheat cropping system as affected by crop residues, fertilizer N and legume green manure

Use of renewable N and C sources such as green manure (GM) and crop residues in rice-wheat cropping systems of South Asia may lead to higher crop productivity and C sequestration. However, information on measurements of gaseous N losses (N₂O+N₂) via denitrification and environmental problems such as N₂O and CO₂ production in rice-wheat cropping systems is not available. An acetylene inhibition-intact soil core technique was employed for direct measurement of denitrification losses, N₂O and CO₂ production, in an irrigated field planted to rice (Oryza sativa L.) and wheat (Triticum aestivum L.) in an annual rotation. The soil was a coarse-textured Tolewal sandy loam soil (Typic Ustochrept) and the site a semi-arid subtropical Punjab region of India. Wheat residue (WR, C:N=94) was incorporated at 6 t ha^-1 and sesbania (Sesbania aculeata L.) was grown as GM crop for 60 days during the pre-rice fallow period. Fresh biomass of GM (C:N.=18) at 20 or 40 t ha^-1 was incorporated into the soil 2 days before transplanting rice. Results of this study reveal that (1) denitrification is a significant N loss process under wetland rice amounting to 33% of the prescribed dose of 120 kg N ha^-1 applied as fertilizer urea-N (FN); (2) integrated management of 6 t WR ha^-1 and 20 t GM ha^-1 supplying 88 kg N ha^-1 and 32 kg FN ha^-1 significantly reduced cumulative gaseous N losses to 51.6 kg N ha^-1 as compared with 58.2 kg N ha^-1 for 120 kg FN ha^-1 alone; (3) application of excessive N and C through applying 40 t GM ha^-1 (176 kg N ha^-1) resulted in the highest gaseous losses of 70 kg N ha^-1; (4) the gaseous N losses under wheat were 0.6% to 2% of the applied 120 kg FN ha^-1 and were eight- to tenfold lower (5-8 kg N ha^-1) than those preceding rice; (5) an interplay between the availability of NO₃^- and organic C largely controlled denitrification and N₂O flux during summer-grown flooded rice whereas temperature and soil aeration status were the primary regulators of the nitrification-denitrification processes and gaseous N losses during winter-grown upland wheat; (6) the irrigated rice-wheat system is a significant source of N₂O as it emits around 15 kg N₂O-N ha^-1 year^-1; (7) incorporation of WR in rice and rice residue (C:N=63) in wheat increased soil respiration, and increased CO₂ production in WR- and GM-amended soils under anaerobic wetland rice coincided with enhanced rates of denitrification; and (8) with adequate soil moisture, most of the decomposable C fraction of added residues was mineralized within one crop-growing season and application of FN and GM further accelerated this process.     

Interactive effects of native and exotic earthworms on resource use and nutrient mineralization in a tropical wet forest soil of Puerto Rico

Investigation of single or mixed assemblages of native Estherella sp. and exotic Pontoscolex corethrurus from a rain forest in Puerto Rico was undertaken to understand resource use patterns, and linkages with C and N mineralization in a 19-day incubation. Resource use was explored with addition of ¹⁵N-enriched leaf litter and ¹³C-enriched glucose to reconstructed organic and mineral soil horizons. Juvenile Estherella sp. became at least 6.06‰ more enriched in ¹³C than sub-adult Estherella sp. or adult P. corethrurus. Sub-adult Estherella sp. became >3.6‰ enriched in ¹³C over P. corethrurus. '¹⁵N acquired by P. corethrurus was greater by 0.83-1.56‰ in the mixed-species than the single-species assemblages. '¹⁵N of sub-adult Estherella sp. was enriched by 0.73-0.81‰ over juvenile Estherella sp. in the single-species assemblage. Net N immobilization occurred in the organic layer of all ¹⁵N-enriched treatments. Net N mineralization in mineral soil layers was significantly greater in microcosms with P. corethrurus than in those containing only Estherella sp.. Cumulative respiration was greatest in P. corethrurus assemblages, however, assemblages with only Estherella sp. released more ¹³C in respiration. P. corethrurus assimilated different N resources when incubated with, as compared to without, native Estherella sp.. '¹³C and '¹⁵N signatures acquired by assimilation of ¹³C and ¹⁵N differed by species, developmental stage, and competitive interactions. The results showed that alone, exotic P. corethrurus induced higher mineralization rates than native Estherella sp., but that the interaction of exotic and native species impinged on resource use by P. corethrurus, reducing the effect of the exotic species on C and N mineralization. Invasion of exotic P. corethrurus may change the mineralization potentials of C and N and their biogeochemical cycling in soils.     

Identification and characterisation of a diuron-degrading bacterium

Isolate D47 has previously been shown to degrade a range of urea-based herbicides. The DNA encoding the16 S rRNA gene of this strain was amplified by polymerase chain reaction (PCR) and sequenced. Database similarity searches indicated the gene was similar to those present in Arthrobacter species. The 16S rRNA gene sequence was compared to eight full-length sequences that have been obtained from related strains in this cluster by both distance and parsimony methods. The analyses confirmed the inferred relationship between D47 and Arthrobacter oxydans-type strains within the Arthrobacter globiformis group. Biochemical tests confirmed this result. Studies with ¹⁴C-carbonyl-labelled diuron indicated that D47 hydrolysed the urea side chain at the carbonyl group. Loss of the parent compound was accompanied by an equal accumulation of 3,4-dichloroaniline and loss of [¹⁴C]-CO₂. Cell-free extracts of D47 indicated a broad temperature optimum for degradative activity between 15°C and 30°C, a broad pH optimum of 6.5-8.0 and a decline in activity with increasing salt concentration beyond 50 mM. This information sets the basic characteristics of the strain and the enzyme for cloning and expression of the gene(s) encoding this activity in a heterologous host.     

Cultivable heterotrophic N<Subscript>2</Subscript>-fixing bacterial diversity in rice fields in the Yangtze River Plain

Heterotrophic N₂-fixing bacteria are a potentially important source of N₂ fixation in rice fields due to the moist soil conditions. This study was conducted at eight sites along a geographic gradient of the Yangtze River Plain in central China. A nitrogen-free solid malate-sucrose medium was used to isolate heterotrophic N₂-fixing bacteria. Numbers of the culturable N₂-fixing bacteria expressed as CFU (colony forming units) ranged between 1.41ǂ.42᎒⁶ and 1.24ǂ.23᎒⁸ in the sampled paddy field sites along the plain. Thirty strains with high ARA (acetylene reduction activity) were isolated and purified; ARA of the strains varied from 0.9 to 537.8 nmol C₂H₄ culture^-1 h^-1, and amounts of ¹⁵N fixed ranged between 0.008 and 0.4866 mg·culture^-1·day^-1. According to morphological and biochemical characteristics, 14 strains were identified as the genus Bacillus, 2 as Burkholderia, 1 as Agrobacterium, 4 as Pseudomonas, 2 as Derxia, 1 as Alcaligenes, 1 as Aeromonas, 2 as Citrobacter, and 3 strains belonged to the corynebacter-form group.     

Nitrous oxide emissions from the wheat-growing season in eighteen Chinese paddy soils: an outdoor pot experiment

To identify the key soil parameters influencing N₂O emission from the wheat-growing season, an outdoor pot experiment with a total of 18 fertilized Chinese soils planted with wheat was conducted in Nanjing, China during the 2000/2001 wheat-growing season. Average seasonal N₂O-N emission for all 18 soils was 610 mg m^-2, ranging from 193 to 1,204 mg m^-2, approximately a 6.2-fold difference between the maximum and the minimum. Correlation analysis indicated that the seasonal N₂O emission was negatively correlated with soil organic C (r²=0.5567, P<0.001), soil total N (r²=0.4684, P<0.01) and the C:N ratio (r²=0.4530, P<0.01), respectively. A positive dependence of N₂O emission on the soil pH (r²=0.3525, P<0.01) was also observed. No clear relationships existed between N₂O emission and soil texture, soil trace elements of Fe, Cu and Mg, and above-ground biomass of the wheat crop at harvest. A further investigation suggested that the seasonal N₂O-N emission (E, mg m^-2) can be quantitatively explained by E=1005-34.2SOC+4.1S_a (R²=0.7703, n=18, P=0.0000). SOC and S_a represent the soil organic C (g kg^-1) and available S (mg kg^-1), respectively.     

Different behavioral patterns of the earthworms Octolasion tyrtaeum and Diplocardia spp. in tallgrass prairie soils: potential influences on plant growth

This study addressed differences between Diplocardia spp. (a native North American earthworm) and Octolasion tyrtaeum (an introduced European species), with respect to behavior, influence on soil microbial biomass, and plant uptake of N in tallgrass prairie soils. We manipulated earthworms in PVC-encased soil cores (20 cm diameter) over a 45-day period under field conditions. Treatments included: (1) control with no earthworms, (2) Diplocardia spp. only, and (3) O. tyrtaeum only. Prior to addition of earthworms, seedlings of Andropogon gerardii (a dominant tallgrass) were established in each core, and a dilute solution of ¹³C-labeled glucose and ¹⁵N-labeled (NH₄)₂SO₄ was added to the soil to facilitate examination of earthworm/microbe/plant interactions. We found that Diplocardia spp. were significantly more active than O. tyrtaeum, and quickly assimilated ¹³C and ¹⁵N from the tracer. Individuals of Diplocardia spp. were present at shallower soil depths than O. tyrtaeum throughout the study. Contrary to expectation, this greater activity of Diplocardia spp. did not result in increased plant productivity. Rather, the activity of Diplocardia spp. was associated with less plant growth and smaller amounts of N acquired by A. gerardii seedlings compared to controls or O. tyrtaeum treatments. We observed few significant influences of earthworm treatments on microbial biomass C or N pool sizes, but the microbial C/N ratio was consistently greater in the presence of Diplocardia spp. relative to O. tyrtaeum. Results of this study indicate that activity of earthworms may enhance competition for N between microbes and plants during the growing season in tallgrass prairie.     

Carbon partitioning in plant and soil,carbon dioxide fluxes and enzyme activities as affected by cutting ryegrass

The effect of a single cut (simulated grazing) and regrowth of Lolium perenne on CO₂ efflux from soil (loamy Haplic Luvisol), on below-ground C translocation and on the distribution of plant C among different soil particle size fractions was investigated under controlled conditions with and without N fertilization by pulse labelling with ¹⁴C 7 times (four before and three after the cutting). The amount of ¹⁴C respired from the rhizosphere of Lolium decreased by a factor of about 3 during 1 month of growth. At the same time the amount of ¹⁴C stored in soil increased. Cut and non-fertilized plants respired less C in the rhizosphere compared to the uncut plants and cut fertilized plants. About 80% of the root-derived CO₂ efflux originated from the C assimilated after defoliation, and 20% originated from the C assimilated before cutting. N fertilization decreased the below-ground C losses (root respiration and exudation) during regrowth. The shoot is the main sink of assimilated C before and after the defoliation. N fertilization led to higher C incorporation into the shoot parts growing after defoliation compared to unfertilized plants. A lower incorporation of ¹⁴C was observed in the roots of N fertilized plants. The relative growth rates (expressed as ¹⁴C specific activity) of roots and stubble were minimal and that of shoot parts growing after defoliation was maximal. Twelve percent of ¹⁴C was found in the newly grown leaves after regrowth; nevertheless, 4.7% and 2.4% of ¹⁴C in the new shoot parts were translocated from the root and shoot reserves of unfertilized and fertilized plants, respectively. Most of the C retranslocated into the new Lolium leaves originates from the stubble and not from the roots. Between 0.5% and 1.7% of ¹⁴C recovered in shoots and below-ground C pools was found in the soil microbial biomass. Cutting and fertilization did not change ¹⁴C incorporation into the microbial biomass and did not affect xylanase, invertase, and protease activities. Tracing the assimilated C in particle size fractions revealed maximal incorporation for the sand and clay fraction.     

Growth stimulation and nitrogen supply to wheat plants inoculated with Azospirillum brasilense

Twelve Azospirillum brasilense strains isolated from wheat (Triticum aestivum L.) roots were compared for root colonization, growth stimulation, and nitrogen (N) supply to young wheat plants cv. Klein Chamaco grown in sterile nutrient solutions without N. All the strains inoculated colonized both the root surface and interior, and most strains stimulated root and shoot growth, although the degree of stimulation was different for the different strains. Some strains increased the total N content of roots and tops at the end of the experiment, in one case up to 80% of the uninoculated plants, while others produced no effect on N content. No correlation could be found between growth stimulation or the amount of N supplied to the plant with the degree of root colonization. When the most efficient strain for N fixation was inoculated to different wheat cultivars, it stimulated growth and supplied N to the five cultivars tested, although the degree of root colonization, growth stimulation and N supply showed differences among the cultivars. Our results suggest that there exists the potential of A. brasilense to supply N to wheat plants in considerable amounts, although an adequate strain are still to be identified.     

Effects of elevated carbon dioxide concentration on biological nitrogen fixation, nitrogen mineralization and carbon decomposition in submerged rice soil

Controlled-environment chambers were used to study the effects of elevated CO₂ concentrations on biological N fixation, N mineralization and C decomposition in rice soil. In three chambers, CO₂ concentration was maintained at 353ᆣ/396ᆫ µmol mol^-1 (day/night; ambient CO₂), while in another three, CO₂ was maintained at 667ᆸ/700ᆽ µmol mol^-1 (day/night; elevated CO₂) throughout the growing season. Rice (var. Nipponbare) seedlings were grown under either ambient or elevated CO₂ concentrations, and then transplanted into the soils in the corresponding chambers. At different growth stages, soil samples were taken from surface (0-1cm) and sub-surface (1-10cm) layers at the centre of four hills, then sieved (<1 mm) to remove root residues. Fresh soil was used to measure N fixation activity (using the acetylene reduction assay), NH₄⁺ content and organic C. Separate sets of soil samples were transferred to serum bottles and anaerobically incubated at 30°C for 30 days to measure potential rates of N mineralization and C decomposition. Under an elevated atmospheric CO₂ concentration, acetylene reduction activity significantly increased in the surface soil layer during the early cultivation stages and in the sub-surface soil layer during the latter part of cultivation. There was no difference in the amount of NH₄⁺ in fresh soils between elevated and ambient CO₂ chambers, while the rate of N mineralization was increased by elevated CO₂ during the latter part of cultivation. Soils from the elevated CO₂ chambers had obviously higher rate of C decomposition than that from the ambient CO₂ chambers. CH₄ production gradually increased with the growth of rice plants. These results suggest that elevated CO₂ affected biological N fixation, N mineralization and C decomposition in submerged rice soil during the different growth stages of rice.     

Nitrogen mineralization potential of dairy manures and its relationship to composition

The objectives of this study were to determine the variability in mineralization of dairy manure N, to determine if N mineralization can be predicted by compositional factors or by near- or mid-infrared reflectance spectroscopy. Dairy manures (n =107) were collected from farms in Maryland, Virginia, Pennsylvania, New York, and Connecticut. The composition of these manures ranged from 14 to 386 g dry matter kg^-1, 0.9 to 9.5 kg total N/m³, and 0.3 to 4.7 kg NH₄⁺-N/m³. Manure-amended soil was aerobically incubated at 25°C and concentrations of NH₄⁺-N and NO₃^--N were determined at day 2 and day 56. The manures were highly variable in their N mineralization characteristics, ranging from a net mineralization of 54.9% to a net immobilization of 29.2% of the organic N. When compositional parameters were individually regressed against percentage mineralized organic N, the highest correlation coefficient (r) was 0.164. A stepwise regression of all 11 variables yielded a maximal r of 0.486. These results suggest that the availability of dairy manure organic N is highly variable and that the availability cannot be predicted from simple compositional parameters. No relationship was found between near-infrared spectral characteristics and N mineralization suggesting that no simple relationship exists between N mineralization and compositional characteristics. There appears to be some potential for the use of mid-infrared for determining the mineralization potential of manures.     

Response of neem (Azadirachta indica A. Juss) to indigenous arbuscular mycorrhizal fungi, phosphate-solubilizing and asymbiotic nitrogen-fixing bacteria under tropical nursery conditions

Neem (Azadirachta indica A. Juss) seedlings were inoculated with arbuscular mycorrhizal (AM) fungi, Glomus intraradices Schenck and Smith and G. geosporum (Nicol. and Gerd.) Walker, Azospirillum brasilense, and phosphate-solubilizing bacteria (PSB) individually or in various combinations in unsterile soil under nursery conditions. Seedlings were harvested at 60 and 120 days after transplantation. Microbial inoculation resulted in increased mycorrhizal colonization, greater plant height, leaf area and number, root collar diameter, biomass, phosphorus, nitrogen and potassium content, and seedling quality. Inoculated seedlings also had low root/shoot ratios and low nutrient utilization efficiencies. Populations of PSB declined with seedling growth; contrarily populations of A. brasilense increased. A. brasilense and PSB populations were related to each other and influenced root colonization by AM fungi. Microbial inoculation effects were greatest when seedlings were inoculated with a combination of microbes rather than individually. This clearly indicates that these microorganisms act synergistically when inoculated simultaneously, with maximum response being when both AM fungi were coinoculated with A. brasilense and PSB. The results emphasize the importance of microbial inoculations for the production of robust, rapidly growing seedlings in nurseries and illustrate the advantage of inoculating soils of a low microbial population with indigenous microbes.     

Cultivar-specific dinitrogen fixation in Vicia faba studied with the nitrogen-15 natural abundance method

N fixation by different faba bean (Vicia faba) cultivars was studied using the natural abundance method. The delta ¹⁵N ('¹⁵N) values of the faba beans and the reference plants differed by 4.6-7.0‰. The non-nodulating V. faba cv. F48 seems to be the best reference plant for nodulated and N₂-fixing V. faba. Significant differences occurred in the quantity of N₂ fixation of six V. faba cultivars. The average fraction of N derived from air (FNdfa) estimated from leaf material ranged between 69 and 80%. Shoot-based estimates of N fixation varied between 200 and 360 kg N ha^-1. N fixation was affected more by differences in FNdfa than by differences in total N accumulation. Fixation data calculated with the non-nodulated reference plant V. faba cv. F48 were lower than those calculated with cabbage (Brassica oleracea) and ryegrass (Lolium perenne) as reference plants. Of all reference plants, non-N₂-fixing V. faba cv. F48 has a root system and temporal pattern of N assimilation that is the one most similar to that of N₂-fixing V. faba plants. Cv. F48 showed senescence as did the other V. faba cultivars after pod-fill was complete, whereas cabbage, ryegrass and camomile had a later senescence period. N fixation during pod-filling appears more important for a good yield than N₂ fixation abilities in the earlier growth period. The best V. faba cultivars left about 100 kg N ha^-1 in residual material on the field as fertilization for the following crops.     

Decomposition and nitrogen release of understorey plant residues in biological and integrated apple orchards under field conditions in New Zealand

In grassed-down apple orchards in New Zealand, the understorey vegetation is usually mown and the plant residues are returned to the orchard floors as a source of nutrients. It is, therefore, important to determine the decomposition pattern and the rate of N release from understorey plant residues. In this study, the decomposition and N release of surface-placed understorey plant residues were determined in the field and compared across treatments of grassed-down biological (BFP) and integrated fruit production (IFP) orchards in two different locations (Lincoln and Clyde) in New Zealand using the litterbag technique. At Lincoln, the field experiment was a randomised complete block design with three different treatments (two BFPs, one IFP) each with three replicates; while at Clyde, the field experiment consisted of non-replicated apple orchard plots with three treatments (two IFPs, one BFP). A comparison was also made between surface-placed and soil-buried understorey plant residues in a BFP orchard at one location. Samples of understorey plant residues collected from orchard mowings in the respective treatments were returned to the same treatment plot in litterbags and retrieved at intervals of 90 days for 360-450 days. Results showed that the single exponential decay model, Y=A₀ e^-kt, accounted significantly (PА.001) for 97-99% of the variation in the decomposition and N release patterns, which ranged from 6.0᎒^-3-9.6᎒^-3 day^-1 and 7.0᎒^-3-13.0᎒^-3 day^-1, respectively. Half-lives for C and N of residues were approximately 70-120 days and 50-110 days, respectively. Soil-buried plant residues showed more rapid decomposition and N release compared with those of surface-placed plant residues (80% vs. 54% in 90 days). In general, plant residue decomposition and N release were significantly more rapid in IFP than in BFP treatments (13.0᎒^-3 vs 7.0᎒^-3 day^-1 for N release). Overall, differences in plant residue decomposition and N release rates related to understorey plant residue quality and treeline management practices rather than the orchard system as a whole.     

Macronutrient accumulation in wheat crop (Triticum aestivum L.) with Azospirillum brasilense associated with nitrogen doses and sources

Abstract

New studies are needed to optimize the nitrogen (N) amount that can be applied to utilize the Azospirillum brasilense benefits. In addition, information regarding the interaction between the urease inhibitor and biological nitrogen fixation (BNF) and how they affect the macronutrients accumulation are also needed. We evaluate the effect of N sources and doses associated with A. brasilense regarding the macronutrients accumulation in straw and grains and wheat grain yield in tropical conditions. A randomized block experimental design was used with four replications in a 2?×?5?×?2 factorial arrangement as follows: two N sources (urea and urea with urease enzyme inhibitor NBPT; five N doses (0, 50, 100, 150, and 200?kg ha^?1) applied in topdressing; with and without A. brasilense inoculation. We found that an increase in N doses positively influenced the accumulation of macronutrients in straw and grains and the wheat grain yield. N sources have similar effects. Inoculation with A. brasilense increased accumulation of Mg and S in straw and P, Ca, and Mg in grains, regardless of the N dose. The inoculation with A. brasilense associated with 140?kg ha^?1 of N increased wheat grain yield. The inoculation can contribute in a more sustainable way to wheat nutrition and optimizing N fertilization.     

Inoculation of wheat with Azospirillum brasilense and Pseudomonas fluorescens: Impact on the production and culturable rhizosphere microflora

Scientific evidence recognizes that the operation of a terrestrial ecosystem depends on soil microbial activity. Some Azospirillum strains produce plant growth regulators, increase the development of roots, and fix atmospheric nitrogen (N₂). Some Pseudomonas strains are capable of producing cytokinins and solubilizing organic phosphorus. A sustainability analysis requires a detailed knowledge of the interrelationships between the microorganisms added to the system and those present in the soil. This study examines the effect of three commercial inoculants Azospirillum brasilense Az1 and Az2 as well as Pseudomonas fluorescens Pf on biomass production, grain yield and rhizosphere colonization of wheat, combined with two levels of N-addition. Plate counts of rhizosphere soil showed that the inoculation and N-addition did not affect the number of P. fluorescens, whereas it significantly affected the number of Azospirillum. N-addition and inoculation did not change the communities of actinomycetes and bacteria but they changed the number of fungi at the rhizosphere of wheat plants. Community-level physiological profiles of carbon-source utilization of rhizosphere soil microbial communities were changed after inoculation with Az1, Az2 and Pf depending on the phenological stage of the crop. Although no significant responses were observed, in average, PGPB inoculation increased aerial biomass by 12%, root biomass by 40% and grain yield by 16%. These increases represent important earnings for the farmer and they may help to obtain a greater sustainability of the agroecosystems.