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.     

Cadaverine production by Azospirillum brasilense and its possible role in plant growth promotion and osmotic stress mitigation

Polyamines are considered as plant growth regulating compounds; among them, cadaverine has been correlated with root growth promotion or osmotic stress mitigation in some plant species. The purpose of this study was to evaluate the capacity of bacterial Azospirillum brasilense Az39 strain to produce cadaverine in chemically defined medium and inoculated plants, and to correlate this capacity with root growth promotion or osmotic stress mitigation in hydroponics conditions. To evaluate cadaverine production in chemically defined medium A. brasilense Az39 was cultivated aerobically at 30 °C and 80 rpm in NFb medium or NFb-l supplemented with the precursor l-lysine. To evaluate the bacterial cadaverine production and growth promotion in plants, rice (Oryza sativa L.) cv. El Paso 144 seedlings were inoculated and hydroponically cultured under optimal conditions in growth chamber. In both, cadaverine was identified and quantified by dansyl-derivative method using a fluorescence-HPLC system, and lysine decarboxylase (LDC) activity was determined by ¹⁴CO₂ production in a closed tube system fed with [¹⁴C]-lysine. To evaluate the possible role of bacterial cadaverine in osmotic stress conditions, abscisic acid (ABA) production was analyzed in rice seedlings hydroponically cultured under 0 (no stress), ?0.47 (stress) or ?0.82 (severe stress) MPa osmotic potential generated by mannitol, with the addition of 1 nM or 1 μM cadaverine or A. brasilense Az39 inoculation. Our results indicate that A. brasilense Az39 promoted root growth and helped mitigate osmotic stress in rice seedlings, due in part to cadaverine production.     

Potential for enhancement of root growth and nodulation of soybean co-inoculated with Azospirillum and Bradyrhizobium in laboratory systems

The potential enhancement of root growth and nodulation in vegetable soybean (AGS190) was studied with application of Azospirillum brasilense (Sp7) and A. lipoferum (CCM3863) co-inoculated with two Bradyrhizobium japonicum strains (TAL102 and UPMR48). Significant root growth stimulation and nodulation were observed in Azospirillum as well as during its co-inoculation with Bradyrhizobium. Nodule formation is linked with the initiation of new roots; nodules were almost absent even in Bradyrhizobium inoculated plant due to the absence of new roots development in clipped rooted seedlings. Total root length, root number, specific root length, root dry matter, root hair development and shoot dry matter were significantly increased by Azospirillum alone and its co-inoculum. Co-inoculated plants significantly influenced the number of nodules and its fresh weight. A. brasilense seemed to perform better in root growth and nodule development compared to A. lipoferum.     

Azospirillum inoculation and nitrogen fertilization effect on grain yield and on the diversity of endophytic bacteria in the phyllosphere of rice rainfed crop

We assessed the Azospirillum inoculation and N-fertilization effect on grain yield and on the phyllosphere endophytic diversity of nitrogen-fixing bacteria in a rice rainfed crop. We used cultivation-based techniques and cultivation-independent methods involving PCR-16S rRNA and denaturing gradient gel electrophoresis (DGGE). In general, we observed that grain yield was improved when inoculated with Azospirillum (depending on the genotype) and/or fertilized with urea. A similar behavior was observed in total N-content in grain and the MPN determination, as the highest values occurred when seeds were inoculated with A. brasilense REC3 (S1) than with A brasilense 13-2C (S2). A positive nitrogenase activity and PCR-nifH amplification suggests that the bacteria associated to inner tissues of rice phyllosphere could have contributed to the different N-contents detected. The bacterial diversity, observed in the number and intensity of DGGE profiles, showed a higher number of bands when total DNA was obtained using only CTAB than with CTAB + PVP. The DGGE profiles revealed great stability in the dominating bands, which presumably represent numerically dominant species. Application of A. brasilense strains as inoculants did not influence the dominant members of the endophytic microbial communities in the phyllosphere, but improved N-content and production of rainfed rice crop.     

Isotopic evidence of significant assimilation of atmospheric-derived nitrogen fixed by Azospirillum brasilense co-inoculated with phosphate-solubilising Pantoea dispersa in pepper seedling

The contribution of plant growth-promoting bacteria (PGPB) of the genus Azospirillum to the plant N budget through biological nitrogen fixation (BNF) is still controversial. The aim of this study was to determine the contribution of BNF by Azospirillum brasilense on pepper grown at different N levels, attained using the ¹⁵N natural abundance method. To this end, pepper plants were grown in a growth chamber and treated with A. brasilense combined with Pantoea dispersa and then irrigated at four different N levels (0, 1, 3 and 7 mM NO₃⁻). The assimilation of fixed N was clear from the lower δ¹⁵N values observed in bacteria-treated plants compared with those of non-bacteria treated plants. The percentage of BNF-derived N decreased with decreasing NO₃⁻ levels in the growth medium. BNF contribution to the total nitrogen content of plants was found to be as high as 46%. The results suggest that the bacteria have a potential to supply a considerable amount of N to pepper seedlings, as well as to stimulate plant growth and N uptake when Azospirillum–Pantoea treatment is combined with low NO₃⁻ levels.     

Implications of Azospirillum brasilense inoculation and nutrient addition on maize in soils of the Brazilian Cerrado under greenhouse and field conditions

The combination of nitrogen and Azospirillum can ensure greater nutrient absorption and crop yield in agricultural areas using high technology. Thus, the objective was to evaluate maize response to Azopirillum brasilense (AZ) inoculation and nutrient (macronutrients and micronutrients) application under greenhouse and field conditions in clay and sandy soils of the Brazilian Cerrado. In the greenhouse assays, the following parameters were measured: shoot dry weight (SDW), root dry weight (RDW), and root volume (RV). In the field experiments, the maize yield was determined after drying the grains at 60 °C for 48 h. In clay soil, there was a significant increase in the SDW, RDW and RV in the treatment with AZ concentrated (10¹¹ cells ml⁻¹ of inoculum) when compared with the control treatment and the treatment with AZ diluted (10⁶ cells ml⁻¹ of inoculum). In this soil, adding micronutrients did not affect the maize response under greenhouse conditions. In sandy soil, there was no difference between the AZ treatment and the control, except for treatments where nutrients and AZ were both added leading to a significant increase in the maize response. In both soils, the RV:RDW ratio was higher in the treatment with AZ concentrated compared to that in the treatment with AZ diluted, but the yield response depended on the addition of nutrients. Inoculation with A. brasilense gave comparable yield to the nitrogen treatment. The grain production was increased by 29% in the treatment with A. brasilense and nitrogen compared to nitrogen fertilization alone. In this study, the yield response was affected significantly when maize was inoculated with A. brasilense, but this response was dependent on the soil type under greenhouse conditions.     

Resistance to oxidation products of caffeic acid is important for efficient colonization of wheat seedlings by Pseudomonas proteolytica strain PSR114

The interrelationships between plants and rhizosphere bacteria are strongly dependent on the quality and quantity of root exudates. The ability to colonize roots is crucial for pseudomonads to function as biological control agents of root- and soil-borne pathogenic microbes. The multiplication of rhizosphere bacteria is restricted in the presence of simple phenolic compounds, which are components of the resistance mechanisms of plants to pathogens. Caffeic acid is a phenolic compound, which is commonly found in wheat tissues. It is prone to oxidation into o-quinones, which are toxic to microorganisms. The aim of the present study was to determine whether the ability of microorganisms to resist caffeic acid and its oxidation products could play a role in the early colonization of wheat seedlings. Among the fluorescent pseudomonads that we have studied, strain PSR114 is one of the most efficient colonizers of wheat seedlings during the first 48 h after seed germination, and it is particularly resistant to products resulting from the spontaneous oxidation of caffeic acid. This strain was isolated from the rhizosphere of oilseed rape and identified as being closely related to Pseudomonas proteolytica through the analysis of 16S rRNA and rpoB gene sequences. At pH 7.0, this strain grew intensively in the presence of 1.50 mg mL⁻¹ of caffeic acid. Its multiplication was partially reduced in the presence of oxidized caffeic acid at concentrations above 0.21 mg mL⁻¹, and completely inhibited at concentrations above 0.38 mg mL⁻¹. A Tn5 transposon mutant of PSR114 had lower level of resistance to the oxidation products of caffeic acid, as well as reduced capacity to colonize wheat seedlings when compared to the wild type strain. This work demonstrates that resistance to oxidation products of caffeic acid can be important for successful bacterial colonization of wheat seedlings.     

Improved soil structure and citrus growth after inoculation with three arbuscular mycorrhizal fungi under drought stress

In a controlled potted experiment, citrus (Poncirus trifoliata) seedlings were inoculated with three species of arbuscular mycorrhizal (AM) fungi, Glomus mosseae, G. versiforme or G. diaphanum. Two soil-water levels (ample water, −0.10 MPa; drought stress, −0.44 MPa) were applied to the pots 4 months after transplantation. Eighty days after water treatments, the soils and the citrus seedlings were well colonized by the three AM fungi. Mycorrhizal fungus inoculation improved plant biomass regardless of soil-water status but decreased the concentrations of hot water-extractable and hydrolyzable carbohydrates of soils. Mycorrhizal soils exhibited higher Bradford-reactive soil protein concentrations than non-mycorrhizal soils. Mycorrhizas enhanced >2 mm, 1–2 mm and >0.25 mm water-stable aggregate fractions but reduced 0.25–0.5 mm water-stable aggregates. Peroxidase activity was higher in AM than in non-AM soils whether drought stressed or not, whereas catalase activity was lower in AM than non-AM soils. Drought stress and AM fungus inoculation did not affect polyphenol oxidase activity of soils. A positive correlation between the Bradford-reactive soil protein concentrations, soil hyphal length densities, and water-stable aggregates (only >2 mm, 1–2 mm and >0.25 mm) suggests beneficial effects of the AM symbiosis on soil structure. It concluded that AM fungus colonization enhanced plant growth under drought stress indirectly through affecting the soil moisture retention via glomalin's effect on soil water-stable aggregates, although direct mineral nutritional effects could not be excluded.     

Azospirillum brasilense Az39 and Bradyrhizobium japonicum E109, inoculated singly or in combination,promote seed germination and early seedling growth in corn (Zea mays L.) and soybean (Glycine max L.)

Inoculants are biological formulations that combine a stable microorganism population and various types of compounds produced and released during fermentation, such as phytohormones and plant growth regulators. Azospirillum brasilense strain Az39 and Brayrhizobium japonicum strain E109 were previously shown to produce indole 3-acetic acid (IAA), gibberellic acid (GA₃) and zeatin (Z). We tested the hypothesis that such compounds are responsible for early growth promotion in inoculated corn (Zea mays L.) and soybean (Glycine max L.) seedlings. Seeds were inoculated with Az39, E109, or both, and kept in a chamber at 20–30 °C under a controlled photoperiod to evaluate seed germination. To evaluate root and shoot length and dry weight, and number of nodules and percentage of nodulated seedlings, in soybean, seedlings were kept in a growth chamber for 14 days under similar photoperiod and temperature conditions. Az39 and E109, singly or in combination, showed the capacity to promote seed germination, nodule formation, and early development of corn and soybean seedlings. Both strains were able to excrete IAA, GA₃ and Z into the culture medium, at a concentration sufficient to produce morphological and physiological changes in young seed tissues.     

Impact of allelochemical exuded from allelopathic rice on soil microbial community

Allelopathic rice releases allelochemicals from its roots to paddy soils at early growth stages to inhibit neighboring weeds. However, little is currently known about the effects of allelochemicals on soil microbes. In this study, we show that allelopathic rice can have great impact on the population and community structure of soil microbes. Allelopathic rice PI312777 seedlings reduced the culturable microbial population and total PLFA when compared to non-allelopathic rice Liaojing-9. Similar results were observed when, instead of growing seedlings, soils were incubated with plant root exudates. This result demonstrates that the composition of root exudates from the rice varieties tested contributes to the soil microbial community. Further experiments showed that the microbial community was affected by the allelochemical 5,4′-dihydroxy-3′,5′-dimethoxy-7-O-β-glucopyranosylflavone exuded from allelopathic rice roots, through immediately hydrolyzing glucose with stimulation on soil bacteria and aglycone (5,7,4′-trihydroxy-3′,5′-dimethoxyflavone) with inhibition on soil fungi. This result indicates that the flavone O-glycoside can provide carbon and interact with soil microbes. PC analysis of the fatty acid data clearly separated the allelopathic PI312777 and the non-allelopathic Liaojing-9 variety (PC1 = 46.4%, PC2 = 20.3%). Similarly, the first principal component (PC1 = 37.4%) together with the second principal component (PC2 = 17.3%) explained 54.7% of the variation between the allelopathic and non-allelopathic root exudates. Furthermore, the canonical correlation between allelopathic root exudates and the flavone O-glycoside was statistically significant (Canonical R = 0.889, χ² (25) = 69.72, p = 0.0041). Although the data generated in this study were not completely consistent between culturable microbes and PLFA profile, it is a fact that variation in soil microbial populations and community structures could be distinguished by the allelopathic and non-allelopathic rice varieties tested. Our results suggest that individual components of rice root exudates, such as allelochemicals from allelopathic rice, can modify the soil microbial community.     

Effects of arbuscular mycorrhizal inoculation and phosphorus supply on the growth and nutrient uptake of Kandelia obovata (Sheue,Liu & Yong) seedlings in autoclaved soil

This study evaluated the interactive effect of arbuscular mycorrhizal fungi (AMF) inoculation and exogenous phosphorus supply on soil phosphotases, plant growth, and nutrient uptake of Kandelia obovata (Sheue, Liu & Yong). We aimed to explore the ecophysiological function of AMF in mangrove wetland ecosystems, and to clarify the possible survival mechanism of mangrove species against nutrient deficiency. K. obovata seedlings with or without AMF inoculation (mixed mangrove AMF), were cultivated for six months in autoclaved sediment medium which was supplemented with KH₂PO₄ (0, 15, 30, 60, 120 mg kg−¹). Then the plant growth, nitrogen and phosphorus content, root vitality, AMF colonization and soil phosphatase activity were analyzed. The inoculated AMF successfully infected K. obovata roots, developed intercellular hyphae, arbuscular (Arum-type), and vesicle structures. Arbuscular mycorrhizal fungi colonization ranged from 9.04 to 24.48%, with the highest value observed under 30 and 60 mg kg⁻¹ P treatments. Soil P supply, in the form of KH₂PO₄, significantly promoted the height and biomass of K. obovata, enhanced root vitality and P uptake, while partially inhibiting soil acid (ACP) and alkaline phosphotase (ALP) activities. Without enhancing plant height, the biomass, root vitality and P uptake were further increased when inoculated with AMF, and the reduction on ACP and ALP activities were alleviated. Phosphorus supply resulted in the decrease of leaf N–P ratio in K. obovata, and AMF inoculation strengthened the reduction, thus alleviating P limitation in plant growth. Arbuscular mycorrhizal fungi inoculation and adequate P supply (30 mg kg⁻¹ KH₂PO₄) enhanced root vitality, maintained soil ACP and ALP activities, increased plant N and P uptake, and resulted in greater biomass of K. obovata. Mutualistic symbiosis with AMF could explain the survival strategies of mangrove plants under a stressed environment (waterlogging and nutrient limitation) from a new perspective.     

Inoculation of field-grown wheat (Triticum aestivum) with Azospirillum spp. in Brazil

Summary Three field experiments with wheat were conducted in 1983, 1984, and 1985 in Terra Roxa soil in Paraná, the major Brazilian wheat-growing region, to study inoculation effects of various strains of Azospirillum brasilense and A. amazonense. In all three experiments inoculation with A. brasilense Sp 245 isolated from surface-sterilized wheat roots in Paraná produced the highest plant dry weights and highest N% in plant tops and grain. Grain yield increases with this strain were up to 31 % but were not significant. The application of 60 or 100 kg N ha^–1 to the controls increased N accumulation and produced yields less than inoculation with this strain. Another A. brasilense strain from surface-sterilized wheat roots (Sp 107st) also produced increased N assimilation at the lower N fertilizer level but reduced dry weights at the high N level, while strain Sp 7 + Cd reduced dry weights and N% in the straw at both N levels. The A. amazonense strain isolated from washed roots and a nitrate reductase negative mutant of strain Sp 245 were ineffective. Strains Sp 245 and Sp 107st showed the best establishment within roots while strain Cd established only in the soil.     

Interactions of assemblages of mycorrhizal fungi with two Florida wetland plants

Arbuscular mycorrhizal (AM) fungi are known to exist in wetlands, but little is known about their function in these environments. We conducted greenhouse experiments to study the effects of AM fungal assemblages—collected from different vegetation communities in a Florida wetland-under free-drained and flooded conditions, and at three phosphorus (P) levels on growth and P nutrition of Typha latifolia L. and Panicum hemitomon Schult. We also studied the effects of flooding on the spread of extraradical hyphae from P. hemitomon roots. For both plants no AM fungal assemblage had a consistent effect on plant growth and P nutrition. For T. latifolia, flooding nearly eliminated AM fungal colonization and, in the free-drained treatments, P amendment suppressed colonization. Furthermore, colonization by some mycorrhizal assemblages increased shoot- and root-P concentrations, but there were no significant plant growth responses. For P. hemitomon, the mycorrhizal association was suppressed by flooding and P amendment but, among the fungal assemblages, there were differences in root colonization. Mycorrhizal colonization improved some plant-growth and P-nutrition parameters at lower P levels relative to nonmycorrhizal controls, but generally conferred no benefit or was detrimental at higher P levels. Extraradical hyphae of most assemblages were restricted by flooding to 2.5 cm, though differences among AM fungal assemblages occurred with a maximum observed extension of 16.5 cm. We conclude that the impact of the mycorrhizal association on these wetland plants was a function of the complex interactions among the AM fungal assemblages, plant species, water condition, and P level. Future studies should focus on understanding the species composition of the assemblages, and potential adaptation to wetland conditions among these fungal species.     

Chemotaxis of deleterious rhizobacteria to birdsfoot trefoil

Intact seeds and seed and seedling root exudates of birdsfoot trefoil (Lotus corniculatus L.) were used as chemoattractants in experiments to determine the relative importance of chemotaxis in spermosphere and rhizosphere colonization by selected rhizobacteria. Results for soft-agar, capillary tube and soil chemotaxis assays indicated that selected deleterious rhizobacteria were attracted to seed and seedling root exudates. Several sugars and phenolic fractions detected in exudates were chemoattractants for these rhizobacteria. Using soil-chemotaxis assemblies, migration of rhizobacterial isolates through 2 cm distances of soil toward birdsfoot trefoil seeds was detected within 24 h. Isolates were not detected at the same site in soils without seeds until 72 h after inoculation. These results suggest that attraction of deleterious rhizobacteria toward seeds and seedling roots mediated by exudates (chemotaxis) might be the first step in the establishment and subsequent colonization of bacteria involved in soilborne disease complexes of birdsfoot trefoil.     

Plant growth promoting potential of Pontibacter niistensis in cowpea (Vigna unguiculata (L.) Walp.)

During the past couple of decades, understanding of rhizosphere biology has progressed with the discovery of a special group of microorganisms known as plant growth promoting rhizobacteria (PGPR) and its application for sustainable agriculture has increased tremendously in various parts of the world. The search for microorganisms that improve soil fertility and enhance plant nutrition has continued to attract attention due to the increasing cost of fertilizers and some of their negative environmental impacts. In this study we demonstrated, a novel bacterial species Pontibacter niistensis NII-0905 isolated from forest soil in Western ghat forest soil with potential plant growth promoting ability (PGP) such as phosphate solubilization, indole acetic acid (IAA), siderophore and hydrogen cyanide (HCN) production. The activity varies with different growth temperatures, strain solubilize 28.5 ± 0.9, 48.02 ± 1.9 and 65.07 ± 2.1 μg mL⁻¹ at 4, 15 and 30 °C respectively and produced 24.8 μg mL⁻¹ day⁻¹ of indole acetic acid (IAA) in tryptophan amended media. Qualitative detection of siderophore production and HCN were also detected at all temperature tested. At a lower temperature (4 °C) strain NII-0905 retained all the plant growth promotion attributes. A significant increase in the growth of cow pea was recorded with inoculations of strain NII-0905 in pot experiments. Scanning electron microscopic study revealed the root colonization on cow pea seedlings against the untreated one. These results demonstrate that, the isolate NII-0905 has the promising PGPR attributes for both in cold as well as in humid condition. It has potential as a biofertilizer to enhance soil fertility and promote the plant growth.     

Botanical survey and screening of plant species which accumulate 226Ra from contaminated soil of uranium waste depot

A geobotanical study was performed of the wild plants growing in the area of the old uranium mill tailings waste depot of a former uranium ore reprocessing factory in South Bohemia and the distribution of ²²⁶Ra in selected plants was determined. The distribution of ²²⁶Ra in contaminated soil was found to be extremely variable (from 7 to 32 Bq ²²⁶Ra g⁻¹ of DW). The differences in plant distribution were caused by factors of disturbance, soil properties (nutrients and salt content, water supply), and successional stage. No direct relation was proved between plant species characteristics and their radioactivity content. The results showed a great range of variation in the accumulation of ²²⁶Ra by the plant species found. The highest activity of ²²⁶Ra was found in Potentilla reptans (4.09 Bq ²²⁶Ra g⁻¹ of DW), Mentha arvensis (4.00 Bq ²²⁶Ra g⁻¹ of DW), and Daucus carota (3.70 Bq ²²⁶Ra g⁻¹ of DW). About half of the plant species are used as medicinal plants and some of them are accumulators of ²²⁶Ra. However, no plants suitable for phytoextraction of ²²⁶Ra contaminated substrates were discovered.     

Arbuscular mycorrhizal abundance in contaminated soils around a zinc and lead deposit

In order to study the variations in spore abundance and root colonization parameters of arbuscular mycorrhizal (AM) fungi in a naturally heavy metals polluted site and their relationships with soil properties, 35 plots in the Anguran Zn and Pb mining region were selected along a transect from the mine to 4500 m away. Within each plot, a composite sample of root and rhizospheric soil from a dominant indigenous plant was collected. The soil samples were analyzed for their physico-chemical characteristics. Spores were extracted, counted and identified at genus level. The roots were examined for colonization, arbuscular abundance, mycorrhizal frequency and intensity. Along the transect, the total and available (DTPA-extractable) concentration of Zn decreased from 6472 to 45 mg kg⁻¹ and 75 to 5 mg kg⁻¹, respectively. For Pb the values varied from 5203 to 0 mg kg⁻¹ and 32 to 0 mg kg⁻¹, respectively. In parallel, root colonization rate in the dominant native plants (except Alyssum sp.) varied from 35% to 85% and the spore numbers from 80 to 1306 per 200 g dry soil along the transect. Spores of Glomus were abundantly found in all plots as dominant, while Acaulospora spores were observed only in some moderately polluted and in control plots. AM fungal propagules never disappeared completely even in soils with the highest rates of both heavy metals. Spore numbers were more affected by Zn and Pb concentrations than root colonization. The variations of AM fungi propagules were better related to available than to total concentration of both metals. Spore numbers were positively correlated with mycorrhizal colonization parameters, particularly with arbuscular abundance.     

Enhancement of arbuscular mycorrhizal fungus (Glomus versiforme) on the growth and Cd uptake by Cd-hyperaccumulator Solanum nigrum

Arbuscular mycorrhizal fungus (AMF) can enhance plant growth and resistance to toxicity produced by heavy metals (HMs), affect the bioavailability of HMs in soil and the uptake of HMs by plants, and thus has been emerged as the most prominent symbiotic fungus for contribution to phytoremediation. A greenhouse pot experiment was conducted to assess the effect of Glomus versiforme BGC GD01C (Gv) on the growth and Cd accumulation of Cd-hyperaccumulator Solanum nigrum in different Cd-added soils (0, 25, 50, 100 mg Cd kg⁻¹ soil). Mycorrhizal colonization rates were generally high (from 71% to 82%) in Gv-inoculated treatments at all Cd levels. Gv colonization enhanced soil acid phosphatase activity, and hence elevated P acquisition and growth of S. nigrum at all Cd levels. Moreover, the presence of Gv significantly increased DTPA-extractable (phytoavailable) Cd concentrations in 25 and 50 mg Cd kg⁻¹ soils, but did not affect phytoavailable Cd in 100 mg Cd kg⁻¹ soil. Similarly, inoculation with Gv significantly increased Cd concentrations of S. nigrum in 25 and 50 mg Cd kg⁻¹ soils, but decreased Cd concentrations of the plants in 100 mg Cd kg⁻¹ soil. Overall, inoculation with Gv greatly improved the total Cd uptakes in all plant tissues at all Cd levels. The present results indicated that S. nigrum associated with Gv effectively improved the Cd uptake by plant and would be a new strategy in microbe-assisted phytoremediation for Cd-contaminated soils.     

Influence of AM fungi on the growth and physiological status of Erythrina variegata Linn. grown under different water stress conditions

The present study investigated the effects of arbuscular mycorrhizal (AM) fungus, Glomus mosseae on the growth and physiology state of Erythrina variegata Linn, grown in sandy loam soil with four water stress levels viz. ?0.06 MPa (well watered/control), ?1.20 MPa (mild), ?2.20 MPa (moderate) and ?3.20 MPa (severe) in a completely randomized design. Plants were harvested after 90 days (60 days after stress induction) of growth. Growth parameters (root &, shoot, dry weight and, leaf area); physiological parameters (chlorophyll content, carotenoids, soluble starch, sugar, protein and proline in shoots); and microbiological parameter (percentage of mycorrhizal infection) were determined. AM fungal plants had significantly higher plant biomass, higher chlorophyll content (chlorophyll a and b), carotenoids and protein content in shoots than non-AM-plants. The AM-inoculation in stressed plants significantly declined the soluble sugar and starch in shoots. Moreover, AM-inoculation also reduced the proline accumulation in shoots and the reduction was significant when plants were severely stressed (?3.2 MPa). Mycorrhizal colonization in roots of E. variegata depressed significantly due to increased water stress. However, the AM colonization did not decline below 11% and enabled the plants to maintain osmotic adjustments and enhanced the plants tolerance against water stress.     

Improvement of Brassica napus growth under cadmium stress by cadmium-resistant rhizobacteria

This study focuses on the characterization of four bacterial isolates from heavy metal-polluted rhizosphere in order to examine their plant growth promoting (PGP) activity. The PGP activity on the canola (Brassica napus) of the strains which showed cadmium resistance and multiple PGP traits was assessed in the presence and in the absence of Cd²⁺. The strains, Pseudomonas tolaasii ACC23, Pseudomonas fluorescens ACC9, Alcaligenes sp. ZN4 and Mycobacterium sp. ACC14 showed 1-aminocyclopropane-1-carboxylate deaminase (ACCD) activity. They also synthesized ACCD enzyme in vitro when 0.4 mM Cd²⁺ was added to the growth medium. The presence of the metal, however, reduced the ACCD activity in Alcaligenes sp. ZN4 and Mycobacterium sp. ACC14, while it did not affect the ACCD activity of P. tolaasii ACC23 and P. fluorescens ACC9. ACC9 and ACC23 produced indole acetic acid (IAA) and siderophores, while ACC14 produced only IAA. IAA and siderophores were produced more actively under Cd-stress.Root elongation assays conducted on B. napus under gnotobiotic conditions demonstrated increases (from 34% up to 97%) in root elongation of inoculated canola seedlings compared to the control plants. Subsequently, the effect of inoculation with these strains on growth and uptake of Cd²⁺ in roots and shoots of canola was studied in pot experiments using Cd-free and Cd-treated (15 μg Cd²⁺ g^?1 dw) soil. Inoculation with P. tolaasii ACC23, P. fluorescens ACC9 and Mycobacterium sp. ACC14 promoted the growth of plants at concentrations of 0 and 15 μg Cd²⁺ g^?1 soil. The maximum growth was observed in the plants inoculated with P. tolaasii ACC23. The strains did not influence the specific accumulation of cadmium in the root and shoot systems, but all increased the plant biomass and consequently the total cadmium accumulation.The present observations showed that the bacterial strains used in this study protect the plants against the inhibitory effects of cadmium, probably due to the production of IAA, siderophores and ACCD activity.