M.V. Martinez-Toledo

Isolation and characterization of Azotobacter and Azospirillum strains from the sugarcane rhizosphere

Bacteria with the ability to grow on nitrogen-free media and with nitrogenase activity under aerobic or microaerobic conditions were isolated from sugarcane roots collected from four different agricultural locations in Granada (Spain). Isolates were Gram negative rods and were identified as Azotobacter chroococcum and Azospirillum brasilense. Our results suggest that Azotobacter isolates do not have a particular affinity for sugarcane rhizospheres and that, on the contrary, Azospirillum isolates show specific association and perhaps endophytic colonization of sugarcane. However, obligate endophytes (Gluconacetobacter diazotrophicus) were not found in the apoplastic fluid of the stems and macerates extracts of sugarcane tissues with the procedure applied. Population of this microorganism might be in low number in the Spanish sugarcane varieties studied which is also discussed.

Effects of culture conditions on the production of polyhydroxyalkanoates by Azotobacter chroococcum H23 in media containing a high concentration of alpechín (wastewater from olive oil mills) as primary carbon source.

Large amounts of homopolymers containing beta-hydroxybutyrate (PHB) and copolymers containing beta-hydroxyvalerate (P[HB-co-HV]) are produced by Azotobacter chroococcum strain H23 when growing in culture media amended with alpechín (wastewater from olive oil mills) as the sole carbon source. Copolymer was formed when valerate (pentanoate) was added as a precursor to the alpechín medium, but it was not formed with the addition of propionate as a precursor. A. chroococcum formed homo- and copolymers of polyhydroxyalkanoates (PHAs) up to 80% of the cell dry weight, when grown on NH(4)(+)-medium supplemented with 60% (v/v) alpechín, after 48 h of incubation at 100 rev min(-1) and 30 degrees C. Production of PHAs by strain H23 using alpechín looks promising, as the use of a cheap substrate for the production of these materials is essential if bioplastics are to become competitive products.

Production of B-group vitamins by two Azotobacter strains with phenolic compounds as sole carbon source under diazotrophic and adiazotrophic conditions

Azotobacter vinelandii strain ATCC 12837 and A. chroococcum strain H23 (CECT 4435) were able to grow on N‐free or NH4Cl‐amended chemically‐defined (Burks) media, with protocatechuic acid (1–2 mmol l−1) or sodium p‐hydroxybenzoate (1–10 mmol l−1) as sole carbon (C) sources. At a concentration of 2 mmol l−1, both substrates supported nitrogen fixation (acetylene reduction assay) at similar or higher rates than bacteria grown in control media amended with 2 mmol l−1 sodium succinate as C source. The two strains produced the B‐group vitamins niacin, pantothenic acid, thiamine, riboflavin and biotin after 72 h of growth in chemically‐defined media with 2 mmol l−1 protocatechuic acid, sodium p‐hydroxybenzoate or sodium succinate as sole C source, either in N‐free media or in media amended with 0·1% NH4Cl. Quantitative production of all vitamins was affected by the use of the different C and N substrates.

Effects of the fungicide Captan on some functional groups of soil microflora

Abstract The effects of Captan at rates of 2.0, 3.5, 5.0 and 10.0 kg ha−1 on microbial function were studied in four agricultural soils under aerobic conditions. Parameters monitored included total culturable populations, numbers of aerobic N2-fixing bacteria, denitrifying bacteria, nitrifying bacteria and nitrogenase activity. Total culturable fungal populations, nitrifying bacteria, aerobic N2-fixing bacteria and nitrogenase activity were significantly decreased at dose rates of 2.0 to 10.0 kg ha−1. However, the presence of Captan enhanced denitrifying and total culturable bacteria, showing that some microbial groups can tolerate high doses of this fungicide.

Production of vitamins by Azospirillum brasilense in chemically-defined media

The production of vitamins by Azospirillum brasilense was studied in chemically-defined media amended with malate, gluconate and fructose. The liberation of vitamins was significantly affected by the presence of different carbon sources and the age of the culture. Thiamine, niacin and pantothenic acid were produced in large amounts. Thiamine and riboflavin were produced only in culture containing malate or fructose. Biotin was not detected in the supernatants of the culture media.

Influence of Rhizobium/Azotobacter and Rhizobium/Azospirillum combined inoculation on mineral composition of faba bean (Vicia faba L.)

Abstract Mixed inoculation of Vicia faba L. with four different Rhizobium/Azospirillum and Rhizobium/Azotobacter combinations led to changes in total content, concentration and/or distribution of the mineral macro- and micronutrients, K, P, Ca, Mg, Fe, B, Mn, Zn and Cu, when compared with plants inoculated with Rhizobium only. The effects varied to a great extent among the Azotobacter and Azospirillum strains selected for combined inoculation.

Bacterial community structure and enzyme activities in a membrane bioreactor (MBR) using pure oxygen as an aeration source

A pilot-scale membrane bioreactor was used to treat urban wastewater using pure oxygen instead of air as a source of aeration, to study its influence on bacterial diversity and levels of enzyme activities (acid and alkaline phosphatases, glucosidase, protease, and esterase) in the sludge. The experimental work was developed in two stages influenced by seasonal temperature. Operational parameters (temperature, pH, BOD5, COD, total and volatile suspended solids) were daily monitored, and enzyme activities measured twice a week. Redundancy analysis (RDA) was used to reveal relationships between the level of enzyme activities and the variation of operational parameters, demonstrating a significant effect of temperature and volatile suspended solids. Bacterial diversity was analyzed by temperature-gradient gel electrophoresis of PCR-amplified partial 16S rRNA genes. Significant differences in community structure were observed between both stages. Sequence analysis revealed that the prevalent Bacteria populations were evolutively close to Alphaproteobacteria (44%), Betaproteobacteria (25%) and Firmicutes (17%).

Effect of an organophosphorus insecticide, profenofos, on agricultural soil microflora

Abstract A study was made of the effects of insecticide profenofos at concentrations of 10, 50, 100, 200 and 300 μg per gram of soil on total bacteria populations, fungi, dinitrogen fixer, denitrifying bacteria, nitrifying bacteria (phase I and II) and nitrogenase activity (C2H2 reduction) in an agricultural soil. The presence of 10 to 300 μg/g significantly increased the total number of bacteria and denitrifying bacteria. However, the population of aerobia dinitrogen fixing bacteria and dinitrogen fixation were suppressed significantly. Nitrifying bacteria and fungal populations decreased initially at concentrations of 100 to 300 μg/g but recovered rapidly to levels similar than those in the control.

Root exudates ofZea mays and production of auxins, gibberellins and cytokinins byAzotobacter chroococcum

Growth ofAzotobacter chroococcum in N-free medium was stimulated in the presence of maize root exudates. Our results show that the production of auxins, gibberellins and cytokinins were significantly stimulated when maize root exudates from 7 to 30 d old plants were added to the culture media.

Liberation of amino acids by heterotrophic nitrogen fixing bacteria

Summary.Large amounts of amino acids are produced by nitrogen-fixing bacteria such as Azotobacter, Azospirillum, Rhizobium, Mesorhizobium and Sinorhizobium when growing in culture media amended with different carbon and nitrogen sources. This kind of bacteria live in close association with plant roots enhanced plant growth mainly as a result of their ability to fix nitrogen, improving shoot and root development suppression of pathogenic bacteria and fungi, and increase of available P concentration. Also, it has been strongly evidenced that production of biologically substances such as amino acids by these rhizobacteria are involved in many of the processes that explain plant-grown promotion. This paper reviews literature concerning amino acids production by nitrogen-fixing bacteria. The role of amino acids in microbial interactions in the rhizosphere and establishment of plant bacterial association is also discussed.