Modified diatomite for soil remediation and its implications for heavy metal absorption in Calendula officinalis.

BACKGROUND: Among different adsorbents, natural and inorganic compounds such as diatomite are important and advantageous in terms of high efficiency and cost-effectiveness, and function in stabilizing heavy metals in the environment. Calendula officinalis, a plant known as a high accumulator of heavy metals, was cultivated in soil treated with varying concentrations of modified diatomite to demonstrate the efficiency of modified diatomite in stabilizating of heavy metals in soils, RESULTS: The modification of diatomite aimed to enhance Calendula officinalis adsorptive properties, particularly towards heavy metals such as lead (Pb), Zinc (Zn), Chromium (Cr), Nickle (Ni), and Copper (Cu), common contaminants in industrial soils. The experimental design included both control and treated soil samples, with assessments at regular intervals. Modified diatomite significantly decreased the bioaccumulation of heavy metals in contaminated soils except Zn, evidenced by decreased DTPA extractable heavy metals in soil and also heavy metal concentrations in plant tissues. Using 10% modified diatomite decreased 91% Pb and Cu, 78% Cr, and 79% Ni concentration of plants compared to the control treatment. The highest concentration of Zn in plant tissue was observed in 2.5% modified diatomite treatment. Remarkably, the application of modified diatomite also appeared to improve the nutrient profile of the soil, leading to enhanced uptake of key nutrients like phosphorus (P) 1.18%, and potassium (K) 79.6% in shoots and 82.3% in roots in Calendula officinalis. Consequently, treated plants exhibited improved growth characteristics, including shoots and roots height of 16.98% and 12.8% respectively, and shoots fresh and dry weight of 48.5% and 50.2% respectively., compared to those in untreated, contaminated soil. CONCLUSION: The findings suggest promising implications for using such amendments in ecological restoration and sustainable agriculture, particularly in areas impacted by industrial pollution.

Microplastics pollution in rice fields: a case study of Pir Bazar rural district of Gilan, Iran.

Various stressors threaten rice fields' productivity. Microplastics (MPs) are ubiquitous pollutants that accumulate in agricultural soils, effectively impairing agroecosystem functioning. The study investigates the MPs pollution status of rice fields and compares it with that of non-paddy vegetable farms under contrasting management practices. Possible sources of MPs in the fields are identified. Additionally, the relationships between MPs abundance and soil characteristics are investigated. This provides innovative insights into the possible impact of MPs on soil health and functioning. Density separation using saturated NaCl solution and oxidative organic matter digestion using Fenton's reagent were employed to extract the MPs. The extracted MPs were categorized according to shape, size, and color. The results indicated that the paddies (1952.86±114.36 particles/kg) contained significantly more MPs than did the non-paddies (1134.44±221.52 particles/kg). Beads (53.75%) and fibers (28.46%) were the most common MPs. More than 90% of all MPs recovered from the fields were less than 1 mm in size. Of the 16 color groups identified, the colors silver, white, and black were the most abundant. Sewage sludge application and mulching were recognized as the primary sources of MPs in the paddies, with sludge contributing more than mulching. Microplastics were shown to potentially alter vital soil characteristics. Rice fields are otherwise overlooked reservoirs of MPs. More attention should be paid to raising awareness of their role as MPs accumulation hotspots among governmental bodies, researchers, producers, and citizens. Contributing MPs sources need to be identified, and managerial decisions should consider the polluting capacity of different practices.

Biodegradation of diethyl phthalate and phthalic acid by a new indigenous Pseudomonas putida.

Diethyl phthalate (DEP) is one of the extensively used plasticizers which has been considered a priority hazardous pollutant due to its carcinogenic, endocrine disrupter, and multi-toxic effects on humans. The identification of DEP in different parts of the ecosphere has increased the global community's attention to the elimination of this pollutant in a bio-eco-friendly way. In this research, a novel aerobic bacterial strain nominates as ShA (GenBank accession number: MN298858) capable of consuming DEP as carbon and energy sources, was isolated from the upper phase (0-10 cm) of Anzali international wetland sediments by enrichment culture method. Morphological characteristics and 16S rRNA gene sequence analysis demonstrated that strain ShA belonged to Pseudomonas putida. The substrate utilization test demonstrated that strain ShA was able to grow in mineral salt medium containing dimethyl phthalate (DMP) and phthalic acid (PA) isomers including terephthalic and isophthalic acid. Degradation assay showed strain ShA completely degraded 200 mg/L DEP within 22 h (pH 7.0, 30 °C). Surprisingly, PA as the main intermediate of DEP biodegradation was identified by GC-FID. Moreover, the rapid degradation of 2000 mg/L PA to CO2 and H2O was viewed in 22 h by strain ShA. The possible route of DEP degradation was DEP directly to PA and then PA consumption for growth. This study obtained results that provide a great contribution to applying strain ShA in the biodegradation of low molecular weight of PAEs and PA isomers in natural ecosystems. This is the first report of a P. putida strain able to degrade DEP and PA.

Effects of foliar application of humic acid extracts and indole acetic acid on important growth indices of canola (Brassica napus L.).

Vermicompost (VC) is a rich source of HA that improves plant growth and yield indices such as fresh and dry weights, plant height, stem diameter, leaf area, and chlorophyll index value. In this study, the effect of foliar application of HA extracted from different types of VC enriched with bacteria and/or fertilizers, commercial HA (CHA) and indole acetic acid (IAA) on the growth characteristics of canola (Brassica napus) in greenhouse conditions were compared. According to the results, the foliar application of HA extracted from VC had complete superiority over CHA and IAA in most traits except for the leaf number. Furthermore, the highest level of foliar application of HA (600 mg L-1) enriched with Azotobacter chroococcum (21Az) + Pseudomonas fluorescens (Ps 59) (HA-AS) generated the highest height, diameter, leaf area, and chlorophyll index value. Also, the highest stomatal conductance and photosynthesis rate were observed with the application of 600 mg L-1 HA extracted from VC enriched with nitrogen, sulfur, and phosphorus (HA-NSP) compared to the other treatments. Besides, dry and fresh weights and seed yield under HA-NSP and HA-AS treatments were at their highest rate. Among the extracted HAs, the one extracted from the nitrogen enriched VC had the lowest efficiency. Based on the present study, the HA extracted from VC enriched with Azotobacter, Pseudomonas and NSP is recommended to increase canola growth and production.

Formulation of a Culture Medium to Optimize the Production of Lipopeptide Biosurfactant by a New Isolate of Bacillus sp.: A Soil Heavy Metal Mitigation Approach.

This research aimed to optimize a lipopeptide biosurfactant produced from Bacillus sp. SHA302 due to its high efficiency of heavy metal release in soil. The results demonstrated that the metal release capacity of the lipopeptide biosurfactant alone increased with increasing the biosurfactant concentration. Among treatments with different biosurfactant concentrations plus acid, the highest metal release rates of 53.8% ± 1.4 and 39.3% ± 1.7 for Zn and Pb, respectively, were observed in the critical micelle concentration (CMC) + HCl treatment. The results of a factorial experiment designed for optimizing biosurfactant production showed that among five inexpensive carbon sources and six mineral nitrogen sources, sugar beet molasses (1%) and ammonium chloride (0.1%) were the most efficient sources in lowering the surface tension (ST) of the culture media to 32.2 ± 0.76 mN/m. The second step of the experiment was a Plackett-Burman design with 11 factors and showed that the four factors of pH, ammonium chloride, magnesium sulfate, and molasses significantly affected (P < 0.05) the changes in ST and biosurfactant production. The third step of the experiment was done using the response surface methodology (RSM) with a central composite design. The results showed that a pH of 7.3, 1.5 g/l of ammonium chloride, 0.3 g/l of magnesium sulfate, and 10% of sugar beet molasses yielded values of 29.2 ± 0.71 mN/m and 5.74 ± 0.52 g/l for the two variables of ST and biosurfactant production, respectively, which reached their most optimal levels.

Effect of Different Enriched Vermicomposts, Humic Acid Extract and Indole-3-Acetic Acid Amendments on the Growth of Brassica napus.

Humic acid (HA) is a specific and stable component of humus materials that behaves similarly to growth stimulants, esp. auxin hormones, contributing to improving growth indices and performance of plants. As a rich source of HA, vermicompost (VC) is also a plant growth stimulating bio-fertilizer that can enhance growth indices and performance in plants. The purpose of the present study is to compare the influence of VC enriched with bacterial and/or fertilizer, commercial humic acid (CHA) extract, and indole-3-acetic acid (IAA) on improving growth characteristics and performance of rapeseed under greenhouse conditions. The results showed the complete superiority of VC over the CHA and IAA (approximately 8% increase in the dry weights of root and aerial organ and nearly three times increase in seed weight). The highest values of these indices were obtained with VC enriched with Nitrogen, Sulfur, and Phosphorus, Azotobacter chroococcum and Pseudomonas fluorescens; the lowest value was obtained with VC enriched with urea. Additionally, the application of 3% VC and the control involved the highest and lowest values in all traits, respectively. The SPAD (chlorophyll index) value and stem diameter were not significantly affected by different application levels of VC. Overall, the applications of IAA and the CHA were not found to be suitable and therefore not recommended.

A Combinational Strategy Mitigated Old-Aged Petroleum Contaminants: Ineffectiveness of Biostimulation as a Bioremediation Technique.

Hydrocarbon contamination emerging from the crude oil industrial-related activities has led to severe environmental issues. Prolonged contamination with the constant infiltration of crude oil into the soil is a severe problem in remediating contaminated soils. Hence, the current study focuses on comparing various bioremediation strategies, thereby isolating native bacteria competent to reduce TPH in both liquid and microcosm environments in an old-aged petroleum hydrocarbon contaminated soil. Assays in the modified 6SW-Vit medium after 7 days of incubation revealed that Bacillus altitudinis strain HRG-1 was highly hydrophobic and had a suitable ability to decrease surface tension (40.98%) and TPH (73.3%). The results of biodegradation in the microcosm proved that among the designated treatments, including bio-stimulated microcosm (SM), bacterialized microcosm (BM), a combined bio-stimulated microcosm and bacterialized microcosm (SB), and natural attenuation (NA), the SB treatment was the most effective in mitigating TPH (38.2%). However, the SM treatment indicated the lowest TPH biodegradation (18%). Pearson correlation coefficient among microcosm biological indicators under investigation revealed that soil basal respiration had the highest correlation with the amount of residual TPH (r = -0.73915, P < 0.0001), followed by the microbial population (r = -0.65218, P < 0.0001), catalase activity (r = 0.48323, P = 0.0028), polyphenol oxidase activity (r = -0.43842, P = 0.0075), and dehydrogenase activity (r = -0.34990, P = 0.0364), respectively. Nevertheless, considering the capability of strain HRG-1 and the higher efficiency of the combined technique, their use is recommended to diminish the concentration of petroleum hydrocarbons in hot and dry contaminated areas.

Investigation of Different Selenium Sources and Supplying Methods for Selenium Enrichment of Basil Vegetable (A Case Study under Calcareous and Non-calcareous Soil Systems).

BACKGROUND: Selenium (Se) is a crucial component of selenoaminoacids and selenoproteins. Therefore, Se-enriched agricultural products can reduce health complications induced by Se deficiency. OBJECTIVE: This research was carried out to investigate the effects of Se bio-enrichment on Basil grown in calcareous and non-calcareous soil systems and also to evaluate the changes in Se concentration in the soil after harvesting. METHODS: The experiment executed in two calcareous and one non-calcareous soil systems, and different Se application methods (control, soil application, seed inoculation, foliar application, and soil + foliar application) were administered. Selenobacteria, a plant growth-promoting rhizobacteria (PGPR), derived from the soil was used as a biofertilizer, compared to the other Se sources. RESULTS: The results showed that both soil types and the methods of Se application had significant effects (P ˂ 0.01) on root and shoot dry weights and concentrations of P, K, Zn, Fe, and Se in both of the root and shoot. Shoot dry weight of plants treated with foliar Se was maximum in the calcareous soil. Compared to the control treatment, foliar application of Se increased shoot Se content in both calcareous and non-calcareous soils by 242% and 204%, respectively. Furthermore, the increase in shoot Se concentration in calcareous soil induced by Se application increased the concentration of other nutrients in the shoot and root. Plant growth parameters and concentrations of nutrients were significantly increased by using selenobacter inoculum. CONCLUSION: The application of Se-containing compounds can improve vegetable quality. Considering the daily requirement of the human body for minerals and nutrients, enriching basil with Se can play an important role in community health. Moreover, some patents have reported the effectiveness of endophyte bacteria.

The potential use of Cordia myxa in the remediation of crude oil pollution.

This study investigated the effects of hydrocarbon-degrading bacteria and organic matter on a crude oil-polluted soil by Cordia myxa. The treatments consisted of crude oil at two levels (3 and 6% w/w), municipal waste compost at two levels (5 and 10% v/v), and two different bacterial strains (Pseudomonas sp.141 and Pseudomonas sp. 27ps). At the end of the growth period, the plants were harvested and prepared for the laboratory analyses. The greatest population of oil degrading-bacteria (4.6 × 106 CFU/g soil) was observed in the treatment containing 10% compost, 6% crude oil, and Pseudomonas sp.141. The highest crude oil degradation (76.49%) was recorded in the soil polluted with 6% crude oil, amended with 10% compost, and inoculated with Pseudomonas sp.141. The investigation on the degradation of the chains of C10-C35 compounds indicated that, in various treatments, the most abundant compound was among those with fewer carbon atoms (C12-C25), so the application of organic matter boosted the degradation of crude oil. In conclusion, C. myxa seedlings has an acceptable efficiency in the remediation of the oil-contaminated soil affected by biological factors (compost and Pseudomonas bacteria), which is because of their high tolerance to the pollution and their ability to penetrate deeper soil layers.

Combined use of municipal solid waste biochar and bacterial biosorbent synergistically decreases Cd(II) and Pb(II) concentration in edible tissue of forage maize irrigated with heavy metal-spiked water.

A pot experiment was carried out to evaluate the effect of a municipal solid waste (MSW) biochar and a bacterial strain on the forage maize growth and the concentration of lead (Pb) and cadmium (Cd) in the edible tissue of maize irrigated with water contaminated with Cd (5 mg L-1) and Pb (100 mg L-1). Experimental treatments included (i) bacterial strain at two levels: no bacterial strain and Enterobacter cloacae R7; (ii) MSW biochar at three levels: 0, 1, and 3% (w/w); and (iii) irrigation water quality at five levels: plants irrigated with 100% freshwater (FW), plants irrigated with 75%FW + 25% contaminated water (CW), plants irrigated with 50%FW + 50% CW, plants irrigated with 25%FW + 75% CW, and plants irrigated with 100% CW. The effect of various treatments on maize growth indices and concentration of Pb(II) and Cd(II) in the plant was significant at 5% level. The concentration of these metals in the shoot of plants irrigated with 75 and 100% CW was higher than the permissible limits for Cd(II) and Pb(II) in livestock feed. However, the concentration of these metals in the shoot of the plants irrigated with 25 and 50% CW was lower than the permissible limit for this use. In this study, the combined application of 3%biochar and E. cloacae R7 had a significant effect on increased root dry weight (ranging from 29 to 33%), shoot dry weight (ranging from 32 to 43%) and bacterial root colonization (ranging from 33 to 53%) and on reduced concentration of Pb (ranging from 78 to 80%) and Cd (ranging from 72 to 76%) of the shoot of maize plant (edible tissues used by livestock), which was below the permissible limits for livestock feed, compared to corresponding controls. According to the results of this study, to reduce the concentration of the heavy metals in forage maize shoot (below the permissible limits for livestock feed), it is suggested using heavy metal-contaminated water either in combination with freshwater (50 or 75% FW) or in combination with biochar and bacterial biosorbent, averting human/animal health risk.

Mining the roots of various species of the halophyte Suaeda for halotolerant nitrogen-fixing endophytic bacteria with the potential for promoting plant growth.

Saline area may tend to be a productive land; however, many of salt-affected soils have nitrogen limitation and depend on plant-associated diazotrophs as their source of 'new' nitrogen. Herein, a total of 316 salinity tolerant nitrogen-fixing endophytic bacteria were isolated from roots of the halophyte Suaeda sp. sampled from 22 different areas of Iran to prepare the collection of nitrogen-fixing bacterial endophytes and evaluate the plant growth-promoting effect of effective isolates on growth of the halophyte Suaeda maritima. All of the identified nitrogen-fixing endophytes were classified to Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes phylum while we did not detect common nitrogen-fixing endophyte of glycophytes like Azospirillum. The genera Pseudomonas and Microbacterium were both encountered in high abundance in all samples, indicating that they might play an advanced role in the micro-ecosystem of the halophyte Suaeda. In addition, the results also showed that not only soil salinity can affect halophyte endophytic composition but also other factors such as geographical location, plant species, and other soil properties may be involved. Interestingly, only Zhihengliuella halotolerans and Brachybacterium sp. belonging to Actinobacteria could grow in semi-solid N-free (NFb) medium supplemented with 6% NaCl and highly enhanced growth of S. maritima in vitro. Overall, this study offers useful new resources for nitrogen-fixing endophytic bacteria which may be utilized to improve approaches for providing bio-fertilizer useful in saline-based agriculture.

Effect of rhizospheric and endophytic bacteria with multiple plant growth promoting traits on wheat growth.

The present study focused on the characterization of plant growth promoting rhizospheric (R) and endophytic (E) bacteria and their impact on wheat cultivars growth. In this study, 400 strains were isolated from the rhizosphere soil (250 isolates) and surface-sterilized roots (150 isolates) of wheat and screened for their ability to plant growth promotion (PGP) traits. Four R isolates and four E isolates with different ability were selected to investigate the interaction between R and B bacteria associated with wheat cultivars under in vitro and greenhouse conditions. Plant growth parameters were found to be enhanced by the combined inoculation of two groups of R and E bacteria compared to individual inoculations (respectively 33.7 and 37.8% increase in root and shoot dry weight), suggesting that PGP rhizobacteria acted synergistically with PGP endophytes in phosphate solubilization. Compared to inoculation with phosphate-solubilizing bacteria (PSB) or indole-3-acetic acid producer bacteria (IAA-PB), inoculation by bacteria with multiple PGP properties (PSB and IAA-PS) showed higher promotion capacity. Also, in greenhouse assay, bacterial inoculation had a positive effect on the soil dehydrogenase (70.2%) and phosphatase (52.2%) activity. It seems PGP traits do not work independently of each other but additively as it was suggested in the "synergistic hypothesis" that multiple mechanisms are responsible for the plant growth promotion and increased yield. Findings of this study could improve the current bio-fertilizer production procedure in research and related industries.

Characterization of rhizosphere and endophytic bacteria from roots of maize (Zea mays L.) plant irrigated with wastewater with biotechnological potential in agriculture.

The aim of this study was to characterize culturable rhizosphere and endophytic bacterial isolates isolated from rhizosphere soil and roots of maize plant irrigated with industrial and municipal wastewater in terms of resistance to heavy metals and salinity and plant growth promoting (PGP) traits. Results illustrated that both rhizosphere isolates and endophytic ones had various PGP characteristics in terms of both the number and the production amount of these characteristics. A substantial number of the bacterial isolates (both endophytic isolates and rhizosphere isolates) were tolerant to heavy metals (multi-metal resistant bacteria). Compared to endophytic isolates, rhizosphere isolates had greater resistance to heavy metals. Both endophytic isolates and rhizosphere ones showed remarkable resistance to salinity (7% NaCl). Based on comparison of 16S rRNA sequences and biochemical tests, the effective isolates, based on having multiple PGP characteristics and higher resistance to heavy metals and salinity, were identified. Isolates N5 and R7 were closely related to Bacillus cereus and Enterobacter cloacae, respectively. In addition, the ability of rhizosphere strain R7, as a multi-metal resistant bacterium, in the removal of cadmium (Cd) and lead (Pb) by its biomass and colonization of maize roots in the presence of these metals was evaluated. This strain could remove these metals from the solution (46.5-88.95%) and colonize both root surface and inside root of maize (4-7 Log10 CFU (colony-forming unit) g-1 fresh root weight) under heavy metal stress. Therefore, it can be concluded that maize plant irrigated with industrial and municipal wastewater harbors salinity and heavy metals-resistant bacteria and may be potential reservoirs for isolating bacteria effective at alleviating heavy metal stress in the plant, reducing accumulation of heavy metals in crops such as maize, and removing heavy metals in aqueous media (bioremediation of heavy metal-contaminated wastewater system).

Presence of Eisenia fetida enhanced phytoremediation of anthracene by Lolium perenne / Presença de Eisenia fetida aumentou a fitorremediação do antraceno por Lolium perene

The efficiency of co-application of Eisenia fetida and ryegrass was evaluated in a process called earthworm-assisted phytoremediation. Anthracene was used as a model compound for polycyclic aromatic hydrocarbons (PAHs). The experiments were conducted on a loamy soil in greenhouse conditions. At the end of the experiment, the soil samples were analyzed for residual anthracene by HPLC. Results showed that, phytoremediation using ryegrass could remove 81% of anthracene; however, the rate of removal was 92% when E. fetida was applied simultaneously. E. fetida alone could also remove the initial concentration of anthracene by 40%. Although ryegrass itself could remove anthracene significantly, the employment of earthworm, together with plant was more efficient than each of them individually. The application of E. fetida could also enhance the growth parameters of ryegrass significantly. In comparison to the control, the presence of E. fetida increased plant dry weight (7.8%), root length (47%), shoots length (32%), and root volume (12%). The number of live earthworms was also increased in the planted pots, indicating the helpfulness of the plant for survival of the earthworm in the PAH-contaminated soil. Although plant and earthworm use completely different mechanisms for anthracene degradation, they improve efficiency and survival of the three-component-system.

A eficiência da co-aplicação de Eisenia fetida e azevém foi avaliada em um processo denominado fitorremediação assistida por minhocas. O antraceno foi usado como um composto modelo para hidrocarbonetos aromáticos policíclicos (PAHs). Os experimentos foram conduzidos em um solo argiloso em condições de estufa. No final da experiência, as amostras de solo foram analisadas quanto ao antraceno residual por HPLC. Os resultados mostraram que, a fitorremediação com azevém pode remover 81% do antraceno; no entanto, a taxa de remoção foi de 92% quando E. fetida foi aplicada simultaneamente. E. fetida sozinha também foi capaz de remover a concentração inicial de antraceno em 40%. Embora o próprio azevém pudesse remover significativamente o antraceno, o emprego da minhoca, juntamente com a planta, foi mais eficiente do que cada um deles individualmente. A aplicação de E. fetida também pode melhorar significativamente os parâmetros de crescimento do azevém. Em comparação com o controle, a presença de E. fetida aumentou o peso seco da planta (7,8%), o comprimento da raiz (47%), o comprimento da parte aérea (32%) e o volume radicular (12%). O número de minhocas vivas também aumentou nos vasos plantados, indicando a utilidade da planta para a sobrevivência da minhoca no solo contaminado com PAH. Embora plantas e minhocas usem mecanismos completamente diferentes para a degradação do antraceno, eles melhoram a eficiência e a sobrevivência do sistema de três componentes.

Effect of different biochars amendment on soil biological indicators in a calcareous soil.

Previous studies suggest that biochar has potential to benefit soil when used as an amendment, but only few studies have investigated how the different biochars affect the microbial activity of soil in a calcareous soil. Hence, to study the effect of the biochars obtained from wheat straw and cow manure and produced under different production conditions on two biological soil indicators, dehydrogenase activity and soil respiration, after 0, 60, and 120 days of incubation (DOI), an incubation experiment as a completely randomized design with factorial arrangement in three replicates was conducted in a calcareous soil. The results of the study showed that with increasing the pyrolysis temperature (300 and 500 °C) and pyrolysis residence times (1, 3, and 6 h) of biochars, regardless of feedstock source, the dehydrogenase activity and soil respiration decreased. Both maximum activity of dehydrogenase (20.93 µg TPF g-1 24 h-1) and maximum soil respiration (0.26 mg CO2 g-1 24 h-1) were found in the biochar produced from wheat straw at 300 °C, and the residence time of 1 h at the level of 10 t ha-1 and minimum of these soil biological traits was observed in control treatments (soil). Moreover, the maximum activity of dehydrogenase and soil respiration was observed in 60 DOI. Therefore, when applying biochar as an amendment for increasing microbial activity in calcareous soil, the production conditions of biochar, type of biochar, and long- and short-term effects of different biochars should be taken into consideration.

Indole-3-acetic acid (IAA) production trait, a useful screening to select endophytic and rhizosphere competent bacteria for rice growth promoting agents.

Plants select plant growth promoting rhizobacteria (PGPR) that are competitively fit to occupy compatible niches without causing pathological stress on them. However, when screening bacteria for plant growth promoting (PGP) agents, it is better to select bacteria for achieving the most promising isolates having suitable colonization and PGP traits. In most researches, it has been seen that following incubation, bacterial flora are taken at random from petri dishes for further study. However, this type of selection may remove some superior bacteria in terms of PGP traits and high colonization ability. Therefore, it is essential to study all the isolated bacteria in an economic way and select the best bacteria in terms of PGP traits and high colonization rate. A simple screening method to detect endophytic and rhizosphere bacteria, isolated from the plants in rotation with rice, for rice PGP agents based on a root colonization bioassay and a PGP trait is characterized. •Selected bacterial isolates based on their IAA producing trait have the potential for more PGP and colonization of rice plant.•IAA may be the first PGP trait for screening bacteria isolated from plant rotated with rice for rice PGP agents.•The screening procedure appears to be very effective and less time consuming.

Bacterial biosynthesis of 1-aminocyclopropane-1-caboxylate (ACC) deaminase, a useful trait to elongation and endophytic colonization of the roots of rice under constant flooded conditions.

This study was conducted to investigate the role of 1-aminocyclopropane-1-carboxylate (ACC) deaminase in Pseudomonas fluorescens strain REN1 and its ability to reduce ethylene levels produced during stress, endophytically colonize and promote the elongation of the roots of rice seedlings under gnotobiotic conditions. We isolated 80 bacteria from inside roots of rice plants grown in the farmers' fields in Guilan, Iran. All of the isolates were characterized for plant growth promoting (PGP) traits. In addition, the colonization assay of these isolates on rice seedlings was carried out to screen for competent endophytes. The best bacterial isolate, based on ACC deaminase production, was identified and used for further study. 16S rDNA sequence analysis revealed that the endophyte was closely related to Pseudomonas fluorescens. The results of this study showed ACC deaminase containing P. fluorescens REN1 increased in vitro root elongation and endophytically colonized the root of rice seedlings significantly, as compared to control under constant flooded conditions. The trait of low amount of indole-3-acetic acid (IAA) production (<15 µg mL(-1)) and the high production of ACC deaminase by bacteria may be main factors in colonizing rice seedling roots compared to other PGP traits (siderophore production and phosphate solubilization) in this study. Endophytic IAA and ACC deaminase-producing bacteria may be preferential selections by rice seedlings. Therefore, it may be suggested that the utilization of ACC as a nutrient gives the isolates advantages in more endophytic colonization and increase of root length of rice seedlings.

Biodegradation of low-density polyethylene (LDPE) by mixed culture of Lysinibacillus xylanilyticus and Aspergillus niger in soil.

In this study, two strains of Aspergillus sp. and Lysinibacillus sp. with remarkable abilities to degrade low-density polyethylene (LDPE) were isolated from landfill soils in Tehran using enrichment culture and screening procedures. The biodegradation process was performed for 126 days in soil using UV- and non-UV-irradiated pure LDPE films without pro-oxidant additives in the presence and absence of mixed cultures of selected microorganisms. The process was monitored by measuring the microbial population, the biomass carbon, pH and respiration in the soil, and the mechanical properties of the films. The carbon dioxide measurements in the soil showed that the biodegradation in the un-inoculated treatments were slow and were about 7.6% and 8.6% of the mineralisation measured for the non-UV-irradiated and UV-irradiated LDPE, respectively, after 126 days. In contrast, in the presence of the selected microorganisms, biodegradation was much more efficient and the percentages of biodegradation were 29.5% and 15.8% for the UV-irradiated and non-UV-irradiated films, respectively. The percentage decrease in the carbonyl index was higher for the UV-irradiated LDPE when the biodegradation was performed in soil inoculated with the selected microorganisms. The percentage elongation of the films decreased during the biodegradation process. The Fourier transform infra-red (FT-IR), x-ray diffraction (XRD) and scanning electron microscopy (SEM) were used to determine structural, morphological and surface changes on polyethylene. These analyses showed that the selected microorganisms could modify and colonise both types of polyethylene. This study also confirmed the ability of these isolates to utilise virgin polyethylene without pro-oxidant additives and oxidation pretreatment, as the carbon source.

Identification and determination of extracellular phytate-degrading activity in actinomycetes.

In this study 97 soil samples from different soil ecosystems were collected. The initial screening was performed on modified glycerol arginine agar (MGAA) to isolate common actinomycetes and on modified MGA-SE (MMGA-SE) to isolate rare actinomycetes. Sixty-seven isolates potentially producing extracellular phytate-degrading activity were identified. The potential to dephosphorylate phytate was confirmed in liquid culture for 46.3 % of the isolates. 12 strains were selected for a direct determination of their phytate-degrading capacity. The results highlighted that the selected isolates produced extracellular phytate-degrading activity; however their capacity in InsP(6) degradation was different. In addition the fermentation medium had an effect on the extent of phytate degradation. Some enzymatic properties of the phytases from isolate No. 43 and isolate No. 63 were determined after obtaining phytase-enriched samples. The enzymes had maximum phytate-degrading capability at 55 °C and pH 5 (isolate No. 43) and 37 °C and pH 7 (isolates No. 63), respectively. Due to their properties, the phytase of isolate No. 43 behaves like a histidine acid phytase, whereas the phytase of No. 63 showed similar enzymatic properties to the phytase of lily. To our knowledge, the results from this study demonstrated for the first time that actinomycetes produce extracellular phytate-degrading activity. By 16SrRNA sequencing, the more closely studied phytase producers were identified as Streptomyces sp. Isolate No. 43 showed 98 % identity to Streptomyces alboniger and S. venezuelae, while isolate No. 63 exhibited 98 % sequence identity to S. ambofaciens and S. lienomycini.