Microbial consortium inoculant increases pasture grasses yield in low-phosphorus soil by influencing root morphology, rhizosphere carboxylate exudation and mycorrhizal colonisation.

BACKGROUND: Pasture farming in south-western Australia is challenged by nutrient-poor soils. We assessed the impact of microbial consortium inoculant (MI) and rock mineral fertiliser (MF) on growth, nutrient uptake, root morphology, rhizosphere carboxylate exudation and mycorrhizal colonisation in three pasture grasses - tall fescue (Festuca arundinacea L.), veldt grass (Ehrharta calycina Sm.) and tall wheatgrass (Thinopyrum ponticum L.) grown in low-phosphorus (P) sandy soil in a glasshouse for 30 and 60 days after sowing (DAS). RESULTS: Veldt grass produced the highest specific root length and smallest average root diameter in both growth periods, and had similar shoot weight, root surface area and fine root length (except at 30 DAS) to tall fescue. Compared with the control, MI alone or combined with MF significantly increased shoot and root biomass (except root biomass at 30 DAS), likely due to the significant increases in root surface area and fine root length. Plants supplied with MI + MF had higher shoot N and P contents than those in the MI and the control treatments at 60 DAS. Malate, citrate and trans-aconitate were the major rhizosphere carboxylates exuded at both 30 and 60 DAS. Malate exudation varied among species and treatments in both growth periods, but citrate exudation was consistently higher in the low-P treatments (control and MI) than the MF and MI + MF treatments. CONCLUSION: Microbial consortium inoculant can positively influence pasture production in low-P soil by increasing root surface area and fine root length, whereas exudation of nutrient-mobilising carboxylates (citrate) is dependent more on soil P supply than microbial consortium inoculant. © 2021 Society of Chemical Industry.

Inoculation of Mimosa Pudica with Paraburkholderia phymatum Results in Changes to the Rhizoplane Microbial Community Structure.

Nitrogen fixing symbiosis between rhizobia and legumes contributes significant amounts of N to agricultural and natural environments. In natural soils, rhizobia compete with indigenous bacterial communities to colonize legume roots, which leads to symbiotic interactions. However, limited information is currently available on the effects of the rhizobial symbiont on the resident microbial community in the legume rhizosphere, rhizoplane, and endosphere, which is partly due to the presence of native nodulating rhizobial strains. In the present study, we used a symbiotic system comprised of Paraburkholderia phymatum and Mimosa pudica to examine the interaction of an inoculant strain with indigenous soil bacteria. The effects of a symbiont inoculation on the native bacterial community was investigated using high throughput sequencing and an analysis of 16S rRNA gene amplicons. The results obtained revealed that the inoculation induced significant alterations in the microbial community present in the rhizoplane+endosphere of the roots, with 13 different taxa showing significant changes in abundance. No significant changes were observed in the rhizospheric soil. The relative abundance of P. phymatum significantly increased in the rhizoplane+endosphere of the root, but significant decreased in the rhizospheric soil. While the rhizosphere, rhizoplane, and root endosphere contained a wide diversity of bacteria, the nodules were predominantly colonized by P. phymatum. A network analysis revealed that the operational taxonomic units of Streptomyces and Phycicoccus were positively associated with P. phymatum as potential keystone taxa. Collectively, these results suggest that the success of an inoculated symbiont depends on its ability to colonize the roots in the face of competition by other soil bacteria. A more detailed understanding of the mechanisms by which an inoculated strain colonizes its plant host is crucial for realizing the full potential of microbial inoculants in sustainable agriculture.

Phragmites australis in combination with hydrocarbons degrading bacteria is a suitable option for remediation of diesel-contaminated water in floating wetlands.

The presence of diesel in the water could reduce the growth of plant and thus phytoremediation efficacy. The toxicity of diesel to plant is commonly explained; because of hydrocarbons in diesel accumulate in various parts of plants, where they disrupt the plant cell especially, the epidemis, leaves, stem and roots of the plant. This study investigated the effect of bacterial augmentation in floating treatment wetlands (FTWs) on remediation of diesel oil contaminated water. A helophytic plant, Phragmites australis (P. australis), was vegetated on a floating mat to establish FTWs for the remediation of diesel (1%, w/v) contaminated water. The FTWs was inoculated with three bacterial strains (Acinetobacter sp. BRRH61, Bacillus megaterium RGR14 and Acinetobacter iwoffii AKR1), possessing hydrocarbon degradation and plant growth-enhancing capabilities. It was observed that the FTWs efficiently removed hydrocarbons from water, and bacterial inoculation further enhanced its hydrocarbons degradation efficacy. Diesel contaminated water samples collected after fifteen days of time interval for three months and were analyzed for pollution parameters. The maximum reduction in hydrocarbons (95.8%), chemical oxygen demand (98.6%), biochemical oxygen demand (97.7%), total organic carbon (95.2%), phenol (98.9%) and toxicity was examined when both plant and bacteria were employed in combination. Likewise, an increase in plant growth was seen in the presence of bacteria. The inoculated bacteria showed persistence in the water, root and shoot of P. australis. The study concluded that the augmentation of hydrocarbons degrading bacteria in FTWs is a better option for treatment of diesel polluted water.

Enhanced tomato plant growth in soil under reduced P supply through microbial inoculants and microbiome shifts.

Soil microbial communities interact with roots, affecting plant growth and nutrient acquisition. In the present study, we aimed to decipher the effects of the inoculants Trichoderma harzianum T-22, Pseudomonas sp. DSMZ 13134, Bacillus amyloliquefaciens FZB42 or Pseudomonas sp. RU47 on the rhizosphere microbial community and their beneficial effects on tomato plants grown in moderately low phosphorous soil under greenhouse conditions. We analyzed the plant mass, inoculant colony forming units and rhizosphere communities on 15, 22, 29 and 43 days after sowing. Selective plating showed that the bacterial inoculants had a good rhizocompetence and accelerated shoot and root growth and nutrient accumulation. 16S rRNA gene fingerprints indicated changes in the rhizosphere bacterial community composition. Amplicon sequencing revealed that rhizosphere bacterial communities from plants treated with bacterial inoculants were more similar to each other and distinct from those of the control and the Trichoderma inoculated plants at harvest time, and numerous dynamic taxa were identified. In conclusion, likely both, inoculants and the rhizosphere microbiome shifts, stimulated early plant growth mainly by improved spatial acquisition of available nutrients via root growth promotion. At harvest, all tomato plants were P-deficient, suggesting a limited contribution of inoculants and the microbiome shifts to the solubilization of sparingly soluble soil P.

Comparative study of plant growth-promoting bacteria on the physiology, growth and fruit quality of strawberry.

BACKGROUND: The strawberry (Fragaria × ananassa Duch.) is, among small fruits, the most cultivated and commercialized in Portugal. Recent studies have evidenced the positive effect of plant growth-promoting bacteria (PGPB) inoculation on strawberry production and, at the same time, provided an alternative strategy to reduce the use of fertilizers. In this study the effects of root inoculation with three PGPB strains (Pedobacter sp. CC1, Bacillus safensis B106 and Bacillus subtilis B167A) on the physiology, growth, fruit production and quality of strawberry cv. Camarosa are presented. RESULTS: PGPB inoculation significantly accelerated crop maturation, with inoculated plants fruiting about 2 weeks earlier than non-inoculated plants. Inoculated plants with Pedobacter sp. CC1 and Bacillus safensis B106 influenced the gas exchange parameters of strawberry plants. The contents of total phenolics and flavonoids in strawberry leaves were found to be greater with Pedobacter sp. CC1, when compared with non-inoculated plants. Furthermore, plants inoculated with the same bacterial strain showed enhancement in the dimensions of fruits, especially fruit length, and shape as well as in the total soluble solids content (°Brix). CONCLUSIONS: The results showed that the PGPB Pedobacter sp. CC1 improved performance of strawberry plants, suggesting that it could be a potential biofertilizer for strawberry plant nutrition. © 2019 Society of Chemical Industry.

Trichoderma-Azotobacter biofilm inoculation improves soil nutrient availability and plant growth in wheat and cotton.

Microbial biofilms are gaining importance in agriculture, due to their multifaceted agronomic benefits and resilience to environmental fluctuations. This study focuses on comparing the influence of single inoculation-Azotobacter chroococcum (Az) or Trichoderma viride (Tv) and their biofilm (Tv-Az), on soil and plant metabolic activities in wheat and cotton grown under Phytotron conditions. Tv-Az proved superior to all the other treatments in terms of better colonisation, plant growth attributes and 10-40% enhanced availability of macronutrients and micronutrients in the soil, over control. Confocal and scanning electron microscopy showed that the cells attached to the root tips initially, followed by their proliferation along the surface of the roots. Soil polysaccharides, proteins and dehydrogenase activity showed several fold enhancement in Tv-Az biofilm inoculated samples. Time course studies revealed that the population of Az and Tv in the rhizoplane and rhizosphere was significantly higher with a 0.14-0.31 log colony-forming unit (CFU) increase in the biofilm-inoculated treatment in both crops. Enhancement in soil biological activities was facilitated by the improved colonisation of the biofilm, due to the synergistic association between Tv and Az. This demonstrates the utility of Tv-Az biofilm as a multifunctional plant growth promoting and soil fertility enhancing option in agriculture.

Effect of Vicia faba L. var. minor and Sulla coronaria (L.) Medik associated with plant growth-promoting bacteria on lettuce cropping system and heavy metal phytoremediation under field conditions.

Researches involving the use of association between legumes and PGPBs (plant growth-promoting bacteria) in heavy metal phytoremediation process were mainly performed for soils highly contaminated. However, even in agriculture soils, with moderate or low contamination levels, plants can accumulate high rates of heavy metals. So, food chain contamination by these metals presents a real threat to animal and human health. This work aimed to evaluate the use of two legumes/PGPB symbioses; Vicia faba var. minor and Sulla coronaria have been inoculated with specific heavy metal-resistant inocula in a crop rotation system with Lactuca sativa as a following crop, in order to assess their effects on soil fertility, lettuce yield, and heavy metal content. Our results showed that legume inoculation significantly enhanced their biomass production, nitrogen and phosphorus content. The use of our symbioses as green manure before lettuce cultivation, as a rotation cropping system, affected positively soil fertility. In fact, we recorded a higher organic matter content, with rapid decomposition in the soil of inoculated plots. Besides, results demonstrated a greater nitrogen and phosphorus content in this soil, especially in the plot cultivated with inoculated V. faba var. minor. The improvement of soil fertility enhanced lettuce yield and its nitrogen and phosphorus content. Moreover, inoculated legumes extracted and accumulated more heavy metals than non-inoculated legumes. Our symbioses play the role of organic trap for heavy metals, making them unavailable for following crops. These facts were supported by lettuce heavy metal content, showing a significant decrease in metal accumulation, mainly zinc and cadmium, in edible parts. Results showed the usefulness of the studied symbioses, as a main part of a rotation system with lettuce. Our symbioses can be suggested for agriculture soil phytoremediation, aiming to enhance non-legume crop yield and limit heavy metal translocation to food chain.

Colonization and Competition Dynamics of Plant Growth-Promoting/Inhibiting Bacteria in the Phytosphere of the Duckweed Lemna minor.

Despite the considerable role of aquatic plant-associated bacteria in host plant growth and nutrient cycling in aquatic environments, the mode of their plant colonization has hardly been understood. This study examined the colonization and competition dynamics of a plant growth-promoting bacterium (PGPB) and two plant growth-inhibiting bacteria (PGIB) in the aquatic plant Lemna minor (common duckweed). When inoculated separately to L. minor, each bacterial strain quickly colonized at approximately 106 cells per milligram (plant fresh weight) and kept similar populations throughout the 7-day cultivation time. The results of two-membered co-inoculation assays revealed that the PGPB strain Aquitalea magnusonii H3 consistently competitively excluded the PGIB strain Acinetobacter ursingii M3, and strain H3 co-existed at almost 1:1 proportion with another PGIB strain, Asticcacaulis excentricus M6, regardless of the inoculation ratios (99:1-1:99) and inoculation order. We also found that A. magnusonii H3 exerted its growth-promoting effect over the negative effects of the two PGIB strains even when only a small amount was inoculated, probably due to its excellent competitive colonization ability. These experimental results demonstrate that there is a constant ecological equilibrium state involved in the bacterial colonization of aquatic plants.

Soybean inoculants in Brazil: an overview of quality control.

The bacterial strains SEMIA 587 and 5019 (Bradyrhizobium elkanii), 5079 (Bradyrhizobium japonicum), and 5080 (Bradyrhizobium diazoefficiens) are recommended for soybean inoculants in Brazil. In several countries, the current regulations are insufficient to induce companies for improving the quality of their products, leading to low performance and subsequent abandonment of inoculant use. From 2010 to 2014, 1086 samples coming mainly from Argentina and the southern region of Brazil were analyzed for viable cells counting, strains identification, and purity analysis according to the SDA/MAPA no. 30/2010 Normative Instruction. Most products were imported and formulated in liquid carriers with 5.0 × 109 colony-forming units (CFU)/mL. The strains most frequently used were SEMIA 5079/5080. Only 2.21% of samples had contaminants. The guaranteed concentration of viable cells in inoculants mostly ranged from 4.1 × 109 to 5.0 × 109 CFU/mL or CFU/g. The most frequently found concentration was above 1.1 × 1010 CFU/mL or CFU/g, which was higher than the product guarantee. The inoculants used for soybean crop in Brazil have excellent quality, leading the country to the leadership in taking advantage of the biological nitrogen fixation benefits for a productive and sustainable agriculture.

Developing tools for evaluating inoculation methods of biocontrol Streptomyces sp. strains into grapevine plants.

The endophytic Streptomyces sp. VV/E1, and rhizosphere Streptomyces sp. VV/R4 strains, isolated from grapevine plants were shown in a previous work to reduce the infection rate of fungal pathogens involved in young grapevine decline. In this study we cloned fragments from randomly amplified polymorphic DNA (RAPD), and developed two stably diagnostic sequence-characterized amplified region (SCAR) markers of 182 and 160 bp for the VV/E1 and VV/R4 strains, respectively. The SCAR markers were not found in another 50 actinobacterial strains isolated from grapevine plants. Quantitative real-time PCR protocols based on the amplification of these SCAR markers were used for the detection and quantification of both strains in plant material. These strains were applied on young potted plants using two methods: perforation of the rootstock followed by injection of the microorganisms or soaking the root system in a bacterial suspension. Both methods were combined with a booster treatment by direct addition of a bacterial suspension to the soil near the root system. Analysis of uprooted plants showed that those inoculated by injection exhibited the highest rate of colonization. In contrast, direct addition of either strain to the soil did not lead to reliable colonization. This study has developed molecular tools for analyzing different methods for inoculating grapevine plants with selected Streptomyces sp. strains which protect them from fungal infections that enter through their root system. These tools are of great applied interest since they could easily be established in nurseries to produce grafted grapevine plants that are protected against fungal pathogens. Finally, this methodology might also be applied to other vascular plants for their colonization with beneficial biological control agents.

In vitro assessment of the factors that determine the activity of the rumen microbiota for further applications as inoculum.

BACKGROUND: The rumen microbiota has been used as inoculum for in vitro studies and as a probiotic to improve productivity in young animals. However, great variability across studies has been noted depending on the inoculum considered. The present study aims to assess the relevance of different factors (microbial fraction, collection time, donor animal diet, fermentation substrate and inoculum preservation method) to maximize the rumen inoculum activity and set the standards for further in vitro and in vivo applications. RESULTS: Rumen inoculum sampled at 3 h after feeding led to greater microbial growth and activity [+12% volatile fatty acid (VFA), +17% ammonia] compared to before feeding. Similar results were noted when rumen liquid or rumen content were used as inocula. Rumen inoculum adapted to concentrate diets increased microbial activity (+19% VFA) independently of the substrate used in vitro. Freezing-thawing the inoculum, in comparison to fresh inoculum, decreased microbial activity (-14% VFA, -96% ammonia), anaerobic fungi and protozoa, with holotrichs protozoa being particularly vulnerable. Inoculum lyophilization had a stronger negative effect on microbial activity (-51% VFA) and delayed re-activation of the microbes, leading to lower levels of methanogens and anaerobic fungi, as well as almost complete wipe out of rumen protozoa. CONCLUSIONS: Fresh rumen fluid sampled at 3 h after feeding from donor animals that were fed concentrate diets should be chosen when the aim is to provide the most diverse and active rumen microbial inoculum. © 2018 Society of Chemical Industry.

Aqueous peat extract exposes rhizobia to sub-lethal stress which may prime cells for improved desiccation tolerance.

Inoculation of legume seed with rhizobia is an efficient and cost-effective means of distributing elite rhizobial strains to broad-acre crops and pastures. However, necessary drying steps after coating seed expose rhizobia to desiccation stress reducing survival and limiting potential nitrogen fixation by legumes. Rhizobial tolerance to desiccation varies with strain and with growth conditions prior to drying. Cells grown in peat generally survive desiccation better than cells grown in liquid broth. We aimed to identify peat-induced proteomic changes in rhizobia that may be linked to desiccation tolerance. Proteins expressed differentially after growth in peat extract when compared with a minimal defined medium were measured in four rhizobial strains. Proteins showing the greatest increase in abundance were those involved in amino acid and carbohydrate transport and metabolism. Proteins involved in posttranslational modification and cell defence mechanisms were also upregulated. Many of the proteins identified in this study have been previously linked to stress responses. In addition, analysis using nucleic acid stains SYTO9 and propidium iodide indicated that membranes had been compromised after growth in peat extract. We targeted the membrane repair protein PspA (ΔRL3579) which was upregulated in Rhizobium leguminosarum bv. viceae 3841 after growth in peat extract to validate whether the inability to repair membrane damage after growth in peat extract reduced desiccation tolerance. The ΔRL3579 mutant grown in peat extract had significantly lower survival under desiccation stress, whereas no difference in survival between wild-type and mutant strains was observed after growth in tryptone yeast (TY) or minimal medium (JMM) media. Staining mutant and wild-type strains with SYTO9 and propidium iodide indicated that membranes of the mutant were compromised after growth in peat extract and to a lesser extent in TY. This study shows that growth in peat extract causes damage to cell membranes and exposes rhizobia to sub-lethal stress resulting in differential expression of several stress-induced proteins. The induction of these proteins may prime and protect the cells when subjected to subsequent stress such as desiccation. Identifying the key proteins involved in desiccation tolerance and properties of peat that stimulate this response will be important to inform development of new inoculant technology that maximises survival of rhizobia during delivery to legume crops and pastures.

Colonization on Cucumber Root and Enhancement of Chlorimuron-ethyl Degradation in the Rhizosphere by Hansschlegelia zhihuaiae S113 and Root Exudates.

The colonization of Hansschlegelia zhihuaiae S113 and its degradation of the herbicide chlorimuron-ethyl in the cucumber rhizosphere was investigated. The results reveal that S113 colonized the cucumber roots (2.14 × 105cells per gram of roots) and were able to survive in the rhizosphere (maintained for 20 d). The root exudates promoted colonization on roots and increased the degradation of chlorimuron-ethyl by S113. Five organic acids in cucumber-root exudates were detected and identified by HPLC. Citric acid and fumaric acid significantly stimulated S113 colonization on cucumber roots, with 18.4 and 15.5% increases, respectively, compared with the control. After irrigation with an S113 solution for 10 days, chlorimuron-ethyl could not be detected in the roots, seedlings, or rhizosphere soil, which allowed for improved cucumber growth. Therefore, the degradation mechanism of chlorimuron-ethyl residues by S113 in the rhizosphere could be applied in situ for the bioremediation of chlorimuron-ethyl contaminated soil to ensure crop safety.

Compost biofortification with diazotrophic and P-solubilizing bacteria improves maturation process and P availability.

BACKGROUND: Phosphorus-containing fertilizers play an important role in tropical agriculture owing to the well documented shortage of plant-available P in soils. Traditional P fertilizer production is based on chemical processing of insoluble rock phosphate (RP), which includes an acid treatment at high temperature. Processing the RP increases fertilizer costs, making it unavailable for undercapitalized and typically family-based farmers. Biotechnological methods have been proposed as an alternative to increase phosphate availability in RP. In this study, Burkholderia silvatlantica and Herbaspirillum seropedicae were co-inoculated into an RP-enriched compost with the aim of determining the effects of this technology on the levels of phosphatase activities and release of plant-available P. RESULTS: Inoculation of both microorganisms resulted in higher organic matter decomposition and higher humic acid formation in composting. Herbaspirillum seropedicae was the most promising microorganism for the production of acid and alkaline phosphatase enzymes. Both microorganisms presented potential to increase the supply of P from poorly soluble sources owing to increased levels of water-soluble P and citric acid P. CONCLUSION: Burkholderia silvatlantica and H. seropedicae in RP-enriched compost may represent an important biotechnological tool to reduce the overall time required for composting and increase the supply of P from poorly soluble sources. © 2016 Society of Chemical Industry.

New formulation and delivery method of Cryphonectria parasitica for biological control of chestnut blight.

AIMS: This study aimed to develop a new formulation of Cryphonectria parasitica hypovirulent mycelium suitable for inoculations of tall trees from the ground. Cryphonectria parasitica hypovirulent strains are widely used for biological control of chestnut blight. However, it is often inconsistent and ineffective not only for biological reasons but also because the current manual application of hypovirulent strains on adult plants is difficult, time-consuming and expensive. Here, we propose an improved formulation and more effective mode of application of hypovirulent strains, which could boost chestnut blight biocontrol. METHODS AND RESULTS: The Cp 4.2H hypovirulent strain was formulated as mycelium discs with polyethylene glycol and hydroxypropyl methylcellulose, loaded into lead-free pellets that are used as carriers to inoculate cankers on chestnut stems by shooting. The formulation of mycelium did not hamper its viability which was stable, with an estimated shelf life of 72 days at 6 ± 1°C. The inoculum effectiveness was confirmed ex planta and in planta in a small-scale pilot study in field, where formulated mycelium discs of hypovirulent strain Cp 4.2H were inoculated by airgun shot method into the chestnut bark. In planta, Cp 4.2H was recovered in 37% of bark samples taken around the inoculated points 1 year after the treatment. CONCLUSIONS: We demonstrated that the proposed airgun shooting inoculation method of C. parasitica hypovirulent strain formulated as mycelium discs is suitable for treatment of adult chestnut trees. SIGNIFICANCE AND IMPACT OF THE STUDY: The proposed method could be a valid alternative to the traditional manual technique of chestnut biocontrol. The main advantages are the cost-effectiveness and the ease to treat high-positioned, otherwise unreachable cankers both in orchards and forests.

Aislados de Bacillus provenientes de la rizósfera de cactus incrementan la germinación y la floración en Mammillaria spp. (Cactaceae) / Bacillus isolates from rhizosphere of cacti improve germination and bloom in Mammillaria spp. (Cactaceae)

Las cactáceas son la vegetación característica de las zonas áridas en México, donde las lluvias son escasas, la evapotranspiración es elevada y la fertilidad de los suelos es baja. Las plantas han desarrollado estrategias fisiológicas como la asociación con microorganismos en la zona de la rizósfera para incrementar la captación de nutrientes. En el presente trabajo se obtuvieron 4 aislados bacterianos de la rizósfera de Mammillaria magnimamma y Coryphantha radians, los que fueron nombrados como QAP3, QAP19, QAP22 y QAP24 e identificados genéticamente como pertenecientes al género Bacillus. Estos aislados exhibieron in vitro propiedades bioquímicas como solubilización de fosfatos, producción de ácido indolacético y actividad ACC deaminasa, que se relacionan con la promoción del crecimiento de las plantas. Dicha promoción fue ensayada inoculando semillas de M. magnimamma y evaluando luego algunos parámetros. Se encontró que todos los aislados incrementaron la germinación desde un 17% hasta un 34,3% (con respecto a las semillas testigo sin inocular); el aislado QAP24 fue el que presentó el mayor efecto en este sentido y permitió la germinación de todas las semillas viables (84,7%) 3 días antes que en el testigo. La inoculación de este aislado en plantas de Mammillaria zeilmanniana mostró un efecto positivo sobre la floración: en 2 meses dentro del período de un año se detectó un incremento en el número de plantas en floración con respecto a las plantas testigo, de hasta el 31,0% en uno de ellos. Se concluye que los aislados de Bacillus spp. caracterizados poseen potencial para ser empleados en programas de conservación de especies vegetales de zonas áridas.

Cacti are the most representative vegetation of arid zones in Mexico where rainfall is scarce, evapotranspiration is high and soil fertility is low. Plants have developed physiological strategies such as the association with microorganisms in the rhizosphere zone to increase nutrient uptake. In the present work, four bacterial isolates from the rhizosphere of Mammillaria magnimamma and Coryphantha radians were obtained and named as QAP3, QAP19, QAP22 and QAP24, and were genetically identified as belonging to the genus Bacillus, exhibiting in vitro biochemical properties such as phosphate solubilization, indoleacetic acid production and ACC deaminase activity related to plant growth promotion, which was tested by inoculating M. magnimamma seeds. It was found that all isolates increased germination from 17 to 34.3% with respect to the uninoculated control seeds, being QAP24 the one having the greatest effect, accomplishing the germination of viable seeds (84.7%) three days before the control seeds. Subsequently, the inoculation of Mammillari zeilmanniana plants with this isolate showed a positive effect on bloom, registering during two months from a one year period, an increase of up to 31.0% in the number of flowering plants compared to control plants. The characterized Bacillus spp. isolates have potential to be used in conservation programs of plant species from arid zones.

Optimization of fertilization characteristics of urine by addition of Nitrosomonas europaea bio-seed.

BACKGROUND: Because of the high concentration of nutrients in human urine, its utilization as an organic fertilizer has been notable throughout history. However, the nitrogen compounds in urine are not stable. Therefore, to convert urine into a suitable fertilizer, it is important to stabilize and adjust unstable nitrogen compounds such as ammonia. Because nitrification can influence the nitrogen profile, the use of nitrifying microorganisms can be useful for stabilizing the nitrogen profile of urine. This study investigated the changes in nitrogen compounds in pure urine and examined the effect of adding Nitrosomonas europaea bio-seed solution on these changes. RESULTS: It was found that the addition of bio-seed could reduce nitrogen loss as well as the time required to stabilize the nitrogen profile. Furthermore, the optimum concentration of bio-seed (6 × 10(5) N. europaea cells L(-1) ) that not only leads to the least nutrient loss but also results in an adequate nitrate/ammonium ratio and regulates the amount of nitrate produced, thereby preventing over-fertilization, was determined. CONCLUSION: At this concentration, no dilution or dewatering is required, thus minimizing water and energy consumption. Usage of the optimum of concentration of bio-seed will also eliminate the need for inorganic chemical additives. © 2016 Society of Chemical Industry.

Effect of Trichoderma on horticultural seedlings' growth promotion depending on inoculum and substrate type.

AIMS: The biostimulant effect of Trichoderma spp. on horticultural crops are highly variable. Thus, practical use of Trichoderma sp. requires feasible formulated products and suitable substrates. METHODS AND RESULTS: This study evaluates the survival and the growth-promotion effect of a Trichoderma saturnisporum rice formulation compared with a nonformulated conidia suspension (seven treatments in total), on tomato, pepper and cucumber seedlings grown in two substrates: (i) rich in organic matter (OM) and (ii) mineral substrate without OM. The results showed beneficial effects on seedling growth in the OM-rich substrate when T. saturnisporum rice formulation (mainly at maximum concentration) was applied, but the effects were opposite when the mineral substrate without OM was used. The effects were closely linked to the level of inoculum in the substrate, which was greater upon application of the formulated inoculum as opposed to the nonformulated one. CONCLUSIONS: The use of rice to prepare the inoculum of T. saturnisporum seems to be promising for seedling growth in the nursery when it is applied in a substrate that is rich in organic matter, but it must be considered that under certain conditions of food shortage, Trichoderma sp. could show pathogenicity to seedlings. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides evidence of the complexity inherent in the use of micro-organisms in agriculture, while also confirming that the activity of the biofertilizers based on Trichoderma depends on the type of inoculum and its concentration, as well as the properties of the medium in which the fungi develop. Further studies assessing the effectiveness or possible pathogenicity of Trichoderma in different soils under greenhouse conditions must be addressed.

A composite microbial agent containing bacterial and fungal species: Optimization of the preparation process, analysis of characteristics, and use in the purification for volatile organic compounds.

Proper preservation of microbial activity over long periods poses a considerable challenge for pollutant biopurification. A composite microbial agent, mainly composed of bacteria and fungi isolated by the current research team, was constructed in this study and its performance in the removal of mixed waste gases (containing α-pinene, n-butyl acetate and o-xylene) was investigated. According to the removal efficiency in the first 24h and the response to starvation, the optimal ratio of selected carriers (activated carbon, wheat bran and sawdust) was found to be 1:2:1. In some cases of storages, the removal capability of the microbial agent was more than twice that of the suspension. Microbial analysis showed that the inoculated bacterial and fungal strains dominated the agent preparation and utilization. These results indicated that the agent has potential for use in biopurification of mixed waste gas, favoring the reduction of environmental passives and longer retention of microbial activity.

Phosphorus supply, arbuscular mycorrhizal fungal species, and plant genotype impact on the protective efficacy of mycorrhizal inoculation against wheat powdery mildew.

A potential alternative strategy to chemical control of plant diseases could be the stimulation of plant defense by arbuscular mycorrhizal fungi (AMF). In the present study, the influence of three parameters (phosphorus supply, mycorrhizal inoculation, and wheat cultivar) on AMF protective efficiency against Blumeria graminis f. sp. tritici, responsible for powdery mildew, was investigated under controlled conditions. A 5-fold reduction (P/5) in the level of phosphorus supply commonly recommended for wheat in France improved Funneliformis mosseae colonization and promoted protection against B. graminis f. sp. tritici in a more susceptible wheat cultivar. However, a further decrease in P affected plant growth, even under mycorrhizal conditions. Two commercially available AMF inocula (F. mosseae, Solrize®) and one laboratory inoculum (Rhizophagus irregularis) were tested for mycorrhizal development and protection against B. graminis f. sp. tritici of two moderately susceptible and resistant wheat cultivars at P/5. Mycorrhizal levels were the highest with F. mosseae (38 %), followed by R. irregularis (19 %) and Solrize® (SZE, 8 %). On the other hand, the highest protection level against B. graminis f. sp. tritici was obtained with F. mosseae (74 %), followed by SZE (58 %) and R. irregularis (34 %), suggesting that inoculum type rather than mycorrhizal levels determines the protection level of wheat against B. graminis f. sp. tritici. The mycorrhizal protective effect was associated with a reduction in the number of conidia with haustorium and with an accumulation of polyphenolic compounds at B. graminis f. sp. tritici infection sites. Both the moderately susceptible and the most resistant wheat cultivar were protected against B. graminis f. sp. tritici infection by F. mosseae inoculation at P/5, although the underlying mechanisms appear rather different between the two cultivars. This study emphasizes the importance of taking into account the considered parameters when considering the use of AMF as biocontrol agents.