Plant Growth-Promoting Rhizobacteria Alleviate High Salinity Impact on the Halophyte Suaeda fruticosa by Modulating Antioxidant Defense and Soil Biological Activity.

Plant growth-promoting rhizobacteria (PGPR) are considered as bio-ameliorators that confer better salt resistance to host plants while improving soil biological activity. Despite their importance, data about the likely synergisms between PGPR and halophytes in their native environments are scarce. The objective of this study was to assess the effect of PGPR (Glutamicibacter sp. and Pseudomonas sp.) inoculation on biomass, nutrient uptake, and antioxidant enzymes of Suaeda fruticosa, an obligate halophyte native in salt marshes and arid areas in Tunisia. Besides, the activity of rhizospheric soil enzyme activities upon plant inoculation was determined. Plants were grown in pots filled with soil and irrigated with 600 mM NaCl for 1 month. Inoculation (either with Pseudomonas sp. or Glutamicibacter sp.) resulted in significantly higher shoot dry weight and less accumulation of Na+ and Cl- in shoots of salt-treated plants. Glutamicibacter sp. inoculation significantly reduced malondialdehyde (MDA) concentration, while increasing the activity of antioxidant enzymes (superoxide dismutase; catalase; ascorbate peroxidase; and glutathione reductase) by up to 100%. This provides strong arguments in favor of a boosting effect of this strain on S. fruticosa challenged with high salinity. Pseudomonas sp. inoculation increased shoot K+ and Ca2+ content and lowered shoot MDA concentration. Regarding the soil biological activity, Pseudomonas sp. significantly enhanced the activities of three rhizospheric soil enzymes (urease, ß-glucosidase, and dehydrogenase) as compared to their respective non-inoculated saline treatment. Hence, Pseudomonas sp. could have a great potential to be used as bio-inoculants in order to improve plant growth and soil nutrient uptake under salt stress. Indole-3-acetic acid concentration in the soil increased in both bacterial treatments under saline conditions, especially with Glutamicibacter sp. (up to +214%). As a whole, Glutamicibacter sp. and Pseudomonas sp. strains are promising candidates as part of biological solutions aiming at the phytoremediation and reclamation of saline-degraded areas.

Showing their mettle: extraradical mycelia of arbuscular mycorrhizae form a metal filter to improve host Al tolerance and P nutrition.

BACKGROUND: New evidence has shown that arbuscular mycorrhizal (AM) fungi can contribute to the aluminum (Al3+ ) tolerance of host plants growing in acidic soils with phytotoxic levels of Al3+ . The aim of this study was to investigate the role of AM fungi isolated from naturally occurring Al3+ acidic soils in conferring host tolerance to Al3+ toxicity in three wheat cultivars differing in Al3+ sensitivity. The experiment was conducted in a soilless substrate (vermiculite/perlite, 2:1 v/v) using two Al3+ -tolerant wheat genotypes and one Al3+ -sensitive wheat genotype. The wheat was colonized with a consortium of AM fungi isolated from an Andisol, with or without Al3+ at a concentration of 200 µmol L-1 . RESULTS: The response of wheat to Al3+ in the medium was dependent on both the plant genotype and AM colonization. The benefits of the AM fungi to the wheat cultivars included an increased P concentration and relatively low Al3+ accumulation in the plants. This was achieved through two mechanisms. First, the metal-chelating capacity of the AM fungi was clear in two of the cultivars ('Tukan' and 'Porfiado'), in which the enhanced extraradical mycelium development was able to retain Al3+ in the glomalin and hyphae. Second, the increased AM-induced acid phosphatase activity in the rhizosphere of the other cultivar ('Atlas 66') increased host nutrition possibly by hyphae-mediated nutrient uptake and glomalin-related soil protein. CONCLUSION: The results suggest that the role of AM fungi in cultivar-specific Al3+ detoxification can be achieved by increased extraradical mycelial filters and enhanced bioavailability of P in the host rhizosphere. © 2019 Society of Chemical Industry.

Phenotypic and molecular traits determine the tolerance of olive trees to drought stress.

Olive trees are known for their capacity to adapt to drought through several phenotypic and molecular variations, although this can vary according to the different provenances of the same olive cultivar. We confronted the same olive cultivar from two different location in Spain: Freila, in the Granada province, with low annual precipitation, and Grazalema, in the Cadiz province, with high annual precipitation, and subjected them to five weeks of severe drought stress. We found distinctive physiological and developmental adaptations among the two provenances. Thus, trees from Freila subjected to drought stress exhibited increasing root dry weights and decreasing leaf numbers and relative stem heights. On the other hand, the treatment with drought in Grazalema trees reduced their leaf chlorophyll contents, but increased their relative stem diameter and their root hydraulic conductivity. The physiological responses of Freila tree roots to drought were linked to different molecular adaptations that involved the regulation of genes related to transcription factors induced by ABA, auxin and ethylene signaling, as well as, the action of a predicted membrane intrinsic protein (MIP). On the other hand, the responses of Grazalema trees were related with different root genes related to oxidation-reduction, ATP synthesis, transduction and posttranslational regulation, with a special mention to the cytokinins signaling through the transcript predicted as a histidine-containing phosphotransfer protein. Our results show that olive trees adapted to dry environments will adjust their growth and water uptake capacity through transcription factors regulation, and this will influence the different physiological responses to drought stress.

Rhizobial symbiosis modifies root hydraulic properties in bean plants under non-stressed and salinity-stressed conditions.

MAIN CONCLUSION: Rhizobial symbiosis improved the water status of bean plants under salinity-stress conditions, in part by increasing their osmotic root water flow. One of the main problems for agriculture worldwide is the increasing salinization of farming lands. The use of soil beneficial microorganisms stands up as a way to tackle this problem. One approach is the use of rhizobial N2-fixing, nodule-forming bacteria. Salinity-stress causes leaf dehydration due to an imbalance between water lost through stomata and water absorbed by roots. The aim of the present study was to elucidate how rhizobial symbiosis modulates the water status of bean (Phaseolus vulgaris) plants under salinity-stress conditions, by assessing the effects on root hydraulic properties. Bean plants were inoculated or not with a Rhizobium leguminosarum strain and subjected to moderate salinity-stress. The rhizobial symbiosis was found to improve leaf water status and root osmotic water flow under such conditions. Higher content of nitrogen and lower values of sodium concentration in root tissues were detected when compared to not inoculated plants. In addition, a drop in the osmotic potential of xylem sap and increased amount of PIP aquaporins could favour higher root osmotic water flow in the inoculated plants. Therefore, it was found that rhizobial symbiosis may also improve root osmotic water flow of the host plants under salinity stress.

Native bacteria promote plant growth under drought stress condition without impacting the rhizomicrobiome.

Inoculation of plants with beneficial plant growth-promoting bacteria (PGPB) emerges a valuable strategy for ecosystem recovery. However, drought conditions might compromise plant-microbe interactions especially in semiarid regions. This study highlights the effect of native PGPB after 1 year inoculation on autochthonous shrubs growth and rhizosphere microbial community composition and activity under drought stress conditions. We inoculated three plant species of semiarid Mediterranean zones, Thymus vulgaris, Santolina chamaecyparissus and Lavandula dentata with a Bacillus thuringiensis strain IAM 12077 and evaluated the impact on plant biomass, plant nutrient contents, arbuscular mycorrhiza fungi (AMF) colonization, soil rhizosphere microbial activity and both the bacterial and fungal communities. Inoculation with strain IAM 12077 improved the ability of all three plants species to uptake nutrients from the soil, promoted L. dentata shoot growth (>65.8%), and doubled the AMF root colonization of S. chamaecyparissus. Inoculation did not change the rhizosphere microbial community. Moreover, changes in rhizosphere microbial activity were mainly plant species-specific and strongly associated with plant nutrients. In conclusion, the strain IAM 12077 induced positive effects on plant growth and nutrient acquisition with no impact on the rhizosphere microbiome, indicating a rhizosphere microbial community resilient to native bacteria inoculation.

Impact of microbial inoculation on biomass accumulation by Sulla carnosa provenances, and in regulating nutrition, physiological and antioxidant activities of this species under non-saline and saline conditions.

Bacteria (Pseudomonas sp. and Bacillus sp.) and/or the arbuscular mycorrhizal (AM) fungus Rhizophagus intraradices were able to improve growth, physiological and biochemical characteristics of four Sulla carnosa Desf. provenances (Sidi khlif, Thelja, Kalbia and Kerker) from Tunisia under both saline and non-saline conditions. S. carnosa is a salt-tolerant legume plant, native from North Africa. The intrinsic bacterial characteristics evidenced the fitness of these bacteria to support salt stress and to stimulate plant growth. Bacillus sp. produced more indol acetic acid (IAA) than Pseudomonas sp. and showed a great surviving capacity under salt conditions supporting its capacity to improve plant growth under stress conditions. The microorganisms applied also have a different potential to increase the nutritional and related plant growth parameters. It is noticeable that some provenances reached the highest level of growth when inoculated with Bacillus sp. in Sidi khlif or by Bacillus plus AMF in Kalbia, which increased shoot by 318% and root by 774%. In contrast, in Thelja and Kerker the impact of the test microorganisms was mainly evidenced at increasing nutritional and physiological functions. Salinity reduced some growth and physiological variables as stomatal conductance, photosynthetic pigments and photosynthetic efficiency and increased electrolyte leakage. However, the microbial inoculants compensated these detrimental effects in a degree depending on the S. carnosa provenance. These microorganisms also orchestrate antioxidant activities involved in adaptative responses in S. carnosa provenances. The intrinsic ability of inoculants allow us to select the provenance/microorganism combination which maximizes S. carnosa growth, nutrition and physiological/biochemical responses under salt and non-salt conditions. The results obtained support that the target microbial inocula are beneficial for the ecological stability if this Mediterranean legume.

Effects of different arbuscular mycorrhizal fungal backgrounds and soils on olive plants growth and water relation properties under well-watered and drought conditions.

The adaptation capacity of olive trees to different environments is well recognized. However, the presence of microorganisms in the soil is also a key factor in the response of these trees to drought. The objective of the present study was to elucidate the effects of different arbuscular mycorrhizal (AM) fungi coming from diverse soils on olive plant growth and water relations. Olive plants were inoculated with native AM fungal populations from two contrasting environments, that is, semi-arid - Freila (FL) and humid - Grazalema (GZ) regions, and subjected to drought stress. Results showed that plants grew better on GZ soil inoculated with GZ fungi, indicating a preference of AM fungi for their corresponding soil. Furthermore, under these conditions, the highest AM fungal diversity was found. However, the highest root hydraulic conductivity (Lpr ) value was achieved by plants inoculated with GZ fungi and growing in FL soil under drought conditions. So, this AM inoculum also functioned in soils from different origins. Nine novel aquaporin genes were also cloned from olive roots. Diverse correlation and association values were found among different aquaporin expressions and abundances and Lpr , indicating how the interaction of different aquaporins may render diverse Lpr values.

Regulation of cation transporter genes by the arbuscular mycorrhizal symbiosis in rice plants subjected to salinity suggests improved salt tolerance due to reduced Na(+) root-to-shoot distribution.

Rice is a salt-sensitive crop whose productivity is strongly reduced by salinity around the world. Plants growing in saline soils are subjected to the toxicity of specific ions such as sodium, which damage cell organelles and disrupt metabolism. Plants have evolved biochemical and molecular mechanisms to cope with the negative effects of salinity. These include the regulation of genes with a role in the uptake, transport or compartmentation of Na(+) and/or K(+). Studies have shown that the arbuscular mycorrhizal (AM) symbiosis alleviates salt stress in several host plant species. However, despite the abundant literature showing mitigation of ionic imbalance by the AM symbiosis, the molecular mechanisms involved are barely explored. The objective of this study was to elucidate the effects of the AM symbiosis on the expression of several well-known rice transporters involved in Na(+)/K(+) homeostasis and measure Na(+) and K(+) contents and their ratios in different plant tissues. Results showed that OsNHX3, OsSOS1, OsHKT2;1 and OsHKT1;5 genes were considerably upregulated in AM plants under saline conditions as compared to non-AM plants. Results suggest that the AM symbiosis favours Na(+) extrusion from the cytoplasm, its sequestration into the vacuole, the unloading of Na(+) from the xylem and its recirculation from photosynthetic organs to roots. As a result, there is a decrease of Na(+) root-to-shoot distribution and an increase of Na(+) accumulation in rice roots which seems to enhance the plant tolerance to salinity and allows AM rice plants to maintain their growing processes under salt conditions.

Autochthonous arbuscular mycorrhizal fungi and Bacillus thuringiensis from a degraded Mediterranean area can be used to improve physiological traits and performance of a plant of agronomic interest under drought conditions.

Studies have shown that some microorganisms autochthonous from stressful environments are beneficial when used with autochthonous plants, but these microorganisms rarely have been tested with allochthonous plants of agronomic interest. This study investigates the effectiveness of drought-adapted autochthonous microorganisms [Bacillus thuringiensis (Bt) and a consortium of arbuscular mycorrhizal (AM) fungi] from a degraded Mediterranean area to improve plant growth and physiology in Zea mays under drought stress. Maize plants were inoculated or not with B. thuringiensis, a consortium of AM fungi or a combination of both microorganisms. Plants were cultivated under well-watered conditions or subjected to drought stress. Several physiological parameters were measured, including among others, plant growth, photosynthetic efficiency, nutrients content, oxidative damage to lipids, accumulation of proline and antioxidant compounds, root hydraulic conductivity and the expression of plant aquaporin genes. Under drought conditions, the inoculation of Bt increased significantly the accumulation of nutrients. The combined inoculation of both microorganisms decreased the oxidative damage to lipids and accumulation of proline induced by drought. Several maize aquaporins able to transport water, CO2 and other compounds were regulated by the microbial inoculants. The impact of these microorganisms on plant drought tolerance was complementary, since Bt increased mainly plant nutrition and AM fungi were more active improving stress tolerance/homeostatic mechanisms, including regulation of plant aquaporins with several putative physiological functions. Thus, the use of autochthonous beneficial microorganisms from a degraded Mediterranean area is useful to protect not only native plants against drought, but also an agronomically important plant such as maize.

Microbial inoculants and organic amendment improves plant establishment and soil rehabilitation under semiarid conditions.

The re-establishment of autochthonous shrub species is an essential strategy for recovering degraded soils under semiarid Mediterranean conditions. A field assay was carried out to determine the combined effects of the inoculation with native rhizobacteria (Bacillus megaterium, Enterobacter sp, Bacillus thuringiensis and Bacillus sp) and the addition of composted sugar beet (SB) residue on physicochemical soil properties and Lavandula dentata L. establishment. One year after planting, Bacillus sp. and B. megaterium + SB were the most effective treatments for increasing shoot dry biomass (by 5-fold with respect to control) and Enterobacter sp + SB was the most effective treatments for increasing dry root biomass. All the treatments evaluated significantly increased the foliar nutrient content (NPK) compared to control values (except B. thuringiensis + SB). The organic amendment had significantly increased available phosphorus content in rhizosphere soil by 29% respect to the control. Enterobacter sp combined with sugar beet residue improved total N content in soil (by 46% respect to the control) as well as microbiological and biochemical properties. The selection of the most efficient rhizobacteria strains and their combined effect with organic residue seems to be a critical point that drives the effectiveness of using these biotechnological tools for the revegetation and rehabilitation of degraded soils under semiarid conditions.

Differential activity of autochthonous bacteria in controlling drought stress in native Lavandula and Salvia plants species under drought conditions in natural arid soil.

The effectiveness of autochthonous plant growth-promoting rhizobacteria was studied in Lavandula dentata and Salvia officinalis growing in a natural arid Mediterranean soil under drought conditions. These bacteria identified as Bacillus megaterium (Bm), Enterobacter sp. (E), Bacillus thuringiensis (Bt), and Bacillus sp. (Bsp). Each bacteria has different potential to meliorate water limitation and alleviating drought stress in these two plant species. B. thuringiensis promoted growth and drought avoidance in Lavandula by increasing K content, by depressing stomatal conductance, and it controlled shoot proline accumulation. This bacterial effect on increasing drought tolerance was related to the decrease of glutathione reductase (GR) and ascorbate peroxidase (APX) that resulted sensitive indexes of lower cellular oxidative damage involved in the adaptative drought response in B. thuringiensis-inoculated Lavandula plants. In contrast, in Salvia, having intrinsic lower shoot/root ratio, higher stomatal conductance and lower APX and GR activities than Lavandula, the bacterial effects on nutritional, physiological and antioxidant enzymatic systems were lower. The benefit of bacteria depended on intrinsic stress tolerance of plant involved. Lavadula demonstrated a greater benefit than Salvia to control drought stress when inoculated with B. thuringiensis. The bacterial drought tolerance assessed as survival, proline, and indolacetic acid production showed the potential of this bacteria to help plants to grow under drought conditions. B. thuringiensis may be used for Lavandula plant establishment in arid environments. Particular characteristic of the plant species as low shoot/root ratio and high stomatal conductance are important factors controlling the bacterial effectiveness improving nutritional, physiological, and metabolic plant activities.

Plant potassium content modifies the effects of arbuscular mycorrhizal symbiosis on root hydraulic properties in maize plants.

It is well known that the arbuscular mycorrhizal (AM) symbiosis helps the host plant to overcome several abiotic stresses including drought. One of the mechanisms for this drought tolerance enhancement is the higher water uptake capacity of the mycorrhizal plants. However, the effects of the AM symbiosis on processes regulating root hydraulic properties of the host plant, such as root hydraulic conductivity and plasma membrane aquaporin gene expression, and protein abundance, are not well defined. Since it is known that K(+) status is modified by AM and that it regulates root hydraulic properties, it has been tested how plant K(+) status could modify the effects of the symbiosis on root hydraulic conductivity and plasma membrane aquaporin gene expression and protein abundance, using maize (Zea mays L.) plants and Glomus intraradices as a model. It was observed that the supply of extra K(+) increased root hydraulic conductivity only in AM plants. Also, the different pattern of plasma membrane aquaporin gene expression and protein abundance between AM and non-AM plants changed with the application of extra K(+). Thus, plant K(+) status could be one of the causes of the different observed effects of the AM symbiosis on root hydraulic properties. The present study also highlights the critical importance of AM fungal aquaporins in regulating root hydraulic properties of the host plant.

Azospirillum and arbuscular mycorrhizal colonization enhance rice growth and physiological traits under well-watered and drought conditions.

The response of rice plants to inoculation with an arbuscular mycorrhizal (AM) fungus, Azospirillum brasilense, or combination of both microorganisms, was assayed under well-watered or drought stress conditions. Water deficit treatment was imposed by reducing the amount of water added, but AM plants, with a significantly higher biomass, received the same amount of water as non-AM plants, with a poor biomass. Thus, the water stress treatment was more severe for AM plants than for non-AM plants. The results showed that AM colonization significantly enhanced rice growth under both water conditions, although the greatest rice development was reached in plants dually inoculated under well-watered conditions. Water level did not affect the efficiency of photosystem II, but both AM and A. brasilense inoculations increased this value. AM colonization increased stomatal conductance, particularly when associated with A. brasilense, which enhanced this parameter by 80% under drought conditions and by 35% under well-watered conditions as compared to single AM plants. Exposure of AM rice to drought stress decreased the high levels of glutathione that AM plants exhibited under well-watered conditions, while drought had no effect on the ascorbate content. The decrease of glutathione content in AM plants under drought stress conditions led to enhance lipid peroxidation. On the other hand, inoculation with the AM fungus itself increased ascorbate and proline as protective compounds to cope with the harmful effects of water limitation. Inoculation with A. brasilense also enhanced ascorbate accumulation, reaching a similar level as in AM plants. These results showed that, in spite of the fact that drought stress imposed by AM treatments was considerably more severe than non-AM treatments, rice plants benefited not only from the AM symbiosis but also from A. brasilense root colonization, regardless of the watering level. However, the beneficial effects of A. brasilense on most of the physiological and biochemical traits of rice plants were only clearly visible when the plants were mycorrhized. This microbial consortium was effective for rice plants as an acceptable and ecofriendly technology to improve plant performance and development.

Enhancement of clover growth by inoculation of P-solubilizing fungi and arbuscular mycorrhizal fungi

This study evaluated the synergism between several P-solubilizing fungi isolates and arbuscular mycorrhizal fungi to improve clover ( Trifolium pratense) growth in the presence of Araxá apatite. Clover was sown directly in plastic pots with 300g of sterilized washed sand, vermiculite and sepiolite 1:1:1 (v:v:v) as substrate, and grown in a controlled environment chamber. The substrate was fertilized with 3 g L-1 of Araxá apatite. A completely randomized design, in 8×2 factorial scheme (eight P-solubilizing fungi treatments with or without arbuscular mycorrhizal fungi)and four replicates were used. The P-solubilizing fungi treatments consisted of five Brazilian P-solubilizing fungi isolates (PSF 7, 9, 20, 21 and 22), two Spanish isolates ( Aspergillus niger and the yeast Yarowia lipolytica) and control (non-inoculated treatment). The greatest clover growth rate was recorded when Aspergillus niger and PSF 21 were co-inoculated with arbuscular mycorrhizal fungi. Aspergillus niger, PSF 7 and PSF 21 were the most effective isolates on increasing clover growth in the presence of arbuscular mycorrhizal fungi. Greater mycorrhizal colonization resulted in greater clover growth rate in most PSF treatments. PSF 7 was the best isolate to improve the establishment of mycorrhizal and rhizobia symbiosis.

Este estudo avaliou o sinergismo entre diversos isolados defungos solubilizadores de fosfato e micorrízicos arbusculares para beneficiar o crescimento de trevo ( Trifolium pratense) na presença de apatita de Araxá. A cultura foi semeada diretamente em potes plásticos com 300 g de substrato esterilizado formado por areia lavada, vermiculita e sepiolita 1:1:1 (v:v:v) e cultivada em câmara climática. O substrato foi fertilizado com 3 g L-1 de apatita de Araxá. O experimento foi instalado em delineamento completamente casualizado, esquema fatorial 8×2 (oito tratamentos de inoculação de fungos solubilizadores de fosfato com ou sem fungos micorrízicos arbusculares) e quatro repetições. Os tratamentos de fungos solubilizadores de fosfato consistiram em cinco isolados brasileiros de fungos solubilizadores de fosfato (FSF 7, 9, 20, 21 e 22), dois isolados procedentes da Espanha ( Aspergillus niger e a levedura Yarowia lipolytica) e o controle (tratamento não inoculado). A maior taxa de crescimento da cultura foi obtida quando Aspergillus niger e FSF 21 foram co-inoculados com fungos micorrízicos arbusculares. Aspergillus niger, FSF 7 e o FSF 21 foram os isolados mais efetivos para incrementar o crescimento de trevo na presença de fungos micorrízicosarbusculares. A maior taxa de colonização micorrízica resultou em alta taxa de crescimento de trevo na maioria dos tratamentos com fungos solubilizadores de fosfato. O isolado FSF 7 foi o melhor para favorecer o estabelecimento das simbioses com fungos micorrízicos e com rizóbio.

Regulation of plasma membrane aquaporins by inoculation with a Bacillus megaterium strain in maize (Zea mays L.) plants under unstressed and salt-stressed conditions.

It is documented that some plant-growth-promoting rhizobacteria (PGPR) enhance plant salt tolerance. However, as to how PGPR may influence two crucial components of plant salt tolerance such as, root hydraulic characteristics and aquaporin regulation has been almost unexplored. Here, maize (Zea mays L.) plants were inoculated with a Bacillus megaterium strain previously isolated from a degraded soil and characterized as PGPR. Inoculated plants were found to exhibit higher root hydraulic conductance (L) values under both unstressed and salt-stressed conditions. These higher L values in inoculated plants correlated with higher plasma membrane type two (PIP2) aquaporin amount in their roots under salt-stressed conditions. Also, ZmPIP1;1 protein amount under salt-stressed conditions was higher in inoculated leaves than in non-inoculated ones. Hence, the different regulation of PIP aquaporin expression and abundance by the inoculation with the B. megaterium strain could be one of the causes of the different salt response in terms of root growth, necrotic leaf area, leaf relative water content and L by the inoculation treatment.

Interactions between Glomus species and Rhizobium strains affect the nutritional physiology of drought-stressed legume hosts.

The growth of legume plants is usually enhanced by the dual symbiosis of arbuscular mycorrhizal (AM) fungi and Rhizobium bacteria. However, most reports on this topic have been carried out under optimal water regime conditions. Here, four Phaseolus vulgaris varieties were single or dual inoculated with two different AM fungus and/or two different Rhizobium strains. All plants were grown under moderate drought conditions. Surprisingly, most of the biological treatments involving one fungus and one Rhizobium together caused a deleterious effect on plant growth. However, these negative effects were dependent on the P. vulgaris variety used as well as on the symbionts implicated. The results showed that AM symbiosis inhibited nodule development and N(2) fixation, causing diminution of plant growth. Therefore, under moderate drought conditions, the dual symbiosis formed by AM fungi and Rhizobium can be deleterious to P. vulgaris growth depending on the plant variety and the symbionts involved. Thus, under these common stress conditions, selection for the appropriated symbionts to each P. vulgaris variety is needed.

Enhancement of clover growth by inoculation of P-solubilizing fungi and arbuscular mycorrhizal fungi.

This study evaluated the synergism between several P-solubilizing fungi isolates and arbuscular mycorrhizal fungi to improve clover ( Trifolium pratense) growth in the presence of Araxá apatite. Clover was sown directly in plastic pots with 300g of sterilized washed sand, vermiculite and sepiolite 1:1:1 (v:v:v) as substrate, and grown in a controlled environment chamber. The substrate was fertilized with 3 g L(-1) of Araxá apatite. A completely randomized design, in 8×2 factorial scheme (eight P-solubilizing fungi treatments with or without arbuscular mycorrhizal fungi)and four replicates were used. The P-solubilizing fungi treatments consisted of five Brazilian P-solubilizing fungi isolates (PSF 7, 9, 20, 21 and 22), two Spanish isolates ( Aspergillus niger and the yeast Yarowia lipolytica) and control (non-inoculated treatment). The greatest clover growth rate was recorded when Aspergillus niger and PSF 21 were co-inoculated with arbuscular mycorrhizal fungi. Aspergillus niger, PSF 7 and PSF 21 were the most effective isolates on increasing clover growth in the presence of arbuscular mycorrhizal fungi. Greater mycorrhizal colonization resulted in greater clover growth rate in most PSF treatments. PSF 7 was the best isolate to improve the establishment of mycorrhizal and rhizobia symbiosis.

Arbuscular mycorrhizal fungi, Bacillus cereus, and Candida parapsilosis from a multicontaminated soil alleviate metal toxicity in plants.

We investigated if the limited development of Trifolium repens growing in a heavy metal (HM) multicontaminated soil was increased by selected native microorganisms, bacteria (Bacillus cereus (Bc)), yeast (Candida parapsilosis (Cp)), or arbuscular mycorrhizal fungi (AMF), used either as single or dual inoculants. These microbial inoculants were assayed to ascertain whether the selection of HM-tolerant microorganisms can benefit plant growth and nutrient uptake and depress HM acquisition. The inoculated microorganisms, particularly in dual associations, increased plant biomass by 148% (Bc), 162%, (Cp), and 204% (AMF), concomitantly producing the highest symbiotic (AMF colonisation and nodulation) rates. The lack of AMF colonisation and nodulation in plants growing in this natural, polluted soil was compensated by adapted microbial inoculants. The metal bioaccumulation abilities of the inoculated microorganisms and particularly the microbial effect on decreasing metal concentrations in shoot biomass seem to be involved in such effects. Regarding microbial HM tolerance, the activities of antioxidant enzymes known to play an important role in cell protection by alleviating cellular oxidative damage, such as superoxide dismutase, catalase, glutathione reductase, and ascorbate peroxidase, were here considered as an index of microbial metal tolerance. Enzymatic mechanisms slightly changed in the HM-adapted B. cereus or C. parapsilosis in the presence of metals. Antioxidants seem to be directly involved in the adaptative microbial response and survival in HM-polluted sites. Microbial inoculations showed a bioremediation potential and helped plants to develop in the multicontaminated soil. Thus, they could be used as a biotechnological tool to improve plant development in HM-contaminated environments.

Arbuscular mycorrhizal fungi increased growth, nutrient uptake and tolerance to salinity in olive trees under nursery conditions.

Inoculating olive plantlets with the arbuscular mycorrhizal fungi (AMF) Glomus mosseae, Glomus intraradices or Glomus claroideum increased plant growth and the ability to acquire nitrogen, phosphorus, and potassium from non-saline as well as saline media. AMF-colonized plants also increased in survival rate after transplant. Osmotic stress caused by NaCl supply reduced stem diameter, number of shoots, shoot length and nutrients in olive plants, but AMF colonization alleviated all of these negative effects on growth. G. mosseae was the most efficient fungus in reducing the detrimental effects of salinity; it increased shoot growth by 163% and root growth by 295% in the non-saline medium, and by 239% (shoot) and by 468% (root) under the saline conditions. AMF colonization enhanced salt tolerance in terms of olive growth and nutrient acquisition. Mycorrhizal olive plants showed the lowest biomass reduction under salinity (34%), while growth was reduced by 78% in control plants. This G. mosseae effect seems to be due to increased K acquisition; K content was enhanced under salt conditions by 6.4-fold with G. mosseae, 3.4-fold with G. intraradices, and 3.7-fold with G. claroideum. Potassium, as the most prominent inorganic solute, plays a key role in the osmoregulation processes and the highest salinity tolerance of G. mosseae-colonized olive trees was concomitant with an enhanced K concentration in olive plants.

Significance of treated agrowaste residue and autochthonous inoculates (Arbuscular mycorrhizal fungi and Bacillus cereus) on bacterial community structure and phytoextraction to remediate soils contaminated with heavy metals.

In this study, we analyzed the impact of treatments such as Aspergillus niger-treated sugar beet waste (SB), PO4(3-) fertilization and autochthonous inoculants [arbuscular mycorrhizal (AM) fungi and Bacillus cereus], on the bacterial community structure in a soils contaminated with heavy metals as well as, the effectiveness on plant growth (Trifolium repens). The inoculation with AM fungi in SB amended soil, increased plant growth similarly to PO4(3-) addition, and both treatments matched in P acquisition but bacterial biodiversity estimated by denaturing gradient gel electrophoresis of amplified 16S rDNA sequences, was more stimulated by the presence of the AM fungus than by PO4(3-) fertilization. The SB amendment plus AM inoculation increased the microbial diversity by 233% and also changed (by 215%) the structure of the bacterial community. The microbial inoculants and amendment used favoured plant growth and the phytoextraction process and concomitantly modified bacterial community in the rhizosphere; thus they can be used for remediation. Therefore, the understanding of such microbial ecological aspects is important for phytoremediation and the recovery of contaminated soils.