Nitrogen fixation by Azospirillium brasilense in soil and the rhizosphere under controlled environmental conditions

Summary Acetylene reduction activity by Azospirillum brasilense, either free-living in soils or associated with wheat roots, was determined in a sterilised root environment at controlled levels of O₂ tension and with different concentrations of mineral N. In an unplanted, inoculated soil nitrogenase activity remained low, at approximately 40 nmol C₂H₄ h^-1 per 2kg fresh soil, increasing to 300 nmol C₂H₄ h^-1 when malic acid was added as a C source via a dialyse tubing system. The N₂ fixation by A. brasilense in the rhizosphere of an actively growing plant was much less sensitive to the repressing influence of free O₂ than the free-living bacteria were. An optimum nitrogenase activity was observed at 10 kPa O₂, with a relatively high level of activity remaining even at an O₂ concentration of 20 kPa. Both NO _inf3 ^sup- and NH _inf4 ^sup+ repressed nitrogenase activity, which was less pronounced in the presence than in the absence of plants. The highest survival rates of inoculated A. brasilense and the highest rates of acetylene reduction were found in plants treated with azospirilli immediately after seedling emergence. Plants inoculated at a later stage of growth showed a lower bacterial density in the rhizosphere and, as a consequence, a lower N₂-fixing potential. Subsequent inoculations with A. brasilense during plant development did not increase root colonisation and did not stimulate the associated acetylene reduction. By using the ¹⁵N dilution method, the affect of inoculation with A. brasilense in terms of plant N was calculated as 0.067 mg N₂ fixed per plant, i.e., 3.3% of the N in the root and 1.6% in the plant shoot were of atmospheric origin. This ¹⁵N dilution was comparable to that seen in plants inoculated with non-N₂-fixing Psudomonas fluorescens.     

Application of compost and clay under water-stressed conditions influences functional diversity of rhizosphere bacteria

Applications of compost and clay to ameliorate soil constraints such as water stress are potential management strategies for sandy agricultural soils. Water repellent sandy soils in rain-fed agricultural systems limit production and have negative environmental effects associated with leaching and soil erosion. The aim was to determine whether compost and clay amendments in a sandy agricultural soil influenced the rhizosphere microbiome of Trifolium subterraneum under differing water regimes. Soil was amended with compost (2% w/w), clay (5% w/w) and a combination of both, in a glasshouse experiment with well-watered and water-stressed (70 and 35% field capacity) treatments. Ion Torrent 16S rRNA sequencing and Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) analysis of functional gene prediction were used to characterise the rhizosphere bacterial community and its functional component involved in nitrogen (N) cycling and soil carbon (C) degradation. Compost soil treatments increased the relative abundance of copiotrophic bacteria, decreased labile C and increased the abundance of recalcitrant C degrading genes. Predicted N cycling genes increased with the addition of clay (N₂ fixation, nitrification, denitrification) and compost + clay (N₂ fixation, denitrification) and decreased with compost (for denitrification) amendment. Water stress did not alter the relative abundance of phylum level taxa in the presence of compost, although copiotrophic Actinobacteria increased in relative abundance with addition of clay and with compost + clay. A significant role of compost and clay under water stress in influencing the composition of rhizosphere bacteria and their implications for N cycling and C degradation was demonstrated.     

膜下滴灌对棉花根际土壤环境的影响研究

试验研究田间膜下滴灌棉花根际土壤的水、热、气变化表明,覆膜滴灌使土壤增温,膜下滴灌使根系浅层土壤含水量高于深层土壤含水量,加剧棉花根系浅层水平的分布使田间土壤干、湿间隔排列,有利于膜下土壤气体交换。棉田膜下滴灌技术节约灌溉水40％～50％,且使棉花增产20％左右。     

Effect of shale fragments on alfalfa growth under controlled conditions

Abstract

The effect of shale fragments as soil conditioner was investigated in two greenhouse experiments using alfalfa as the test plant. Four soils differing by the texture and the shale content were used, the < 2 mm material alone or in a mixture with coarser fragments, shale or gravel. In the first experiment, cuts 1–4 were taken under adequate watering before water stress was applied for cuts 5–8. Water stress was applied since the beginning in the second experiment.

Soil‐shale substrates generally produced the highest yields. Soil‐gravel substrates followed next in the first experiment but sustained generally the lowest yield in the second experiment.

The results were related to the effect of the coarse fragments on soil structure and water retention of the substrates.     

Effect of starter fertilizer in seed-row on emergence,biomass and nutrient uptake by six pulse crops grown under controlled environment conditions

Abstract

Legumes have a unique ability to obtain a significant portion of atmospheric nitrogen (N₂) through a symbiotic relationship with Rhizobia spp of bacteria but it takes time, thus, an early supply of N to the plant may positively influence growth and development. However, too much fertilizer in close proximity to the seed can damage the seedling. Therefore, this study was conducted to determine the maximum safe rates for starter seed-row fertilizer application under low seedbed utilization conditions (15%). Emergence, biomass yield and nitrogen (N), phosphorus (P) and sulfur (S) uptake responses to starter fertilizer products and blends applied at 0, 10, 20 and 30?kg?N?ha^?1 in the seed-row were investigated for six different pulse crops: soybean, pea, faba bean, black bean, lentil and chickpea. The general sensitivity (injury potential) for starter N, P, S fertilizer was lentil?≥?pea?≥?chickpea?>?soybean?≥?black bean?>?faba bean. Lentil, pea and chickpea could generally only tolerate the 10?kg?N?ha^?1 rates while soybean and black bean could tolerate 10–20?kg?N?ha^?1. Faba bean emergence appeared relatively unaffected by all three rates of N and showed least sensitivity to seed row placed fertilizer. In terms of 30-day biomass response, soybean and black bean were most responsive to fertilization, while pea, faba bean, lentil and chickpea were least responsive to the starter fertilizer applications, with no benefit increasing above the 10?kg?N?ha^?1 rate.     

Cadmium and zinc accumulation by the hyperaccumulator Thlaspi caerulescens from soils enriched with insoluble metal compounds

Abstract

The clay mineralogy of seven Dystrandepts developed on basalts in Northland (New Zealand), the French Massif Central and Western Oregon (U.S.A.) was determined by selective dissolution—differential infrared spectroscopy in combination with chemical, X-ray diffrac tion, electron microscopic and thermogravimetric analyses. Of 14 soil samples, 6 from Northland and Cantal (French Massif Central) contained allophane and imogolite, whereas the remaining 8 did not. Allophane-like constituents and/or “alumina” were found in all the samples, and opaline silica was present in three A₁ horizons. The contents of 2 : 1–2 : 1 : 1 layer silicates and their intergrades varied somewhat among the samples. Predominant volcanic glass shards in the Northland Dystrandepts and quartz in the Western Oregon Dystrandepts indicated that their parent materials were not restricted to basalt.     

Deleterious properties of certain rhizosphere bacteria on field pea (Pisum sativum) under gnotobiotic and non-sterile conditions

Three strains of Pseudomonas putida, one non-fluorescent and two fluorescent, were investigated in a series of complementary experiments to characterise their inhibitory effects on peas under different environmental conditions. Firstly, a gnotobiotic growth pouch system was developed to observe the deleterious effects of the strains on pea root development. A negative impact of the strains was observed on the development of the root morphology using these pouches which was accompanied with a subsequent reduction in root biomass. By using this method it was concluded that the deleterious effect of one of the strains was dependent on the inoculum density. Secondly, two complementary studies in non-sterile growth systems where pea seeds/seedlings were inoculated with the bacteria, showed deleterious effects on plant biomass by two of the strains. Thirdly, by using a sterile plant growth system allowing microscopic observations on root hair development, all strains were found to be able to induce root hair deformations on pea seedlings. The results showed that the mode of action for the deleterious effect differ between the strains. Based on our findings, we would like to emphasise the necessity to include a palette of different sterile and non-sterile growth systems to be able to identify characteristics of importance for deleterious rhizosphere bacteria.     

Effect of manganese-reducing rhizosphere bacteria on the growth of Gaeumannomyces graminis var. tritici and on manganese uptake by wheat (Triticum aestivum L.)

Summary Five bacterial strains capable of Mn reduction were isolated from the rhizosphere of plants growing in different South Australian soils. They differed in their Mn-reducing capacity. The antagonism of these strains compared to the imported strain 2–79 (from the United States) against Gaeumannomyces graminis var. tritici was tested in agar and in a soil sandwich experiment at different Mn²⁺ concentrations in the soil. In addition, wheat seeds were coated with the different strains and with MnSO₄ or with MnSO₄ only in order to investigate their effect on plant growth and Mn uptake. With one exception, all strains inhibited the growth of G. graminis in agar, but to different degrees. In contrast, only two strains significantly inhibited the growth of the fungus in the soil. The hyphal density was decreased more than the hyphal length. The Mn²⁺ concentration in the soil also had a marked effect on fungal growth; low Mn concentrations slightly increased while high Mn concentrations strongly decreased the fungal growth. Seed treatment with MnSO₄ only (+Mn) increased Mn uptake above that of the control (no seed treatment). Only the weakest Mn reducer on agar significantly increased plant growth and Mn uptake from soil in comparison with the Mn treatment. One strain was tested as seed coating without adding MnSO₄; it increased the plant growth to an extent similar to the Mn treatment. Increasing the Mn uptake by plants may be one of the growth-promoting effects exerted by rhizosphere bacteria.     

Brassica genotypes differ in growth, phosphorus uptake and rhizosphere properties under P-limiting conditions

Compared to other crops, Brassicas are generally considered to grow well in soils with low P availability, however, little is known about genotypic differences within Brassicas in this respect. To assess the role of rhizosphere properties in growth and P uptake by Brassicas, three Brassica genotypes (mustard, Brassica juncea cv Chinese greens and canola, Brassica napus cvs Drum and Outback) were grown in an acidic soil with low P availability at two treatments of added P: 25 and 100 mg P kg⁻¹ as FePO₄ (P25 and P100). The plants were harvested at the 6-leaf stage, at flowering and at maturity. Shoot and root dry weight (dry weight) and root length increased with time and were lower in P25 than in P100. In P25, shoot dry weight was lowest in Outback and highest in Chinese greens. In the P100 treatment, Chinese greens had a higher shoot dry weight than the two canola cultivars. Chinese greens had a lower root dry weight and root length at flowering and maturity than the canola genotypes in both P treatments. Irrespective of P treatment, shoot P concentration was lower in Chinese greens than in the two canola genotypes. Specific P uptake (μg P m⁻¹ root length) decreased with time. In P25, Chinese greens had the lowest specific P uptake at the 6-leaf stage but it was higher than in the two canola genotypes at flowering and maturity. In P100, Outback had the lowest specific P uptake. Available P in the rhizosphere (resin P) decreased over time with the greatest decrease from the 6-leaf stage to flowering. In P25, resin P in the rhizosphere was greatest in Chinese greens at the 6-leaf stage and flowering and smallest in Outback at flowering. Microbial P and acid phosphatase activity changed little over time, were not affected by P treatment and there were only small differences between the genotypes. The rhizosphere microbial community composition [assessed by fatty acid methyl ester (FAME) analysis] of Outback and Chinese greens differed from that of the other two genotypes at the 6-leaf stage and flowering, respectively. At maturity, all three genotypes had distinct microbial communities. Plant traits such as production of high biomass at low shoot P concentrations as well as the capacity to maintain high P availability in the rhizosphere by P mobilisation can explain the observed differences in plant growth and P uptake among the Brassica genotypes.     

Impact of controlled redox conditions on nickel in a serpentine soil

Purpose  

Serpentine soils exist in many regions around the world; they are naturally enriched with nickel (Ni). An adequate understanding of soil processes determining Ni solubility is a special need particularly since less research has been addressed to Ni behavior under dynamic and controlled redox conditions. Our aim was (1) to characterize the properties of a serpentine soil and (2) to determine the impact of predefined redox windows on the mobility and dynamics of Ni in a serpentine soil.     

Infectivity and acetylene reduction of diazotrophic rhizosphere bacteria in wheat (Triticum aestivum) seedlings under gnotobiotic conditions

Summary Wheat seedlings were inoculated with rhizosphere nitrogen-fixing bacteria and grown gnotobiotically for 15 days. The growth medium consisted of semisolid agar with or without plant nutrients. The bacteria, isolated from roots of field-grown wheat, were three unidentified Gram-negative rods (A1, A2, E1), one Enterobacter agglomerans (C1) and two Bacillus polymyxa (B1, B2). A strain of Azospirillum brasilense (USA 10) was included for comparison.Nitrogenase activity (acetylene reduction activity, ARA) was tested on intact plants after 8 and 15 days of growth. In semisolid agar without plant nutrients, five isolates showed ARA of 0.01–0.9 nmol C₂H₄ plant^–1 h^–1, while the two strains of B. polymyxa had higher ARA of 3.3–10.6 nmol C₂H₄ plant^–1 h^–1.Plant development was not affected by inoculation with bacteria, except that inoculation with B. polymyxa resulted in shorter shoots and lower root weight.Transmission electronmicroscopy of roots revealed different degrees of infection. A. brasilense, A1 and A2, occurred mainly in the mucilage on the root surface and between outer epidermal cells (low infectivity). B. polymyxa strains and E1 were found in and between epidermal cells (intermediate infectivity) while E. agglomerans invaded the cortex and was occasionally found within the stele (high infectivity).     

Chemical changes in the rhizosphere of metal hyperaccumulator and excluder Thlaspi species

Rhizosphere processes involved in hyperaccumulation and exclusion of metals are still largely unknown. Therefore, we conducted a rhizobag experiment on contaminated and non‐contaminated soils to investigate the chemical changes in the rhizosphere of the hyperaccumulators Thlaspi goesingense and T. caerulescens, and the metal‐excluder T. arvense. Root growth was restricted to the rhizobags separated by a nylon membrane (7 μm \x 25 μm mesh size) from surrounding bulk soil. Depletion of labile Zn in rhizosphere could not explain the amount of metals accumulated in T. caerulescens, whereas the difference in EDTA‐extractable Zn even exceeded total plant uptake. Substantially increased DOC in T. arvense rhizosphere indicates alleviation of phytotoxicity by formation of metal‐organic complexes. Hyperaccumulation and depletion of labile Zn in the rhizosphere was observed for T. goesingense grown on contaminated soil. On non‐contaminated soil, Zn was accumulated but labile Zn in rhizosphere was not changed. Nickel present in background concentrations in both soils was accumulated by T. goesingense only when grown on non‐contaminated soil. In contrast, labile Ni in the rhizosphere was increased in both soils, suggesting a general tendency of Ni mobilization by T. goesingense.     

Effect of plant growth-promoting bacteria and soil compaction on barley seedling growth, nutrient uptake, soil properties and rhizosphere microflora

Inoculants are of great importance in sustainable and/or organic agriculture. In the present study, plant growth of barley (Hordeum vulgare) has been studied in sterile soil inoculated with four plant growth-promoting bacteria and mineral fertilizers at three different soil bulk densities and in three harvests of plants. Three bacterial species were isolated from the rhizosphere of barley and wheat. These bacteria fixed N₂, dissolved P and significantly increased growth of barley seedlings. Available phosphate in soil was significantly increased by seed inoculation of Bacillus M-13 and Bacillus RC01. Total culturable bacteria, fungi and P-solubilizing bacteria count increased with time. Data suggest that seed inoculation of barley with Bacillus RC01, Bacillus RC02, Bacillus RC03 and Bacillus M-13 increased root weight by 16.7, 12.5, 8.9 and 12.5% as compared to the control (without bacteria inoculation and mineral fertilizers) and shoot weight by 34.7, 34.7, 28.6 and 32.7%, respectively. Bacterial inoculation gave increases of 20.3–25.7% over the control as compared with 18.9 and 35.1% total biomass weight increases by P and NP application. The concentration of N and P in soil was decreased by increasing soil compaction. In contrast to macronutrients, the concentration of Fe, Cu and Mn was lower in plants grown in the loosest soil. Soil compaction induced a limitation in root and shoot growth that was reflected by a decrease in the microbial population and activity. Our results show that bacterial population was stimulated by the decrease in soil bulk density. The results suggest that the N₂-fixing and P-solubilizing bacterial strains tested have a potential on plant growth activity of barley.     

Growth and nutrient uptake by cotton roots under field conditions

Abstract

The relationship between nutrient uptake and root growth of cotton (Gossypium hirsutum L.) was studied under field conditions. This basic information could be beneficial when making best management decisions concerning the time of application and placement of fertilizer. A field study was conducted in North Alabama on a fertile Dewey silt loam (clayey, kaolinitic, thermic Typic Paleudult). Aboveground whole plants were harvested at approximately 10‐day intervals beginning at 211 cumulative heat units (CHU) after planting (37 days after planting: 4‐true leaves). Root length of harvested plants was also measured by depth and distance from the plant. Maximum root length was obtained at 1174 CHU (117 days after planting), while dry matter continued to increase until a maximum was obtained at 1317 CHU (128 days after planting). Maximum root length density of 1.60 cm cm³ was obtained in the surface 0–15 cm layer in the in‐row position at 912 CHU (99 days after planting). After first bloom approximately 70% of the cotton root system was in the surface 30 cm of soil. Average daily influx of S per m of root length increased with plant age until 1317 CHU (near cut‐out), after which influx declined. Nitrogen (N), calcium (Ca), and iron (Fe) influx peaked very early in the season (291–469 CHU) followed by a general decrease with plant age. Maximum daily influx of potassium (K), phosphorus (P), magnesium (Mg), copper (Cu), manganese (Mn), and zinc (Zn) per meter of root occurred at approximately peak‐bloom (764–912 CHU, 87–99 days after planting) and decreased with plant age. Copper, Fe, Mn, and Zn influx rates were ～ 1000 times lower as compared to the other nutrients.     

Effect of extracellular polysaccharides of rhizosphere bacteria on the concentration of molybdenum in plants

Extracellular polysaccharides produced by bacteria have been shown to bind molybdenum (Mo) with the consequence that less Mo entered plants. The polysaccharides are produced by a variety of bacteria but those which contain uronic acids appear to be reponsible for binding the Mo. The amount of Mo bound is affected by pH.     

Acetylene reduction by bacteria isolated from the rhizosphere of wheat

An asymbiotic bacillus possessing N₂-ase activity and capable of growth on low-N media was consistently isolated from the rhizosphere of a chromosome substitution line of spring wheat in which a pair of chromosomes 5D from the cultivar Rescue had been substituted for those of ‘Cadet’. None of the bacilli were isolated from either of the two parent cultivars, Rescue and Cadet, or from the corresponding substitution line involving a homoeologous chromosome 5B. Maximum C₂H₂ reduction to C₂H₄ by pure culture isolates was found to occur at a partial pressure of 0.2 atm C₂H₂. C₂H₂ reduction was diminished in the presence of O₂ and nearly ceased at a partial pressure of 0.08 atm O₂.The data presented suggest that altering the genetics of the wheat plant can change the root environment to favor the establishment of bacilli that exhibit N₂-ase activity in pure culture.     

Effect of ectomycorrhizae on the growth and uptake and transport of 15N-labeled compounds by Pinus tabulaeformis seedlings under water-stressed conditions

 Seedlings of Chinese pine (Pinus tabulaeformis Carr.) were grown in a growth chamber using a sterilized soil/sand/vermiculite mixture, and were inoculated with the ectomycorrhizal fungus Cenococcum graniforme (Sow.) Ferd. and Winge. The function of external mycelium of ectomycorrhiza for uptake and transport of N from ¹⁵N-labeled NH₄ ⁺ and NO₃ ^– for plant nutrition was evaluated under well-watered conditions and water stress. The pots comprised two-compartments, whereby the penetration of roots was prevented by a nylon mesh bag which the mycorrhizal hyphae were allowed to pass in order to colonize the rest of the pot. ¹⁵N-labeled NH₄ ⁺ or NO₃ ^– was applied to the area of the pot to which the root had no access. At harvest, the ¹⁵NH₄ ⁺ concentration in plant tissues was significantly promoted by the formation of mycorrhizae both under well-watered conditions and water stress. The ¹⁵NO₃ ^– concentration was reduced by water stress and increased by mycorrhizal formation. The enhancement of ¹⁵NO₃ ^– uptake caused by mycorrhizal formation was more evident under water stress than under well-watered conditions. The external mycelia of the ectomycorrhizae took up and transported NH₄ ⁺ and NO₃ ^– from the soil to the plant, thereby improving plant nutrition and growth, in addition to helping the plants to avoid the effects of water stress. Received: 31 October 1997     

Diversity of rhizosphere bacteria associated with different soybean cultivars in two soil conditions

Aiming at learning the effects of soil conditions and cultivar on the bacterial diversity in the rhizosphere of soybean (Glycine max(L.) Merr.), bacterial communities associated with four soybean cultivars grown in two soils were revealed by terminal-restriction fragment length polymorphism (T-RFLP) combined with sequencing analysis of a 16S rDNA clone library. Lower bacteria diversity was found in soil A which has higher salinity and nutrient contents, while the highest bacterial diversity was found in the rhizosphere of cv. Jidou 12 in both soils. These results revealed that both the soil conditions and soybean cultivar affected the community composition of rhizosphere bacteria, but the effect of soil conditions was greater than that of soybean cultivar as demonstrated by the principal component analysis. It also revealed that the abundant rhizosphere bacteria may also the main symbiotic or non-symbiotic nodule endophytes.     

Effects of heavy metals on the growth and mineral composition of a nickel hyperaccumulator

Alyssum pintodasilvae Dudley is a nickel (Ni) hyperaccumulator endemic to serpentine soils of north‐east Portugal. In one experiment, the effects of cadmium (Cd), chromium (Cr), copper (Cu), Ni, lead (Pb), and zinc (Zn) on the growth and mineral composition of this species were evaluated. The growth of A. pintodasilvae, measured by dry matter accumulation, was not influenced by the presence of Cr, Cu, Pb, Ni, or Zn in the soil, but Cd applications led to significant decreases in dry matter yield. The addition of heavy metals to the soil resulted in increased uptake and translocation by A. pintodasilvae but only Ni was accumulated to high levels. In a second experiment, two cuts of A. pintodasilvae, grown on a Ni‐enriched soil, were compared. Nickel concentrations were higher in the second cut, suggesting the possibility of continued growth and harvest of this plant to detoxify Ni‐contaminated soils.