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Fulltext Antifungal Activity of the Extract of a Macroalgae, Gracilariopsis persica, against Four Plant Pathogenic Fungi

Pourakbar L, Moghaddam SS, Enshasy HAE, Sayyed RZ

Plants (Basel), 2021 Aug 26;10(9).
PMID: 34579314 DOI: 10.3390/plants10091781

Nowadays, the extract of seaweeds has drawn attention as a rich source of bioactive metabolites. Seaweeds are known for their biologically active compounds whose antibacterial and antifungal activities have been documented. This research aimed to study the profile of phenolic compounds using the HPLC method and determine biologically active compounds using the GC-MS method and the antifungal activity of Gracilariopsis persica against plant pathogenic fungi. G. persica was collected from its natural habitat in Suru of Bandar Abbas, Iran, dried, and extracted by methanol. The quantitative results on phenolic compounds using the HPLC method showed that the most abundant compounds in G. persica were rosmarinic acid (20.9 ± 0.41 mg/kg DW) and quercetin (11.21 ± 0.20 mg/kg DW), and the least abundant was cinnamic acid (1.4 ± 0.10 mg/kg DW). The GC-MS chromatography revealed 50 peaks in the methanolic extract of G. persica, implying 50 compounds. The most abundant components included cholest-5-en-3-ol (3 beta) (27.64%), palmitic acid (17.11%), heptadecane (7.71%), and palmitic acid methyl ester (6.66%). The antifungal activity of different concentrations of the extract was determined in vitro. The results as to the effect of the alga extract at the rates of 200, 400, 600, 800, and 1000 μL on the mycelial growth of four important plant pathogenic fungi, including Botrytis cinerea, Aspergillus niger, Penicillium expansum, and Pyricularia oryzae, revealed that the mycelial growth of all four fungi was lower at higher concentrations of the alga extract. However, the extract concentration of 1000 μL completely inhibited their mycelial growth. The antifungal activity of this alga may be related to the phenolic compounds, e.g., rosmarinic acid and quercetin, as well as compounds such as palmitic acid, oleic acid, and other components identified using the GC-MS method whose antifungal effects have already been confirmed.
Recent advances in PGPR-mediated resilience toward interactive effects of drought and salt stress in plants

Al-Turki A, Murali M, Omar AF, Rehan M, Sayyed RZ

Front Microbiol, 2023;14:1214845.
PMID: 37829451 DOI: 10.3389/fmicb.2023.1214845

The present crisis at hand revolves around the need to enhance plant resilience to various environmental stresses, including abiotic and biotic stresses, to ensure sustainable agriculture and mitigate the impact of climate change on crop production. One such promising approach is the utilization of plant growth-promoting rhizobacteria (PGPR) to mediate plant resilience to these stresses. Plants are constantly exposed to various stress factors, such as drought, salinity, pathogens, and nutrient deficiencies, which can significantly reduce crop yield and quality. The PGPR are beneficial microbes that reside in the rhizosphere of plants and have been shown to positively influence plant growth and stress tolerance through various mechanisms, including nutrient solubilization, phytohormone production, and induction of systemic resistance. The review comprehensively examines the various mechanisms through which PGPR promotes plant resilience, including nutrient acquisition, hormonal regulation, and defense induction, focusing on recent research findings. The advancements made in the field of PGPR-mediated resilience through multi-omics approaches (viz., genomics, transcriptomics, proteomics, and metabolomics) to unravel the intricate interactions between PGPR and plants have been discussed including their molecular pathways involved in stress tolerance. Besides, the review also emphasizes the importance of continued research and implementation of PGPR-based strategies to address the pressing challenges facing global food security including commercialization of PGPR-based bio-formulations for sustainable agricultural.
Fulltext Recent Understanding of Soil Acidobacteria and Their Ecological Significance: A Critical Review

Kalam S, Basu A, Ahmad I, Sayyed RZ, El-Enshasy HA, Dailin DJ, et al.

Front Microbiol, 2020;11:580024.
PMID: 33193209 DOI: 10.3389/fmicb.2020.580024

Acidobacteria represents an underrepresented soil bacterial phylum whose members are pervasive and copiously distributed across nearly all ecosystems. Acidobacterial sequences are abundant in soils and represent a significant fraction of soil microbial community. Being recalcitrant and difficult-to-cultivate under laboratory conditions, holistic, polyphasic approaches are required to study these refractive bacteria extensively. Acidobacteria possesses an inventory of genes involved in diverse metabolic pathways, as evidenced by their pan-genomic profiles. Because of their preponderance and ubiquity in the soil, speculations have been made regarding their dynamic roles in vital ecological processes viz., regulation of biogeochemical cycles, decomposition of biopolymers, exopolysaccharide secretion, and plant growth promotion. These bacteria are expected to have genes that might help in survival and competitive colonization in the rhizosphere, leading to the establishment of beneficial relationships with plants. Exploration of these genetic attributes and more in-depth insights into the belowground mechanics and dynamics would lead to a better understanding of the functions and ecological significance of this enigmatic phylum in the soil-plant environment. This review is an effort to provide a recent update into the diversity of genes in Acidobacteria useful for characterization, understanding ecological roles, and future biotechnological perspectives.
Fulltext Pythium Damping-Off and Root Rot of Capsicum annuum L.: Impacts, Diagnosis, and Management

Arora H, Sharma A, Sharma S, Haron FF, Gafur A, Sayyed RZ, et al.

Microorganisms, 2021 Apr 13;9(4).
PMID: 33924471 DOI: 10.3390/microorganisms9040823

Capsicum annuum L. is a significant horticulture crop known for its pungent varieties and used as a spice. The pungent character in the plant, known as capsaicinoid, has been discovered to have various health benefits. However, its production has been affected due to various exogenous stresses, including diseases caused by a soil-borne pathogen, Pythium spp. predominantly affecting the Capsicum plant in younger stages and causing damping-off, this pathogen can incite root rot in later plant growth stages. Due to the involvement of multiple Pythium spp. and their capability to disperse through various routes, their detection and diagnosis have become crucial. However, the quest for a point-of-care technology is still far from over. The use of an integrated approach with cultural and biological techniques for the management of Pythium spp. can be the best and most sustainable alternative to the traditionally used and hazardous chemical approach. The lack of race-specific resistance genes against Pythium spp. can be compensated with the candidate quantitative trait loci (QTL) genes in C. annuum L. This review will focus on the epidemiological factors playing a major role in disease spread, the currently available diagnostics in species identification, and the management strategies with a special emphasis on Pythium spp. causing damping-off and root rot in different cultivars of C. annuum L.
Fulltext Silver nanoparticles from insect wing extract: Biosynthesis and evaluation for antioxidant and antimicrobial potential

Jakinala P, Lingampally N, Hameeda B, Sayyed RZ, Khan M Y, Elsayed EA, et al.

PLoS One, 2021;16(3):e0241729.
PMID: 33735177 DOI: 10.1371/journal.pone.0241729

Silver nanoparticles (AgNPs) are among the most widely synthesized and used nanoparticles (NPs). AgNPs have been traditionally synthesized from plant extracts, cobwebs, microorganisms, etc. However, their synthesis from wing extracts of common insect; Mang mao which is abundantly available in most of the Asian countries has not been explored yet. We report the synthesis of AgNPs from M. mao wings extract and its antioxidant and antimicrobial activity. The synthesized AgNPs were spherical, 40-60 nm in size and revealed strong absorption plasmon band around at 430 nm. Highly crystalline nature of these particles as determined by Energy-dispersive X-ray analysis and X-ray diffraction further confirmed the presence of AgNPs. Hydrodynamic size and zeta potential of AgNPs were observed to be 43.9 nm and -7.12 mV, respectively. Fourier-transform infrared spectroscopy analysis revealed the presence of characteristic amide proteins and aromatic functional groups. Thin-layer chromatography (TLC) and Gas chromatography-mass spectroscopy (GC-MS) analysis revealed the presence of fatty acids in the wings extract that may be responsible for biosynthesis and stabilization of AgNPs. Further, SDS-PAGE of the insect wing extract protein showed the molecular weight of 49 kDa. M. mao silver nanoparticles (MMAgNPs) exhibit strong antioxidant, broad-range antibacterial and antifungal activities, (66.8 to 87.0%), broad-range antibacterial and antifungal activities was found with maximum zone of inhibition against Staphylococcus aureus MTCC 96 (35±0.4 mm) and Fusarium oxysporum f. sp. ricini (86.6±0.4) which signifies their biomedical and agricultural potential.
Fulltext Genetic assessment of the internal transcribed spacer region (ITS1.2) in Mangifera indica L. landraces

Fazeli-Nasab B, Sayyed RZ, Farsi M, Ansari S, El-Enshasy HA

Physiol Mol Biol Plants, 2020 Jan;26(1):107-117.
PMID: 32158124 DOI: 10.1007/s12298-019-00732-x

Mango (Mangifera indica) is one of the most important tropical fruits in the world. Twenty-two genotypes of native mangoes from different regions of southern Iran (Hormozgan and Kerman) were collected and analyzed for the ribosomal genes. GC content was found to be 55.5%. Fu and Li's D* test statistic (0.437), Fu and Li's F* test statistic (0.500) and Tajima's D (1.801) were positive and nonsignificant. A total of 769 positions were identified (319 with insertion or deletion including 250 polymorphic and 69 monomorphic loci; 450 loci without any insertion or deletion including 35 Singletons and 22 haplotypes). Nucleotide diversity of 0.309 and a high genetic differentiation including Chi square of 79.8; P value of 0.3605 and df value of 76 was observed among mango genotypes studied. The numerical value of the ratio dN/dS (0.45) indicated a pure selection in the examined gene and the absence of any key changes. Cluster analysis differentiated the mango used in this research (M. indica L.) into two genotypes but could not differentiate their geographical locations. The results of this study indicated that a high genetic distance exists between HajiGholam (Manojan) and Arbabi (Rodan) genotypes and showed higher genetic diversity in mango of Rodan region. Results of present study suggested that for successful breeding, the genotypes of Rodan region mango especially Arbabi mango can be used as a gene donor and ITS can be a suitable tool for genetic evaluations of inter and intra species.
Fulltext Purification and kinetics of the PHB depolymerase of Microbacterium paraoxydans RZS6 isolated from a dumping yard

Sayyed RZ, Wani SJ, Alyousef AA, Alqasim A, Syed A, El-Enshasy HA

PLoS One, 2019;14(6):e0212324.
PMID: 31211775 DOI: 10.1371/journal.pone.0212324

Poly-β-hydroxybutyrate (PHB) depolymerase is known to decompose PHB, biodegradable polymers and therefore has great commercial significance in the bioplastic sector. However, reports on PHB depolymerases from isolates obtained from plastic-contaminated sites that reflect the potential of the source organism is scarce. In this study, we evaluated the production of extracellular PHB depolymerase from Microbacterium paraoxydans RZS6 isolated from the plastic-contaminated site in the municipal area of Shahada, Maharashtra, India, for the first time. The isolate was identified using 16S rRNA gene sequencing, gas chromatographic analysis of fatty acid methyl esters (GC-FAME), and BIOLOG method. Ithydrolyzed PHB on minimal salt medium (MSM) containing PHB as the only source of carbon. The isolate produced PHB depolymerase at 45°C during 48 h of incubation. The enzyme was purified most efficiently using octyl-sepharose CL-4B column, with the highest purification yield of 6.675 Umg-1mL-1. The activity of the enzyme was enhanced in the presence of Ca2+ and Mg2+ ions but inhibited by Fe2+ (1 mM) ions and mercaptoethanol (1000 rpm). the nzyme kinetic analysis revealed that the enzyme was a metalloenzyme; requiring Mg2+ ions, that showed optimum enzyme activity at 30°C (mesophilic) and under neutrophilic (pH 7) conditions. Scale-up from the shake-flask level to a laboratory-scale bioreactor further enhanced the enzyme yield by 0.809 UmL-1. The molecular weight of the enzyme (40 kDa), as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, closely resembled the PHB depolymerase of Aureobacterium saperdae. Our findings highlighted the applicability of M. paraoxydans as a producer of extracellular PHB depolymerase having potential of degrading PHB under diverse conditions.
Fulltext Eco-friendly soil amendments improve growth, antioxidant activities, and root colonization in lingrain (Linum Usitatissimum L.) under drought conditions

Fallah M, Hadi H, Amirnia R, Hassanzadeh-Ghorttapeh A, Zuan ATK, Sayyed RZ

PLoS One, 2021;16(12):e0261225.
PMID: 34941919 DOI: 10.1371/journal.pone.0261225

This study's primary purpose was to investigate the possible amelioration of limited irrigation conditions by mycorrhiza (AMF), vermicompost, and green manure for lingrain plants. This experiment was accomplished as a factorial based on the completely randomized design with three replications. The first factor was green manure (without green manure and with Trifolium pratense as green manure); the second factor consisted of Rhizophagus irregularis mycorrhiza, vermicompost, a combination of mycorrhiza and vermicompost and none of them, and also the third factor was irrigation regime (full irrigation and late-season water limitation). Green manure, vermicompost, and mycorrhiza single-use enhanced the plant's underwater limitation conditions compared to the control. However, vermicompost and green manure or mycorrhiza developed a positive synergistic effect on most traits. Combining green manure with the dual fertilizer (mycorrhiza + vermicompost) resulted in the vermicompost and mycorrhiza synergistic effects, especially under limited irrigation. Consequently, the combination of green manure, mycorrhiza, and vermicompost experienced the highest amount of leaf relative water content, root colonization, leaf nitrogen, chlorophyll a, chlorophyll b, carotenoids, antioxidant enzymes activity, grain yield, and oil yield, which would lead to more resistance of plants to limited irrigation conditions.
Fulltext Optimization and identification of siderophores produced by Pseudomonas monteilii strain MN759447 and its antagonism toward fungi associated with mortality in Dalbergia sissoo plantation forests

Srivastava P, Sahgal M, Sharma K, Enshasy HAE, Gafur A, Alfarraj S, et al.

Front Plant Sci, 2022;13:984522.
PMID: 36438130 DOI: 10.3389/fpls.2022.984522

Siderophore-positive bacteria present in the rhizosphere and in bulk soil assist plants by either inhibiting phytopathogen proliferation or increasing plant growth. The bacterial diversity of the Shisham forest ecosystem in the Tarai region of the Western Himalayas was studied and used for siderophore production, taking into account the large-scale dieback and wilt-induced mortality in Dalbergia sissoo (common name: shisham) plantation forests and the importance of soil microbes in tree health. In addition, Pseudomonas, Burkholderia, and Streptomyces were prominent siderophore-positive bacteria in Shisham forests. Pseudomonas species are known for their remarkable siderophore-producing ability. Bacterial siderophores inhibit pathogen growth by rapidly lowering the number of ferric ions in the rhizosphere. The Pseudomonas monteilii strain MN759447 was isolated from a D. sissoo plantation forest at the Agroforestry Research Centre, Pantnagar, Uttarakhand (28°58'N 79°25'E/28.97°N 79.41°E). It produces a significant number of siderophore units (80.36% in total). A two-stage optimization of growth factors was attempted in the strain MN759447 for better siderophore recovery. In the first-stage single-factor experiment, among the five variables studied, only pH, NH4NO3 concentration, and Fe concentration affected siderophore synthesis. In the second stage, an optimization of pH, NH4NO3 concentration, and Fe concentration for improved growth and enhanced siderophore production was carried out using a Box-Behnken design with response surface methodology. By using LC-MS, two derivatives of pseudomonine, salicylic acid, and kynurenic acid were detected as siderophores in the purified XAD-2 methanol extract of the P. monteilii strain MN759447. In addition to siderophore production, the P. monteilii strain MN759447 also exhibited a broad range of antagonistic activity against Aspergillus calidoustus (65%), Fusarium oxysporum (41.66%), Talaromyces pinophilus (65%), and Talaromyces verruculosus (65.1%) that are linked to sissoo mortality. To our knowledge, this is the first report on siderophore-producing bacteria isolated, identified, and characterized from the D. sissoo Roxb. forest habitat. This strain can also be developed as a commercial product.
Fulltext Production, purification and evaluation of biodegradation potential of PHB depolymerase of Stenotrophomonas sp. RZS7

Sayyed RZ, Wani SJ, Alarfaj AA, Syed A, El-Enshasy HA

PLoS One, 2020;15(1):e0220095.
PMID: 31910206 DOI: 10.1371/journal.pone.0220095

There are numerous reports on poly-β-hydroxybutyrate (PHB) depolymerases produced by various microorganisms isolated from various habitats, however, reports on PHB depolymerase production by an isolate from plastic rich sites scares. Although PHB has attracted commercial significance, the inefficient production and recovery methods, inefficient purification of PHB depolymerase and lack of ample knowledge on PHB degradation by PHB depolymerase have hampered its large scale commercialization. Therefore, to ensure the biodegradability of biopolymers, it becomes imperative to study the purification of the biodegrading enzyme system. We report the production, purification, and characterization of extracellular PHB depolymerase from Stenotrophomonas sp. RZS7 isolated from a dumping yard rich in plastic waste. The isolate produced extracellular PHB depolymerase in the mineral salt medium (MSM) at 30°C during 4 days of incubation under shaking. The enzyme was purified by three methods namely ammonium salt precipitation, column chromatography, and solvent purification. Among these purification methods, the enzyme was best purified by column chromatography on the Octyl-Sepharose CL-4B column giving optimum yield (0.7993 Umg-1mL-1). The molecular weight of purified PHB depolymerase was 40 kDa. Studies on the assessment of biodegradation of PHB in liquid culture medium and under natural soil conditions confirmed PHB biodegradation potential of Stenotrophomonas sp. RZS7. The results obtained in Fourier-Transform Infrared (FTIR) analysis, High-Performance Liquid Chromatography (HPLC) study and Gas Chromatography Mass-Spectrometry (GC-MS) analysis confirmed the biodegradation of PHB in liquid medium by Stenotrophomonas sp. RZS7. Changes in surface morphology of PHB film in soil burial as observed in Field Emission Scanning Electron Microscopy (FESEM) analysis confirmed the biodegradation of PHB under natural soil environment. The isolate was capable of degrading PHB and it resulted in 87.74% biodegradation. A higher rate of degradation under the natural soil condition is the result of the activity of soil microbes that complemented the biodegradation of PHB by Stenotrophomonas sp. RZS7.
Fulltext Nuts as a Part of Dietary Strategy to Improve Metabolic Biomarkers: A Narrative Review

Khalili L, A-Elgadir TME, Mallick AK, El Enshasy HA, Sayyed RZ

Front Nutr, 2022;9:881843.
PMID: 35425791 DOI: 10.3389/fnut.2022.881843

BACKGROUND: Nuts are in the spotlight because of their association with improved health outcomes. We aimed to summarize the findings of previous studies to evaluate the impact of nuts consumption on glycaemic and lipid profile, inflammation, and oxidative stress.
METHODS: Electronic searches for observational and intervention studies were undertaken in PubMed, Embase, Web of Science, and Science Direct until 2022 for searching the studies aiming the application of different types of nuts and the beneficial effects of nuts in improving glycemia, dyslipidemia, inflammation, and oxidative stress.
RESULTS: Results from 56 interventional, 9 narrative and 3 systematic reviews, and 12 meta-analysis studies, aiming at the evaluating beneficial effects of different types of nuts on metabolic markers, showed that nut consumption could improve metabolic markers, including glycaemic factors, lipid profile, and inflammatory and oxidative stress parameters in both healthy and individuals with metabolic disorders in a type-, dose- and duration-dependent manner. According to their unique nutrient components, nuts can be known as a part of a healthy diet, resulting in improved metabolic biomarkers.
CONCLUSION: Considering the efficacy of nuts in improving metabolic markers, incorporation of, incorporating nuts the effectiveness of nuts in improving metabolic markers, incorporating nuts in the diet may prevent the incidence or aggravation of chronic metabolic diseases. Considering the health benefits of the nuts' components, including essential micronutrients, if consumed in the appropriate dose and duration to provide the necessary amount of effective micronutrients to improve health, we will see an improvement in metabolic factors. At the same time, more research is required to determine the optimal type, dose, and duration of nut intervention with regards to metabolic control and reducing the risk of developing metabolic disorders.
Fulltext Role of Bacillus cereus in Improving the Growth and Phytoextractability of Brassica nigra (L.) K. Koch in Chromium Contaminated Soil

Akhtar N, Ilyas N, Yasmin H, Sayyed RZ, Hasnain Z, A Elsayed E, et al.

Molecules, 2021 Mar 12;26(6).
PMID: 33809305 DOI: 10.3390/molecules26061569

Plant growth-promoting rhizobacteria (PGPR) mediate heavy metal tolerance and improve phytoextraction potential in plants. The present research was conducted to find the potential of bacterial strains in improving the growth and phytoextraction abilities of Brassica nigra (L.) K. Koch. in chromium contaminated soil. In this study, a total of 15 bacterial strains were isolated from heavy metal polluted soil and were screened for their heavy metal tolerance and plant growth promotion potential. The most efficient strain was identified by 16S rRNA gene sequencing and was identified as Bacillus cereus. The isolate also showed the potential to solubilize phosphate and synthesize siderophore, phytohormones (indole acetic acid, cytokinin, and abscisic acid), and osmolyte (proline and sugar) in chromium (Cr+3) supplemented medium. The results of the present study showed that chromium stress has negative effects on seed germination and plant growth in B. nigra while inoculation of B. cereus improved plant growth and reduced chromium toxicity. The increase in seed germination percentage, shoot length, and root length was 28.07%, 35.86%, 19.11% while the fresh and dry biomass of the plant increased by 48.00% and 62.16%, respectively, as compared to the uninoculated/control plants. The photosynthetic pigments were also improved by bacterial inoculation as compared to untreated stress-exposed plants, i.e., increase in chlorophyll a, chlorophyll b, chlorophyll a + b, and carotenoid was d 25.94%, 10.65%, 20.35%, and 44.30%, respectively. Bacterial inoculation also resulted in osmotic adjustment (proline 8.76% and sugar 28.71%) and maintained the membrane stability (51.39%) which was also indicated by reduced malondialdehyde content (59.53% decrease). The antioxidant enzyme activities were also improved to 35.90% (superoxide dismutase), 59.61% (peroxide), and 33.33% (catalase) in inoculated stress-exposed plants as compared to the control plants. B. cereus inoculation also improved the uptake, bioaccumulation, and translocation of Cr in the plant. Data showed that B. cereus also increased Cr content in the root (2.71-fold) and shoot (4.01-fold), its bioaccumulation (2.71-fold in root and 4.03-fold in the shoot) and translocation (40%) was also high in B. nigra. The data revealed that B. cereus is a multifarious PGPR that efficiently tolerates heavy metal ions (Cr+3) and it can be used to enhance the growth and phytoextraction potential of B. nigra in heavy metal contaminated soil.
Purification and Characterization of Desferrioxamine B of Pseudomonas fluorescens and Its Application to Improve Oil Content, Nutrient Uptake, and Plant Growth in Peanuts

Nithyapriya S, Sundaram L, Eswaran SUD, Perveen K, Alshaikh NA, Sayyed RZ, et al.

Microb Ecol, 2024 Apr 17;87(1):60.
PMID: 38630182 DOI: 10.1007/s00248-024-02377-0

Microorganisms produce siderophores, which are low-molecular-weight iron chelators when iron availability is limited. The present analyzed the role of LNPF1 as multifarious PGPR for improving growth parameters and nutrient content in peanut and soil nutrients. Such multifarious PGPR strains can be used as effective bioinoculants for peanut farming. In this work, rhizosphere bacteria from Zea mays and Arachis hypogaea plants in the Salem area of Tamil Nadu, India, were isolated and tested for biochemical attributes and characteristics that stimulate plant growth, such as the production of hydrogen cyanide, ammonia (6 µg/mL), indole acetic acid (76.35 µg/mL), and solubilizing phosphate (520 µg/mL). The 16S rRNA gene sequences identified the isolate LNPF1 as Pseudomonas fluorescens with a similarity percentage of 99% with Pseudomonas sp. Isolate LNPF1 was evaluated for the production of siderophore. Siderophore-rich supernatant using a Sep Pack C18 column and Amberlite-400 Resin Column (λmax 264) produced 298 mg/L and 50 mg/L of siderophore, respectively. The characterization of purified siderophore by TLC, HPLC, FTIR, and 2D-NMR analysis identified the compound as desferrioxamine, a hydroxamate siderophore. A pot culture experiment determined the potential of LNPF1 to improve iron and oil content and photosynthetic pigments in Arachis hypogaea L. and improve soil nutrient content. Inoculation of A. hypogea seeds with LNPF1 improved plant growth parameters such as leaf length (60%), shoot length (22%), root length (54.68%), fresh weight (47.28%), dry weight (37%), and number of nuts (66.66) compared to the control (untreated seeds). This inoculation also improved leaf iron content (43.42), short iron content (38.38%), seed iron (46.72%), seed oil (31.68%), carotenoid (64.40%), and total chlorophyll content (98.%) compared to control (untreated seeds). Bacterized seeds showed a substantial increase in nodulation (61.65%) and weight of individual nodules (95.97) vis-à-vis control. The results of the present study indicated that P. fluorescens might be utilized as a potential bioinoculant to improve growth, iron content, oil content, number of nuts and nodules of Arachishypogaea L., and enrich soil nutrients.
Fulltext Tree bark scrape fungus: A potential source of laccase for application in bioremediation of non-textile dyes

Sayyed RZ, Bhamare HM, Sapna, Marraiki N, Elgorban AM, Syed A, et al.

PLoS One, 2020;15(6):e0229968.
PMID: 32497077 DOI: 10.1371/journal.pone.0229968

Although laccase has been recognized as a wonder molecule and green enzyme, the use of low yielding fungal strains, poor production, purification, and low enzyme kinetics have hampered its large-scale application. Thus,this study aims to select high yielding fungal strains and optimize the production, purification, and kinetics of laccase of Aspergillus sp. HB_RZ4. The results obtained indicated that Aspergillus sp. HB_RZ4 produced a significantly large amount of laccase under meso-acidophilic shaking conditions in a medium containing glucose and yeast extract. A 25 μM CuSO4 was observed to enhance the enzyme yield. The enzyme was best purified on a Sephadex G-100 column. The purified enzyme resembled laccase of A. flavus. The kinetics of the purified enzyme revealed high substrate specificity and good velocity of reaction,using ABTS as a substrate. The enzyme was observed to be stable over various pH values and temperatures. The peptide structure of the purified enzyme was found to resemble laccase of A. kawachii IFO 4308. The fungus was observed to decolorize various dyes independent of the requirement of a laccase mediator system.Aspergillus sp. HB_RZ4 was observed to be a potent natural producer of laccase, and it decolorized the dyes even in the absence of a laccase mediator system. Thus, it can be used for bioremediation of effluent that contains non-textile dyes.
Fulltext Production of Biodegradable Polymer from Agro-Wastes in Alcaligenes sp. and Pseudomonas sp

Sayyed RZ, Shaikh SS, Wani SJ, Rehman MT, Al Ajmi MF, Haque S, et al.

Molecules, 2021 Apr 22;26(9).
PMID: 33922162 DOI: 10.3390/molecules26092443

The present study was aimed to evaluate the suitability of agro-wastes and crude vegetable oils for the cost-effective production of poly-β-hydroxybutyrate (PHB), to evaluate growth kinetics and PHB production in Alcaligenes faecalis RZS4 and Pseudomonas sp. RZS1 with these carbon substrates and to study the biodegradation of PHB accumulated by these cultures. Alcaligenes faecalis RZS4 and Pseudomonas sp. RZS1 accumulates higher amounts of PHB corn (79.90% of dry cell mass) and rice straw (66.22% of dry cell mass) medium respectively. The kinetic model suggests that the Pseudomonas sp. RZS1 follows the Monod model more closely than A. faecalis RZS4. Both the cultures degrade their PHB extract under the influence of PHB depolymerase. Corn waste and rice straw appear as the best and cost-effective substrates for the sustainable production of PHB from Alcaligenes faecalis RZS4 and Pseudomonas sp. RZS1. The biopolymer accumulated by these organisms is biodegradable in nature. The agro-wastes and crude vegetable oils are good and low-cost sources of nutrients for the growth and production of PHB and other metabolites. Their use would lower the production cost of PHB and the low-cost production will reduce the sailing price of PHB-based products. This would promote the large-scale commercialization and popularization of PHB as an ecofriendly bioplastic/biopolymer.
Fulltext Psychrotolerant Mesorhizobium sp. Isolated from Temperate and Cold Desert Regions Solubilizes Potassium and Produces Multiple Plant Growth Promoting Metabolites

Baba ZA, Hamid B, Sheikh TA, Alotaibi SH, El Enshasy HA, Ansari MJ, et al.

Molecules, 2021 Sep 23;26(19).
PMID: 34641302 DOI: 10.3390/molecules26195758

Soil potassium (K) supplement depends intensively on the application of chemical fertilizers, which have substantial harmful environmental effects. However, some bacteria can act as inoculants by converting unavailable and insoluble K forms into plant-accessible forms. Such bacteria are an eco-friendly approach for enhancing plant K absorption and consequently reducing utilization of chemical fertilization. Therefore, the present research was undertaken to isolate, screen, and characterize the K solubilizing bacteria (KSB) from the rhizosphere soils of northern India. Overall, 110 strains were isolated, but only 13 isolates showed significant K solubilizing ability by forming a halo zone on solid media. They were further screened for K solubilizing activity at 0 °C, 1 °C, 3 °C, 5 °C, 7 °C, 15 °C, and 20 °C for 5, 10, and 20 days. All the bacterial isolates showed mineral K solubilization activity at these different temperatures. However, the content of K solubilization increased with the upsurge in temperature and period of incubation. The isolate KSB (Grz) showed the highest K solubilization index of 462.28% after 48 h of incubation at 20 °C. The maximum of 23.38 µg K/mL broth was solubilized by the isolate KSB (Grz) at 20 °C after 20 days of incubation. Based on morphological, biochemical, and molecular characterization (through the 16S rDNA approach), the isolate KSB (Grz) was identified as Mesorhizobium sp. The majority of the strains produced HCN and ammonia. The maximum indole acetic acid (IAA) (31.54 µM/mL) and cellulase (390 µM/mL) were produced by the isolate KSB (Grz). In contrast, the highest protease (525.12 µM/mL) and chitinase (5.20 µM/mL) activities were shown by standard strain Bacillus mucilaginosus and KSB (Gmr) isolate, respectively.
Pan-genome analysis and molecular docking unveil the biocontrol potential of Bacillus velezensis VB7 against Phytophthora infestans

Kadiri M, Sevugapperumal N, Nallusamy S, Ragunathan J, Ganesan MV, Alfarraj S, et al.

Microbiol Res, 2023 Mar;268:127277.
PMID: 36577205 DOI: 10.1016/j.micres.2022.127277

Management of late blight of potato incited by Phytophthora infestans remains a major challenge. Coevolution of pathogen with resistant strains and the rise of fungicide resistance have made it more challenging to prevent the spread of P. infestans. Here, the anti-oomycete potential of Bacillus velezensis VB7 against P. infestans through pan-genome analysis and molecular docking were explored. The Biocontrol potential of VB7 against P. infestans was assessed using a confrontational assay. The biomolecules from the inhibition zone were identified and subjected to in silico analysis against P. infestans target proteins. Nucleotide sequences for 54 B. velezensis strains from different geographical locations were used for pan-genome analysis. The confrontational assay revealed the anti-oomycetes potential of VB7 against P. infestans. Molecular docking confirmed that the penicillamine disulfide had the maximum binding energy with eight effector proteins of P. infestans. Besides, scanning electron microscopic observations of P. infestans interaction with VB7 revealed structural changes in hypha and sporangia. Pan-genome analysis between 54 strains of B. velezensis confirmed that the core genome had 2226 genes, and it has an open pan-genome. The present study confirmed the anti-oomycete potential of B. velezensis VB7 against P. infestans and paved the way to explore the genetic potential of VB7.
Fulltext Evaluation of Selenium Nanoparticles in Inducing Disease Resistance against Spot Blotch Disease and Promoting Growth in Wheat under Biotic Stress

Shahbaz M, Akram A, Mehak A, Haq EU, Fatima N, Wareen G, et al.

Plants (Basel), 2023 Feb 08;12(4).
PMID: 36840109 DOI: 10.3390/plants12040761

In the present study, SeNPs were synthesized using Melia azedarach leaf extracts and investigated for growth promotion in wheat under the biotic stress of spot blotch disease. The phytosynthesized SeNPs were characterized using UV-visible spectroscopy, scanning electron microscopy (SEM), energy-dispersive X-ray (EDX), and Fourier-transformed infrared spectroscopy (FTIR). The in vitro efficacy of different concentrations of phytosynthesized SeNPs (i.e., 100 μg/mL, 150 μg/mL, 200 μg/mL, 250 μg/mL, and 300 μg/mL) was evaluated using the well diffusion method, which reported that 300 μg/mL showed maximum fungus growth inhibition. For in vivo study, different concentrations (10, 20, 30, and 40 mg/L) of SeNPs were applied exogenously to evaluate the morphological, physiological, and biochemical parameters under control conditions and determine when infection was induced. Among all treatments, 30 mg/L of SeNPs performed well and increased the plant height by 2.34% compared to the control and 30.7% more than fungus-inoculated wheat. Similarly, fresh plant weight and dry weight increased by 17.35% and 13.43% over the control and 20.34% and 52.48% over the fungus-treated wheat, respectively. In leaf surface area and root length, our findings were 50.11% and 10.37% higher than the control and 40% and 71% higher than diseased wheat, respectively. Plant physiological parameters i.e., chlorophyll a, chlorophyll b, and total chlorophyll content, were increased 14, 133, and 16.1 times over the control and 157, 253, and 42 times over the pathogen-inoculated wheat, respectively. Our findings regarding carotenoid content, relative water content, and the membrane stability index were 29-, 49-, and 81-fold higher than the control and 187-, 63-, and 48-fold higher than the negative control, respectively. In the case of plant biochemical parameters, proline, sugar, flavonoids, and phenolic contents were recorded at 6, 287, 11, and 34 times higher than the control and 32, 107, 33, and 4 times more than fungus-inoculated wheat, respectively. This study is considered the first biocompatible approach to evaluate the potential of green-synthesized SeNPs as growth-promoting substances in wheat under the spot blotch stress and effective management strategy to inhibit fungal growth.
Nanovesicle and extracellular polymeric substance synthesis from the remediation of heavy metal ions from soil

Budamagunta V, Shameem N, Irusappan S, Parray JA, Thomas M, Marimuthu S, et al.

Environ Res, 2023 Feb 15;219:114997.
PMID: 36529326 DOI: 10.1016/j.envres.2022.114997

Heavy metal toxicity affects aquatic plants and animals, disturbing biodiversity and ecological balance causing bioaccumulation of heavy metals. Industrialization and urbanization are inevitable in modern-day life, and control and detoxification methods need to be accorded to meet the hazardous environment. Microorganisms and plants have been widely used in the bioremediation of heavy metals. Sporosarcina pasteurii, a gram-positive bacterium that is widely known for its calcite precipitation property in bio-cementing applications has been explored in the study for its metal tolerance ability for the first time. S. pasteurii SRMNP1 (KF214757) can tolerate silver stress to form nanoparticles and can remediate multiple heavy metals to promote the growth of various plants. This astounding property of the isolate warranted extensive examinations to comprehend the physiological changes during an external heavy metal stress condition. The present study aimed to understand various physiological responses occurring in S. pasteuriiSRMNP1 during the metal tolerance phenomenon using electron microscopy. The isolate was subjected to heavy metal stress, and a transmission electron microscope examination was used to analyze the physiological changes in bacteria to evade the metal stress. S. pasteurii SRMNP1 was tolerant against a wide range of heavy metal ions and can withstand a broad pH range (5-9). Transmission Electron Microscopy (TEM) examination of S. pasteurii SRMNP1 followed by 5 mM nickel sulfate treatment revealed the presence of nanovesicles encapsulating nanosized particles in intra and extracellular spaces. This suggests that the bacteria evade the metal stress by converting the metal ions into nanosized particles and encapsulating them within nanovesicles to efflux them through the vesicle budding mechanism. Moreover, the TEM images revealed an excessive secretion of extracellular polymeric substances by the strain to discharge the metal particles outside the bacterial system. S. pasteurii can be foreseen as an effective bioremediation agent with the potential to produce nanosized particles, nanovesicles, and extracellular polymeric substances. This study provides physiological evidence that, besides calcium precipitation applications, S. pasteurii can further be explored for its multidimensional roles in the fields of drug delivery and environmental engineering.
Development of novel kinetic model based on microbiome and biochar for in-situ remediation of total petroleum hydrocarbons (TPHs) contaminated soil

Saeed M, Ilyas N, Bibi F, Shabir S, Jayachandran K, Sayyed RZ, et al.

Chemosphere, 2023 May;324:138311.
PMID: 36878368 DOI: 10.1016/j.chemosphere.2023.138311

A novel kinetic model has been developed to explain the degradation of total petroleum hydrocarbons. Microbiome engineered biochar amendment may result in a synergistic impact on degradation of total petroleum hydrocarbons (TPHs). Therefore, the present study analyzed the potential of hydrocarbon-degrading bacteria A designated as Aeromonas hydrophila YL17 and B as Shewanella putrefaciens Pdp11 morphological characterized as rod shaped, anaerobic and gram-negative immobilized on biochar, and the degradation efficiency was measured by gravimetric analysis and gas chromatography-mass spectrometry (GC-MS). Whole genome sequencing of both strains revealed the existence of genes responsible for hydrocarbon degradation. In 60 days remediation setup, the treatment consisting of immobilization of both strains on biochar proved more efficient with less half-life and better biodegradation potentials compared to biochar without strains for decreasing the content of TPHs and n-alkanes (C12-C18). Enzymatic content and microbiological respiration showed that biochar acted as a soil fertilizer and carbon reservoir and enhanced microbial activities. The removal efficiency of hydrocarbons was found to be a maximum of 67% in soil samples treated with biochar immobilized with both strains (A + B), followed by biochar immobilized with strain B 34%, biochar immobilized with strain A 29% and with biochar 24%, respectively. A 39%, 36%, and 41% increase was observed in fluorescein diacetate (FDA) hydrolysis, polyphenol oxidase and dehydrogenase activities in immobilized biochar with both strains as compared to control and individual treatment of biochar and strains. An increase of 35% was observed in the respiration rate with the immobilization of both strains on biochar. While a maximum colony forming unit (CFU/g) was found 9.25 with immobilization of both strains on biochar at 40 days of remediation. The degradation efficiency was due to synergistic effect of both biochar and bacteria based amendment on the soil enzymatic activity and microbial respiration.

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