Electron microscopy of some rock phosphate dissolving bacteria and fungi

BacteriaPseudomonas striata, Bacillus polymyxa, B. megaterium andB. pulvifaciens, and fungiAspergillus awamori, A. niger andPenicillium digitatum dissolve tricalcium phosphate and, much less, Mussorie and Udaipur rock phosphate. The solubilizing power of fungi was higher than that of bacteria, the highest being withA. awamori andA. niger, and withP. striata. Electron microscopy of the various cultures showed an electron-dense layer on the bacterial surface after negative staining. The size of phosphate particles decreased by the microbial action, with tricalcium phosphate from 140 — 250 to 30 — 90 nm after three weeks of incubation.     

Rhizosphere mediated nutrient management in Allium hookeri Thwaites by using phosphate solubilizing rhizobacteria and tricalcium phosphate amended soil

This work describes integrated nutrient management for cultivation of Allium hookeri by using phosphate solubilizing bacteria (PSB) applied in rhizosphere, along with tricalcium phosphate (TCP). Arthrobacter luteolus S4C7, Enterobacter asburiae S5C7, Klebsiella pneumoniae S4C9, S4C10 and S6C1, and K. quasipneumoniae S6C2, were isolated from rhizosphere of Allium hookeri Thwaites, and were found to release substantial amount of soluble phosphate (124.8–266.4?μg/mL) from TCP in vitro conditions. These isolates were experimented for plant growth promoting attributes, including IAA, siderophore, and nitrogen-fixation. Treatment with PSB resulted in enhanced growth of A. hookeri Th., which was even better with TCP amendment with PSB. K.quasipneumoniae  S6C2 resulted in 39.1% and 533.3% increase (p?≤?0.05) of root length and weight respectively. The treatment with these isolates, in TCP amended soil also resulted in 200–250% increase in available P in soil, which was maximum for K. quasipneumoniae (1.866?mg/g).     

Uranium(VI) bioprecipitation mediated by a phosphate solubilizing Acinetobacter sp. YU-SS-SB-29 isolated from a high natural background radiation site

Bioprecipitation of uranium (U) into uranyl phosphate (U-P) mediated by soluble ortho-phosphate is an attractive proposition for U bioremediation. As an alternative to the microbial phosphatase, we have investigated the dissolution of phosphate by the organic acids produced by bacteria to aid in U precipitation. The bacterium Acinetobacter sp. YU-SS-SB-29, isolated from monazite sand of natural background radiation site solubilized 952.0 ± 46.7 mg L⁻¹ phosphate from tri-calcium phosphate (TCP) in the Pikovskaya's medium and showed tolerance to 120 ppm U(VI). U(VI) bioprecipitation was investigated by adding different concentrations of U(VI) to a cell-free culture supernatant containing ortho-phosphate released from TCP by the bacterium. A yellow precipitate was immediately formed following which there was a reduction in U(VI) concentration. A strong positive correlation (R² = 0.98) was observed between % decrease in phosphate and U(VI) concentration (up to 750 ppm U) added. FTIR and EDX spectra of the yellow precipitate demonstrated the involvement of phosphate groups in U(VI) binding. Furthermore, the XRD pattern of the precipitate agrees well with that of chernikovite, a uranyl phosphate mineral. The results from this study demonstrate the potential of the U tolerant, phosphate solubilizing bacterium Acinetobacter sp. YU-SS-SB-29 for non-reductive in situ bioprecipitation of uranium.     

Comparative in situ remediation potential of Pseudomonas putida 710A and Commamonas aquatica 710B using plant (Vigna radiata (L.) wilczek) assay

A greenhouse study was conducted to determine the effect of cadmium (Cd)-resistant Pseudomonas putida strain 710A and Commamonas aquatica strain 710B, used either alone or as a binary mixture on the toxicity of cadmium to mungbean [Vigna radiata (L.) wilczek] plants. The bacterial strains 710A and 710B, isolated from the Semra mines in Ranchi, India, were resistant to 0.5 mM and 1 mM, CdCl₂ respectively. Moreover, both strains grew well at 10 °C and 30 °C. Of the two bacterial strains, strain 710B showed 129.6 and 83.5 times higher P solubilizing activity than strain 710A, when grown in liquid broth supplemented with cadmium at 10 °C and 30 °C, respectively. Furthermore, 16S ribosomal DNA (rDNA) sequencing identified 710A as a Pseudomonas putida strain and 710B as a Commamonas aquatica strain. The strain 710A significantly (p?<?0.05) stimulated the growth of roots (35.2 %) and shoots (30 %) of mungbean plants grown in soil treated with 110 μM CdCl₂. However, the increase in the case of 710B was found to be 15.4 % and 5.17 %, respectively. The loss of chlorophyll content was replenished by up to 80 %, 66 %, and 77.3 % by inoculation of 710A, 710B and binary mixture, respectively. Moreover the strains were able to reduce Cd accumulation in roots and shoots. Most significantly, residual Cd concentration in soil was reduced in the presence of bioinoculants. However, in terms of efficiency, the strains were found to be in the following order: 710A?>?710A +710B?>?710B. The results suggest that P. putida 710A strain is a potentially effective candidate metal to be used for sequestering and as a growth-promoting bioinoculant in Cd polluted soil.     

Isolation and identification of phosphate solubilizing bacteria able to enhance the growth and aloin-A biosynthesis of Aloe barbadensis Miller

The effect of four phosphate solubilizing bacteria (PSB) was studied on growth and aloin-A content of Aloe barbadensis in soil containing tricalcium phosphate (TCP). PSB were identified based on 16S rRNA gene sequencing as Pseudomonas synxantha, Burkholderia gladioli, Enterobacter hormaechei and Serratia marcescens. These PSB solubilized 25-340 μg ml(-1) of TCP into the liquid phase. The treatment of plants with individual PSB or mixture of these increased soil available P, P uptake in plants and plant growth. The increase in aloin-A content due to higher plant biomass and unit biomass production was 673%, 294%, 276%, 119% and 108% in plants treated with a PSB consortium, P. synxantha, S. marcescens, B. gladioli, and E. hormaechei in TCP amended soil, respectively.     

Expression and Secretion of Defined Cutinase Variants by Aspergillus awamori

Several cutinase variants derived by molecular modelling and site-directed mutagenesis of a cutinase gene from Fusarium solani pisi are poorly secreted by Saccharomyces cerevisiae. The majority of these variants are successfully produced by the filamentous fungus Aspergillus awamori. However, the L51S and T179Y mutations caused reductions in the levels of extracellular production of two cutinase variants by A. awamori. Metabolic labelling studies were performed to analyze the bottleneck in enzyme production by the fungus in detail. These studies showed that because of the single L51S substitution, rapid extracellular degradation of cutinase occurred. The T179Y substitution did not result in enhanced sensitivity towards extracellular proteases. Presumably, the delay in the extracellular accumulation of this cutinase variant is caused by the enhanced hydrophobicity of the molecule. Overexpression of the A. awamori gene encoding the chaperone BiP in the cutinase-producing A. awamori strains had no significant effect on the secretion efficiency of the cutinases. A cutinase variant with the amino acid changes G28A, A85F, V184I, A185L, and L189F that was known to aggregate in the endoplasmic reticulum of S. cerevisiae, resulting in low extracellular protein levels, was successfully produced by A. awamori. An initial bottleneck in secretion occurred before or during translocation into the endoplasmic reticulum but was rapidly overcome by the fungus.     

Cellulose amendment promotes P solubilization by Penicillium aculeatum in non-sterilized soil

Successful application of microbial biofertilizers, such as phosphorus (P) solubilizing fungi to agroecosystems, is constrained from the lack of knowledge about their ecology; for example in terms of how they respond to an external input of carbon (C) to get established in the soil. In two soil incubation experiments we examined the performance of the P solubilizing fungus Penicillium aculeatum in non-sterile and semi-sterile (γ-irradiated) soil with different C and P sources. Results from the first experiment with C sources showed that starch and cellulose generally improved P solubilization by P. aculeatum measured as water extractable P (P_wep), though only significantly in non-sterile soil. This coincided with an increased population density of P. aculeatum measured with a hygromycin B resistant strain of this fungus. Soil respiration used to measure soil microbial activity was overall much higher in treatments with C compounds than without C in both non-sterile and semi-sterile soil. However, soil respiration was highest with cellulose in semi-sterile soil, especially in combination with P. aculeatum. Hence, for the second experiment with P sources (tricalcium phosphate (TCP) and sewage sludge ash) cellulose was used as a C source for P. aculeatum growth in all treatments. Main results showed that P. aculeatum in combination with cellulose soil amendment increased soil P_wep independent of soil sterilization and P source treatments. Soil resin P (P_res) and microbial P (P_mic), which represents stocks of potentially plant available P, were also affected from P. aculeatum inoculation. Increased soil P_res from TCP and sewage sludge ash was observed with P. aculeatum independent of soil type. On the other hand soil P_mic was higher after P. aculeatum inoculation only in semi-sterile soil. Population density of P. aculeatum measured with qPCR was maintained or increased in non-sterile and semi-sterile soil, respectively, compared to the original inoculum load of P. aculeatum. In conclusion, our results underline the importance of C source addition for P. aculeatum if used as a biofertilizer. For this, cellulose seems to be a promising option promoting P. aculeatum growth and P solubilization also in non-sterilized soil.     

Enhanced stability of a rumen-derived xylanase using SpyTag/SpyCatcher cyclization

A phosphate solubilizing bacterium ZB was isolated from the rhizosphere soil of Araucaria, which falls into the species Pantoea agglomerans. Optimization for phosphate solubilization by strain ZB was performed. At optimum culture conditions, the isolate showed great ability of solubilizing different insoluble inorganic phosphate sources viz. Ca₃(PO₄)₂ (TCP), Hydroxyapatite (HP), CaHPO₄, AlPO₄, FePO₄ along with rock phosphates (RPs). Inoculation with planktonic cells was found to enhance dissolved phosphorous as compared to that achieved by symplasma inoculation. Besides inoculation with different status of cells, pre-incubation could also exert a great effect on phosphate solubilization ability of P. agglomerans. When isolate ZB was cultured with glucose as carbon sources, phosphorous was more efficiently dissolved from HP and RP without pre-incubation in comparison to that obtained with pre-cultivation. Pre-cultivation, however, was more suitable for P solubilization than no pre-cultivation when bacteria were grown with xylose. A positive correlation was detected between the production of organic acids and phosphate solubilization. P. agglomerans ZB possessed many plant growth promotion traits such as N₂ fixation and production of indole 3-acetic acid, phytase, alkaline phosphatase. Pot experiment showed inoculation with single isolate ZB or biofertilizer prepared from semi-solid fermentation of isolate ZB with spent mushroom substrate (SMS) compost could enhance plant growth with respect to number of leaves, plant leave area, stem diameter, root length, root dry mass, shoot dry mass and biomass when compared to the abiotic control, revealing strain ZB could be a promising environmental-friendly biofertilizer to apply for agricultural field.

     

Identification,characterization and optimization of phosphate solubilizing rhizobacteria (PSRB) from rice rhizosphere

Two billion people worldwide take rice (Oryza sativa L.) as a staple food. Phosphorus (P) and Nitrogen (N) are the major requirements of rice; although these are available in limited concentrations within rice growing regions. Among different types of Plant growth-promoting rhizobacteria (PGPR), Phosphate solubilizing rhizobacteria (PSRB) constitute an important class. These are known for plant growth promotion by enhancing P and N uptake. PSRB are nowadays used as biofertilizers to restore the soil health. Under the present investigation identification, characterization and optimization of phosphate solubilizing activity of these microbes at different pH, temperature and salt concentrations was carried out. Thirty-seven isolates were recovered from different regions of rice rhizosphere on Pikovskaya (PVK) agar among which 15 isolates were recovered from R.S. Pura, 12 isolates from Bishnah and 10 isolates were recovered from Akhnoor sector of Jammu, India. A prominent halo zone of clearance was developed around the colonies of 12 different isolates, indicating phosphate solubilization activity. Four distinct isolates were amplified, cloned and sequenced for taxonomic identification using 16S primers. The results indicated that PS 1, PS 2, PS 3, PS 4 were related to Pseudomonas aeruginosa, Bacillus subtilis strain 1, B. subtilis strain 2, B. subtilis strain 3, respectively. These strains when grown at a wide range of ecological factors showed maximum growth at pH between 6.8 and 8.8, temperature between 28 °C and 37 °C and salinity between 1% and 2%. Screening for phosphate solubilization activity revealed that the halo zone diameter formed by these isolates extended from 2.1 to 3.2 mm. The phosphate solubilizing efficiency (SE) ranged from 35.4 to 50.9 with highest value of 50.9 by PS4 and maximum P solubilization of 10.22 µg/ml was recorded by PS4 at 7th day. Phosphate solubilization activity of these identified PSRB strains can be utilized and explored in the rice growing belts of Jammu region which are deficient in phosphorus. MIC value for zinc sulphate heptahydrate in 12 isolates varied from 1 mg/ml to 6 mg/ml. Phosphate solubilization activity and MIC of these identified PSRB strains can be utilized and explored in the rice growing belts of Jammu region which are deficient in phosphorus.     

Repression of mineral phosphate solubilizing phenotype in the presence of weak organic acids in plant growth promoting fluorescent pseudomonads

Two phosphate solubilizing bacteria (PSB), M3 and SP1, were obtained from the rhizosphere of mungbean and sweet potato, respectively and identified as strains of Pseudomonas aeruginosa. Their rock phosphate (RP) solubilizing abilities were found to be due to secretion high amount of gluconic acid. In the presence of malate and succinate, individually and as mixture, the P solubilizing ability of both the strains was considerably reduced. This was correlated with a nearly 80% decrease in the activity of the glucose dehydrogenase (GDH) but not gluconate dehydrogenase (GAD) in both the isolates. Thus, GDH enzyme, catalyzing the periplasmic production of gluconic acid, is under reverse catabolite repression control by organic acids in P. aeruginosa M3 and SP1. This is of relevance in rhizospheric conditions and is a new explanation for the lack of field efficacy of such PSB.     

Effect of Abiotic Stress on Phosphate Solubilization by Biocontrol Fungus <Emphasis Type="Italic">Trichoderma</Emphasis> sp.

This study was undertaken to explore the role of Trichoderma sp. in phosphate (P) solubilization and antagonism against fungal phytopathogens. All fungal isolates (SE₆, KT₆, KT₂₈, and BRT₁₁) and a standard culture of T. harzianum (Th-std) were able to antagonize two fungal phytopathogens (Sclerotium rolfsii and Rhizoctonia solani) of chickpea (Cicer arietinum L.) wilt complex. Transmission electron microscopic studies (TEM) further confirmed ultra-cytological changes in the sclerotia of S. rolfsii parasitized by Trichoderma sp. All fungal cultures exhibited production of NH₃ and siderophore, but only BRT₁₁, SE₆, and Th-std could produce HCN. Among all the cultures tested, isolate KT₆ was found to be most effective for solubilization of ferric phosphate releasing 398.4 μg ml⁻¹ phosphate while isolates BRT₁₁ and SE₆ showed more potential for tricalcium phosphate (TCP) solubilization releasing 449.05 and 412.64 μg ml⁻¹ phosphate, respectively, in their culture filtrates. Part of this study focused on the influence of abiotic stress conditions such as pH, temperature, and heavy metal (cadmium) on phosphate (TCP) solubilizing efficiency. Two selected cultures KT₆ and T. harzianum retained their P solubilizing potential at varying concentrations of cadmium (0–1000 μg ml⁻¹). Isolate KT₆ and standard culture of T. harzianum released 278.4 and 287.6 μg ml⁻¹ phosphate, respectively, at 1000 μg ml⁻¹cadmium. Maximum solubilization of TCP was obtained at alkaline pH and at 28°C temperature. Isolate BRT₁₁ was found most alkalo-tolerant releasing 448.0 μg ml⁻¹ phosphate at pH 9.     

Solubilization of organic and inorganic phosphates by three highly efficient soil bacterial isolates

Screening soil samples collected from a diverse range of slightly alkaline soil types, we have isolated 22 competent phosphate solubilizing bacteria (PSB). Three isolates identified as Pantoea agglomerans strain P5, Microbacterium laevaniformans strain P7 and Pseudomonas putida strain P13 hydrolyzed inorganic and organic phosphate compounds effectively. Bacterial growth rates and phosphate solubilization activities were measured quantitatively under various environmental conditions. In general, a close association was evident between phosphate solubilizing ability and growth rate which is an indicator of active metabolism. All three PSB were able to withstand temperature as high as 42°C, high concentration of NaCl upto 5% and a wide range of initial pH from 5 to 11 while hydrolyzing phosphate compounds actively. Such criteria make these isolates superior candidates for biofertilizers that are capable of utilizing both organic and mineral phosphate substrates to release absorbable phosphate ion for plants.     

Characterisation of Phosphate Solubilising Bacteria in Sandy Loam Soil Under Chickpea Cropping System

With the aim to explore the possible role of phosphate-solubilizing bacteria (PSB) in phosphorus (P) cycling in agricultural soils, we isolated PSB inhabiting naturally in the sandy loam soils under chickpea cropping of Patiala (Punjab State). A total of 31 bacterial isolates showing solubilizing activities were isolated on Pikovskaya agar plates. The potent phosphate solubilizers were selected for further characterization. These isolates were shown to belong to the genera Pseudomonas and Serratia by partial sequencing analysis of their respective 16S rDNA genes. ERIC-PCR based fingerprinting was done for tracking the survival of introduced populations of the PSB during mass inoculation of these strains under chickpea plots. The results showed positive correlation (r² = 0.853) among soil phosphatase activity and phosphate solubilizers population, which was also positively correlated (r² = 0.730) to available phosphorus. Identification and characterization of soil PSB for the effective plant growth-promotion broadens the spectrum of phosphate solubilizers available for field application.     

Exopolysaccharide: a novel important factor in the microbial dissolution of tricalcium phosphate

Four strains (Enterobacter sp. EnHy-401, Arthrobacter sp.ArHy-505, Azotobacter sp.AzHy-510 and Enterobacter sp.EnHy-402) which have the ability to solubilize tricalcium phosphate (TCP) were used to study the mechanism of P-solubilization. It was found that three phosphate solubilizing bacteria (EnHy-401, ArHy-505 and AzHy-510) producing exopolysaccharide (EPS) have a stronger ability for P-solubilization than isolate EnHy-402 without EPS production, of those, the strain EnHy-401 with the highest EPS production and efficient organic acids on P-solubilization had a stronger capacity for P-solubilization than the others. Further studies demonstrated that addition of EPS into medium could increase the amount of phosphorus solubilized by organic acid, but failed to release phosphorus from TCP alone. The synergistic effects of EPS and organic acid on TCP solubilization varied with the origin and the concentration of EPS in medium. EPS produced by EnHy-401 was most effective in promoting phosphorus release at an optimal concentration in medium. The increase of P-solubilization brought by EPS attributed to the participation of EPS led to the change in homeostasis of P-solubilization, pushing it towards P dissolved by holding free phosphorus in the medium, consequently resulting in greater phosphorus released from insoluble phosphate. We therefore suggest that EPS with ability of phosphorus-holding may be a novel important factor in the microbial dissolution of TCP except for organic acid.     

Stabilization of dextransucrase from <Emphasis Type="Italic">Leuconostoc mesenteroides</Emphasis> NRRL B-640

Phosphate solubilizing yeast (PSY) were isolated from rhizosphere, non-rhizosphere and fruits from Bhavnagar district. The potential of 25 yeasts were analyzed on the basis of phosphate solubilizing zone to growth on solid medium denoted as solubilization index (SI) which ranged from 1.10 to 1.50. Among 25 yeast isolates, 6 yeast belonging to genus Saccharomyces (2), Hansenula, Klockera, Rhodotorula and Debaryomyces exhibited highest SI (1.33–1.50) were further examined for in vitro tricalcium phosphate (TCP) and low grade rock phosphate (RP) solubilization. TCP proved superior to RP with all the yeasts. Within low grade RPs tested, except isolate Y5, all isolates showed maximum solubilization with Hirapur RP (HRP) ranging from 7.24 to 19.30 mg% P₂O₅. Among six PSY screened, Debaryomyces hansenii showing maximal HRP solubilization was chosen for further physiological studies. Maximum HRP solubilization was expressed in following condition: pH optima 7.0, temperature optima 28°C and optimal period of incubation were 15 days. Acidic pH of the spent media was a constant feature in all the cases. No correlation could be established between final acidity produced by yeasts and the quantity of phosphate liberated.     

Rhizosphere mediated growth enhancement using phosphate solubilizing rhizobacteria and their tri-calcium phosphate solubilization activity under pot culture assays in Rice (Oryza sativa.)

Phosphate solubilizing rhizobacteria are considered as an important alternative to increase the availability of accumulated phosphates through solubilization. These increase the growth of plant by enhancing the efficiency of fixing biological nitrogen. This was studied through a pot experiment involving two Phosphate Solubilizing Rhizobacteria (PSRB) isolates, Pseudomonas aeruginosa and Bacillus subtilis along with Tri-calcium phosphate (TCP) on availibity of nutrients, biological composition of soil and yield attributes of rice crop at its growth stages. Experiment was laid in factorial completely randomized design (CRD) comprising of eight treatments replicated thrice with two factors viz. factor 1 with or without TCP (1 g^?1soil) and factor 2 with single or combined inoculation of PSRB isolates. Considerable enhancement in available content of potassium (K), phosphorous (P), nitrogen (N) in soil was found with TCP 1 g^?1soil (P₁) and consortium of Pseudomonas aeruginosa and Bacillus subtilis broth culture at crop growth stages. Highest increase in available N (17.13% and 19.1%), available P (232% and 265%), available K (19.6% and 29.2%) over control were recorded in B₃ (consortium of Pseudomonas aeruginosa and Bacillus subtilis broth culture). Similarly, maximum nutrient uptake N (6.4%), P (15.8%) and K (8.9%) were recorded with same treatment. A considerable growth in soil microbial biomass carbon and dehydrogenase activity at crop growth stages was recorded on application of TCP 1 g^?1soil (P₁) and consortium of PSRB isolates' Pseudomonas aeruginosa and Bacillus subtilis (B₃). Highest increase in microbial biomass carbon (16.4% and 16.5%) and dehydrogenase activity 34.7% and 43.8% over control were recorded in B₃ (consortium of PSRB isolates Pseudomonas aeruginosa and Bacillus subtilis) and was found best among all treatments in terms of yield (63.2%) and yield attributes; number of panicles^?1plant (54.8%), number of grains^?1panicle (156%) and average panicle length (63.9%).     

Nymphal and adult performance of Apolygus lucorum (Hemiptera: Miridae) on a preferred host plant, mungbean Vigna radiata

Mungbean, Vigna radiata (L.) Wilczek, a preferred host plant of Apolygus lucorum (Meyer-Dür) (Hemiptera: Miridae), is used in China as a trap crop for this pest in cotton fields. Little is known, though, about the relationship between adult oviposition preference and offspring performance of A. lucorum on mungbean. In this study, we compared the oviposition preference of A. lucorum on mungbean versus cotton plants in both open-field plots and field cages. Field-plot experiments demonstrated that the population density of first instar nymphs of A. lucorum and the ratio of first instar nymphs to adults were significantly higher on mungbean than on cotton. Choice and no-choice cage experiments revealed that the number of eggs laid on mungbean was significantly higher than on cotton. We also compared adult longevity and fecundity, as well as offspring survival and development on these two host plants. We found that A. lucorum adult females survived longer and laid more eggs on mungbean than on cotton (P?<?0.05). Mungbean also promoted higher rates of development than did cotton (P?<?0.05), but did not affect nymphal survival (P?>?0.05). In summary, the preference of A. lucorum adults for mungbean is consistent with both adult and nymphal performance. This study furthers the understanding of the relationship between the polyphagous species A. lucorum and its host plants, which will help improve trap-cropping strategies for the control of A. lucorum that are based on the use of mungbean.     

Genetic diversity of phosphate-solubilizing peanut (Arachis hypogaea L.) associated bacteria and mechanisms involved in this ability

In this study, attempts were made to analyze mechanisms involved in the bacterial phosphate-solubilizing ability of peanut isolates. Bacteria were taxonomically identified by analysis of 16S rDNA sequence. Levels of soluble P released by the isolates in unbuffered or buffered with Tris–HCl or MES NBRIP-BPB medium as well as the production of D-gluconic acid were determined in their culture. Presence of two of the genes encoding the cofactor PQQ of GDH enzyme was analyzed in the genome of this bacterial collection. 16S rDNA sequence analysis indicated that isolates belong to genera Serratia, Enterobacter, Pantoea, Acinetobacter, Bacillus and Enterococcus. All bacteria showed ability to solubilize tricalcium phosphate either in unbuffered or buffered medium. Nevertheless, addition of buffer solutions reduced levels of Pi liberated by the isolates. Although almost all isolates produced detectable amounts of D-gluconic acid, no correlation with levels of P soluble released were observed. The presence of pqqE and pqqC genes was detected only in Gram negative bacteria. It was concluded from this study that the mechanism involved in phosphate solubilization is organic acids production and, presence of pqq genes in all Gram negative bacteria analyzed encourages to confirm their role in bacterial phosphate solubilizing ability as well to identify genes involved in this PGP trait in Gram positive bacteria.     

Phosphorus Recovery from Phosphate Rocks Using Phosphate-Solubilizing Bacteria

In this paper, we are presenting a biological process to recover phosphorus by solubilizing low-grade phosphate rocks. To this end, the efficiency of different phosphate-solubilizing microorganism (PSM) species for solubilizing P from phosphate rocks using both pure cultures and associations. Nutritional conditions, phosphate rock concentrations, and reactor designs were tested. The genus Bacillus, especially Bacillus megaterium (ATCC 14581), was found to be the most promising PSM for solubilizing P. Production of organic acids and acidic pH values were shown to be directly related to P solubilizing. However, associations between tested microorganisms did not significantly enhance process efficiency. We conclude that nutritional factors of the medium are important to solubilization, and lower phosphate rock concentrations lead to better solubilization. The Air Lift reactor was promising for B. megaterium (ATCC 14581), but adaptations are needed for further tests.     

Comparative study of biochemical properties of glucoamylases from the filamentous fungi Penicillium and Aspergillus

Here we report the first isolation to homogeneous forms of two glucoamylases from the fungus Penicillium verruculosum and their study in comparison with known glucoamylases from Aspergillus awamori and Aspergillus niger. Genes that encode glucoamylases from P. verruculosum were cloned and expressed in the fungus Penicillium canescens, and the recombinant glucoamylases were obtained with subsequent study of their molecular weights, isoelectric points, optimal temperature and pH values, and stability. The catalytic activities of the recombinant glucoamylases were determined in relation to soluble potato starch. Changes in molecular mass distribution and content of low molecular weight products during starch hydrolysis by glucoamylases from P. verruculosum, A. awamori, and A. niger were studied. An exo-depolymerization mechanism was established to be the pathway for destruction of starch by the glucoamylases.