Induction of Paenibacillus lentimorbus biofilm by sodium alginate and CaCl2 alleviates drought stress in chickpea

Drought is a major environmental factor that limits chickpea production. An improvement in the adaption of crop to the fluctuating environmental conditions is therefore a major aim in chickpea breeding. However, the complexity of the trait has allowed only marginal progress. Our findings provide a solution to the current situation in the form of improved plant‐growth‐promoting effects caused by the biofilm formation of Paenibacillus lentimorbus B‐30488 (B‐30488) under water‐limiting conditions. In vitro assays demonstrating the biofilm‐forming ability of B‐30488 and the factors enhancing it were studied. Greenhouse experiments were conducted for validating the in vitro results and assessing the effect of seed coating supplements in alleviating drought stress effects in chickpea seedlings. The chickpea seed bacterisation with B‐30488 along with sodium alginate (1%) and CaCl₂ (1 mM) caused an increase in germination percent and increased colony‐forming units (CFU) of B‐30488 in rhizosphere, resulting in amelioration of drought stress by positively influencing the dehydration‐induced physiological responses. The whole study reflects a prospective role of sodium alginate and CaCl₂ in influencing the biofilm formation of B‐30488, and depicts the assistance of seed coating supplements in stress adaptation and protection of plants by alleviation of drought stress effects in chickpea without causing any major changes in the functional diversity of soil micro‐organisms.     

Tripartite interactions among <Emphasis Type="Italic">Paenibacillus lentimorbus</Emphasis> NRRL B-30488, <Emphasis Type="Italic">Piriformospora indica</Emphasis> DSM 11827, and <Emphasis Type="Italic">Cicer arietinum</Emphasis> L.

Tripartite interactions among Paenibacillus lentimorbus NRRL B-30488 (B-30488), Piriformospora indica DSM 11827 (DSM 11827) and their consortia (B-30488:DSM 11827:: 1:1) with native rhizobial population in the rhizosphere of Cicer arietinum L. (Chick pea) was tested for enhancing nodulations and plant growth promotion. Number of nodules and dry weight per plant significantly enhanced (P = 0.05), which is further evident by N, P, and K uptake by plants and were found to be maximum in B-30488 treated followed by B-30488: DSM 11827 and DSM 11827, as compared with uninoculated control, in 60 days grown chickpea plants. Microbial community structure in the rhizosphere of the four treatments was assessed, using Biolog Eco and MT plates. Principal component analysis (PCA) of carbon source utilization pattern on Biolog Eco plates did not show any clustering among the four samples indicating that in case of individually DSM 11827 and B-30488 treated chickpea rhizosphere there was significant change in microbial community structure, compared with lesser changes in un-inoculated and B-30488 and DSM 11827 consortium treated chickpea rhizosphere microflora. Additional carbon sources tested using Biolog MT plates, higher activity of lignin, chitin, and cellulose utilizing microbial communities in the rhizosphere being stimulated by root exudates treated with B-30488 alone or in consortia with DSM 11827, and, in turn, should encourage beneficial symbiotic or mutualistic microorganisms that can act as plant growth promoting and biocontrol agents.     

Biologic Control Ability of Plant Growth–Promoting Paenibacillus lentimorbus NRRL B-30488 Isolated from Milk

A plant growth–promoting Paenibacillus lentimorbus NRRL B-30488 (B-30488) was isolated from cows’ milk. Bacterial colonization and growth responses of different plant species after inoculation with B-30488 were evaluated in a controlled environment and in microplot assays. Survival and colonization of B-30488 in the phytosphere of plants and soil was monitored using a chromosomally located rifampicin-marked mutant B-30488 (B-30488R). The strain showed variable ability to invade plants. The interaction between B-30488R and Fusarium oxysporum f. sp. ciceri was studied by scanning electron microscopy. Chitinase and β-1,3-glucanase enzymes were produced when B-30488R was grown in the presence of colloidal chitin as sole carbon source. Deliberate dilution of B-30488R with field soil offers a reliable process for decreasing the cost of bacterial inoculants in developing countries. Seed treatment of chickpea demonstrated significantly (P = 0.05) greater seedling mortality in nonbacterized compared with bacterized seedlings. Bacterization significantly (P = 0.05) improved seed germination, plant height, number of pods/plant^–1, and seed dry weight.     

Plant growth promotion by a hexavalent chromium reducing bacterial strain, <Emphasis Type="Italic">Cellulosimicrobium</Emphasis><Emphasis Type="Italic">cellulans</Emphasis> KUCr3

This article reports on the isolation and characterization of a Cr(VI) resistant bacterial strain, having plant growth promoting properties to improve general growth of plant in chromium-contaminated soil through rhizosphere colonization. The strain was isolated from the sludge of waste canal carrying industrial effluents. The minimum inhibitory concentration of chromium to this strain was found to be 450 and 400 mM in complex and minimal media, respectively. The strain also showed varied degree of resistance to Cd, Co, As, Ni and Zn. It exhibited potential Cr(VI) reducing ability under aerobic culture conditions, and the factors affecting Cr(VI) reduction by this strain were evaluated. The optimum pH and temperature required to achieve maximum Cr(VI) reduction values were 7 and 35°C, respectively. Higher concentration of Cr(VI) slowed down the reduction, but with longer incubation time it reduced nearly all detectable amount of Cr(VI). The strain showed positive response to IAA production and phosphate solubilization. It promoted the growth of chilli plants in waste-fed soil with or without additional Cr through its establishment in rhizosphere. The successful establishment of KUCr3 in the rhizosphere of chilli plants helped to reduce Cr uptake by the test plant. This strain shows a promise that the multifarious role of this strain would be useful in the Cr-contaminated rhizosphere soil as a good bioremediation and plant growth promoting agent as well. Through biochemical characterization and 16S rDNA sequence analysis, the strain KUCr3, as the name given to it, was identified as a strain of Cellulosimicrobium cellulans.     

Paenibacillus lentimorbus Inoculation Enhances Tobacco Growth and Extenuates the Virulence of Cucumber mosaic virus

Previous studies with Paenibacillus lentimorbus B-30488” (hereafter referred as B-30488), a plant growth promoting rhizobacteria (PGPR) isolated from cow’s milk, revealed its capabilities to improve plant quality under normal and stress conditions. Present study investigates its potential as a biocontrol agent against an economically important virus, Cucumber mosaic virus (CMV), in Nicotiana tabacum cv. White Burley plants and delineates the physical, biophysical, biochemical and molecular perturbations due to the trilateral interactions of PGPR-host-CMV. Soil inoculation of B-30488 enhanced the plant vigor while significantly decreased the virulence and virus RNA accumulation by ~12 fold (91%) in systemic leaves of CMV infected tobacco plants as compared to the control ones. Histology of these leaves revealed the improved tissue’s health and least aging signs in B-30488 inoculated tobacco plants, with or without CMV infection, and showed lesser intercellular spaces between collenchyma cells, reduced amount of xyloglucans and pectins in connecting primary cells, and higher polyphenol accumulation in hypodermis layer extending to collenchyma cells. B-30488 inoculation has favorably maneuvered the essential biophysical (ion leakage and photosynthetic efficiency) and biochemical (sugar, proline, chlorophyll, malondialdehyde, acid phosphatase and alkaline phosphatase) attributes of tobacco plants to positively regulate and release the virus stress. Moreover, activities of defense related enzymes (ascorbate peroxidase, guaiacol peroxidase, superoxide dismutase and catalase) induced due to CMV-infection were ameliorated with inoculation of B-30488, suggesting systemic induced resistance mediated protection against CMV in tobacco. The quantitative RT-PCR analyses of the genes related to normal plant development, stress and pathogenesis also corroborate well with the biochemical data and revealed the regulation (either up or down) of these genes in favor of plant to combat the CMV mediated stress. These improvements led tobacco plant to produce more flowers and seeds with no negative impact on plant health. The present study may advocate the applicability of B-30488 for crop yield improvement in virus infested areas.     

Paenibacillus lentimorbus Enhanced Abiotic Stress Tolerance Through Lateral Root Formation and Phytohormone Regulation

Under the stressed conditions plant growth-promoting rhizobacteria (PGPR) are able to stimulate plant growth through several mechanisms, including antioxidants alleviation, regulation of stress responsive genes and phytohormones etc. Present study is conducted to investigate the impact of Paenibacillus lentimorbus B-30488 inoculation on salinity and drought stress mitigation in Arabidopsis thaliana through modulation in defense enzymes, phyto-hormones and root system architecture associated gene expression profiling. In vitro experiments clearly demonstrated the role of B-30488 in stimulating the root length, branches, lateral root formation and biomass under salinity and drought stress. The inoculation of B-30488 modulated the phytohormones levels to protect the plants from salinity and drought stress. Similarly, defence enzymes were also activated under the stressed conditions, but B-30488 inoculation reduced the antioxidants content during salinity and drought stress as compared to their respective controls. Microscopy results showed decrease in lateral roots hair formation under both stresses and B-30488 inoculation not only mitigate but also enhanced the lateral root formation. Gene expression analysis through real time polymerase chain reaction (RT-PCR) showed modulated expression of several genes related to root development, stress and lateral root formation in B-30488 inoculated seedlings. Results based on the present study, B-30488 is also involved in alteration root architecture, its growth regulation via modulation in phytohormones and genes expression and overall significant improvement in plant growth under stress conditions.

     

Hexavalent chromium-reducing plant growth-promoting rhizobacteria are utilized to bio-fortify trivalent chromium in fenugreek by promoting plant development and decreasing the toxicity of hexavalent chromium in the soil

BackgroundFenugreek is known to have good anti-diabetes properties. Moreover, several studies accounted that the trivalent form of chromium [Cr(III)] also have anti-diabetic properties. However, its hexavalent form i.e., Cr(VI) is known to be highly toxic and carcinogenic to living beings and retarded plant growth even if it is present in low concentration in soil. Many plant growth-promoting rhizobacteria (PGPR) are reported to have the potential to reduce the Cr(VI) into Cr(III) in soil. In view of the above, the present objective was designed to effectively utilize Cr(VI) reducing PGPRs for the growth and development of fenugreek plant in Cr(VI) amended soil, apart from reducing Cr(VI) in soil and fortification of Cr(III) in the aerial part of plants.MethodsThe experiment was carried out to evaluate the effect of Cr(VI)-reducing PGPRs viz. Bacillus cereus (SUCR44); Microbacterium sp. (SUCR140); Bacillus thuringiensis (SUCR186) and B. subtilis (SUCR188) on growth, uptake and translocation of Cr as well as other physiological parameters in fenugreek grown under artificially Cr(VI) amended soil (100 mg kg⁻¹ of Cr(VI) in soil).ResultsThe aforementioned concentration of Cr(VI) in soil cause severe reduction in root length (41 %), plant height (43 %), dry root (38 %) and herb biomass (48 %), when compared with control negative (CN; uninoculated plant not grown in Cr(VI) contaminated soil). However, the presence of Microbacterium sp.˗SURC140 (MB) mitigates the Cr toxicity resulting in improved root length (92 %), plant height (86 %), dry root (74 %) and herb biomass (99 %) as compared with control positive (CP; uninoculated plants grown in Cr(VI) contaminated soil). The maximum reduction in bioavailability (82 %) of Cr(VI) in soil and its uptake (50 %) by the plant were also observed in MB-treated plants. However, All Cr(VI)-reducing PGPRs failed to decrease the translocation of Cr to the aerial parts. Moreover, the plant treated with MB observed diminution in relative water content (13 %), electrolyte leakage (16%) and lipid peroxidation (38 %) as well as higher chlorophyll (37 %) carotenoids (17 %) contents and antioxidants (18%) potential.ConclusionThis study demonstrates that MB can lower the Cr(VI) toxicity to the plant by reducing the bioavailable Cr(VI), consequently reducing the Cr(VI) toxicity level in soil and helping in improving the growth and yield of fenugreek. Additionally, Cr(III) uptakes and translocation may improve the effectiveness of fenugreek in treating diabetes.     

The use of different soil nitrogen sources by young Norway spruce plants

 Three-year-old Norway spruce trees were planted into a low-nitrogen mineral forest soil and supplied either with two different levels of mineral nitrogen (NH₄NO₃) or with a slow-release form of organic nitrogen (keratin). Supply of mineral nitrogen increased the concentrations of ammonium and nitrate in the soil solution and in CaCl₂-extracts of the rhizosphere and bulk soil. In the soil solution, in all treatments nitrate concentrations were higher than ammonium concentrations, while in the soil extracts ammonium concentrations were often higher than nitrate concentrations. After 7 months of growth, ¹⁵N labelled ammonium or nitrate was added to the soil. Plants were harvested 2 weeks later. Keratin supply to the soil did not affect growth and nitrogen accumulation of the trees. In contrast, supply of mineral nitrogen increased shoot growth and increased the ratio of above-ground to below-ground growth. The proportion of needle biomass to total above-ground biomass was not increased by mineral N supply. The atom-% ¹⁵N was higher in younger needles than in older needles, and in younger needles higher in plants supplied with ¹⁵N-nitrate than in plants supplied with ¹⁵N-ammonium. The present data show that young Norway spruce plants take up nitrate even under conditions of high plant internal N levels. Received: 1 April 1998 / Accepted: 9 October 1998     

Mycorrhizal fungi enhance accumulation and tolerance of chromium in sunflower (Helianthus annuus)

Chromium (Cr) is a heavy metal risk to human health, and a contaminant found in agricultural soils and industrial sites. Phytoremediation, which relies on phytoextraction of Cr with biological organisms, is an important alternative to costly physical and chemical methods of treating contaminated sites. The ability of the arbuscular mycorrhizal fungus (AM),Glomus intraradices, to enhance Cr uptake and plant tolerance was tested on the growth and gas exchange of sunflower (Helianthus annuus L.). Mycorrhizal-colonized (AM) and non-inoculated (Non-AM) sunflower plants were subjected to two Cr species [trivalent cation (Cr³⁺) Cr(III) , and divalent dichromate anion (Cr₂O₇⁻) Cr(VI) ]. Both Cr species depressed plant growth, decreased net photosynthesis (A) and increased the vapor pressure difference; however, Cr(VI) was more toxic. Chromium accumulation was greatest in roots, intermediate in stems and leaves, and lowest in flowers. Greater Cr accumulation occurred with Cr(VI) than Cr(III). AM enhanced the ability of sunflower plants to tolerate and hyperaccumulate Cr. At higher Cr levels greater mycorrhizal dependency occurred, as indicated by proportionally greater growth, higherA and reduced visual symptoms of stress, compared to Non-AM plants. AM plants had greater Cr-accumulating ability than Non-AM plants at the highest concentrations of Cr(III) and Cr(VI), as indicated by the greater Cr phytoextraction coefficient. Mycorrhizal colonization (arbuscule, vesicle, and hyphae formation) was more adversely affected by Cr(VI) than Cr(III), however high levels of colonization still occurred at even the most toxic levels. Arbuscules, which play an important role in mineral ion exchange in root cortical cells, had the greatest sensitivity to Cr toxicity. Higher levels of both Cr species reduced leaf tissue phosphorus (P). While tissue P was higher in AM plants at the highest Cr(III) level, tissue P did not account for mycorrhizal benefits observed with Cr(VI) plants.     

Effects of litter incorporation and nitrogen fertilization on the contents of extractable aluminium in the rhizosphere soil of tea plant (Camallia sinensis (L.) O. Kuntze)

The effects of litter incorporation and nitrogen application on the properties of rhizosphere and bulk soils of tea plants (Camellia sinensis (L.) O. Kuntze) were examined in a pot experiment. Total of 8 treatments included four levels of tea litter additions at 0, 4.9, 9.8, and 24.5 g kg^–1 in combination with two N levels (154.6 mg kg^–1 and without). After 18 months of growth the rhizosphere soil was collected by removing the soil adhering to plant roots and other soil was referred to as bulk soil. The dry matter productions of tea plants were significantly increased by N fertilization and litter incorporation. The effect of litter was time-depending and significantly decreased the content of exchangeable Al (Al_ex, by 1 mol L^–1 KCl) and Al saturation at 9 months after litter incorporation whereas soil pH was not affected, although the litter contained high Al content. After 18 months, the contents of extractable Al by dilute CaCl₂, CuCl₂ + KCl, NH₄OAC, ammonium oxalate and sodium citrate (Al_CaCl2, Al_Cu/KCl, Al_NH4OAC, Al_Oxal, and Al_Cit respectively) and Al_ex, were not affected by litter application, except that of Al_CaCl2 in the rhizosphere soil which was decreased following litter additions. Nitrogen fertilization with NH₄ ⁺ (urea and (NH₄)₂SO₄) significantly reduced soil pH, the contents of exchangeable Ca, K, Mg and base saturation while raised extractable Al levels (Al_CaCl2, Al_Cu/KCl, Al_NH4OAC, and Al_ex). In the rhizosphere soils exchangeable K accumulated in all treatments while exchangeable Ca and Mg depleted in treatments without litter application. The depletions of Ca and Mg were no longer observed following litter incorporation. This change of distribution gradients in rhizosphere was possibly due to the increase of nutrient supplies from litter decomposition and/or preferable root growth in soil microsites rich in organic matter. Lower pH and higher extractable Al (Al_CaCl2, Al_ex, and Al_NH4OAC) in the rhizosphere soils, regardless of N and litter treatments, were distinct and consistent in all treatments. Such enrichments of extractable Al in the rhizosphere soil might be of importance for tea plants capable of taking up large amounts of Al.     

Immobilization and mobilization of labelled sulphur in relation to soil arylsulphatase activity in rhizosphere soil of field-grown rape,barley and fallow

During plant growth, rhizosphere soils from fallow, barley (Hordeum vulgare L. cv Esterel) and rape (Brassica napus L. cv Capitole) grown in a calcareous soil were sampled 5 times (every fortnight) from May to July 2001 at plant maturity. In order to estimate the impact of C derived from photosynthesis, the aerial parts of rape and barley in an area of 1 m² were cut off about 2 cm from the soil surface, and left a fortnight before each sampling. Both soil arylsulphatase activity and a 1-week immobilization of S fertilizer in the sampled soils were then measured. The immobilization of S fertilizer was higher in fallow, followed by barley and rape rhizosphere soil. A strong positive linear correlation (r ²=0.71, P<0.001) was found between soil arylsulphatase activity and S fertilizer immobilized. Conversely, the mobilization of endogenous organic ³⁵S (obtained after leaching free and adsorbed SO₄ ^2–-³⁵S by 0.009 M Ca(H₂PO₄)₂) in the rhizosphere soil of each plant cover pooled at the end of the 5 samplings and materialized by ryegrass (Lolium perenne L. cv Massa) ³⁵S uptake, was about 3 and 2 times higher, respectively, in rape and barley than in fallow rhizosphere soil. Accordingly, strong inverse polynomial relationships were observed between soil arylsulphatase activity and ³⁵S uptake by the whole plant (r ²=0.904, P<0.02) and roots (r ²=0.970, P<0.01) of ryegrass. Plant cuttings affected both the immobilization and mobilization of S. It is concluded that the turnover of S freshly immobilized in rape rhizosphere soil was relatively high. Therefore, rape as a preceding crop in the rotations may have a beneficial effect by increasing S availability on the succeeding crop.     

Plant induced alteration in the rhizosphere and the utilisation of soil heterogeneity

Plant-soil interactions result in a special rhizosphere soil chemistry, differing from that of the bulk soil found only a few mm from the root. The aim of this study was to investigate adaptation mechanisms of herbs growing in acid soils through studying their rhizosphere chemistry in a greenhouse experiment and in a field study. Ten herbs were grown in acid soil (pH 4.2 in the soil solution) in the greenhouse. The concentrations of NO₃ ^-, SO₄ ^2-, phosphates, Ca²⁺, Mg²⁺, Mn²⁺, K⁺, Na⁺, NH₄ ⁺ and pH were analysed in soil solutions obtained by centrifugation. The general pattern found was a depletion of nutrients in the rhizosphere compared with their concentrations in the bulk soil. The pH increase (up to 0.7 units) in the rhizosphere soil appeared to be caused by plant uptake of NO₃ ^- (r²=0.88). The ion concentrations in the soil solution of the rhizosphere were dependent on plant species and biomass increase. Although species with a larger biomass and higher growth rates showed a higher degree of ion depletion (except for Na⁺, SO₄ ^2-) in the rhizosphere, there were also species specific responses. A field study of five herbs at five oak forest sites in Southern Sweden (Scania) was also carried out. In addition to the soil solution concentrations, the loss on ignition (LOI) and the concentrations of 0.1 M BaCl₂ extractable K⁺, Mg²⁺, Mn²⁺, Ca²⁺, and Al ions were measured. The amount of soil solution Al was determined as free ionic (quickly reacting) Al. For all species and sites, the LOI and the concentrations of exchangeable cations were higher in the rhizosphere than in the bulk soil, apparently due to the roots preferably growing at organic-rich microsites. The concentrations of the ions as measured in the centrifuged soil solution, were either higher in the rhizosphere than in the bulk soil or were the same in both, except for NO₃ ^- and quickly reacting Al. The lower concentrations of quickly reacting Al in the rhizosphere, compared with the bulk soil could indicate the uptake of Al by the plant or the exudation of complexing substances. The pH differences were only small and mostly non-significant. Plant-soil interactions and the ability of plants to utilise heterogeneity of the soil appear to be more important for plant growth in acid soils than recognised heretofore. Rhizosphere studies provide an important means of understanding plant strategies in acid soils. This revised version was published online in June 2006 with corrections to the Cover Date.     

A Method for Detection of Pseudobactin, the Siderophore Produced by a Plant-Growth-Promoting Pseudomonas Strain, in the Barley Rhizosphere

Detection in the rhizosphere of the siderophore produced by an inoculated microorganism is critical to determining the role of microbial iron chelators on plant growth promotion. We previously reported the development of monoclonal antibodies (MAb) to ferric pseudobactin, the siderophore of plant-growth-promoting Pseudomonas strain B10. One of these MAb reacted less strongly to pseudobactin than to ferric pseudobactin. The MAb reacted to Al(III), Cr(III), Cu(II), and Mn(II) complexes of pseudobactin at a level similar to the level at which it reacted to ferric pseudobactin and reacted less to the Zn(II) complex, but these metals would make up only a small fraction of chelated pseudobactin in soil on the basis of relative abundance of metals and relative binding constants. Fourteen-day-old barley plants grown in limed and autoclaved soil were inoculated with 10⁹ CFU of Pseudomonas strain Sm1-3, a strain of Pseudomonas B10 Rif^r Nal^r selected for enhanced colonization, and sampled 3 days later. Extraction and analysis of the roots and surrounding soil using the MAb in an immunoassay indicated a concentration of 3.5 × 10^-10 mol of ferric pseudobacting g^-1 (wet weight). This is the first direct measurement of a pseudobactin siderophore in soil or rhizosphere samples.     

Using the Selective Ion Exchange Resin Extraction and XANES Methods to Evaluate the Effect of Compost Amendments on Soil Chromium(VI) Phytotoxicity

The Cu-saturated selective ion exchange resin (DOWEX M4195) extraction method was used to investigate the effects of two amendments, 5 and 15% organic matter in the form of hog-dung compost (HC) or cattle-dung compost (CC), on Cr(VI) bioavailability in three soils spiked with various levels of Cr(VI). The results showed that addition of composts could decrease the amounts of resin-extractable Cr(VI) in Cr(VI)-spiked soils, and the CC amendment decreased resin-extractable Cr(VI) more than the HC amendment. The X-ray Absorption Near-edge Structure spectroscopy (XANES) method was used to examine the distribution of Cr(III) and Cr(VI) species in Cr(VI)-spiked soils that were affected by compost amendments, and to elucidate the mechanisms for the decrease of resin-extractable Cr(VI) due to the application of composts. The XANES results suggested that the decrease in the amounts of resin-extractable Cr(VI) after compost addition was mainly due to the reduction of Cr(VI) to Cr(III). The amounts of soil resin-extractable Cr(VI) were also correlated with wheat seedling growth in order to evaluate the effect of compost amendments on decreasing the phytotoxicity of soil Cr(VI). The results showed that there was a sigmoidal relationship between soil resin-extractable Cr(VI) and the plant height of wheat seedlings and the obtained effective concentrations of resin-extractable Cr(VI) resulting in 10 and 50% growth inhibition (EC₁₀ and EC₅₀) were 76 and 191 mg kg⁻¹ respectively. The above results suggested that the resin extraction method was a useful tool for assessing Cr(VI) phytotoxicity and that addition of composts would enhance Cr(VI) reduction to Cr(III) in soils and thus relieve Cr(VI) phytotoxicity.     

Depletion of Selenium in Soil Solution due to its Enhanced Sorption in the Rhizosphere of Soybean

The effect of plant roots on selenium (Se) mobility in soil was studied by a large-scale pot experiment in order to understand the environmental behavior of Se in agricultural soils under plant growth conditions. Soybean plants (Glycine max (L.) Merrill) were grown in a greenhouse for 84 d. The concentrations of Se and major elements (K, Ca, Mg, Na, and Al) in the soil solutions and in the plants were measured at different growth periods. Concentrations of Se and major cations in soil solution decreased as the soybean plants grew, while the concentrations of Al increased. It was assumed that the soybean roots released H⁺ with the uptake of cations; consequently, due to the acidification of the rhizosphere, Al³⁺ was released starting from the soil solid phase. The decreased Se concentration in the soil solution should be due to the enhancement of Se sorption onto the soil solid phase. The increase of Se sorption level in the rhizosphere was examined in a small-scale pot experiment. The soil–soil solution distribution coefficient of Se (K _d-Se) was observed as an index of Se sorption level. K _d-Se clearly increased in the rhizosphere soil after cultivation. The effects of pH and Al³⁺ in the rhizosphere on Se sorption were assessed by K _d-Se measurements at different levels of HCl and AlCl₃. In this third experiment, a decrease in pH increased K _d-Se values, but no specific effect was observed on Se sorption due to increased Al³⁺. These results show that the Se mobility in agricultural soil could be decreased by plant roots under plant growth conditions due to enhanced Se sorption in the rhizosphere.     

Copper uptake and phytotoxicity as assessed in situ for durum wheat (Triticum turgidum durum L.) cultivated in Cu-contaminated, former vineyard soils

This work assessed in situ, copper (Cu) uptake and phytotoxicity for durum wheat (Triticum turgidum durum L.) cropped in a range of Cu-contaminated, former vineyard soils (pH 4.2–7.8 and total Cu concentration 32–1,030 mg Cu kg⁻¹) and identified the underlying soil chemical properties and related root-induced chemical changes in the rhizosphere. Copper concentrations in plants were significantly and positively correlated to soil Cu concentration (total and EDTA). In addition, Cu concentration in roots which was positively correlated to soil pH tended to be larger in calcareous soils than in non-calcareous soils. Symptoms of Cu phytotoxicity (interveinal chlorosis) were observed in some calcareous soils. Iron (Fe)–Cu antagonism was found in calcareous soils. Rhizosphere alkalisation in the most acidic soils was related to decreased CaCl₂-extractable Cu. Conversely, water-extractable Cu increased in the rhizosphere of both non-calcareous and calcareous soils. This work suggests that plant Cu uptake and risks of Cu phytotoxicity in situ might be greater in calcareous soils due to interaction with Fe nutrition. Larger water extractability of Cu in the rhizosphere might relate to greater Cu uptake in plants exhibiting Cu phytotoxic symptoms.     

Rhizoreduction of arsenate and chromate in Australian native grass, shrub and tree vegetation

Background and aims

Rhizosphere influences the dynamics of nutrients and contaminants through increased microbial activity, release of root exudates and alteration of pH. The objective of this study was to evaluate the rhizosphere-induced reduction (i.e. rhizoreduction) and redistribution of arsenate [As(V)] and chromate [Cr(VI)] in Australian native vegetation in relation to their bioavailability.

Methods

The reduction of As(V) and Cr(VI) was examined using rhizosphere soils from a number of Australian native vegetation (Acacia pubescens, Eucalyptus camaldulensis, Enchylaena tomentosa, Templetonia retusa, Dichantheum sericeum and Austrodanthonia richardsonii). Naturally contaminated As and Cr soils were used to examine the effect of Dichantheum sericeum on the redistribution and bioavailability of these metal(loid)s.

Results

The rhizosphere soil contained higher levels of microbial activity, dissolved organic carbon and organic acid content than the non-rhizosphere soil. The rhizosphere soil caused up to 2.4 and 5.1 fold increases in the rate of reduction of As(V) and Cr(VI), respectively. There was a significant relationship between rhizosphere-induced increases in microbial activity (Δ basal respiration) and As(V) and Cr(VI) reduction (Δ rate of reduction), indicating the role of increased microbial activity in rhizosphere soil on metal(loid) reduction. In the plant growth experiment, Dichantheum sericeum enhanced the reduction of metal(loid)s in the naturally contaminated soils, thereby increasing the bioavailability of As but decreasing that of Cr.

Conclusions

Depending on the nature of metal(loid)s present in soil, the rhizosphere-induced reduction by plant species such as Dichantheum sericeum and Templetonia retusa has implications to both their bioavailability to higher plants and microorganisms, and remediation of contaminated soils. While rhizoreduction decreases Cr bioavailability it increases that of As.     

Does Seed Priming Induce Changes in the Levels of Some Endogenous Plant Hormones in Hexaploid Wheat Plants Under Salt Stress？

In order to assess whether salt tolerance could be Improved In spring wheat （Triticum aestivum L.）, the present study was performed by soaking the seeds of two cultlvars, namely MH-97 （salt sensitive） and Inqlab-91 （salt tolerant）, for 12 h In distilled water or 100 mol/m^3 CaCl2, KCI, or NaCI. Primed seeds from each treatment group and non-primed seeds were sown In a field In which NaCI salinity of 15 dS/m was developed. Priming of seeds with CaCl2, followed by priming with KCI and NaCI, was found to be effective In alleviating the adverse effects of salt stress on both wheat cultivars In terms of shoot fresh and dry weights and grain yield. Priming with CaCl2 alleviated the adverse effects of salt stress on hormonal balance In plants of both cultlvars. In MH-97 plants, CaCl2 pretreatment considerably reduced leaf absclslc acid （ABA） concentrations and Increased leaf free salicylic acid （SA） concentrations under both saline and non-saline conditions. In contrast, In the Inqlab-91 plant, CaCl2 Increased free Indoleacetic acid （IAA） and indolebutyrlc acid （IBA） content. However, priming of seeds with CaCl2 did not alter free polyamlne levels in either cultlvar, although spermldlne levels were considerably lower In plants raised from seeds treated with CaCl2 for both cultlvars under saline conditions. Priming with KCI Increased growth In Inqlab-91 plants, but not In MH-97 plants, under saline conditions. The salinity Induced reducUon In auxins （IAA and IBA） was alleviated by NaCI priming In both cultlvars under saline conditions. However, NaCI Increased leaf free ABA content and lowered leaf SA and putresclne levels In Inqlab-91 plants under saline conditions. In conclusion, although all three priming agents （I.e. CaCl2, KCI, and NaCI） were effective In alleviating the adverse effects of salt stress on wheat plants, their effects on altering the levels of different plant hormones were different In the two cuItlvars.     

Biotoxic Effect of Chromium (VI) on Plant Growth-Promoting Traits of Novel Cellulosimicrobium funkei Strain AR8 Isolated from Phaseolus vulgaris Rhizosphere

The aim of this study was to investigate the effect of Cr(VI) on the plant growth-promoting traits of potential rhizobacterial strain isolated from Phaseolus vulgaris rhizosphere. A total of 36 rhizobacterial strains were recovered from the rhizosphere of P. vulgaris. Among these strains, the strain AR8 was specifically selected due to the highest resistance against heavy metals and the maximum production of plant growth-promoting substances. The rhizobacterial strain AR8 was identified as Cellulosimicrobium funkei (KM263188) following 16S rDNA gene sequencing. Strain AR8 solubilized phosphate and produced indole-3-acetic acid (32.57 µg/ml), exopolysaccharide (17.23 µg/ml), ammonia (54.16 µg/ml), catalase, biosurfactant, protease, amylase, and lipase. Under Cr(VI) stress, Cr(VI) concentration-dependent progressive decline in all plant growth-promoting traits of the C. funkei exposed was observed except for exopolysaccharide production, which consistently increased with increasing concentrations of Cr(VI). The root elongation assay resulted that the application of C. funkei strain AR8 significantly increased root length of test crops both in the presence and absence of Cr(VI) compared to uninoculated Cr(VI) treated plants. Moreover, AR8 generated a large number of colonies in diverse agricultural crops. Due to these intrinsic abilities, strain AR8 could be utilized for growth promotion as well as for the remediation of chromium in chromium-contaminated soil.     

Salicylic acid induces amelioration of chromium toxicity and affects antioxidant enzyme activity in Sorghum bicolor L.

Aim: Chromium (Cr(VI)) would inflict serious morphological, metabolic, and physiological anomalies in plants ranging from chlorosis of shoot to lipid peroxidation and protein degradation. Cr(VI) toxicity is often associated with oxidative stress, caused by the excessive formation of reactive oxygen species (ROS). In response, plants are equipped with a repertoire of mechanisms to counteract heavy metal (HM) toxicity. Salicylic acid (SA) plays a key role in the signal transduction pathways of various stress responses, demonstrating the protective effect of SA against abiotic stress factors. So, the present investigation was carried out to study the amelioration of pernicious effects of different concentration of Cr(VI) (0.0, 2.0, and 4.0?mg Cr(VI) kg^?1 soil in the form of potassium dichromate) by treatments of salicylic acid solution viz. pretreatment and foliar spray via antioxidative enzymes and their metabolites.

Results: With different treatments of salicylic acid solution, the reinstatement from ill effects of Cr(VI) toxicity was contemplated but the most conspicuous effect was observed when salicylic acid solution was supplied through the foliar spray (0.50?mM). This was accompanied with an increase in ascorbate peroxidase activity and hydrogen peroxide content and decrease in peroxidase activity and ascorbic acid content.

Significance of the study: This study suggests that salicylic acid when applied through pre-treatment of seeds or through a foliar spray can be used to ameliorate the toxic effects of chromium (VI). Salicylic acid has the great potential for reducing the toxicity of heavy metals without negatively impacting the growth of the plants.