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1.
Appl Biochem Biotechnol ; 191(4): 1638-1652, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32198600

RESUMEN

Microbes are often subjected to oxidative stress in nature that badly affects their growth rate and viability. Although the response of microbes against oxidative stress has been characterized at the chemical, physiological, and molecular levels, the mechanism of gene-regulation network adaptations of bacteria in response to oxidative stress remains largely unknown. In this study, transcriptomic profiling of glyphosate-tolerant Enterobacter strain NRS-1 was analyzed under 9 mM H2O2 stress using RNA-seq and qRT-PCR. The lag period in the growth of NRS-1 was very short compared with wild-type strain under H2O2 treatment. A total of 113 genes are identified as differentially expressed genes (DEGs) under H2O2 that include 38 upregulated and 75 downregulated transcripts. But not any genes regulated by major oxidative regulons, viz., oxyR, soxR, rpoS, perR, ohrR, and σв, have been reported in DEGs, hence potentially reflecting that specific changes have occurred in NRS-1 for adaptation to oxidative stress. Based on the functions of the DEGs, six elements namely formate dehydrogenase, processes associated with iron ions, repair programs, multidrug resistance, antioxidant defense, and energy generation (mqo, sdhC) might have contributed for stress tolerance in NRS-1. These elements are proposed to form a molecular network explaining gene response of NRS-1 to stress, and ensure global cell protection and growth recovery of NRS-1. These findings enrich the view of gene regulation in bacteria in response to H2O2 oxidative stress.


Asunto(s)
Enterobacter/genética , Regulación Bacteriana de la Expresión Génica , Peróxido de Hidrógeno/química , Estrés Oxidativo , Proteínas Bacterianas/genética , Secuencia de Bases , Enterobacter/efectos de los fármacos , Perfilación de la Expresión Génica , Glicina/análogos & derivados , Secuenciación de Nucleótidos de Alto Rendimiento , Proteínas Represoras/genética , Factores de Transcripción/genética , Transcriptoma , Glifosato
2.
3 Biotech ; 8(10): 422, 2018 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-30305993

RESUMEN

Knowledge of biological evolution and genetic mechanisms is gained by studying the adaptation of bacteria to survive in adverse environmental conditions. In this regard, transcriptomic profiling of a glyphosate-tolerant Enterobacter strain NRS-1 was studied under four different treatments to investigate the gene-regulatory system for glyphosate tolerance. A total of 83, 83, 60 and 74 genes were up-regulated and 108, 87, 178 and 117 genes down-regulated under 60-NPG, 110-NPG, NaCl (355 mM) and HCl (pH 4.46) stress treatments, respectively. Complex gene network was identified to be involved in regulating tolerance to glyphosate. This study revealed that NRS-1 has gained glyphosate tolerance at the cost of osmotic and acidic resistance. The 25 differentially expressed genes are reported to may have partly changed the function for providing resistance to glyphosate directly, among them genes metK, mtbK, fdnG and wzb that might detoxify/degrade the glyphosate. However, under 110-NPG condition, NRS-1 might have utilized economical and efficient ways by depressing its metabolism and activity to pass through this stress. Hence, the present study provides insights into the genes involved in glyphosate tolerance, which can be effectively utilized to engineer herbicide-resistant crop varieties after their proper validation to manage weed growth.

3.
FEMS Microbiol Lett ; 349(2): 135-43, 2013 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-24237416

RESUMEN

Glyphosate is a widely used herbicide that inhibits 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) activity. Most plants and microbes are sensitive to glyphosate. However, transgenic-resistant crops that contain a modified epsps obtained from the resistant microbes have been commercially successful and therefore, new resistance genes and their adaptive regulatory mechanisms are of great interest. In this study, a soil-borne, glyphosate-resistant bacterium was selected and identified as Enterobacter. The EPSPS in this strain was found to have been altered to a resistant one. A total of 42 differentially expressed genes (DEGs) in the glyphosate were screened using microarray techniques. Under treatment, argF, sdhA, ivbL, rrfA-H were downregulated, whereas the transcripts of speA, osmY, pflB, ahpC, fusA, deoA, uxaC, rpoD and a few ribosomal protein genes were upregulated. Data were verified by quantitative real-time PCR on selected genes. All transcriptional changes appeared to protect the bacteria from glyphosate and associated osmotic, acidic and oxidative stresses. Many DEGs may have the potential to confer resistance to glyphosate alone, and some may be closely related to the shikimate pathway, reflecting the complex gene interaction network for glyphosate resistance.


Asunto(s)
Farmacorresistencia Bacteriana/genética , Enterobacter/efectos de los fármacos , Enterobacter/genética , Regulación Bacteriana de la Expresión Génica , Genes Bacterianos , Glicina/análogos & derivados , Herbicidas/farmacología , Secuencia de Aminoácidos , Enterobacter/clasificación , Enterobacter/crecimiento & desarrollo , Enterobacter/aislamiento & purificación , Redes Reguladoras de Genes , Glicina/farmacología , Datos de Secuencia Molecular , Fenotipo , Filogenia , Alineación de Secuencia , Análisis de Secuencia de ADN , Estrés Fisiológico , Glifosato
4.
Mol Biol Rep ; 40(8): 4737-45, 2013 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-23666055

RESUMEN

Carotenoid oxygenase is a key enzyme in carotenoid metabolism leading to the synthesis of two phytohormones, abscisic acid (ABA) and strigolactone, as well as norisoprenoids. Few studies have analyzed inter-relationship of the metabolic networks of these three substances. In this present paper, soybean carotenoid oxygenase genes were identified to reveal their phylogenetic relationships, and the transcriptional response of these genes to four abiotic stresses (NaCl, PEG, high and low temperature) and ABA treatment were investigated to characterize their potential roles in plant resistance. Positive selection was found in the branches of carotenoid cleavage dioxygenase (CCD1), CCD8 and NCED (9-cis-epoxycarotenoid oxygenase), indicating an adaptive evolution in these clades. In soybean eight carotenoid oxygenase genes were identified. The transcriptional responses of almost all of them under stress and ABA conditions were significantly altered when assessed by quantitative polymerase chain reaction. Notably, CCD1 and CCD4, previously known as the key genes in norisoprenoids metabolism, showed especially strong responses to the abiotic stresses and ABA treatment. Furthermore, transcription levels of CCD7 and CCD8, key genes for the strigolactone pathway, highly increased during ABA treatment providing further evidence that ABA is involved in regulating strigolactone metabolism. All of the carotenoid oxygenase genes in soybean are involved in plant abiotic stress physiology, and ABA is presumed to be a core regulatory substance. These findings provide some insights into the mechanisms that underlie the regulation of tolerance response to abiotic stresses in soybean.


Asunto(s)
Adaptación Biológica/genética , Regulación de la Expresión Génica de las Plantas/genética , Glycine max/enzimología , Oxigenasas/genética , Filogenia , Estrés Fisiológico/genética , Ácido Abscísico/toxicidad , Teorema de Bayes , Biología Computacional , Cartilla de ADN/genética , Dioxigenasas/genética , Dioxigenasas/metabolismo , Genoma de Planta/genética , Modelos Genéticos , Polietilenglicoles/toxicidad , Selección Genética , Cloruro de Sodio/toxicidad , Temperatura
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