Search results for: rhizosphere microbiome
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 142

Search results for: rhizosphere microbiome

52 Potential Application of Selected Halotolerant PSB Isolated from Rhizospheric Soil of Chenopodium quinoa in Plant Growth Promotion

Authors: Ismail Mahdi, Nidal Fahsi, Mohamed Hafidi, Abdelmounaim Allaoui, Latefa Biskri

Abstract:

To meet the worldwide demand for food, smart management of arable lands is needed. This could be achieved through sustainable approaches such as the use of plant growth-promoting microorganisms including bacteria. Phosphate (P) solubilization is one of the major mechanisms of plant growth promotion by associated bacteria. In the present study, we isolated and screened 14 strains from the rhizosphere of Chenopodium quinoa wild grown in the experimental farm of UM6P and assessed their plant growth promoting properties. Next, they were identified by using 16S rRNA and Cpn60 genes sequencing as Bacillus, Pseudomonas and Enterobacter. These strains showed dispersed capacities to solubilize P (up to 346 mg L−1) following five days of incubation in NBRIP broth. We also assessed their abilities for indole acetic acid (IAA) production (up to 795,3 µg ml−1) and in vitro salt tolerance. Three Bacillus strains QA1, QA2, and S8 tolerated high salt stress induced by NaCl with a maximum tolerable concentration of 8%. Three performant isolates, QA1, S6 and QF11, were further selected for seed germination assay because of their pronounced abilities in terms of P solubilization, IAA production and salt tolerance. The early plant growth potential of tested strains showed that inoculated quinoa seeds displayed greater germination rate and higher seedlings growth under bacterial treatments. The positive effect on seed germination traits strongly suggests that the tested strains are growth promoting, halotolerant and P solubilizing bacteria which could be exploited as biofertilizers.

Keywords: phosphate solubilizing bacteria, IAA, Seed germination, salt tolerance, quinoa

Procedia PDF Downloads 103
51 Detection and Expression of Peroxidase Genes in Trichoderma harzianum KY488466 and Its Response to Crude Oil Degradation

Authors: Michael Dare Asemoloye, Segun Gbolagade Jonathan, Rafiq Ahmad, Odunayo Joseph Olawuyi, D. O. Adejoye

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Fungi have potentials for degrading hydrocarbons through the secretion of different enzymes. Crude oil tolerance and degradation by Trichoderma harzianum was investigated in this study with its ability to produce peroxidase enzymes (LiP and MnP). Many fungal strains were isolated from rhizosphere of grasses growing on a crude oil spilled site, and the most frequent strain based on percentage incidence was further characterized using morphological and molecular characteristics. Molecular characterization was done through the amplification of Ribosomal-RNA regions of 18s (1609-1627) and 28s (287-266) using ITS1 and ITS4 combinations and it was identified using NCBI BLAST tool. The selected fungus was also subjected to an in-vitro tolerance test at crude oil concentrations of 5, 10, 15, 20 and 25% while 0% served as control. In addition, lignin peroxidase genes (lig1-6) and manganese peroxidase gene (mnp) were detected and expressed in this strain using RT-PCR technique, its peroxidase producing activities was also studied in aliquots (U/ml). This strain had highest incidence of 80%, it was registered in NCBI as Trichoderma harzianum asemoJ KY488466. The strain KY488466 responded to crude oil concentrations as it increase, the dose inhibition response percentage (DIRP) increased from 41.67 to 95.41 at 5 to 25 % crude oil concentrations. All the peroxidase genes are present in KY488466, and expressed with amplified 900-1000 bp through RT-PCR technique. In this strain, lig2, lig4 and mnp genes were over-expressed, lig 6 was moderately expressed, while none of the genes was under-expressed. The strain also produced 90±0.87 U/ml lignin peroxidase and 120±1.23 U/mil manganese peroxidase enzymes in aliquots. These results imply that KY488466 can tolerate and survive high crude oil concentration and could be exploited for bioremediation of oil-spilled soils, the produced peroxidase enzymes could also be exploited for other biotechnological experiments.

Keywords: crude oil, enzymes, expression, peroxidase genes, tolerance, Trichoderma harzianum

Procedia PDF Downloads 191
50 The Mechanism of Parabacteroides goldsteinii on Immune Modulation and Anti-Obsogenicity

Authors: Yu-Ling Tsai, Chih-Jung Chang, Chia-Chen Lu, Eric Wu, Chuan-Sheng Lin, Tzu-Lung Lin, Hsin-Chih Lai

Abstract:

It is urgent that novel anti-obesity measures that are safe, effective and widely available are developed for counteracting the rapidly growing obesity epidemics. In the present study, we show that a probiotic bacterium Parabacteroides goldsteinii screened through culture under the high molecular weight polysaccharides prepared from two iconic medicinal fungi, the Ganoderma lucidum and the Hirsutella sinensis, reduced body weight by ca. 20% in high-fat diet (HFD)-fed mice. The bacterium also decreased intestinal permeability, metabolic endotoxemia, inflammation and insulin resistance. Notably, oral administration of live, but not high temperature-killed, P. goldsteinii to HFD fed mice considerably reduces weight gain and obesity-associated metabolic disorders. A three months feeding of the mice with P. goldsteinii did not show any aberrant side effects, indicating the safety of this bacterium. Transcriptome analysis indicated that P. goldsteinii enhances immunity in resting dendritic cells, but reduces inflammation in lipopolysaccharide (LPS)-induced dendritic cells. On top, Naïve T-cells were skewed towards regulatory T-cells after encountering with dendritic cells (DCs) pretreated with P. goldsteinii. These results indicated P. goldsteinii showed anti-inflammatory effects and can work as a potential probiotic ameliorating obesogenicity and related metabolic syndromes.

Keywords: Parabacteroides goldsteinii, gut microbiome, obesity, immune modulation

Procedia PDF Downloads 140
49 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)

Procedia PDF Downloads 168
48 Deniplant Nutraceuticals for Endometriosis Pain

Authors: Gheorghe Giurgiu, Manole Cojocaru, Mihnea Andrei Nicodin

Abstract:

Background: Inflammation has the main role in the progression of endometriosis. The mechanisms by which endometriosis induces a chronic pain state remain poorly understood. Unfortunately, there is no known cure for endometriosis. But you can manage it with medication and at-home treatments. Some findings have highlighted the main role of inflammation in endometriosis by acting on proliferation, apoptosis, and angiogenesis. The introduction of new agents can be effective in improving the condition of patients; for example, plants are promising sources of bioactive natural components. Objectives: These natural compounds could be interesting strategies in therapy. While there is no absolute cure for this condition, some home remedies can relieve the pain and discomfort it brings. The purpose of this study is to summarize the potential action of Deniplant nutraceuticals in endometriosis by acting on inflammation. Materials and Methods: The primary symptoms of endometriosis are pelvic pain and infertility. The use of Deniplant nutraceuticals could be interesting in disease management for women. Results: Treating pain-related aspects of endometriosis would contribute to the improvement of mental health and daytime function. Because the microbiome can influence inflammation, new therapies can develop through its natural modulation. There are other options, including natural remedies, herbs like cinnamon twigs or licorice root, or supplements such as thiamine, magnesium, or omega-3 fatty acids. Conclusion: Deniplant nutraceuticals can downregulate inflammation in endometriosis. Nevertheless, the limited number of studies focusing on the different interactions of Deniplant nutraceuticals in endometriosis restricts its clear and immediate use in a therapeutic strategy.

Keywords: endometriosis, diet, Deniplant nutraceuticals, pain

Procedia PDF Downloads 39
47 Screening of Rice Genotypes in Methane and Carbon Dioxide Emissions Under Different Water Regimes

Authors: Mthiyane Pretty, Mitsui Toshiake, Nagano Hirohiko, Aycan Murat

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Among the most significant greenhouse gases released from rice fields are methane and carbon dioxide. The primary focus of this research was to quantify CH₄ and CO₂ gas using different 4 rice cultivars, two water regimes, and a recording of soil moisture and temperature. In this study, we hypothesized that paddy field soils may directly affect soil enzymatic activities and physicochemical properties in the rhizosphere soil of paddy fields and subsequently indirectly affect the activity, abundance, diversity, and community composition of methanogens, ultimately affecting CH₄ flux. The experiment was laid out in the randomized block design with two treatments and three replications for each genotype. In two treatments, paddy fields and artificial soil were used. 35 days after planting (DAP), continuous flooding irrigation, Alternate wetting, and drying (AWD) were applied during the vegetative stage. The highest recorded measurements of soil and environmental parameters were soil moisture at 76%, soil temperature at 28.3℃, Bulk EC at 0.99 ds/m, and pore water EC at 1,25, using HydraGO portable soil sensor system. Gas samples were carried out once on a weekly basis at 09:00 am and 12: 00 pm to obtain the mean GHG flux. Gas Chromatography (GC, Shimadzu, GC-2010, Japan) was used for the analysis of CH4 and CO₂. The treatments with paddy field soil had a 1.3℃ higher temperature than artificial soil. The overall changes in Bulk EC were not significant across the treatment. The CH₄ emission patterns were observed in all rice genotypes, although they were less in treatments with AWD with artificial soil. This shows that AWD creates oxic conditions in the rice soil. CO₂ was also quantified, but it was in minute quantities, as rice plants were using CO₂ for photosynthesis. The highest tillering number was 7, and the lowest was 3 in cultivars grown. The rice varieties to be used for breeding are Norin 24, with showed a high number of tillers with less CH₄.

Keywords: greenhouse gases, methane, morphological characterization, alternating wetting and drying

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46 Deciphering the Gut Microbiome's Role in Early-Life Immune Development

Authors: Xia Huo

Abstract:

Children are more vulnerable to environmental toxicants compared to adults, and their developing immune system is among the most sensitive targets regarding toxicity of environmental toxicants. Studies have found that exposure to environmental toxicants is associated with impaired immune function in children, but only a few studies have focused on the relationship between environmental toxicant exposure and vaccine antibody potency and immunoglobulin (Ig) levels in children. These studies investigated the associations of exposure to polychlorinated biphenyls (PCBs), perfluorinated compounds (PFCs), heavy metals (Pb, Cd, As, Hg) and PM2.5 with the serum-specific antibody concentrations and Ig levels against different vaccines, such as anti-Hib, tetanus, diphtheria toxoid, and analyze the possible mechanisms underlying exposure-related alterations of antibody titers and Ig levels against different vaccines. Results suggest that exposure to these toxicants is generally associated with decreased potency of antibodies produced from childhood immunizations and an overall deficiency in the protection the vaccines provide. Toxicant exposure is associated with vaccination failure and decreased antibody titers, and increased risk of immune-related diseases in children by altering specific immunoglobulin levels. Age, sex, nutritional status, and co-exposure may influence the effects of toxicants on the immune function in children. Epidemiological evidence suggests that exposure-induced changes to humoral immunerelated tissue/cells/molecules response to vaccines may have predominant roles in the inverse associations between antibody responsiveness to vaccines and environmental toxicants. These results help us to conduct better immunization policies for children under environmental toxicant burden.

Keywords: environmental toxicants, immunotoxicity, vaccination, antibodies, children's health

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45 Adaptive Response of Plants to Environmental Stress: Natural Oil Seepage; The Living Laboratory in Tramutola, Basilicata Region

Authors: Maria Francesca Scannone, Martina Bochicchio

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One of the major environmental problems today is hydrocarbon contamination. The promising sustainable technologies for the treatment of these contaminated sites involves the use of biological organisms. In Agri Valley (Basilicata Region) there is a living laboratory (natural oil seeps) where the selective pressure has enriched the environmental matrices with microorganisms, fungi and plant species able to use the hydrocarbons as a source of metabolic energy, to degrade or tolerate hydrocarbons. Observers visiting this area are fascinated by its unspoiled nature, and the condition of the ecosystem does not appear to has been damaged. The amazing resiliency observed in Tramutola site is of key importance to try to bring green remediation technologies, but no research has been done to identify high-performing native species. The aim of this research was to study how natural processes affect the fate of released oil or how individual species or communities of plants and animals are capable of dealing with the burden of otherwise toxic chemicals. The survey of vegetation was carried out, more than 60 species have been identified and divided into tree, shrub and herb layer. Plant data sheets have been completed only for the species that showed the most appropriate properties for phytoremediation. In general, members of the Salicales, Cyperales, Poales, Fagales, Cornales, Equisetales orders were the most commonly identified orders. They are pioneer plants with high adaptive capacity and vegetative propagation. The literature review has highlighted the existence of rhizosphere effect and a green liver model on selected plants. The study provides significant information on the environmental stress adaptation processes of many indigenous plants that are living and growing on a natural leak of crude oil and gas that migrates up through subsurface.

Keywords: green liver, hydrocarbon degradation, oil seeps, phytoremediation

Procedia PDF Downloads 147
44 Identification and Application of Biocontrol Agents against Cotton Leaf Curl Virus Disease in Gossypium hirsutum under Green House Conditions

Authors: Memoona Ramzan, Bushra Tabassum, Anwar Khan, Muhammad Tariq, Mudassar Fareed Awan, Idrees Ahmad Nasir, Zahida Qamar, Naila Shahid, Tayyab Husnain

Abstract:

Biological control is a novel approach being used in crop protection nowadays. Bacteria like Bacillus and Pseudomonas are reported for this purpose and few of their products are commercially available too. Rhizosphere and phyllosphere of healthy cotton plants were used as a source to isolate bacteria capable of exhibiting properties worthy for selection as biocontrol agent. For this purpose all isolated strains were screened for the activities like phosphate solubilization, Indole acetic acid (IAA) production and biocontrol against fungi. Two strains S1HL3 and S1HL4 showed phosphate solubilization and IAA production simultaneously while two other JS2HR4 and JS3HR2 were good inhibitors of fungal pathogens. Through biochemical and molecular characterization these bacteria were identified as P. aeruginosa, Burkholderia and Bacillus respectively. In green house trials of these isolates against Cotton leaf curl virus (CLCuV), seven treatments including individual bacterial isolate and consortia were included. Treated plants were healthy as compared to control plants in which upto 74% CLCuV symptomatic plants exist. Maximum inhibition of CLCuV was observed in T7 treated plants where viral load was only 0.4% as compared to control where viral load was upto 74%. This treatment consortium included Bacillus and Pseudomonas isolates; S1HL3, S1HL4, JS2HR4 and JS3HR2. Principal Component Biplot depicted highly significant correlation between percentage viral load and the disease incidence.

Keywords: cotton leaf curl virus, biological control, bacillus, pseudomonas

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43 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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42 The Effect of Air Injection in Irrigation Water on Sugar Beet Yield

Authors: Yusuf Ersoy Yildirim, Ismail Tas, Ceren Gorgusen, Tugba Yeter, Aysegul Boyacioglu, K. Mehmet Tugrul, Murat Tugrul, Ayten Namli, H. Sabri Ozturk, M. Onur Akca

Abstract:

In recent years, a lot of research has been done for the sustainable use of scarce resources in the world. Especially, effective and sustainable use of water resources has been researched for many years. Sub-surface drip irrigation (SDI) is one of the most effective irrigation methods in which efficient and sustainable use of irrigation water can be achieved. When the literature is taken into consideration, it is often emphasized that, besides its numerous advantages, it also allows the application of irrigation water to the plant root zone along with air. It is stated in different studies that the air applied to the plant root zone with irrigation water has a positive effect on the root zone. Plants need sufficient oxygen for root respiration as well as for the metabolic functions of the roots. Decreased root respiration due to low oxygen content reduces transpiration, disrupts the flow of ions, and increases the ingress of salt reaching toxic levels, seriously affecting plant growth. Lack of oxygen (Hypoxia) can affect the survival of plants. The lack of oxygen in the soil is related to the exchange of gases in the soil with the gases in the atmosphere. Soil aeration is an important physical parameter of a soil. It is highly dynamic and is closely related to the amount of water in the soil and its bulk weight. Subsurface drip irrigation; It has higher water use efficiency compared to irrigation methods such as furrow irrigation and sprinkler irrigation. However, in heavy clay soils, subsurface drip irrigation creates continuous wetting fronts that predispose the rhizosphere region to hypoxia or anoxia. With subsurface drip irrigation, the oxygen is limited for root microbial respiration and root development, with the continuous spreading of water to a certain region of the root zone. In this study, the change in sugar beet yield caused by air application in the SDI system will be explained.

Keywords: sugar beet, subsurface drip irrigation, air application, irrigation efficiency

Procedia PDF Downloads 52
41 Inactivation of Root-Knot Nematode Eggs Meloidogyne enterolobii in Irrigation Water Treated with Ozone

Authors: I. A. Landa-Fernandez, I. Monje-Ramirez, M. T. Orta-Ledesma

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Every year plant-parasitic nematodes diminish the yield of high-value crops worldwide causing important economic losses. Currently, Meloidogyne enterolobii has increased its importance due to its high aggressiveness, increasing geographical distribution and host range. Root-knot nematodes inhabit the rhizosphere soil around plant roots. However, they can come into contact with irrigation water. Thus, plant-parasitic nematodes can be transported by water, as eggs or juveniles. Due to their high resistance, common water disinfection methods are not effective for inactivating these parasites. Ozone is the most effective disinfectant for microbial inactivation. The objective of this study is to demonstrate that ozone treatment is an alternative method control in irrigation water of the root-knot nematode M. enterolobii. It has been shown that ozonation is an effective treatment for the inactivation of protozoan cysts and oocysts (Giardia and Cryptosporidium) and for other species of the genus Meloidogyne (M. incognita), but not for the enterolobii specie. In this study, the strain of M. enterolobii was isolated from tomatoes roots. For the tests, eggs were used and were inoculated in water with similar characteristics of irrigation water. Subsequently, the disinfection process was carried out in an ozonation unit. The performance of the treatments was evaluated through the egg's viability by assessing its structure by optical microscopy. As a result of exposure to ozone, the viability of the nematode eggs was reduced practically in its entirety; with dissolved ozone levels in water close to the standard concentration (equal to 0.4 mgO₃/L), but with high contact times (greater than 4 min): 0.2 mgO₃/L for 15 minutes or 0.55 mgO₃/L for 10 minutes. Additionally, the effect of temperature, alkalinity and organic matter of the water was evaluated. Ozonation is effective and a promising alternative for the inactivation of nematodes in irrigation water, which could contribute to diminish the agricultural losses caused by these organisms.

Keywords: inactivation process, irrigation water treatment, ozonation, plant-parasite nematodes

Procedia PDF Downloads 137
40 Chemotrophic Signal Exchange between the Host Plant Helianthemum sessiliflorum and Terfezia boudieri

Authors: S. Ben-Shabat, T. Turgeman, O. Leubinski, N. Roth-Bejerano, V. Kagan-Zur, Y. Sitrit

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The ectomycorrhizal (ECM) desert truffle Terfezia boudieri produces edible fruit bodies and forms symbiosis with its host plant Helianthemum sessiliflorum (Cistaceae) in the Negev desert of Israel. The symbiosis is vital for both partners' survival under desert conditions. Under desert habitat conditions, ECMs must form symbiosis before entering the dry season. To secure a successful encounter, in the course of evolution, both partners have responded by evolving special signals exchange that facilitates recognition. Members of the Cistaceae family serve as host plants for many important truffles. Conceivably, during evolution a common molecule present in Cistaceae plants was recruited to facilitate successful encounter with ectomycorrhizas. Arbuscular vesicular fungi (AM) are promiscuous in host preferences, in contrast, ECM fungi show specificity to host plants. Accordingly, we hypothesize that H. sessiliflorum secretes a chemotrophic-signaling, which is common to plants hosting ECM fungi belonging to the Pezizales. However, thus far no signaling molecules have been identified in ECM fungi. We developed a bioassay for chemotrophic activity. Fractionation of root exudates revealed a substance with chemotrophic activity and molecular mass of 534. Following the above concept, screening the transcriptome of Terfezia, grown under chemoattraction, discovered genes showing high homology to G proteins-coupled receptors of plant pathogens involved in positive chemotaxis and chemotaxis suppression. This study aimed to identify the active molecule using analytical methods (LC-MS, NMR etc.). This should contribute to our understanding of how ECM fungi communicate with their hosts in the rhizosphere. In line with the ability of Terfezia to form also endomycorrhizal symbiosis like AM fungi, analysis of the mechanisms may likewise be applicable to AM fungi. Developing methods to manipulate fungal growth by the chemoattractant can open new ways to improve inoculation of plants.

Keywords: chemotrophic signal, Helianthemum sessiliflorum, Terfezia boudieri, ECM

Procedia PDF Downloads 379
39 The Effect of Different Metal Nanoparticles on Growth and Survival of Pseudomonas syringae Bacteria

Authors: Omar Alhamd, Peter A. Thomas, Trevor J. Greenhough, Annette K. Shrive

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The Pseudomonas syringae species complex includes many plant pathogenic strains with highly specific interactions with varied host species and cultivars. The rapid spread of these bacteria over the last ten years has become a cause for concern. Nanoparticles have previously shown promise in microbiological action. We have therefore investigated in vitro and in vivo the effects of different types and sizes of nanoparticles in order to provide quantitative information about their effect on the bacteria. The effects of several different nanoparticles against several bacteria strains were investigated. The effect of NP on bacterial growth was studied by measuring the optical density, biochemical and nutritional tests, and transmission electron microscopy (TEM) to determine the shape and size of NP. Our results indicate that their effects varied, with either a negative or a positive impact on both bacterial and plant growth. Additionally, the methods of exposure to nanoparticles have a crucial role in accumulation, translocation, growth response and bacterial growth. The results of our studies on the behaviour and effects of nanoparticles in model plants showed. Cerium oxide (CeO₂) and silver (Ag) NP showed significant antibacterial activity against several pathogenic bacteria. It was found that titanium nanoparticles (TiO₂) can have either a negative or a positive impact, according to concentration and size. It is also thought that environmental conditions can have a major influence on bacterial growth. Studies were therefore also carried out under some environmental stress conditions to test bacterial survival and to assess bacterial virulence. All results will be presented including information about the effects of different nanoparticles on Pseudomonas syringae bacteria.

Keywords: plant microbiome, nanoparticles, 16S rRNA gene sequencing, bacterial survival

Procedia PDF Downloads 178
38 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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37 Microbiota Effect with Cytokine in Hl and NHL Patient Group

Authors: Ekin Ece Gürer, Tarık Onur Tiryaki, Sevgi Kalayoğlu Beşışık, Fatma Savran Oğuz, Uğur Sezerman, Fatma Erdem, Gülşen Günel, Dürdane Serap Kuruca, Zerrin Aktaş, Oral Öncül

Abstract:

Aim: Chemotherapytreatment in HodgkinLymphomaandNon-HodgkinLymphoma (NHL) diseasescausesgastrointestinalepithelialdamage, disruptstheintestinalmicrobiotabalanceandcausesdysbiosis. Inourstudy, it wasaimedtoshowtheeffect of thedamagecausedbychemotherapy on themicrobiotaandtheeffect of thechangingmicrobiota flora on thecourse of thedisease. Materials And Methods: Seven adult HL and seven adult HL patients to be treatedwithchemotherapywereincluded in the study. Stoolsamplesweretakentwice, beforechemotherapytreatmentandafterthe 3th course of treatment. SamplesweresequencedusingNextGenerationSequencing (NGS) methodafternucleicacidisolation. OTU tableswerepreparedusing NCBI blastnversion 2.0.12 accordingtothe NCBI general 16S bacterialtaxonomyreferencedated 10.08.2021. Thegenerated OTU tableswerecalculatedwith R Statistical Computer Language version 4.04 (readr, phyloseq, microbiome, vegan, descrand ggplot2 packages) to calculate Alpha diversityandtheirgraphicswerecreated. Statistical analyzeswerealsoperformedusing R Statistical Computer Language version 4.0.4 and studio IDE 1.4 (tidyverse, readr, xlsxand ggplot2 packages). Expression of IL-12 and IL-17 cytokineswasperformedbyrtPCRtwice, beforeandaftertreatment. Results: InHL patients, a significantdecreasewasobserved in themicrobiota flora of Ruminococcaceae_UCG-014 genus (p:0.036) andUndefined Ruminococcaceae_UCG-014 species (p:0.036) comparedtopre-treatment. When the post-treatment of HL patientswerecomparedwithhealthycontrols, a significantdecreasewasfound in themicrobiota of Prevotella_7 genus (p:0.049) andButyricimonas (p:0.006) in the post-treatmentmicrobiota of HL patients. InNHL patients, a significantdecreasewasobserved in themicrobiota flora of Coprococccus_3 genus (p:0.015) andUndefined Ruminoclostridium_5 (p:0.046) speciescomparedtopre-treatment. When post-treatment of NHL patientswerecomparedwithhealthycontrols, a significantabundance in theBacilliclass (p:0.029) and a significantdecrease in theUndefinedAlistipesspecies (p:0.047) wereobserved in the post-treatmentmicrobiota of NHL patients. While a decreasewasobserved in IL-12 cytokineexpressionuntilbeforetreatment, an increase in IL-17 cytokineexpressionwasdetected. Discussion: Intestinal flora monitoringafterchemotherapytreatmentshowsthat it can be a guide in thetreatment of thedisease. It is thoughtthatincreasingthediversity of commensalbacteria can alsopositivelyaffecttheprognosis of thedisease.

Keywords: hodgkin lymphoma, non-hodgkin, microbiota, cytokines

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36 Phylogenetic Differential Separation of Environmental Samples

Authors: Amber C. W. Vandepoele, Michael A. Marciano

Abstract:

Biological analyses frequently focus on single organisms, however many times, the biological sample consists of more than the target organism; for example, human microbiome research targets bacterial DNA, yet most samples consist largely of human DNA. Therefore, there would be an advantage to removing these contaminating organisms. Conversely, some analyses focus on a single organism but would greatly benefit from the additional information regarding the other organismal components of the sample. Forensic analysis is one such example, wherein most forensic casework, human DNA is targeted; however, it typically exists in complex non-pristine sample substrates such as soil or unclean surfaces. These complex samples are commonly comprised of not just human tissue but also microbial and plant life, where these organisms may help gain more forensically relevant information about a specific location or interaction. This project aims to optimize a ‘phylogenetic’ differential extraction method that will separate mammalian, bacterial and plant cells in a mixed sample. This is accomplished through the use of size exclusion separation, whereby the different cell types are separated through multiple filtrations using 5 μm filters. The components are then lysed via differential enzymatic sensitivities among the cells and extracted with minimal contribution from the preceding component. This extraction method will then allow complex DNA samples to be more easily interpreted through non-targeting sequencing since the data will not be skewed toward the smaller and usually more numerous bacterial DNAs. This research project has demonstrated that this ‘phylogenetic’ differential extraction method successfully separated the epithelial and bacterial cells from each other with minimal cell loss. We will take this one step further, showing that when adding the plant cells into the mixture, they will be separated and extracted from the sample. Research is ongoing, and results are pending.

Keywords: DNA isolation, geolocation, non-human, phylogenetic separation

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35 Scalable and Accurate Detection of Pathogens from Whole-Genome Shotgun Sequencing

Authors: Janos Juhasz, Sandor Pongor, Balazs Ligeti

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Next-generation sequencing, especially whole genome shotgun sequencing, is becoming a common approach to gain insight into the microbiomes in a culture-independent way, even in clinical practice. It does not only give us information about the species composition of an environmental sample but opens the possibility to detect antimicrobial resistance and novel, or currently unknown, pathogens. Accurately and reliably detecting the microbial strains is a challenging task. Here we present a sensitive approach for detecting pathogens in metagenomics samples with special regard to detecting novel variants of known pathogens. We have developed a pipeline that uses fast, short read aligner programs (i.e., Bowtie2/BWA) and comprehensive nucleotide databases. Taxonomic binning is based on the lowest common ancestor (LCA) principle; each read is assigned to a taxon, covering the most significantly hit taxa. This approach helps in balancing between sensitivity and running time. The program was tested both on experimental and synthetic data. The results implicate that our method performs as good as the state-of-the-art BLAST-based ones, furthermore, in some cases, it even proves to be better, while running two orders magnitude faster. It is sensitive and capable of identifying taxa being present only in small abundance. Moreover, it needs two orders of magnitude less reads to complete the identification than MetaPhLan2 does. We analyzed an experimental anthrax dataset (B. anthracis strain BA104). The majority of the reads (96.50%) was classified as Bacillus anthracis, a small portion, 1.2%, was classified as other species from the Bacillus genus. We demonstrate that the evaluation of high-throughput sequencing data is feasible in a reasonable time with good classification accuracy.

Keywords: metagenomics, taxonomy binning, pathogens, microbiome, B. anthracis

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34 Interpersonal Variation of Salivary Microbiota Using Denaturing Gradient Gel Electrophoresis

Authors: Manjula Weerasekera, Chris Sissons, Lisa Wong, Sally Anderson, Ann Holmes, Richard Cannon

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The aim of this study was to characterize bacterial population and yeasts in saliva by Polymerase chain reaction followed by denaturing gradient gel electrophoresis (PCR-DGGE) and measure yeast levels by culture. PCR-DGGE was performed to identify oral bacteria and yeasts in 24 saliva samples. DNA was extracted and used to generate DNA amplicons of the V2–V3 hypervariable region of the bacterial 16S rDNA gene using PCR. Further universal primers targeting the large subunit rDNA gene (25S-28S) of fungi were used to amplify yeasts present in human saliva. Resulting PCR products were subjected to denaturing gradient gel electrophoresis using Universal mutation detection system. DGGE bands were extracted and sequenced using Sanger method. A potential relationship was evaluated between groups of bacteria identified by cluster analysis of DGGE fingerprints with the yeast levels and with their diversity. Significant interpersonal variation of salivary microbiome was observed. Cluster and principal component analysis of the bacterial DGGE patterns yielded three significant major clusters, and outliers. Seventeen of the 24 (71%) saliva samples were yeast positive going up to 10³ cfu/mL. Predominately, C. albicans, and six other species of yeast were detected. The presence, amount and species of yeast showed no clear relationship to the bacterial clusters. Microbial community in saliva showed a significant variation between individuals. A lack of association between yeasts and the bacterial fingerprints in saliva suggests the significant ecological person-specific independence in highly complex oral biofilm systems under normal oral conditions.

Keywords: bacteria, denaturing gradient gel electrophoresis, oral biofilm, yeasts

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33 Physiological and Biochemical Assisted Screening of Wheat Varieties under Partial Rhizosphere Drying

Authors: Muhammad Aown Sammar Raza

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Environmental stresses are one of the major reasons for poor crop yield across the globe. Among the various environmental stresses, drought stress is the most damaging one, especially in arid and semi-arid regions. Wheat is the major staple food of many countries of the world, which is badly affected by drought stress. In order to fulfill the dietary needs of increasing population with depleting water resources there is a need to adopt technologies which result in sufficient crop yield with less water consumption. One of them is partial root zone drying. Keeping in view these conditions, a wire house experiment was conducted at agronomic research area of University College of Agriculture and Environmental Sciences, The Islamia University Bahawalpur during 2015, to screen out the different wheat varieties for partial root zone drying (PRD). Five approved local wheat varieties (V1= Galaxy-2013, V2= Punjab-2011, V3 = Faisalabad-2008, V4 = Lasani-2008 and V5 = V.8200) and two irrigation levels (I1= control irrigation and I2 = PRD irrigation) with completely randomized design having four replications were used in the experiment. Among the varieties, Galaxy-2013 performed the best and attained maximum plant height, leaf area, stomatal conductance, photosynthesis, total sugars, proline contents and antioxidant enzymes activities and minimum values of growth and physiological parameters were recorded in variety V.8200. For irrigation levels, higher values of growth, physiological and water related parameters were recorded in control treatment (I1) except leaf water potential, osmotic potential, total sugars and proline contents. However, enzyme activities were higher under PRD treatment for all varieties. It was concluded that Galaxy-2013 is the most compatible and V.8200 is the most susceptible variety for PRD, respectively and more quality traits and enzymatic activities were recorded under PRD irrigation as compared to control treatment.

Keywords: antioxidant enzymes activities, osmolytes concentration, partial root zone drying, photosynthetic rate, water relations, wheat

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32 Bacterial Interactions of Upper Respiratory Tract Microbiota

Authors: Sarah Almuhayya, Andrew Mcbain, Gavin Humphreys

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Background. The microbiome of the upper respiratory tract (URT) has received less research attention than other body sites. This study aims to investigate the microbial ecology of the human URT with a focus on the antagonism between the corynebacteria and staphylococci. Methods. Mucosal swabs were collected from the anterior nares and nasal turbinates of 20 healthy adult subjects. Genomic DNA amplification targeting the (V4) of the 16Sr RNA gene was conducted and analyzed using QIIME. Nasal swab isolates were cultured and identified using near full-length sequencing of the 16S rRNA gene. Isolates identified as corynebacteria or staphylococci were typed using (rep-PCR). Antagonism was determined using an agar-based inhibition assay. Results. Four major bacterial phyla (Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria) were identified from all volunteers. The typing of cultured staphylococci and corynebacteria suggested that intra-individual strain diversity was limited. Analysis of generated nasal microbiota profiles suggested an inverse correlation in terms of relative abundance between staphylococci and corynebacteria. Despite the apparent antagonism between these genera, it was limited when investigated on agar. Of 1000 pairwise interactions, observable zones of inhibition were only reported between a single strain of C.pseudodiphtheriticum and S.aureus. Imaging under EM revealed this effect to be bactericidal with clear lytic effects on staphylococcal cell morphology. Conclusion. Nasal microbiota is complex, but culturable staphylococci and corynebacteria were limited in terms of clone type. Analysis of generated nasal microbiota profiles suggested an inverse correlation in terms of relative abundance between these genera suggesting an antagonism or competition between these taxonomic groups.

Keywords: nasal, microbiota, S.aureus, microbioal interaction

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31 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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30 Variability of the Arbuscular Mycorrhizal Fungi Communities Associated with Wild Agraz Plants (Vaccinium meridionale Swartz) in the Colombian Andes

Authors: Gabriel Roveda-Hoyos, Margarita Ramirez-Gomez, Adrian Perez, Diana Paola Serralde

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The objective of this study was to determine the variability of arbuscular mycorrhizal fungi (HFMA) communities associated with wild agraz plants (Vaccinium meridionale Swartz) in the Colombian Andes. This species is one of the most promising fruits within the genus Vaccinium because of the high content of anthocyanins and antioxidants in its fruits, and like other species of the Ericaceae family, it depends on the association with HFM for its development in the natural environment. In this study, the presence of mycorrhizae in wild communities of V. meridionale was evaluated, and their relationship with the edaphic and climatic conditions of the study area was analyzed. Sampling was conducted in the rural area of the municipalities of Raquira, and Chiquinquira, Chia, and Tabio in the departments of Cundinamarca and Boyaca, Colombia. Seven sites were selected, and in each site, 5 plants were randomly selected, root and soil samples were taken from each plant in the rhizosphere zone for the quantification of colonization and the presence of spores. The samples were collected on different soils, taxonomic orders Entisols, Inceptisols, and Alfisols, located at altitudes between 2,600 and 3,000 above sea level in the Eastern Cordillera of Colombia. The physicochemical characteristics of the soil were compared with the density of spores and the percentage of presence of mycorrhizae in the roots and variables with the morphometric and physiological characteristics of the plants. Four types of mutual associations were found: arbuscular mycorrhizae, ectendomycorrhiza, ericoid mycorrhizae, and endophytic septate fungi. The main results obtained show a predominance of spores of the genera Glomus and Acaulsopora, in most of the soils analyzed. The spore density of Glomeromycete fungi in the soil varied considerably between the different sites; it was higher ( > 50 spores/g of dry soil) in soil samples with lower bulk density and higher content of organic matter; in these soils a higher cation exchange capacity was found, as well as of nitrogen, calcium, magnesium, manganese and zinc concentration. It can be concluded that Vaccinium meridionale is able to establish in a natural way, association with HFMA.

Keywords: Ericaceae, Arbuscular mycorrhizae, Andes, soils, Glomus sp.

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29 Biomimetic Strategies to Design Non-Toxic Antimicrobial Textiles

Authors: Isabel Gouveia

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Antimicrobial textile materials may significantly reduce the risk of infections and because they are able to absorb substances from the skin and release therapeutic compounds to the skin, they can also find applications as complementary therapy of skin-diseases as part of standard management. Although functional textiles may be a promising area in skin disease/injury management, as part of standard management, few offer complementary treatment even though they are well known to reduce scratching and aiding emollient absorption, reducing infection, and alleviating pruritus. The reason for this may rely on the low quality of supporting evidence and negative effect that antimicrobial agents may exert on skin microbiome, as for example additional irritation of the vulnerable skin, and by causing resistant bacteria. Several antimicrobial agents have been tested in textiles: quaternary ammonium compounds, silver, polyhexamethylene-biguanides and triclosan have been used, with success. They have powerful bactericidal activity but the majority have a reduce spectrum of microbial inhibition and may cause skin irritation, ecotoxicity and bacteria resistance. Furthermore, the rising flow of strains resistant to last-resort antibiotics rekindles interest in alternative strategies. In this regard, new functional textiles incorporating highly specific antimicrobial agents towards pathogenic bacteria, are required. Recent research has been conducted on naturally occurring antimicrobials as novel alternatives to antibiotics. Conscious of this need our team firstly reported new approaches using L-cysteine and antimicrobial peptides (AMP). Briefly, we were able to develop different immobilization processes towards 6 Log Reduction against bacteria such as S. aureus and K. pneumoniae. Therefore, here we present several innovative antimicrobial textiles incorporating AMP and L-Cysteine which may open new avenues for the medical textiles market and biomaterials in general. Team references will be discussed as an overview and for comparison purposes in terms of potential therapeutic applications.

Keywords: Antimicrobials, Antimicrobial Textiles, Biomedical Textiles, Biomimetic surface functionalization

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28 Phytoremediation of Heavy Metals by the Perennial Tussock Chrysopogon Zizanioides Grown on Zn and Cd Contaminated Soil Amended with Biochar

Authors: Dhritilekha Deka, Deepak Patwa, Ravi K., Archana M. Nair

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Bioaccumulation of heavy metal contaminants due to intense anthropogenic interference degrades the environment and ecosystem functions. Conventional physicochemical methods involve energy-intensive and costly methodologies. Phytoremediation, on the other hand, provides an efficient nature-based strategy for the reclamation of heavy metal-contaminated sites. However, the slow process and adaptation to high-concentration contaminant sequestration often limit the efficiency of the method. This necessitates natural amendments such as biochar to improve phytoextraction and stabilize the green cover. Biochar is a highly porous structure with high carbon sequestration potential and containing negatively charged functional groups that provide binding sites for the positively charged metals. This study aims to develop and determine the synergy between sugarcane bagasse biochar content and phytoremediation. A 60-day pot experiment using perennial tussock vetiver grass (Chrysopogon zizanioides) was conducted for different biochar contents of 1%, 2%, and 4% for the removal of cadmium and zinc. A concentration of 500 ppm is maintained for the amended and unamended control (CK) samples. The survival rates of the plants, biomass production, and leaf area index were measured for the plant growth characteristics. Results indicate a visible change in the plant growth and the heavy metal concentration with the biochar content. The bioconcentration factor (BCF) in the plant improved significantly for the 4% biochar content by 57% in comparison to the control CK treatment in Cd-treated soils. The Zn soils indicated the highest reduction in the metal concentration by 50% in the 2% amended samples and an increase in the BCF in all the amended samples. The translocation from the rhizosphere to the shoots was low but not dependent on the amendment content and varied for each contaminant type. The root-to-shoot ratio indicates higher values compared to the control samples. The enhanced tolerance capacities can be attributed to the nutrients released by the biochar in the soil. The study reveals the high potential of biochar as a phytoremediation amendment, but its effect is dependent on the soil and heavy metal and accumulator species.

Keywords: phytoextraction, biochar, heavy metals, chrysopogon zizanioides, bioaccumulation factor

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27 Theory of Negative Trigger: The Contract between Oral Probiotics and Immune System

Authors: Cliff Shunsheng Han

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Identifying the direct allergy cause that can be easily mitigated is the foundation to stop the allergy epidemic that has been started in the seventies. It has confirmed that the personal and social hygiene practices are associated with the allergy prevalence. But direct causes have been found, and proposed translational measures have not been effective. This study, assisted by a particular case of allergies, has seen the direct cause of allergies, developed a valid test resulted in lasting relief for allergies, and constructed theory describing general relationship between microbiota and host immune system. Saliva samples were collected from a subject for three years during which time the person experienced yearlong allergy, seasonal allergy, and remission of allergy symptoms. Bacterial DNA was extracted and 16S rRNA genes were profiled with Illumina sequencing technology. The analyzing results indicate that the possible direct cause of allergy is the lacking probiotic bacteria in the oral cavity, such as genera Streptococcus and Veilonella, that can produce metabolites to pacify immune system. Targeted promotion of those bacteria with a compound designed for them, has led to lasting remissions of allergic rhinitis. During the development of the translational measure, the subject's oral biofilm was completely destructed by a moderate fever due to an unrelated respiratory infection. The incident not only facilitated the development of the heat based microbiota reseeding procedure but also indicated a possible natural switch that subsequently increases the efficacy of the immune system previously restrained by metabolites from microbiota. These results lead to the proposal of a Theory of Negative Trigger (TNT) to describe the relationship between oral probiotics and immune system, in which probiotics are the negative trigger that will release the power of immune system when removed by fever or modern lifestyles. This study could open doors leading to further understanding of how the immune system functions under the influence of microbiota as well as validate simple traditional practices for healthy living.

Keywords: oral microbiome, allergy, immune system, infection

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26 An Assessment of Nodulation and Nitrogen Fixation of Lessertia Frutescens Plants Inoculated with Rhizobial Isolates from the Cape Fynbos

Authors: Mokgadi Miranda Hlongwane, Ntebogeng Sharon Mokgalaka, Felix Dapare Dakora

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Lessertia (L.) frutescens (syn. Sutherlandia frutescens) is a leguminous medicinal plant indigenous to South Africa. Traditionally, L. frutescens has been used to treat cancer, diabetes, epilepsy, fever, HIV, stomach problems, wounds and other ailments. This legume is endemic to the Cape fynbos, with large populations occurring wild and cultivated in the Cape Florist Region. Its widespread distribution in the Western Cape, Northern Cape, Eastern Cape and Kwazulu-Natal is linked to its increased use as a phytomedicine in the treatment of various diseases by traditional healers. The frequent harvesting of field plants for use as a medicine has made it necessary to undertake studies towards the conservation of Lessertia frutescens. As a legume, this species can form root nodules and fix atmospheric N₂ when in symbiosis with soil bacteria called rhizobia. So far, however, few studies (if any) have been done on the efficacy and diversity of native bacterial symbionts nodulating L. frutescens in South Africa. The aim of this project was to isolate and characterize L. frutescens-nodulating bacteria from five different locations in the Western Cape Province. This was done by trapping soil rhizobia using rhizosphere soil suspension to inoculate L. frutescens seedlings growing in sterilized sand and receiving sterile N-free Hoagland nutrient solution under glasshouse conditions. At 60 days after planting, root nodules were harvested from L. frutescens plants, surface-sterilized, macerated, and streaked on yeast mannitol agar (YMA) plates and incubated at 28 ˚C for observation of bacterial growth. The majority of isolates were slow-growers that took 6-14 days to appear on YMA plates. However, seven isolates were fast-growers, taking 2-4 days to appear on YMA plates. Single-colony cultures of the isolates were assessed for their ability to nodulate L. frutescens as a homologous host under glasshouse conditions. Of the 92 bacterial isolates tested, 63 elicited nodule formation on L. frutescens. Symbiotic effectiveness varied markedly between and among test isolates. There were also significant (p≤0.005) differences in nodulation, shoot biomass, photosynthetic rates, leaf transpiration and stomatal conductance of L. frutescens plants inoculated with the test isolates, which is an indication of their functional diversity.

Keywords: lessertia frutescens, nodulating, rhizobia, symbiotic effectiveness

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25 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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24 Exploring Marine Bacteria in the Arabian Gulf Region for Antimicrobial Metabolites

Authors: Julie Connelly, Tanvi Toprani, Xin Xie, Dhinoth Kumar Bangarusamy, Kris C. Gunsalus

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The overuse of antibiotics worldwide has contributed to the development of multi-drug resistant (MDR) pathogenic bacterial strains. There is an increasing urgency to discover antibiotics to combat MDR pathogens. The microbiome of the Arabian Gulf is a largely unexplored and potentially rich source of novel bioactive compounds. Microbes that inhabit the Abu Dhabi coastal regions adapt to extreme environments with high salinity, hot temperatures, large temperature fluctuations, and acute exposure to solar energy. The microbes native to this region may produce unique metabolites with therapeutic potential as antibiotics and antifungals. We have isolated 200 pure bacterial strains from mangrove sediments, cyanobacterial mats, and coral reefs of the Abu Dhabi region. In this project, we aim to screen the marine bacterial strains to identify antibiotics, in particular undocumented compounds that show activity against existing antibiotic-resistant strains. We have acquired the ESKAPE pathogen panel, which consists of six antibiotic-resistant gram-positive and gram-negative bacterial pathogens that collectively cause most clinical infections. Our initial efforts of the primary screen using colony-picking co-culture assay have identified several candidate marine strains producing potential antibiotic compounds. We will next apply different assays, including disk-diffusion and broth turbidity growth assay, to confirm the results. This will be followed by bioactivity-guided purification and characterization of target compounds from the scaled-up volume of candidate strains, including SPE fraction, HPLC fraction, LC-MS, and NMR. For antimicrobial compounds with unknown structures, our final goal is to investigate their mode of action by identifying the molecular target.

Keywords: marine bacteria, natural products, drug discovery, ESKAPE panel

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23 Gut Mycobiome Dysbiosis and Its Impact on Intestinal Permeability in Attention-Deficit/Hyperactivity Disorder

Authors: Liang-Jen Wang, Sung-Chou Li, Yuan-Ming Yeh, Sheng-Yu Lee, Ho-Chang Kuo, Chia-Yu Yang

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Background: Dysbiosis in the gut microbial community might be involved in the pathophysiology of attention deficit/hyperactivity disorder (ADHD). The fungal component of the gut microbiome, namely the mycobiota, is a hyperdiverse group of multicellular eukaryotes that can influence host intestinal permeability. This study therefore aimed to investigate the impact of fungal mycobiome dysbiosis and intestinal permeability on ADHD. Methods: Faecal samples were collected from 35 children with ADHD and from 35 healthy controls. Total DNA was extracted from the faecal samples, and the internal transcribed spacer (ITS) regions were sequenced using high-throughput next-generation sequencing (NGS). The fungal taxonomic classification was analysed using bioinformatics tools, and the differentially expressed fungal species between the ADHD and healthy control groups were identified. An in vitro permeability assay (Caco-2 cell layer) was used to evaluate the biological effects of fungal dysbiosis on intestinal epithelial barrier function. Results: The β-diversity (the species diversity between two communities), but not α-diversity (the species diversity within a community), reflected the differences in fungal community composition between ADHD and control groups. At the phylum level, the ADHD group displayed a significantly higher abundance of Ascomycota and significantly lower abundance of Basidiomycota than the healthy control group. At the genus level, the abundance of Candida (especially Candida albicans) was significantly increased in ADHD patients compared to the healthy controls. In addition, the in vitro cell assay revealed that C. albicans secretions significantly enhanced the permeability of Caco-2 cells. Conclusions: The current study is the first to explore altered gut mycobiome dysbiosis using the NGS platform in ADHD. The findings from this study indicated that dysbiosis of the fungal mycobiome and intestinal permeability might be associated with susceptibility to ADHD.

Keywords: ADHD, fungus, gut–brain axis, biomarker, child psychiatry

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