Search results for: rhizobacteria
Commenced in January 2007
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Edition: International
Paper Count: 32

Search results for: rhizobacteria

32 Biocontrol Potential of Growth Promoting Rhizobacteria against Root Rot of Chili and Enhancement of Plant Growth

Authors: Kiran Nawaz, Waheed Anwar, Sehrish Iftikhar, Muhammad Nasir Subhani, Ahmad Ali Shahid

Abstract:

Plant growth promoting rhizobacteria (PGPR) have been extensively studied and applied for the biocontrol of many soilborne diseases. These rhizobacteria are very efficient against root rot and many other foliar diseases associated with solanaceous plants. These bacteria may inhibit the growth of various pathogens through direct inhibition of target pathogens or indirectly by the initiation of systemic resistance (ISR) which is active all over the complete plant. In the present study, 20 different rhizobacterial isolates were recovered from the root zone of healthy chili plants. All soil samples were collected from various chili-growing areas in Punjab. All isolated rhizobacteria species were evaluated in vitro and in vivo against Phytophthora capsici. Different species of Bacillus and Pseudomonas were tested for the antifungal activity against P. capsici the causal organism of Root rot disease in different crops together with chili. Dual culture and distance culture bioassay were carried out to study the antifungal potential of volatile and diffusible metabolites secreted from rhizobacteria. After seven days of incubation at 22°C, growth inhibition rate was recorded. Growth inhibition rate depended greatly on the tested bacteria and screening methods used. For diffusible metabolites, inhibition rate was 35-62% and 20-45% for volatile metabolites. The screening assay for plant growth promoting and disease inhibition potential of chili associated PGPR indicated 42-100% reduction in disease severity and considerable enhancement in roots fresh weight by 55-87%, aerial parts fresh weight by 35-65% and plant height by 65-76% as compared to untreated control and pathogen-inoculated plants. Pseudomonas flourescene, B. thuringiensis, and B. subtilis were found to be the most efficient isolates in inhibiting P. capsici radial growth, increase plant growth and suppress disease severity.

Keywords: rhizobacteria, chili, phytophthora, root rot

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31 Alleviation of Thermal Stress in Pinus ponderosa by Plant-Growth Promoting Rhizobacteria Isolated from Mixed-Conifer Forests

Authors: Kelli G. Thorup, Kristopher A. Blee

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Climate change enhances the occurrence of extreme weather: wildfires, drought, rising summer temperatures, all of which dramatically decline forest growth and increase tree mortality in the mixed-conifer forests of Sierra Nevada, California. However, microbiota living in mutualistic relations with plant rhizospheres have been found to mitigate the effects of suboptimal environmental conditions. The goal of this research is to isolate native beneficial bacteria, plant-growth promoting rhizobacteria (PGPR), that can alleviate heat stress in Pinus ponderosa seedlings. Bacteria were isolated from the rhizosphere of Pinus ponderosa juveniles located in mixed-conifer stand and further characterized for PGP potential based on their ability to produce key growth regulatory phytohormones including auxin, cytokinin, and gibberellic acid. Out of ten soil samples taken, sixteen colonies were isolated and qualitatively confirmed to produce indole-3-acetic acid (auxin) using Salkowski’s reagent. Future testing will be conducted to quantitatively assess phytohormone production in bacterial isolates. Furthermore, bioassays will be performed to determine isolates abilities to increase tolerance in heat-stressed Pinus ponderosa seedlings. Upon completion of this research, a PGPR could be utilized to support the growth and transplantation of conifer seedlings as summer temperatures continue to rise due to the effects of climate change.

Keywords: conifer, heat-stressed, phytohormones, Pinus ponderosa, plant-growth promoting rhizobacteria

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30 Metabolic Regulation of Rhizobacteria for Cool-Season Grass Tolerance to Heat Stress

Authors: Kashif Jaeel, Bingru Huang

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Stress-induced accumulation of ethylene exacerbates drought damages in plants, and suppressing stress induction of ethylene may promote plant tolerance to heat stress. The objective of this study was to investigate the effects of endophytic bacteria (Paraburkholderia aspalathi) with 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase enzymes in suppressing ethylene production on plant tolerance to heat stress and underlying physiological mechanisms of P. aspalathi-regulation in creeping bentgrass (Agrostis stolonifera). A novel strain of P. aspalathi, ‘WSF23’, with ACC deaminase activity was used to inoculate the roots of plants (cv. ‘Penncross’) subjected to heat stress in controlled-environment chambers. Inoculation with WSF23 bacteria resulted in improved shoot and root growth during heat stress. The differential changes in metabolite regulation due to the bacterial inoculation could contribute to ACC deamination bacteria-improved heat tolerance in cool-season grass species.

Keywords: rhizobacteria, grass, heat, plant metabolism, soil bacteria

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29 Nitrogen-Fixing Rhizobacteria (Rhizobium mililoti 2011) Enhances the Tolerance and the Accumulation of Cadmium in Medicago sativa

Authors: Tahar Ghnaya, Majda Mnasri, Hanen Zaier, Rim Ghabriche, Chedly Abdelly

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It is known that the symbiotic association between plant and microorganisms are beneficial for plant growth and resistance to metal stress. Hence, it was demonstrated that Arbuscular mycorrhizal fungi have a positive effect on host plants growing in metal polluted soils. Legume plants are those which normally associate to rhizobacteria in order to fix atmospheric nitrogen. The aim of this work was to evaluate the effect this type of symbiosis on the tolerance and the accumulation of Cd. We chose Medicago sativa, as a modal for host legume plants and Rhizobium mililoti 2011 as rhizobial strain. Inoculated and non-inoculated plants of M. sativa were submitted during three month to 0, 50, and 100 mgCd/kg dry soil. Results showed that the presence of Cd in the medium induced, in both inoculated and non-inoculated plants, a chlorosis and necrosis. However, these symptoms were more pronounced in non-inoculated plants. The beneficial effect of inoculation of M. sativa with R. meliloti, on plant growth was confirmed by the measurement of biomass production which showed that the symbiotic association between host plant and rhizobacteria alleviates significantly Cd effect on biomass production, so inoculated plants produced more dry weight as compared to non-inoculated ones in the presence of all Cd tretments. On the other hand, under symbiosis conditions, Cd was more accumulated in different plant organs. Hence, in these plants, shoot Cd concentration reached 425 and it was 280 µg/gDW in non-inoculated ones in the presence of 100 ppm Cd. This result suggests that symbiosis enhances the absorption and translocation of Cd in this plant. In nodules and roots, we detected the highest Cd concentrations, demonstrating that these organs are able to concentrate Cd in their tissues. These data confirm that M. sataiva, cultivated in symbiosis with Rhizobium mililoti could be used in phytoextraction of Cd from contaminated soils.

Keywords: Cd, phytoremediation, Medicago sativa, Arbuscular mycorrhizal

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28 Screening of Indigenous Rhizobacteria for Growth Promoting and Antagonistic Activity against Fusarium Oxysporoum in Tomato

Authors: Mohammed H. Abu-Dieyeh, Mohammad M. Zalloum

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Plant growth-promoting rhizobacteria (PGPR) are known to enhance plant growth and/or reduce plant damage due to soil-borne pathogens. Tomato is the highest consumable vegetable world-wide including Jordan. Fusarium oxysporum is a pathogen that causes well-known damages and losses to many vegetable crops including tomato. In this study, purification of 112 isolates of PGPR strains from rhizosphere environment of different regions in Jordan was accomplished. All bacterial isolates were In-vitro screened for antagonistic effects against F. oxysporum. The eleven most effective isolates that caused 30%-50% in-vitro growth reduction of F. oxysporum were selected. 8 out of 11 of these isolates were collected from Al-Halabat (arid-land). 7 isolates of Al-Halabat exerted 40-54% In-vitro growth reduction of F. oxysporum. Four-week-old seedlings of tomato cultivar (Anjara, the most susceptible indigenous cultivar to F. oxysporum) treated with PGPR5 (Bacillus amyloliquefaciens), and exposed to F. oxysporum, showed no disease symptoms and no significant changes in biomasses or chlorophyll contents indicating a non-direct mechanism of action of PGPR on tomato plants. However PGPR3 (Bacillus sp.), PGPR4 (Bacillus cereus), and PGPR38 (Paenibacillus sp.) treated plants or PGPR treated and exposed to F. oxysporum showed a significant increasing growth of shoot and root biomasses as well as chlorophyll contents of leaves compared to control untreated plants or plants exposed to the fungus without PGPR treatment. A significant increase in number of flowers per plant was also recorded in all PGPR treated plants. The characterization of rhizobacterial strains were accomplished using 16S rRNA gene sequence analysis in addition to microscopic characterization. Further research is necessary to explore the potentiality of other collected PGPR isolates on tomato plants in addition to investigate the efficacy of the identified isolates on other plant pathogens and then finding a proper and effective methods of formulation and application of the successful isolates on selected crops.

Keywords: antagonism, arid land, growth promoting, rhizobacteria, tomato

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27 Rejuvenation of Peanut Seedling from Collar Rot Disease by Azotobacter sp. RA2

Authors: Ravi R. Patel, Vasudev R. Thakkar

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Use of plant growth-promoting rhizobacteria (PGPR) to increase the production and decrees disease occurrence is a recent method in agriculture. An RA2 rhizospheric culture was isolated from peanut rhizosphere from Junagadh region of Gujarat, India and showed different direct and indirect plant growth promoting activity like indole acetic acid, gibberellic acid, siderophore, hydrogen cyanide, Ammonia and (1-Aminocyclopropane-1-Carboxylate) deaminase production, N2 fixation, phosphate and potassium solubilization in vitro. RA2 was able to protect peanut germinating seedling from A. niger infection and reduce collar rot disease incidence 60-35% to 72-41% and increase germination percentage from 70-82% to 75-97% in two varieties GG20 and GG2 of peanut. RA2 was found to induce resistance in A. hypogaea L. seedlings via induction of different defense-related enzymes like phenylalanine ammonia lyase, peroxidase, polyphenol oxidase, lipoxygenase and pathogenesis related protein like chitinase, ß – 1,3- glucanase. Jasmonic acid one of the major signaling molecules of inducing systemic resistance was also found to induced due to RA2 treatments. RA2 bacterium was also promoting peanut growth and reduce A. niger infection in pot studies. 16S rDNA sequence of RA2 showed 99 % homology to Azotobacter species.

Keywords: plant growth promoting rhizobacteria, peanut, aspergillus niger, induce systemic resistance

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26 Biohydrogen Production from Rice Water Using Bacteria Isolated from Wetland Sediment

Authors: Jerry John T. M., Sylas V. P., Shijo Joy

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Hydrogen is the most essential gas that can be used for many purposes. During the production of hydrogen using raw materials like Soil and leftover cooked rice water (kanjivellam), the major by-product formed is water. Soil is collected from three different places in kottayam district: Kallara, Meenachilar, and Athirampuzha. Collected samples are mixed with rice water and tested for traces of hydrogen using a biohydrogen sensor after 72 hours. The result was the presence of hydrogen in all the 3 samples. After streaking, PCR and gel electrophoresis detected the bacteria which produced the hydrogen. RGCB Thiruvananthapuram conducted the sequencing of the PCR resultant. And identified the bacterial strains. Five variants of Bacillus bacteria ( (1) Bacillus cereus strain JTM GenBank: OP278839.1 (2) Bacillus toyonensis strain JTM2 GenBank: OP278841.1 (3) Bacillus anthracis strain JTM_SR2989-3-R_H08 GenBank: OP278960.1 (4) Bacillus thuringiensis strain JRY1 GenBank: OP278976.1 (5) Bacillus anthracis strain JTM_SR2989-3-F_H07 GenBank: OP278959.1 ) are identified and successfully registered in NCBI Gen bank. These Bacillus bacteria are major types of Rhizobacteria that can form spores and can survive in the soil for a long time period under harsh environmental conditions. Also, plant growth is enhanced by PGPR (Plant growth promoting rhizobacteria) through the induction of systemic resistance, antibiosis, and competitive omission. The molecular sequencing was submitted to the NCBI Gen Bank, and the accession numbers were allotted for the bacterial cultures.

Keywords: bio hydrogen production, bacterial bio hydrogen production, plant related to bacillus bacteria., bacillus bacteria study

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25 Antifungal Potential of the Plant Growth-Promoting Rhizobacteria Infecting Kidney Beans

Authors: Zhazira Shemsheyeva, Zhanara Suleimenova, Olga Shemshura, Gulnaz Mombekova, Zhanar Rakhmetova

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Bacteria that colonize plant roots and promote plant growth are referred to as plant growth-promoting rhizobacteria (PGPR). They not only provide nutrients to the plants (direct plant growth promotion) and protect plants against the phytopathogens (indirect plant growth promotion) but also increase the soil fertility. Indirectly PGPRs improve the plant growth by becoming a biocontrol agent for a fungal pathogen. The antifungal activities of the PGPrhizobacteria were assayed against different species of phytopathogenic fungi such as Fusarium tricinctum, Fusarium oxysporum, Sclerotiniasclerotiorum, and Botrytis cinerea. Pseudomonas putidaSM-1, Azotobacter sp., and Bacillus thuringiensis AKS/16 strains have been used in experimental tests on growth inhibition of phytopathogenic fungi infecting Kidney beans. Agar well diffusion method was used in this study. Diameters of the zones of inhibition were measured in millimeters. It was found that Bacillus thuringiensis AKS/16 strain showed the lowest antifungal activity against all fungal pathogens tested. Zones of inhibition were 15-18 mm. In contrast, Pseudomonas putida SM-1 exhibited good antifungal activity against Fusarium oxysporum and Fusarium tricinctum by producing 29-30 mm clear zones of inhibition. The moderate inhibitory effect was shown by Azotobacter sp. against all fungal pathogens tested with zones of inhibition from24 to 26 mm. In summary, Pseudomonas putida SM-1 strain demonstrated the potential of controlling root rot diseases in kidney beans.

Keywords: PGPR, pseudomonas putida, kindey beans, antifungal activity

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24 Potential Growth of Tomato Plants in Induced Saline Soil with Rhizobacteria (PGPR)

Authors: Arfan Ali, Idrees Ahmad Nasir

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The critical evaluation of tolerance in tomato plants against the induced saline soil were assessed by transcript analysis of genes coding for products potentially involved in stress tolerance. A reverse transcriptase PCR experiment was performed with Hsp90-1, MT2, and GR1like protein genes using RNA isolated from different tissues of tomato plants. Four strains of Bacillus magisterium were inoculated with 100 Mm & 200 Mm concentrations of salt. Eleven treatments each ten replica pots were installed in green house experiment and the parameters taken into account were morphological (length, weight, number of leaves, leaf surface area), chemical (anthocyanin, chlorophyll-a, chlorophyll-b, carotenoids) and biological (gene expression). Results bare a response i.e. highest response of MT2 like gene was at 24 hpi and the highest levels of GR1 like protein transcript accumulation were detected at 36 hpi. The chemical and morphological parameters at diverse salt concentrations bequeath superlative response amongst strains which candidly flank on Zm7 and Zm4. Therefore, Bacillus magisterium Zm7 strains and somehow Zm4 strain can be used in saline condition to make plants tolerant. The overall performance of strains Zm7, Zm6, and Zm4 was found better for all studied traits under salt stress conditions. Significant correlations among traits root length, shoot length, number of leaves, leaf surface area, carotenoids, anthocyanin, chlorophyll-a and chlorophyll-b were found and suggested that the salt tolerance in tomato may be improved through the use of PGPR strains.

Keywords: Bacillus magisterium, gene expression glutathione reductase, metallothionein, PGPR, Rhizobacteria, saline

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23 Effect of Chemical Fertilizer on Plant Growth-Promoting Rhizobacteria in Wheat

Authors: Tessa E. Reid, Vanessa N. Kavamura, Maider Abadie, Adriana Torres-Ballesteros, Mark Pawlett, Ian M. Clark, Jim Harris, Tim Mauchline

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The deleterious effect of chemical fertilizer on rhizobacterial diversity has been well documented using 16S rRNA gene amplicon sequencing and predictive metagenomics. Biofertilization is a cost-effective and sustainable alternative; improving strategies depends on isolating beneficial soil microorganisms. Although culturing is widespread in biofertilization, it is unknown whether the composition of cultured isolates closely mirrors native beneficial rhizobacterial populations. This study aimed to determine the relative abundance of culturable plant growth-promoting rhizobacteria (PGPR) isolates within total soil DNA and how potential PGPR populations respond to chemical fertilization in a commercial wheat variety. It was hypothesized that PGPR will be reduced in fertilized relative to unfertilized wheat. Triticum aestivum cv. Cadenza seeds were sown in a nutrient depleted agricultural soil in pots treated with and without nitrogen-phosphorous-potassium (NPK) fertilizer. Rhizosphere and rhizoplane samples were collected at flowering stage (10 weeks) and analyzed by culture-independent (amplicon sequence variance (ASV) analysis of total rhizobacterial DNA) and -dependent (isolation using growth media) techniques. Rhizosphere- and rhizoplane-derived microbiota culture collections were tested for plant growth-promoting traits using functional bioassays. In general, fertilizer addition decreased the proportion of nutrient-solubilizing bacteria (nitrate, phosphate, potassium, iron and, zinc) isolated from rhizocompartments in wheat, whereas salt tolerant bacteria were not affected. A PGPR database was created from isolate 16S rRNA gene sequences and searched against total soil DNA, revealing that 1.52% of total community ASVs were identified as culturable PGPR isolates. Bioassays identified a higher proportion of PGPR in non-fertilized samples (rhizosphere (49%) and rhizoplane (91%)) compared to fertilized samples (rhizosphere (21%) and rhizoplane (19%)) which constituted approximately 1.95% and 1.25% in non-fertilized and fertilized total community DNA, respectively. The analyses of 16S rRNA genes and deduced functional profiles provide an in-depth understanding of the responses of bacterial communities to fertilizer; this study suggests that rhizobacteria, which potentially benefit plants by mobilizing insoluble nutrients in soil, are reduced by chemical fertilizer addition. This knowledge will benefit the development of more targeted biofertilization strategies.

Keywords: bacteria, fertilizer, microbiome, rhizoplane, rhizosphere

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22 Effect of Chemical Mutagen on Seeds Germination of Lima Bean

Authors: G. Ultanbekova, Zh. Suleimenova, Zh. Rakhmetova, G. Mombekova, S. Mantieva

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Plant Growth Promoting Rhizobacteria (PGPR) are a group of free-living bacteria that colonize the rhizosphere, enhance plant growth of many cereals and other important agricultural crops and protect plants from disease and abiotic stresses through a wide variety of mechanisms. The use of PGPR has been proven to be an environmentally sound way of increasing crop yields by facilitating plant growth. In the present study, strain improvement of PGPR isolates were carried out by chemical mutagenesis for the improvement of growth and yield of lima bean. Induced mutagenesis is widely used for the selection of microorganisms producing biologically active substances and further improving their activities. Strain improvement is usually done by classical mutagenesis which involves exposing the microbes to chemical or physical mutagens. The strains of Pseudomonas putida 4/1, Azotobacter chroococcum Р-29 and Bacillus subtilis were subjected to mutation process for strain improvement by treatment with a chemical agent (sodium nitrite) to cause mutation and were observed for its consequent action on the seeds germination and plant growth of lima bean (Phaseolus lunatus). Bacterial mutant strains of Pseudomonas putida M-1, Azotobacter chroococcum M-1 and Bacillus subtilis M-1, treated with sodium nitrite in the concentration of 5 mg/ml for 120 min, were found effective to enhance the germination of lima bean seeds compared to parent strains. Moreover, treatment of the lima bean seeds with a mutant strain of Bacillus subtilis M-1 had a significant stimulation effect on plant growth. The length of the stems and roots of lima bean treated with Bacillus subtilis M-1 increased significantly in comparison with parent strain in 1.6 and 1.3 times, respectively.

Keywords: chemical mutagenesis, germination, kidney bean, plant growth promoting rhizobacteria (PGPR)

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21 Screening and Evaluation of Plant Growth Promoting Rhizobacteria of Wheat/Faba Bean for Increasing Productivity and Yield

Authors: Yasir Arafat, Asma Shah, Hua Shao

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Background and Aims: Legume/cereal intercropping is used worldwide for enhancement in biomass and yield of cereal crops. However, because of intercropping, the belowground biological and chemical interactions and their effect on physiological parameters and yield of crops are limited. Methods: Wheat faba bean (WF) intercropping was designed to understand the underlying changes in the soil's chemical environment, soil microbial communities, and effect on growth and yield parameters. Experimental plots were established as having no root partition (NRP), semi-root partition (SRP), complete root partition (CRP), and their sole cropping (CK). Low molecular weight organic acids (LMWOAs) were determined by GC-MS, and high throughput sequencing of the 16S rRNA gene was carried out to screen microbial structure and composition in different root partitions of the WF intercropping system. Results: We show that intercropping induced a shift in the relative abundance of some genera of plant growth promoting rhizobacteria (PGPR) such as Allorhizobium, Neorhizobium, Pararhizobium, and Rhizobium species and resulted in better growth and yield performance of wheat. Moreover, as the plant's distance of wheat from faba beans decreased, the diversity of microbes increased, and a positive effect was observed on physiological traits and crop yield. Furthermore, an abundance and positive correlations of palmitic acid, arachidic acid, stearic acid, and 9-Octadecenoic with PGPR were recorded in the root zone of WF intercropping, which can play an important role in this facilitative mechanism of enhancing growth and yield of cereals. Conclusion: The two treatments clearly affected soil microbial and chemical composition, which can be reflected in growth and yield enhancement.

Keywords: intercropping, microbial community, LMWOAs, PGPR, soil chemical environment

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20 Screening of Plant Growth Promoting Rhizobacteria in the Rhizo- and Endosphere of Sunflower (Helianthus anus) and Their Role in Enhancing Growth and Yield Attriburing Trairs and Colonization Studies

Authors: A. Majeed, M.K. Abbasi, S. Hameed, A. Imran, T. Naqqash, M. K. Hanif

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Plant growth-promoting rhizobacteria (PGPR) are free-living soil bacteria that aggressively colonize the rhizosphere/plant roots, and enhance the growth and yield of plants when applied to seed or crops. Root associated (endophytic and rhizospheric) PGPR were isolated from Sunflower (Helianthus anus) grown in soils collected from 16 different sites of sub division Dhirkot, Poonch, Azad Jammu & Kashmir, Pakistan. A total of 150 bacterial isolates were isolated, purified, screened in vitro for their plant growth promoting (PGP) characteristics. 11 most effective isolates were selected on the basis of biochemical assays (nitrogen fixation, phosphate solubilization, growth hormone production, biocontrol assay, and carbon substrates utilization assay through gas chromatography (GCMS), spectrophotometry, high performance liquid chromatography HPLC, fungal and bacterial dual plate assay and BIOLOG GN2/GP2 microplate assay respectively) and were tested on the crop under controlled and field conditions. From the inoculation assay, the most promising 4 strains (on the basis of increased root/shoot weight, root/shoot length, seed oil content, and seed yield) were than selected for colonization studies through confocal laser scanning and transmission electron microscope. 16Sr RNA gene analysis showed that these bacterial isolates belong to Pseudononas, Enterobacter, Azospirrilum, and Citobacter genera. This study is the clear evident that such isolates have the potential for application as inoculants adapted to poor soils and local crops to minimize the chemical fertilizers harmful for soil and environment

Keywords: PGPR, nitrogen fixation, phosphate solubilization, colonization

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19 Molecular Interactions between Vicia Faba L. Cultivars and Plant Growth Promoting Rhizobacteria (PGPR), Utilized as Yield Enhancing 'Plant Probiotics'

Authors: Eleni Stefanidou, Nikolaos Katsenios, Ioanna Karamichali, Aspasia Efthimiadou, Panagiotis Madesis

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The excessive use of pesticides and fertilizers has significant environmental and human health-related negative effects. In the frame of the development of sustainable agriculture practices, especially in the context of extreme environmental changes (climate change), it is important to develop alternative practices to increase productivity and biotic and abiotic stress tolerance. Beneficial bacteria, such as symbiotic bacteria in legumes (rhizobia) and symbiotic or free-living Plant Growth Promoting Rhizobacteria (PGPR), which could act as "plant probiotics", can promote plant growth and significantly increase the resistance of crops under adverse environmental conditions. In this study, we explored the symbiotic relationships between Faba bean (Vicia faba L.) cultivars with different PGPR bacteria, aiming to identify the possible influence on yield and biotic-abiotic phytoprotection benefits. Transcriptomic analysis of root and whole plant samples was executed for two Vicia faba L. cultivars (Polikarpi and Solon) treated with selected PGPR bacteria (6 treatments: B. subtilis + Rhizobium-mixture, A. chroococcum + Rhizobium-mixture, B. subtilis, A. chroococcum and Rhizobium-mixture). Preliminary results indicate a significant yield (Seed weight and Total number of pods) increase in both varieties, ranging around 25%, in comparison to the control, especially for the Solon cultivar. The increase was observed for all treatments, with the B. subtilis + Rhizobium-mixture treatment being the highest performing. The correlation of the physiological and morphological data with the transcriptome analysis revealed molecular mechanisms and molecular targets underlying the observed yield increase, opening perspectives for the use of nitrogen-fixing bacteria as a natural, more ecological enhancer of legume crop productivity.

Keywords: plant probiotics, PGPR, legumes, sustainable agriculture

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18 Effect of Plant Growth Promoting Rhizobacteria on the Germination and Early Growth of Onion (Allium cepa)

Authors: Dragana R. Stamenov, Simonida S. Djuric, Timea Hajnal Jafari

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Plant growth promoting rhizobacteria (PGPR) are a heterogeneous group of bacteria that can be found in the rhizosphere, at root surfaces and in association with roots, enhancing the growth of the plant either directly and/or indirectly. Increased crop productivity associated with the presence of PGPR has been observed in a broad range of plant species, such as raspberry, chickpeas, legumes, cucumber, eggplant, pea, pepper, radish, tobacco, tomato, lettuce, carrot, corn, cotton, millet, bean, cocoa, etc. However, until now there has not been much research about influences of the PGPR on the growth and yield of onion. Onion (Allium cepa L.), of the Liliaceae family, is a species of great economic importance, widely cultivated all over the world. The aim of this research was to examine the influence of plant growth promoting bacteria Pseudomonas sp. Dragana, Pseudomonas sp. Kiš, Bacillus subtillis and Azotobacter sp. on the seed germination and early growth of onion (Allium cepa). PGPR Azotobacter sp., Bacillus subtilis, Pseudomonas sp. Dragana, Pseudomonas sp. Kiš, from the collection of the Faculty of Agriculture, Novi Sad, Serbia, were used as inoculants. The number of cells in 1 ml of the inoculum was 10⁸ CFU/ml. The control variant was not inoculated. The effect of PGPR on seed germination and hypocotyls length of Allium cepa was evaluated in controlled conditions, on filter paper in the dark at 22°C, while effect on the plant length and mass in semicontrol conditions, in 10 l volume vegetative pots. Seed treated with fungicide and untreated seed were used. After seven days the percentage of germination was determined. After seven and fourteen days hypocotil length was measured. Fourteen days after germination, length and mass of plants were measured. Application of Pseudomonas sp. Dragana and Kiš and Bacillus subtillis had a negative effect on onion seed germination, while the use of Azotobacter sp. gave positive results. On average, application of all investigated inoculants had a positive effect on the measured parameters of plant growth. Azotobacter sp. had the greatest effect on the hypocotyls length, length and mass of the plant. In average, better results were achieved with untreated seeds in compare with treated. Results of this study have shown that PGPR can be used in the production of onion.

Keywords: germination, length, mass, microorganisms, onion

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17 Application of Free Living Nitrogen Fixing Bacteria to Increase Productivity of Potato in Field

Authors: Govinda Pathak

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In modern agriculture, the sustainable enhancement of crop productivity while minimizing environmental impacts remains a paramount challenge. Plant Growth Promoting Rhizobacteria (PGPR) have emerged as a promising solution to address this challenge. The rhizosphere, the dynamic interface between plant roots and soil, hosts intricate microbial interactions crucial for plant health and nutrient acquisition. PGPR, a subset of rhizospheric microorganisms, exhibit multifaceted beneficial effects on plants. Their abilities to stimulate growth, confer stress tolerance, enhance nutrient availability, and suppress pathogens make them invaluable contributors to sustainable agriculture. This work examines the pivotal role of free living nitrogen fixer in optimizing agricultural practices. We delve into the intricate mechanisms underlying PGPR-mediated plant-microbe interactions, encompassing quorum sensing, root exudate modulation, and signaling molecule exchange. Furthermore, we explore the diverse strategies employed by PGPR to enhance plant resilience against abiotic stresses such as drought, salinity, and metal toxicity. Additionally, we highlight the role of PGPR in augmenting nutrient acquisition and soil fertility through mechanisms such as nitrogen fixation, phosphorus solubilization, and mineral mobilization. Furthermore, we discuss the potential of PGPR in minimizing the reliance on chemical fertilizers and pesticides, thereby contributing to environmentally friendly agriculture. However, harnessing the full potential of PGPR requires a comprehensive understanding of their interactions with host plants and the surrounding microbial community. We also address challenges associated with PGPR application, including formulation, compatibility, and field efficacy. As the quest for sustainable agriculture intensifies, harnessing the remarkable attributes of PGPR offers a holistic approach to propel agricultural productivity while maintaining ecological balance. This work underscores the promising prospect of free living nitrogen fixer as a panacea for addressing critical agricultural challenges regarding chemical urea in an era of sustainable and resilient food production.

Keywords: PGPR, nitrogen fixer, quorum sensing, Rhizobacteria, pesticides

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16 The Use of Bituminaria bituminosa (L.) Stirton and Microbial Biotechnologies for Restoration of Degraded Pastoral Lands: The Case of the Middle Atlas of Morocco

Authors: O. Zennouhi, M. El Mderssa, J. Ibijbijen, E. Bouiamrine, L. Nassiri

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Rangelands and silvopastoral systems of the middle Atlas are under a heavy pressure, which led to pasture degradation, invasion by non-palatable and toxic species and edaphic aridification due to the regression of the global vegetation cover. In this situation, the introduction of multipurpose leguminous shrubs, such as Bituminaria bituminosa (L.) Stirton, commonly known as bituminous clover, could be a promising socio-ecological alternative for the rehabilitation of these degraded areas. The application of biofertilizers like plant growth promoting rhizobacteria especially phosphate solubilizing bacteria (PSB) can ensure a successful installation of this plant in the selected degraded areas. The main objective of the present work is to produce well-inoculated seedlings using the best efficient PSB strains in the greenhouse to increase their ability to resist to environmental constraints once transplanted to the field in the central Middle Atlas.

Keywords: biofertilizers, bituminaria bituminosa, phosphate solubilizing bacteria, rehabilitation

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15 Effect of Fertilization and Combined Inoculation with Azospirillum brasilense and Pseudomonas fluorescens on Rhizosphere Microbial Communities of Avena sativa (Oats) and Secale Cereale (Rye) Grown as Cover Crops

Authors: Jhovana Silvia Escobar Ortega, Ines Eugenia Garcia De Salamone

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Cover crops are an agri-technological alternative to improve all properties of soils. Cover crops such as oats and rye could be used to reduce erosion and favor system sustainability when they are grown in the same agricultural cycle of the soybean crop. This crop is very profitable but its low contribution of easily decomposable residues, due to its low C/N ratio, leaves the soil exposed to erosive action and raises the need to reduce its monoculture. Furthermore, inoculation with the plant growth promoting rhizobacteria contributes to the implementation, development and production of several cereal crops. However, there is little information on its effects on forage crops which are often used as cover crops to improve soil quality. In order to evaluate the effect of combined inoculation with Azospirillum brasilense and Pseudomonas fluorescens on rhizosphere microbial communities, field experiments were conducted in the west of Buenos Aires province, Argentina, with a split-split plot randomized complete block factorial design with three replicates. The factors were: type of cover crop, inoculation and fertilization. In the main plot two levels of fertilization 0 and 7 40-0-5 (NPKS) were established at sowing. Rye (Secale cereale cultivar Quehué) and oats (Avena sativa var Aurora.) were sown in the subplots. In the sub-subplots two inoculation treatments are applied without and with application of a combined inoculant with A. brasilense and P. fluorescens. Due to the growth of cover crops has to be stopped usually with the herbicide glyphosate, rhizosphere soil of 0-20 and 20-40 cm layers was sampled at three sampling times which were: before glyphosate application (BG), a month after glyphosate application (AG) and at soybean harvest (SH). Community level of physiological profiles (CLPP) and Shannon index of microbial diversity (H) were obtained by multivariate analysis of Principal Components. Also, the most probable number (MPN) of nitrifiers and cellulolytics were determined using selective liquid media for each functional group. The CLPP of rhizosphere microbial communities showed significant differences between sampling times. There was not interaction between sampling times and both, types of cover crops and inoculation. Rhizosphere microbial communities of samples obtained BG had different CLPP with respect to the samples obtained in the sampling times AG and SH. Fertilizer and depth of sampling also caused changes in the CLPP. The H diversity index of rhizosphere microbial communities of rye in the sampling time BG were higher than those associated with oats. The MPN of both microbial functional types was lower in the deeper layer since these microorganisms are mostly aerobic. The MPN of nitrifiers decreased in rhizosphere of both cover crops only AG. At the sampling time BG, the NMP of both microbial types were larger than those obtained for AG and SH. This may mean that the glyphosate application could cause fairly permanent changes in these microbial communities which can be considered bio-indicators of soil quality. Inoculation and fertilizer inputs could be included to improve management of these cover crops because they can have a significant positive effect on the sustainability of the agro-ecosystem.

Keywords: community level of physiological profiles, microbial diversity, plant growth promoting rhizobacteria, rhizosphere microbial communities, soil quality, system sustainability

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14 Productivity and Profitability of Field Pea as Influenced by Different Levels of Fertility and Bio-Fertilizers under Irrigated Condition

Authors: Akhilesh Mishra, Geeta Rai, Arvind Srivastava, Nalini Tiwari

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A field experiment was conducted during two consecutive Rabi seasons of 2007 and 2008 to study the economics of different bio-fertilizer’s inoculations in fieldpea (cv. Jai) at Chandra Shekhar Azad University of Agriculture and Technology, Kanpur (India). Results indicated that the seed inoculation with Rhizobium + PSB + PGPR improved all the growth; yield attributes and yields of field pea. Fresh and dry weight plant-1, nodules number and dry weight plant-1 were found significantly maximum. Number of grains pod-1, number and weight of pods plant-1 at maturity attributed significantly in increasing the grain yield as well as net return. On pooled basis, maximum net income (Rs.22169 ha-1) was obtained with the use of Rhizobium + PSB + PGPR which was improved by a margin of Rs.1502 (6.77%), 2972 (13.40%), 2672 (12.05%), 5212 (23.51%), 6176 (27.85%), 4666 (21.04%) and 8842/ha (39.88%) over the inoculation of PSB + PGPR, Rhizobium + PGPR, Rhizobium + PSB, PGPR, PSB, Rhizobium and control, respectively. Thus, it can be recommended that to earn the maximum net profit from dwarf field pea, seed should be inoculated with Rhizobium + PSB + PGPR.

Keywords: rhizobium, phosphorus solubilizing bacteria, plant growth promoting rhizobacteria, field pea

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13 Effects of Plant Growth Promoting Rhizobacteria on the Yield and Nutritive Quality of Tomato Fruits

Authors: Narjes Dashti, Nida Ali, Magdy Montasser, Vineetha Cherian

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The influence of two PGPR strains, Pseudomonas aeruginosa and Stenotrophomonas rhizophilia, on fruit yields, pomological traits and chemical contents of tomato (Solanum lycopersicum) fruits were studied. The study was conducted separately on two different cultivar varieties of tomato, namely Supermarmande and UC82B. The results indicated that the presence of the PGPR almost doubled the average yield per plant. There was a significant improvement in the pomological qualities of the PGPR treated tomato fruits compared to the corresponding healthy treatments especially in traits such as the average fruit weight, height, and fruit volume. The chemical analysis of tomato fruits revealed that the presence of the PGPRs increased the total protein, lycopene, alkalinity and phenol content of the tomato fruits compared to the healthy controls. They had no influence on the reduced sugar, total soluble solids or the titerable acid content of fruits. However their presence reduced the amount of ascorbic acid in tomato fruits compared to the healthy controls.

Keywords: PGPR, tomato, fruit quality

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12 Impact of Elevated Temperature on Spot Blotch Development in Wheat and Induction of Resistance by Plant Growth Promoting Rhizobacteria

Authors: Jayanwita Sarkar, Usha Chakraborty, Bishwanath Chakraborty

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Plants are constantly interacting with various abiotic and biotic stresses. In changing climate scenario plants are continuously modifying physiological processes to adapt to changing environmental conditions which profoundly affect plant-pathogen interactions. Spot blotch in wheat is a fast-rising disease in the warmer plains of South Asia where the rise in minimum average temperature over most of the year already affecting wheat production. Hence, the study was undertaken to explore the role of elevated temperature in spot blotch disease development and modulation of antioxidative responses by plant growth promoting rhizobacteria (PGPR) for biocontrol of spot blotch at high temperature. Elevated temperature significantly increases the susceptibility of wheat plants to spot blotch causing pathogen Bipolaris sorokiniana. Two PGPR Bacillus safensis (W10) and Ochrobactrum pseudogrignonense (IP8) isolated from wheat (Triticum aestivum L.) and blady grass (Imperata cylindrical L.) rhizophere respectively, showing in vitro antagonistic activity against Bipolaris sorokiniana were tested for growth promotion and induction of resistance against spot blotch in wheat. GC-MS analysis showed that Bacillus safensis (W10) and Ochrobactrum pseudogrignonense (IP8) produced antifungal and antimicrobial compounds in culture. Seed priming with these two bacteria significantly increase growth, modulate antioxidative signaling and induce resistance and eventually reduce disease incidence in wheat plants at optimum as well as elevated temperature which was further confirmed by indirect immunofluorescence assay using polyclonal antibody raised against Bipolaris sorokiniana. Application of the PGPR led to enhancement in activities of plant defense enzymes- phenylalanine ammonia lyase, peroxidase, chitinase and β-1,3 glucanase in infected leaves. Immunolocalization of chitinase and β-1,3 glucanase in PGPR primed and pathogen inoculated leaf tissue was further confirmed by transmission electron microscopy using PAb of chitinase, β-1,3 glucanase and gold labelled conjugates. Activity of ascorbate-glutathione redox cycle related enzymes such as ascorbate peroxidase, superoxide dismutase and glutathione reductase along with antioxidants such as carotenoids, glutathione and ascorbate and osmolytes like proline and glycine betain accumulation were also increased during disease development in PGPR primed plant in comparison to unprimed plants at high temperature. Real-time PCR analysis revealed enhanced expression of defense genes- chalcone synthase and phenyl alanineammonia lyase. Over expression of heat shock proteins like HSP 70, small HSP 26.3 and heat shock factor HsfA3 in PGPR primed plants effectively protect plants against spot blotch infection at elevated temperature as compared with control plants. Our results revealed dynamic biochemical cross talk between elevated temperature and spot blotch disease development and furthermore highlight PGPR mediated array of antioxidative and molecular alterations responsible for induction of resistance against spot blotch disease at elevated temperature which seems to be associated with up-regulation of defense genes, heat shock proteins and heat shock factors, less ROS production, membrane damage, increased expression of redox enzymes and accumulation of osmolytes and antioxidants.

Keywords: antioxidative enzymes, defense enzymes, elevated temperature, heat shock proteins, PGPR, Real-Time PCR, spot blotch, wheat

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11 Quality and Yield of Potato Seed Tubers as Influenced by Plant Growth Promoting Rhizobacteria

Authors: Muhammad Raqib Rasul, Tavga Sulaiman Rashid

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Fertilization increases efficiency and obtains better quality of product recovery in agricultural activities. However, fertilizer consumption increased exponentially throughout the world, causing severe environmental problems. Biofertilizers can be a practical approach to minimize chemical fertilizer sources and ultimately develop soil fertility. This study was carried out to isolate, identify and characterize bacteria from medicinal plant (Rumex tuberosus L. and Verbascum sp.) rhizosphere for in vivo screening. 25 bacterial isolates were isolated and several biochemical tests were performed. Two isolates that were positive for most biochemical tests were chosen for the field experiment. The isolates were identified as Go1 Alcaligenes faecalis (Accession No. OP001725) and T11 (Bacillus sp.) based on the 16S rRNA sequence analysis that was compared with related bacteria in GenBank database using MEGA 6.1. For the field trial isolate GO1 and T11 (separately and mixed), NPK as a positive control was used. Both isolates increased plant height, chlorophyll content, number of tubers, and tuber’s weight. The results demonstrated that these two isolates of bacteria can potentially replace with chemical fertilizers for potato production.

Keywords: biofertilizer, Bacillus subtilis, Alcaligenes faecalis, potato tubers, in vivo screening

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10 Nature-based Solutions for Mitigating the Impact of Climate Change on Plants: Utilizing Encapsulated Plant Growth Regulators and Associative Microorganisms

Authors: Raana Babadi Fathipour

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Over the past decades, the climatic CO2 concentration and worldwide normal temperature have been expanding, and this drift is anticipated to before long gotten to be more extreme. This situation of climate alter escalate abiotic stretch components (such as dry spell, flooding, saltiness, and bright radiation) that debilitate timberland and related environments as well as trim generation. These variables can contrarily influence plant development and advancement with a ensuing lessening in plant biomass aggregation and surrender, in expansion to expanding plant defenselessness to biotic stresses. As of late, biostimulants have ended up a hotspot as an viable and economical elective to reduce the negative impacts of stresses on plants. In any case, the larger part of biostimulants has destitute solidness beneath natural conditions, which leads to untimely debasement, shortening their organic movement. To unravel these bottlenecks, small scale- and nano-based definitions containing biostimulant atoms and/or microorganisms are picking up consideration as they illustrate a few points of interest over their routine details. In this survey, we center on the embodiment of plant development controllers and plant acquainted microorganisms as a technique to boost their application for plant assurance against abiotic stresses. We moreover address the potential restrictions and challenges confronted for the execution of this innovation, as well as conceivable outcomes with respect to future inquire about.

Keywords: bio stimulants, Seed priming, nano biotechnology, plant growth-promoting, rhizobacteria, plant growth regulators, microencapsulation

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9 Characterization of the Microorganisms Associated with Pleurotus ostractus and Pleurotus tuber-Regium Spent Mushroom Substrate

Authors: Samuel E. Okere, Anthony E. Ataga

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Introduction: The microbial ecology of Pleurotus osteratus and Pleurotus tuber–regium spent mushroom substrate (SMS) were characterized to determine other ways of its utilization. Materials and Methods: The microbiological properties of the spent mushroom substrate were determined using standard methods. This study was carried out at the Microbiology Laboratory University of Port Harcourt, Rivers State, Nigeria. Results: Quantitative microbiological analysis revealed that Pleurotus osteratus spent mushroom substrate (POSMS) contained 7.9x10⁵ and 1.2 x10³ cfu/g of total heterotrophic bacteria and total fungi count respectively while Pleurotus tuber-regium spent mushroom substrate (PTSMS) contained 1.38x10⁶ and 9.0 x10² cfu/g of total heterotrophic bacteria count and total fungi count respectively. The fungi species encountered from Pleurotus tuber-regium spent mushroom substrate (PTSMS) include Aspergillus and Cladosporum species, while Aspergillus and Penicillium species were encountered from Pleurotus osteratus spent mushroom substrate (POSMS). However, the bacteria species encountered from Pleurotus tuber-regium spent mushroom substrate include Bacillus, Acinetobacter, Alcaligenes, Actinobacter, and Pseudomonas species while Bacillus, Actinobacteria, Aeromonas, Lactobacillus and Aerococcus species were encountered from Pleurotus osteratus spent mushroom substrate (POSMS). Conclusion: Therefore based on the findings from this study, it can be concluded that spent mushroom substrate contain microorganisms that can be utilized both in bioremediation of oil-polluted soils as they contain important hydrocarbon utilizing microorganisms such as Penicillium, Aspergillus and Bacillus species and also as sources of plant growth-promoting rhizobacteria (PGPR) such as Pseudomonas and Bacillus species which can induce resistance on plants. However, further studies are recommended, especially to molecularly characterize these microorganisms.

Keywords: characterization, microorganisms, mushroom, spent substrate

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8 Plant Growth and Yield Enhancement of Soybean by Inoculation with Symbiotic and Nonsymbiotic Bacteria

Authors: Timea I. Hajnal-Jafari, Simonida S. Đurić, Dragana R. Stamenov

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Microbial inoculants from the group of symbiotic-nitrogen-fixing rhizobia are well known and widely used in production of legumes. On the other hand, nonsymbiotic plant growth promoting rhizobacteria (PGPR) are not commonly used in practice. The objective of this study was to examine the effects of soybean inoculation with symbiotic and nonsymbiotic bacteria on plant growth and seed yield of soybean. Microbiological activity in rhizospheric soil was also determined. The experiment was set up using a randomized block system in filed conditions with the following treatments: control-no inoculation; treatment 1-Bradyrhizobium japonicum; treatment 2-Azotobacter sp.; treatment 3-Bacillus sp..In the flowering stage of growth (FS) the number of nodules per plant (NPP), root length (RL), plant height (PH) and weight (PW) were measured. The number of pod per plant (PPP), number of seeds per pod (SPP) and seed weight per plant (SWP) were recorded at the end of vegetation period (EV). Microbiological analyses of soil included the determination of total number of bacteria (TNB), number of fungi (FNG), actinomycetes (ACT) and azotobacters (AZB) as well as the activity of the dehydrogenase enzyme (DHA). The results showed that bacterial inoculation led to the formation of root nodules regardless of the treatments with statistically no significant difference. Strong nodulation was also present in control treatment. RL and PH were positively influenced by inoculation with Azotobacter sp. and Bacillus sp., respectively. Statistical analyses of the number of PPP, SPP, and SWP showed no significant differences among investigated treatments. High average number of microorganisms were determined in all treatments. Most abundant were TNB (log No 8,010) and ACT (log No 6,055) than FNG and AZB with log No 4,867 and log No 4,025, respectively. The highest DHA activity was measured in the FS of soybean in treatment 3. The application of nonsymbiotic bacteria in soybean production can alleviate initial plant growth and help the plant to better overcome different stress conditions caused by abiotic and biotic factors.

Keywords: bacteria, inoculation, soybean, microbial activity

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7 Controlling Olive Anthracnose with Antifungal Metabolites from Bacillus Species: A Biological Approach

Authors: Hafiz Husnain Nawaz

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Anthracnose disease in olive, caused by the fungal pathogen Colletotrichum acutatum, is considered one of the most critical issues in olive orchards in Pakistan. This disease poses a significant threat as it results in infections that can lead to the complete damage of olive plants, affecting leaves, stems, and fruits in the field. Controlling this disease is particularly challenging due to the absence of an effective fungicide that does not pose risks to farmer health and the environment. To address this challenge, our study aimed to evaluate the antagonistic activity of a biosurfactant produced by the Bacillus subtilis PE-07 strain against the anthracnose-causing agent in olive plants. This strain was selected after screening sixty rhizobacteria strains. Additionally, we assessed the heat stability, pH range, and toxicity of the biosurfactant produced by strain PE-07. Our results revealed that the biosurfactant exhibited maximum antifungal activity against C. acutatum. In vitro studies indicated that the biosurfactant could reduce fungal activity by inhibiting the spore germination of C. acutatum. Furthermore, the biosurfactant demonstrated a wide pH and temperature range, displaying antifungal activity at pH levels ranging from 5 to 10 and a temperature range from room temperature to 110°C. To evaluate the biosurfactant's safety, we conducted toxicity tests on zebra fish (Danio rerio). The results showed that the biosurfactant had minimal harmful effects, even at maximum concentrations. In conclusion, our study confirmed that the biosurfactant produced by B. subtilis exhibited high pH and heat stability with minimal harmful effects. Therefore, it presents a promising alternative to chemical pesticides for effectively controlling olive anthracnose in Pakistan.

Keywords: biological control, heat stability and PH range, toxicity, Danio rerio

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6 Technological Development of a Biostimulant Bioproduct for Fruit Seedlings: An Engineering Overview

Authors: Andres Diaz Garcia

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The successful technological development of any bioproduct, including those of the biostimulant type, requires to adequately completion of a series of stages allied to different disciplines that are related to microbiological, engineering, pharmaceutical chemistry, legal and market components, among others. Engineering as a discipline has a key contribution in different aspects of fermentation processes such as the design and optimization of culture media, the standardization of operating conditions within the bioreactor and the scaling of the production process of the active ingredient that it will be used in unit operations downstream. However, all aspects mentioned must take into account many biological factors of the microorganism such as the growth rate, the level of assimilation to various organic and inorganic sources and the mechanisms of action associated with its biological activity. This paper focuses on the practical experience within the Colombian Corporation for Agricultural Research (Agrosavia), which led to the development of a biostimulant bioproduct based on native rhizobacteria Bacillus amyloliquefaciens, oriented mainly to plant growth promotion in cape gooseberry nurseries and fruit crops in Colombia, and the challenges that were overcome from the expertise in the area of engineering. Through the application of strategies and engineering tools, a culture medium was optimized to obtain concentrations higher than 1E09 CFU (colony form units)/ml in liquid fermentation, the process of biomass production was standardized and a scale-up strategy was generated based on geometric (H/D of bioreactor relationships), and operational criteria based on a minimum dissolved oxygen concentration and that took into account the differences in the capacity of control of the process in the laboratory and pilot scales. Currently, the bioproduct obtained through this technological process is in stages of registration in Colombia for cape gooseberry fruits for export.

Keywords: biochemical engineering, liquid fermentation, plant growth promoting, scale-up process

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5 The Effect of Durability and Pathogen Strains on the Wheat Induced Resistance against Zymoseptoria tritici as a Response to Paenibacillus sp. Strain B2

Authors: E. Samain, T. Aussenac, D. van Tuinen, S. Selim

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Plant growth promoting rhizobacteria are known as potential biofertilizers and plant resistance inducers. The present work aims to study the durability of the resistance induced as a response to wheat seeds inoculation with PB2 and its influence by Z. tritici strains. The internal and external roots colonization have been determined in vitro, seven days post inoculation, by measuring the colony forming unit (CFU). In planta experimentations were done under controlled conditions included four wheat cultivars with different levels of resistance against Septoria Leaf Blotch (SLB) and four Z. tritici strains with high aggressiveness and resistance levels to fungicides. Plantlets were inoculated with PB2 at sowing and infected with Z. tritici at 3 leaves or tillering growth stages. The infection level with SLB was evaluated at 17 days post inoculation using real-time quantitative polymerase chain reaction (PCR). Results showed that PB2 has a high potential of wheat root external colonization (> 10⁶ CFU/g of root). However, the internal colonization seems to be cultivar dependent. Indeed, PB2 has not been observed as endophytic for one cultivar but has a high level of internal colonization with more than 104 CFU/g of root concerning the three others. Two wheat cultivars (susceptible and moderated resistant) were used to investigate PB2-induced resistance (PB2-IR). After the first infection with Z. tritici, results showed that PB2-IR has conferred a high protection efficiency (40-90%) against SLB in the two tested cultivars. Whereas the PB2-IR was effective against all tested strains with the moderate resistant cultivar, it was higher with the susceptible cultivar (> 64%) but against three of the four tested strains. Concerning the durability of the PB2-IR, after the second infection timing, it has been observed a significant decrease (10-59%) depending strains in the moderate resistant cultivar. Contrarily, the susceptible cultivar showed a stable and high protection level (76-84%) but against three of the four tested strains and interestingly, the strain that overcame PB2-IR was not the same as that of the first infection timing. To conclude, PB2 induces a high and durable resistance against Z. tritici. The PB2-IR is pathogen strain, plant growth stage and genotype dependent. These results may explain the loss of the induced resistance effectiveness under field conditions.

Keywords: induced resistance, Paenibacillus sp. strain B2, wheat genotypes, Zymoseptoria tritici

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4 Rhizobia-Containing Rhizobacterial Consortia and Intercropping Improved Faba Bean and Wheat Performances Under Stress Combining Drought and Phosphorus Deficiency

Authors: Said Cheto, Khawla Oukaltouma, Imane Chamkhi, Ammar Ibn Yasser, Bouchra Benmrid, Ahmed Qaddoury, Lamfeddal Kouisni, Joerg Geistlinger, Youssef Zeroual, Adnane Bargaz, Cherki Ghoulam

Abstract:

Our study aimed to assess, the role of inoculation of faba bean/wheat intercrops with selected rhizobacteria consortia gathering one rhizobia and two phosphate solubilizing bacteria “PSB” to alleviate the effects of combined water deficit and P limitation on Faba bean/ wheat intercrops versus monocrops under greenhouse conditions. One Vicia faba L variety (Aguadulce “Ag”), and one Triticum durum L. variety (Karim “K”) were grown as sole crops or intercrop in pots containing sterilized substrate (sand: peat 4:1v/v) added either with rock phosphate (RP) as the alone P source (P limitation) or with KH₂PO₄ in nutrient solution (P sufficient control). Plant inoculation was done using rhizobacterial consortia composed; C1(Rhizobium laguerreae, Kocuria sp, and Pseudomonas sp) and C2 (R. laguerreae, Rahnella sp, and Kocuria sp). Two weeks after inoculation, the plants were submitted to water deficit consisting of 40% of substrate water holding Capacity (WHC) versus 80% WHC for well-watered plants. At the flowering stage, the trial was assessed, and the results showed that inoculation with both consortia (C1 and C2) improved faba bean biomass in terms of shoots, roots, and nodules compared to inoculation with rhizobia alone, particularly C2 improved these parametres by 19.03, 78.99, and 72.73%, respectively. Leaf relative water content decreased under combined stress, particularly in response to C1 with a significant improvement of this parameter in wheat intercrops. For faba bean under P limitation, inoculation with C2 increased stomatal conductance (gs) by 35.73% compared to plants inoculated with rhizobia alone. Furthermore, the same inoculum C2 improved membrane stability by 44,33% versus 16,16% for C1 compared to inoculation with rhizobia alone under P deficit. For sole cropped faba bean plants, inoculation with both consortia improved N accumulation compared to inoculation with rhizobia alone with an increase of 70.75% under P limitation. Moreover, under the combined stress, intercropping inoculation with C2 improved plant biomass and N content (112.98%) in wheat plants, compared to the sole crop. Our finding revealed that consortium C2 might offer an agronomic advantage under water and P deficit and could be used as inoculum for enhancing faba bean and wheat production under both monocropping and intercropping systems.

Keywords: drought, phosphorus, intercropping, PSB, rhizobia, vicia faba, Triticum durum

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3 Degradation of the Cu-DOM Complex by Bacteria: A Way to Increase Phytoextraction of Copper in a Vineyard Soil

Authors: Justine Garraud, Hervé Capiaux, Cécile Le Guern, Pierre Gaudin, Clémentine Lapie, Samuel Chaffron, Erwan Delage, Thierry Lebeau

Abstract:

The repeated use of Bordeaux mixture (copper sulphate) and other chemical forms of copper (Cu) has led to its accumulation in wine-growing soils for more than a century, to the point of modifying the ecosystem of these soils. Phytoextraction of copper could progressively reduce the Cu load in these soils, and even to recycle copper (e.g. as a micronutrient in animal nutrition) by cultivating the extracting plants in the inter-row of the vineyards. Soil cleaning up usually requires several years because the chemical speciation of Cu in solution is mainly based on forms complexed with dissolved organic matter (DOM) that are not phytoavailable, unlike the "free" forms (Cu2+). Indeed, more than 98% of Cu in the solution is bound to DOM. The selection and inoculation of invineyardsoils in vineyard soils ofbacteria(bioaugmentation) able to degrade Cu-DOM complexes could increase the phytoavailable pool of Cu2+ in the soil solution (in addition to bacteria which first mobilize Cu in solution from the soil bearing phases) in order to increase phytoextraction performance. In this study, sevenCu-accumulating plants potentially usable in inter-row were tested for their Cu phytoextraction capacity in hydroponics (ray-grass, brown mustard, buckwheat, hemp, sunflower, oats, and chicory). Also, a bacterial consortium was tested: Pseudomonas sp. previously studied for its ability to mobilize Cu through the pyoverdine siderophore (complexing agent) and potentially to degrade Cu-DOM complexes, and a second bacterium (to be selected) able to promote the survival of Pseudomonas sp. following its inoculation in soil. Interaction network method was used based on the notions of co-occurrence and, therefore, of bacterial abundance found in the same soils. Bacteria from the EcoVitiSol project (Alsace, France) were targeted. The final step consisted of incoupling the bacterial consortium with the chosen plant in soil pots. The degradation of Cu-DOMcomplexes is measured on the basis of the absorption index at 254nm, which gives insight on the aromaticity of the DOM. The“free” Cu in solution (from the mobilization of Cu and/or the degradation of Cu-MOD complexes) is assessed by measuring pCu. Eventually, Cu accumulation in plants is measured by ICP-AES. The selection of the plant is currently being finalized. The interaction network method targeted the best positive interactions ofFlavobacterium sp. with Pseudomonassp. These bacteria are both PGPR (plant growth promoting rhizobacteria) with the ability to improve the plant growth and to mobilize Cu from the soil bearing phases (siderophores). Also, these bacteria are known to degrade phenolic groups, which are highly present in DOM. They could therefore contribute to the degradation of DOM-Cu. The results of the upcoming bacteria-plant coupling tests in pots will be also presented.

Keywords: complexes Cu-DOM, bioaugmentation, phytoavailability, phytoextraction

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