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Biofertilizers are the eco-friendly bio-input being used to sustain the agriculture by reducing the chemical inputs and improving the soil health. Quality is the major concern of biofertilizer technology which often leads to poor performance in the field and thereby loses the farmers' faith. To authenticate the strain as well as its presumed cell load of a commercial product, sequence characterized amplified region (SCAR) markers were developed for three biofertilizer strains viz., Azospirillum brasilense (Sp7), Bacillus megaterium (Pb1) and Azotobacter chroococcum (Ac1). We evaluated the feasibility of multiplex-PCR and quantitative real-time PCR for SCAR marker-based quality assessment of the product as well as the persistence of the strains during crop growth. We showed that multiplex PCR can concurrently discriminate the strains based on the amplicons' size and detects up to 104 cells per g or per ml of carrier-based or liquid formulation of biofertilizer, respectively. The detection limit of quantitative PCR targeting SCAR markers is 103 cells per g or ml of biofertilizer. Both the PCR methods detected and quantified them in the maize rhizosphere. Hence SCAR marker-based quality assessment would be a sensitive tool to monitor the biofertilizer production as well as its persistence in the inoculated crop rhizosphere.
Asunto(s)
Azospirillum brasilense/aislamiento & purificación , Azotobacter/aislamiento & purificación , Bacillus megaterium/aislamiento & purificación , Técnicas de Tipificación Bacteriana/métodos , Fertilizantes/microbiología , Reacción en Cadena de la Polimerasa Multiplex/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Microbiología del Suelo , Agricultura , Azospirillum brasilense/genética , Azotobacter/genética , Bacillus megaterium/genética , Secuencia de Bases , Dermatoglifia del ADN , Cartilla de ADN/genética , ADN Bacteriano/análisis , Marcadores Genéticos , Raíces de Plantas/microbiología , Rizosfera , Sensibilidad y Especificidad , Zea mays/microbiologíaRESUMEN
BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive and fatal interstitial lung disease of unknown origin. Alveolar epithelial cells (AECs) play an important role in the fibrotic process as they undergo sustained endoplasmic reticulum (ER) stress, and may acquire a mesenchymal phenotype through epithelial-to-mesenchymal transition (EMT), two phenomena that could be induced by localized alveolar hypoxia. Here we investigated the potential links between hypoxia, ER stress and EMT in AECs. METHODS: ER stress and EMT markers were assessed by immunohistochemistry, western blot and qPCR analysis, both in vivo in rat lungs exposed to normoxia or hypoxia (equivalent to 8% O2) for 48 h, and in vitro in primary rat AECs exposed to normoxia or hypoxia (1.5% O2) for 2â»6 days. RESULTS: Hypoxia induced expression of mesenchymal markers, pro-EMT transcription factors, and the activation of ER stress markers both in vivo in rat lungs, and in vitro in AECs. In vitro, pharmacological inhibition of ER stress by 4-PBA limited hypoxia-induced EMT. Calcium chelation or hypoxia-inducible factor (HIF) inhibition also prevented EMT induction under hypoxic condition. CONCLUSIONS: Hypoxia and intracellular calcium are both involved in EMT induction of AECs, mainly through the activation of ER stress and HIF signaling pathways.
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Células Epiteliales Alveolares/citología , Butilaminas/farmacología , Estrés del Retículo Endoplásmico/efectos de los fármacos , Factores de Transcripción/metabolismo , Células Epiteliales Alveolares/efectos de los fármacos , Células Epiteliales Alveolares/metabolismo , Animales , Calcio/metabolismo , Quelantes del Calcio/farmacología , Hipoxia de la Célula/efectos de los fármacos , Células Cultivadas , Modelos Animales de Enfermedad , Transición Epitelial-Mesenquimal/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
Uveal melanoma (UM) is the most common cancer of the eye. The loss of chromosome 3 (M3) is associated with a high risk of metastases. M3 tumors are more infiltrated by T-lymphocytes than low-risk disomic-3 (D3) tumors, contrasting with other tumor types in which T cell infiltration correlates with better prognosis. Whether these T cells represent an antitumor response and how these T cells would be primed in the eye are both unknown. Herein, we characterized the T cells infiltrating primary UMs. CD8+ and Treg cells were more abundant in M3 than in D3 tumors. CD39+PD-1+CD8+ T cells were enriched in M3 tumors, suggesting specific responses to tumor antigen (Ag) as confirmed using HLA-A2:Melan-A tetramers. scRNAseq-VDJ analysis of T cells evidenced high numbers of proliferating CD39+PD1+CD8+ clonal expansions, suggesting in situ antitumor Ag responses. TCRseq and tumor-Ag tetramer staining characterized the recirculation pattern of the antitumor responses in M3 and D3 tumors. Thus, tumor-Ag responses occur in localized UMs, raising the question of the priming mechanisms in the absence of known lymphatic drainage.
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Melanoma , Neoplasias de la Úvea , Humanos , Melanoma/terapia , Linfocitos T CD8-positivos , DrenajeRESUMEN
Low availability of phosphorus (P) in both acidic and alkaline soils is a major problem for sustainable improvement in wheat crops yield. Optimization of crops productivity can be achieved by increasing the bioavailability of P by phosphate solubilizing Actinomycetota (PSA). However, their effectiveness may vary with changing agro-climatic conditions. In this regard, a greenhouse experiment was conducted to assess the interaction inoculation of five potential PSA (P16-P18-BC3-BC10 and BC11) and RPs (RP1- RP2-RP3 and RP4) on the growth and yield of wheat crop in unsterilized P- deficient alkaline and acidic soils. Their performance was compared with single super phosphate (TSP) and reactive RP (BG4). The in-vitro tests showed that all PSA colonize wheat root and form a strong biofilm except Streptomyces anulatus strain P16. Our findings revealed that all PSA significantly improve the shoot/root dry weights, spike biomass, chlorophyll contents as well as nutrients uptake in plants fertilized with RP3 and RP4. However, the combined application of Nocardiopsis alba BC11 along with RP4 in alkaline soil, was effective in optimizing wheat yield attributes and improve the yield biomass up to 19.7% as compared to the triple superphosphate (TSP). This study supports the view that the inoculation with Nocardiopsis alba BC11 has a broad RP solubilization and could alleviate the agricultural losses due to P limitation in acidic and alkaline soils.
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The growing interest in low-input agriculture in recent years has focused the use of microbial biofertilizers to improve plant growth and yield through a better mobilization of indigenous source of key nutrients such as nitrogen, phosphorus, potassium etc. In this context, soil microorganisms especially Actinobacteria might play an important role. With their multifunctional activities, they are involved in nutrient cycling, soil quality and crop productivity as well as plant health which make them not only the eco-friendly alternative for agriculture but also for humankind. Bearing this in mind, it is primordial to further explore the special link between these microorganisms and soil -plant ecosystems. Therefore, this review discusses the importance of Actinobacteria as microbial biofertilizers and highlights the future needs and challenges for using them for sustaining crop. The patents and scientific literature analysis from 2000 to 2020 show that 16 patents claiming Actinobacteria as biocontrol or biofertilizer in agriculture and 949 indexed research articles related to Actinobacteria effect on plant growth and phosphate solubilization have been published. Furthermore, Actinobacteria ability to increase growth and yield of staple crops such as wheat maize, tomato, rice, and chickpea plant have been highlighted. Much more effort and progress are expected in the industrial development of actinobacterial bioinoculants as areas such as synthetic biology and nano-biotechnology advance.
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Actinobacteria , Fertilizantes , Agricultura , Bacterias , Productos Agrícolas , Ecosistema , Fertilizantes/microbiología , Suelo , Microbiología del SueloRESUMEN
For many years, garlic has been used as a condiment in food and traditional medicine. However, the garlic skin, which accounts for 25% of the garlic bulk, is considered agricultural waste. In this study, cellulose nanocrystals (CNCs) and garlic extract (GE) from garlic skin were isolated and used as fillers to manufacture biocomposite films. The films were characterized in terms of UV barrier, thermal, mechanical, biodegradability, and antimicrobial activity. The chitosan-containing films and CNCs have significantly improved the films' tensile strength, Young's modulus, and elongation but decreased the film transparency compared to chitosan films. The combination of the CNCs and GE, on the other hand, slightly reduced the mechanical properties. The addition of CNCs slightly decreased the film transparency, while the addition of GE significantly improved the UV barrier properties. Thermal studies revealed that the incorporation of CNC and GE had minimal effect on the thermal stability of the chitosan films. The degradability rate of the chitosan composite films was found to be higher than that of the neat chitosan films. The antimicrobial properties of films were studied against Escherichia coli, Streptomyces griseorubens, Streptomyces alboviridis, and Staphylococcus aureus, observing that their growth was considerably inhibited by the addition of GE in composite films. Films incorporating both CNCs and GE from garlic skin hold more promise for active food packaging applications due to a combination of enhanced physical characteristics and antibacterial activity.
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Antiinfecciosos , Quitosano , Ajo , Nanopartículas , Antibacterianos/química , Antibacterianos/farmacología , Antiinfecciosos/química , Antiinfecciosos/farmacología , Antioxidantes/química , Celulosa/química , Quitosano/química , Escherichia coli , Embalaje de Alimentos , Nanopartículas/químicaRESUMEN
Soil fertility and plant nutrition require an adequate management of essential macronutrients such as potassium (K) and phosphorus (P), which are mandatory for plant development. Bioleaching of K and P bearing minerals improves their chemical weathering and increases the performance of the biofertilization strategies. In this study, in vitro and greenhouse experiments were carried out to investigate P and K solubilization traits of nine Actinobacteria (P13, P14, P15, P16, P17, P18, BC3, BC10, and BC11) under fertilization with rock phosphate (RP). K and P solubilization were evaluated on Alexandrov and NBRIP media containing mica and six RP samples, respectively. The actinobacterial strains were able to solubilize K in Alexandrov medium supplemented with RP. However, when soluble P was used instead of RP, only four strains of Actinobacteria (Streptomyces alboviridis P18-Streptomyces griseorubens BC3-Streptomyces griseorubens BC10 and Nocardiopsis alba BC11) solubilized K. The solubilization values of K ranged from 2.6 to 41.45 mg/L while those of P varied from 0.1 to 32 mg/L. Moreover, all strains were able to produce IAA, siderophore, HCN, and ammonia and significantly improved the germination rate and the vigor index of wheat. The pot experiments revealed that four strains (Streptomyces alboviridis P18, Streptomyces griseorubens BC3, Streptomyces griseorubens BC10, and Nocardiopsis alba BC11) significantly improved the growth parameters of wheat, namely root length (1.75-23.84%), root volume (41.57-71.46%), root dry weight (46.89-162.41%), shoot length (8.92-23.56%), and shoot dry weight (2.56-65.68%) compared to the uninoculated control. These findings showed that Streptomyces griseorubens BC10 and Nocardiopsis alba BC11 are promising candidates for the implementation of efficient biofertilization strategies to improve soil fertility and plant yield under rock P and rock K fertilization.
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Plant growth promoting microbes (PGPMs) play major roles in diverse ecosystems, including atmospheric nitrogen fixation, water uptake, solubilization, and transport of minerals from the soil to the plant. Different PGPMs are proposed as biofertilizers, biostimulants, and/or biocontrol agents to improve plant growth and productivity and thereby to contribute to agricultural sustainability and food security. However, little information exists regarding the use of PGPMs in micropropagation such as the in vitro plant tissue culture. This review presents an overview of the importance of PGPMs and their potential application in plant micropropagation. Our analysis, based on published articles, reveals that the process of in vitro classical tissue culture techniques, under strictly aseptic conditions, deserves to be reviewed to allow vitroplants to benefit from the positive effect of PGPMs. Furthermore, exploiting the potential benefits of PGPMs will lead to lessen the cost production of vitroplants during micropropagation process and will make the technique of plant tissue culture more efficient. The last part of the review will indicate where research is needed in the future.
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For all living organisms, nitrogen is an essential element, while being the most limiting in ecosystems and for crop production. Despite the significant contribution of synthetic fertilizers, nitrogen requirements for food production increase from year to year, while the overuse of agrochemicals compromise soil health and agricultural sustainability. One alternative to overcome this problem is biological nitrogen fixation (BNF). Indeed, more than 60% of the fixed N on Earth results from BNF. Therefore, optimizing BNF in agriculture is more and more urgent to help meet the demand of the food production needs for the growing world population. This optimization will require a good knowledge of the diversity of nitrogen-fixing microorganisms, the mechanisms of fixation, and the selection and formulation of efficient N-fixing microorganisms as biofertilizers. Good understanding of BNF process may allow the transfer of this ability to other non-fixing microorganisms or to non-leguminous plants with high added value. This minireview covers a brief history on BNF, cycle and mechanisms of nitrogen fixation, biofertilizers market value, and use of biofertilizers in agriculture. The minireview focuses particularly on some of the most effective microbial products marketed to date, their efficiency, and success-limiting in agriculture. It also highlights opportunities and difficulties of transferring nitrogen fixation capacity in cereals.
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The production of biofertilizers at industrial level is a bottleneck because bacterial strains are generally developed and managed by research laboratories and not by production units. A seamless transition from laboratory to field application is, therefore necessary. This review provides an overview of the constraints that limiting the application or the implementation of Actinobacteria based biofertilizers especially in agricultural field and suggests solutions to overcome some of these limits. General processes of making and controlling the quality of the inoculum are briefly described. In addition, the paper underlines the opportunity of biofertilizers alone or in combination with chemical fertilizers. This review also, highlights the latest studies (until June 2019) and focuses on P-solubilization microorganisms mainly Actinobacteria. The biotechnology of these bacteria is a glimmer of hope for rock phosphate (RP) bioformulation. Since direct application of RP fertilizer is not always agronomically effective due to its sparse solubility.
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Endoplasmic Reticulum (ER) stress of alveolar epithelial cells (AECs) is recognized as a key event of cell dysfunction in pulmonary fibrosis (PF). However, the mechanisms leading to AECs ER stress and ensuing unfolded protein response (UPR) pathways in idiopathic PF (IPF) remain unclear. We hypothesized that alveolar hypoxic microenvironment would generate ER stress and AECs apoptosis through the hypoxia-inducible factor-1α (HIF-1α). Combining ex vivo, in vivo and in vitro experiments, we investigated the effects of hypoxia on the UPR pathways and ER stress-mediated apoptosis, and consecutively the mechanisms linking hypoxia, HIF-1α, UPR and apoptosis. HIF-1α and the pro-apoptotic ER stress marker C/EBP homologous protein (CHOP) were co-expressed in hyperplastic AECs from bleomycin-treated mice and IPF lungs, not in controls. Hypoxic exposure of rat lungs or primary rat AECs induced HIF-1α, CHOP and apoptosis markers expression. In primary AECs, hypoxia activated UPR pathways. Pharmacological ER stress inhibitors and pharmacological inhibition or silencing of HIF-1α both prevented hypoxia-induced upregulation of CHOP and apoptosis. Interestingly, overexpression of HIF-1α in normoxic AECs increased UPR pathways transcription factors activities, and CHOP expression. These results indicate that hypoxia and HIF-1α can trigger ER stress and CHOP-mediated apoptosis in AECs, suggesting their potential contribution to the development of IPF.