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.     

Seedlings growth promotion by Azospirillum brasilense under normal and drought conditions remains unaltered in Tebuconazole-treated wheat seeds

The wide use of pesticides in modern agriculture may cause side effects on the non-target microflora. Data on the fungicide Tebuconazole effects on Azospirillum-wheat association are scarce. We analyzed the effects of Tebuconazole on: (a) Azospirillum brasilense Sp245 growth in pure culture, (b) A. brasilense Sp245 colonization of Triticum aestivum cv ProINTA Oasis roots, (c) A. brasilense Sp245-inoculated seedlings growth under normal and water stress conditions in the presence of 20% polyethylene glycol 8000. Seeds were separated in Tebuconazole-free and Tebuconazole-treated lots. Inoculated and non-inoculated seedlings were grown in hydroponics in the dark at 20 °C for 72 h. Root surface, coleoptile length, fresh and dry (DW) weights in both tissues and diazotrophic bacterial most probable number in roots were determined. Water contents and shoot-to-roots DW ratio were calculated. Neither Azospirillum growth nor root colonization was affected by Tebuconazole. Under normal growth conditions most of the growth parameters analyzed, revealed a clear positive effect of A. brasilense on wheat seedlings up to 72 h treatments. The characteristic Azospirillum enhancing effects observed on roots remained unaltered by Tebuconazole. The present study shows that Tebuconazole is compatible with A. brasilense Sp245-wheat inoculation.     

Determination of free and conjugated indole-3-acetic acid, tryptophan, and tryptophan metabolites in grape must and wine.

Tryptophan (Trp) and its metabolites, especially indole-3-acetic acid (IAA), are considered to be potential precursors of 2-aminoacetophenone (AAP), an aroma compound that causes an "untypical aging off-flavor" (UTA) in Vitis vinifera wines. In this study, RP-HPLC with fluorescence detection was used for the qualitative and quantitative analysis of Trp and Trp metabolites in grape musts and wines to which different viticultural measures had been applied (time of harvest, soil treatment, leaf plucking, vine prune). An alkaline hydrolysis was developed to release bound IAA and Trp. A sensitive and selective determination of different Trp metabolites was achieved after solid phase extraction using a strong anion exchange material. In the examined grape musts, more than 95% of the total IAA was bound either as ester conjugate or as amide conjugate. Free IAA and other Trp metabolites were below the detection limit (<3 microg/L) or could be determined only in traces. Their amounts increased significantly during fermentation, whereas the amount of Trp decreased. It could be shown that the different viticultural measures applied (except the vine prune) as well as the climatic conditions of the vintage exhibited significant influences on the amounts of Trp and Trp metabolites in grape musts or wines.     

Arbuscular mycorrhizal fungi inoculation and exogenous indole-3-acetic acid application induce antioxidant defense response to alleviate cadmium toxicity in Broussonetia papyrifera

Cadmium (Cd) contamination in soil poses a huge threat to plants even at low concentrations; Broussonetia papyrifera has great potential in remediation of soil heavy metal contamination. However, whether exogenous indole-3-acetic acid (IAA) application and arbuscular mycorrhizal fungi (AMF) have synergistic effects on Cd tolerance of B. papyrifera remains unclear. To investigate the effects of AMF inoculation and IAA application on the tolerance of B. papyrifera to Cd stress, two experiments were conducted: the first to investigate the effect of AMF (Rhizophagus irregularis) inoculation on the tolerance of B. papyrifera to Cd stress and the second to investigate the combined effects of AMF inoculation and IAA application on the tolerance of B. papyrifera to Cd stress. Parameters including endogenous hormone concentration, antioxidant defense response, malondialdehyde (MDA) content, and gene expression related to antioxidant enzyme system and hormone were measured. The results indicated that AMF alleviated Cd toxicity of B. papyrifera by reducing MDA content and improving antioxidant enzyme activities and Cd absorption capacity. Furthermore, the combination of AMF inoculation and IAA application had a synergetic effect on the tolerance of B. papyrifera to Cd stress through upregulating BpAUX1 and BpAUX2, which might contribute to root growth and root xylem synthesis, and by upregulating BpSOD2 and BpPOD34 to enhance the antioxidant enzyme system. This work provides a new insight into the application of IAA in the remediation of soil Cd pollution by mycorrhizal plants.     

Dual inoculation with strains of Bradyrhizobium japonicum and Azospirillum brasilense to improve growth and biological nitrogen fixation of soybean (Glycine max L.)

The effects of inoculation with Bradyrhizobium japonicum and Azospirillum brasilense strains on the growth of soybean were evaluated with regard to the estimation of N₂ fixation using the ¹⁵N isotope dilution technique. Inoculation, in general, increased the dry mass of soybean as well as nitrogen content. Dual inoculation with a mixture of B. japonicum and A. brasilense strains was superior over single inoculation with B. japonicum. Nitrogen fixed (Ndfa) varied according to inoculant and soil conditions. Percentages of nitrogen derived from air (% Ndfa) using a non-nodulating isoline were 72% and 76% for B. japonicum and B. japonicum plus A. brasilense, respectively, in non-sterile soil. A similar but higher trend was recorded in sterilized soil, in which the percentages of N₂ fixed were 81% and 86% for single and dual inoculation, respectively. The correlation coefficient between N₂ fixed and N uptake (r=0.94) and dry mass (r=0.89) was significant. Application of special bacterial inoculants in agricultural systems of Egypt seems to be a promising technology and could be used for improving soybean growth as well as soil fertility, thus minimizing environmental pollution. Received: 10 January 1996     

腐植酸促进植物生长的机理研究进展

【目的】腐植酸在我国农业生产中发挥了重要作用,许多研究证实,腐植酸具有促进植物生长的功能,本文从腐植酸刺激植物根系生长、调控土壤与肥料养分转化及肥料利用率和影响土壤微生物和酶活性方面,系统总结了国内外施用腐植酸促进植物生长的途径,阐述了腐植酸对植物生长促进作用的机理,旨在梳理腐植酸促进植物生长机理的研究现状,为腐植酸的进一步研究和应用提供参考依据。[主要进展]1）腐植酸能够对植物产生类似生物刺激素的效应。它能够提高植物根系H+-ATP酶等的活性、刺激植物根伸长和侧根生长点的增加,从而增加根系活力及植物根系与土壤养分的接触面积,增加植物对养分的吸收;2）逆境胁迫下,腐植酸能够通过调节植物体内的新陈代谢并改善植物生长环境,缓解甚至消除逆境胁迫对植物的伤害,从而促进植物生长;3）腐植酸能够通过与氮素、磷素和钾素发生结合效应,与磷酸盐产生竞争效应和对钾离子的吸附作用固持与活化土壤与肥料中的养分,提高土壤肥料有效性和缓释性能,提高肥料利用率,从而促进植物生长;4）腐植酸还能够影响土壤中与养分转化相关的酶活性和微生物群落结构及数量,在活化养分的同时,保蓄养分,降低养分的损失,为植物生长保障持久的养分供应;5）腐植酸对植物生长的促进效应受腐植酸结构特征、添加量和供试植物种类等因素的影响。[建议与展望]由于技术手段的限制和研究技术的差异,人们对腐植酸促进植物生长机理的认识还不够系统和深入,因此,腐植酸的基本特征、影响腐植酸作用的主控因子、土壤-植物系统中腐植酸促进植物生长的主要途径和腐植酸对土壤功能性微生物等的影响都将成为未来研究的重要方向。     

Alleviation of salinity stress in rice plant by encapsulated salt tolerant plant growth promoting bacteria Pantoea agglomerans strain KL and its root colonization ability

ABSTRACT

The present investigation indicates the effects of 0 to 8% NaCl stress on plant growth promoting traits such as ACC (1-aminocyclopropane-1-carboxylate) deaminase, phosphate solubilization, IAA (indole acetic acid), ammonia and exopolysaccharide production of Pantoea agglomerans strain KL isolated from salt-stressed soil of Kolhapur, Maharashtra, India. We have studied the effect of encapsulated inoculum (EI) and free inoculum (FI) of P. agglomerans strain KL on the alleviation of salinity stress (100 mM NaCl) and promotion of rice plant growth in the pot experiment. The present study showed significant improvement in plant growth supplemented with EI in terms of increased length, biomass, photosynthetic pigment and decreased level of proline, malondialdehyde. Furthermore, EI supplemented plant exhibited decreased sodium and increased calcium and potassium uptake. Root colonization study revealed the survival of encapsulated organism which was less after 10 days. However, a significant number of colony forming unit were noted after 20 and 30 days. In addition, the scanning electron microscopic analysis of salt-stressed plant root showed tremendous root colonization by EI. Hence, the present study demonstrates the potency of P. agglomerans strain KL in the expression of plant growth promoting traits and amelioration of salt stress by EI.     

"Untypical aging off-flavor" in wine: synthesis of potential degradation compounds of indole-3-acetic acid and kynurenine and their evaluation as precursors of 2-aminoacetophenone

Kynurenine (1) and indole-3-acetic acid (2) are considered as potential precursors of 2-aminoacetophenone (3), which is regarded to be the aroma impact compound causing an "untypical aging off-flavor" (UTA) in Vitis vinifera wines. The mechanism of the formation of 3 was studied using model fermentation and model sulfuration media spiked with 1 or 2 as potential precursors. Possible degradation products such as kynurenamine (4) and kynurenic acid (5), or skatole (6), 2-oxoskatole (7), 2-formamidoacetophenone (8), 2-oxindole-3-acetic acid (9), and 3-(2-formylaminophenyl)-3-oxopropionic acid (10) were evaluated by HPLC-UV of the fermentation and sulfuration media and comparison with synthesized 7, 8, 9, and 10. The synthesis of the possible precursor 4-(2-aminophenyl)-2,4-dioxobutanoic acid (11), a proposed metabolite of 1 failed because a spontaneous cyclization yields 5 and N-oxo-kynurenic acid (12), but not 11. It could be shown that the formation of 3 is triggered by an oxidative degradation of 2 after sulfuration with potassium bisulfite via the intermediates 10 and 8. However, no formation of 3 occurred during sulfuration of a model wine spiked with 1 or during fermentation of a model must spiked with 1 or 2.     

Cell-based and chemical assays of the ability to modulate the production of intracellular Reactive Oxygen Species of eleven Mediterranean plant species related to ethnobotanic traditions

The aim of this study was to compare three tests for the evaluation of the antioxidant potential of eleven plant species traditionally used to cure or to enhance animal and/or human wellbeing. In addition to the ability to modulate the production of intracellular reactive oxygen species the effect of the treatment on cell viability was also considered. In the cell-based experiment DCF (2′,7′dichlorofluorescein diacetate) and in the chemical assays DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2′-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt), Pistacia lentiscus exhibited high antioxidant capacity, while Cistus creticus and Euphorbia characias were ineffective in cell-culture and displayed high activity in the chemical assays, and others (Malva sylvestris, Matricaria chamomilla, Urtica dioica) showed an opposite trend. Olea europaea (sylvestris and europaea) and Umbilicus rupestris displayed highest antioxidant efficacy in both HUVEC and HL-60 cells, and provided also appreciable activity in the chemical assays. The study gives evidence that herbal extracts may have similar antioxidant properties but different effects on cells. Therefore, it is important to combine analytical and biological testing methods in order to measure the antioxidant capacity of natural extracts. Results encourage to explore further the potential nutraceutical and functional properties of Mediterranean plant species related to ethno-botanic traditions for animal health care.     

Impact of plant genotype and nitrogen level on rice growth response to inoculation with Azospirillum sp. strain B510 under paddy field conditions

Abstract

Twenty rice cultivars, including three genetically-distinct groups (japonica, indica-1, and indica-2), were evaluated for their response to inoculation with Azospirillum sp. strain B510 in paddy fields with standard nitrogen (SN) and low nitrogen (LN) fertilization. In the SN field, the tiller numbers in most indica-2 cultivars, 37?days after transplanting (DAT), were significantly increased by the B510 inoculation, whereas those in 4 japonica cultivars were significantly decreased. A similar growth response was observed in the LN field, although the impacts of the B510 inoculation were more varied than in the SN field. At 58 DAT, the tiller numbers in most cultivars were lower or unaffected by the B510 inoculation under both SN and LN conditions, except that the tiller number of the Nipponbare cultivar, which is classified as japonica, was significantly higher in the LN field only. These results suggest that the effects of inoculation with Azospirillum sp. strain B510 on the growth of rice plants, especially on tiller numbers at the early growth stage, vary depending on the rice genotype, as well as nitrogen level. Therefore, the plant genotypes, growth stages, and fertilization managements must be considered when a plant-associated bacterium is evaluated for beneficial effects under field conditions.     

Glutamate wastewater as a culture medium for <Emphasis Type="Italic">Azospirillum rugosum</Emphasis> production and its impact on plant growth

Monosodium glutamate wastewater (MSGW) was used as a culture medium for Azospirillum rugosum strain IMMIB AFH-6 inoculant (In) by optimizing the dilution at 2.5% and pH 7.0 ± 0.1. A. rugosum growth studies were carried out in shake flasks and subsequently tested for plant growth promotion by seed germination bioassay and greenhouse pot experiments. The highest colony-forming units (cfu) 8.58 and 8.87 log cfu mL⁻¹, respectively, at 48 h of growth were recorded in MSGW and MSGW with yeast extract 0.04%. Seed bioassay results revealed that A. rugosum grown on MSGW stimulated the root and shoot elongation of maize (Zea mays L.) and pai tsai (Brassica chinensis L.). Pai tsai recorded significantly higher dry matter production when treated with A. rugosum and chemical fertilizers than control. The MSGW is a suitable culture medium for A. rugosum growth, and it is comparable to commercial media when tested for agricultural production.     

Efficiency of gypsum application to acid Andosols estimated using aluminum release rates and plant root growth

Abstract

A great deal of information on the efficiency of gypsum or phosphogypsum to ameliorate acidity in highly weathered soils is available, but only limited information is available on the efficiency in acid Andosols, which possess large amounts of active aluminum (Al). We examined the effectiveness of gypsum application to non-allophanic Andosols (one humus-rich A horizon and two B horizons poor in humus) using extractable soil Al analyses (batch and continuous extraction methods) and a cultivation test using burdock (Arctium lappa). With gypsum amendment, pH(H₂O) values of the soil decreased from 4.5–4.7 to 4.2–4.4, whereas the treatment made almost no difference to the values of pH(KCl). Total active Al (acid oxalate-extractable Al) was hardly affected by gypsum for all samples. Potassium chloride-extractable Al definitely decreased with the addition of gypsum in all soils; however, the decrease was small (0.1–1.4 cmol_c kg^?1) and the values still exceeded “the threshold of 2 cmol_c kg^?1” for inducing Al toxicity in sensitive plants (4.4–8.6 cmol_c Al kg^?1). The change in Al solubility with gypsum application represented by Al release rates from soils using continuous extraction methods with a dilute acetate buffer solution (10^?3 mol L^?1, pH 3.5) differed greatly among the soil samples: The release rate of one of the B horizon samples decreased by 71%, certainly showing the insolubilization of Al compounds, whereas the release rates of the A horizon sample showed almost no change. These changes in Al solubility were well correlated with the plant root growth. Root growth was improved with gypsum in the B horizon sample, whereas improvement was not observed in the A horizon soil. The decrease in the rate of Al release of another B horizon soil with gypsum treatment was smaller (by 20–34%), possibly because of lower pH values after gypsum application (pH[H₂O] of 4.2–4.3). In the B horizon soil, root growth improved only slightly. Thus, the effectiveness of gypsum application to acid Andosols appeared to be largely influenced by soil humus contents and slight differences in soil pH values, and corresponded to a decrease in Al release rates using the continuous extraction method.     

不同品种菠菜叶柄和叶片的硝态氮含量及其与植株生长的关系

温室盆栽试验研究了我国北方不同菠菜品种叶柄和叶片的硝态氮含量及其与植株生长的关系。结果表明,30个菠菜品种地上部分的生长量和硝态氮含量存在显著差异。叶柄和叶片在反映品种间生长量和硝态氮含量变异方面的作用并不相同。叶片占植株地上部鲜重的比例高于叶柄,品种间叶片生长量的差异亦大于叶柄,叶片与植株生长量的正相关关系更为显著。但与生长量的情况不同,叶柄的硝态氮含量、累积总量均显著高于叶片,是菠菜累积硝态氮的主要器官。叶柄硝态氮含量的品种间差异远大于叶片,与植株地上部硝态氮含量的正相关性更为显著。菠菜不同品种之间,叶柄硝态氮含量与地上部鲜重、干重及水分均表现出显著的正相关关系,而叶片硝态氮含量与植株生物量及其各组分之间却无这种关系。     

Cadmium fractions in an acid sandy soil and Cd in soil solution as affected by plant growth

In a previous experiment, plants were able to immobilize or solubilize Cadmium (Cd) in a sandy acid soil enriched with 40 μmol Cd kg^–1, because Cd solution concentration was decreased by maize (Zea mays) and sunflower (Helianthus annuus), and increased by flax (Linum usitatissimum L. ssp. usitatissimum) and spinach (Spinacia oleracea). It is assumed that the equilibrium with Cd fractions in the soil solid phase and the chemical form of Cd in the soil solution were affected. In the present study, the effect of the four plant species mentioned above on Cd binding in soil was investigated by means of a fractionation of soil Cd with a sequential extraction of seven steps. The seven fractions of Cd are operationally defined by the extraction sequence that follows the order of increasing acidity with extractants of different complexing and redox properties. In the unplanted soil, Cd was predominantly present in the exchangeable Fraction I (F. I) and easily mobilizable Fraction II (F. II) (64%). Significant concentrations of Cd were found in F. III (occluded in Mn oxides; 22%) and F. IV (organically bound; 10%). Fractions V (occluded in poorly crystalline Fe oxides), F. VI (occluded in well crystallized Fe oxides), and F. VII (residual fraction) amounted to less than 5% of the total soil Cd concentration. The plants changed the binding of Cd in soil in a different manner. All plants decreased F. I, but F. II was increased by maize and spinach, decreased by flax or remained unaffected by sunflower. Fraction III was not affected by maize and flax, but decreased by sunflower and spinach, and F. IV was not affected by sunflower and spinach, but was increased by maize and flax. These changes of Cd fractions were not related to the changes the plants had caused in total Cd or Cd²⁺ concentration of the soil solution. These results show that plant species differ in how they affect Cd binding to the soil solid phase, but this effect is not related to how they affect Cd in soil solution. The mechanisms by which plants affect the relationship between the soil solid and liquid phase are still unclear.     

Comparison of in vitro solubilization activity of diverse phosphate-solubilizing bacteria native to acid soil and their ability to promote Phaseolus vulgaris growth

To identify plant growth promotion ability of phosphorus-solubilizing native bacteria, we have examined a collection of isolates representing the diversity of culturable phosphate-solubilizing bacteria from acid soils of the northeast of Argentina. Assays in growth medium supplemented with tricalcium phosphate revealed different phosphorus solubilization activity and temporal patterns of solubilization. Acidification of the broth medium coincided with phosphorus solubilization. The isolates were grouped according to their Rep fingerprinting profiles and phylogenetically classified by 16S rDNA and biochemical analyses. These isolates were assigned to the genera Enterobacter, Pantoea, Pseudomonas, Acinetobacter, Burkholderia, and Exiguobacterium. Four isolates showing high phosphorus solubilizing activity in in vitro assays were inoculated on common beans (Phaseolus vulgaris); some of them promoted plant growth and increased photosynthesis and the P and N content of leaves. The results indicated that the ability to in vitro solubilize P is not necessarily associated to the promotion of plant growth.