16 results on '"Diego Herman Sauka"'
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2. Bacillus thuringiensis Bt_UNVM-84, a Novel Strain Showing Insecticidal Activity against Anthonomus grandis Boheman (Coleoptera: Curculionidae)
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Diego Herman Sauka, Cecilia Peralta, Melisa Paula Pérez, Antonella Molla, Tadeo Fernandez-Göbel, Federico Ocampo, and Leopoldo Palma
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Bacillus thuringiensis ,insect pests ,insecticidal proteins ,Vpb1/Vpa2 proteins ,Cry8 proteins ,biological control ,Medicine - Abstract
Bacillus thuringiensis is a Gram-positive bacterium known for its insecticidal proteins effective against various insect pests. However, limited strains and proteins target coleopteran pests like Anthonomous grandis Boheman, causing substantial economic losses in the cotton industry. This study focuses on characterizing a Bacillus sp. strain, isolated from Oncativo (Argentina), which exhibits ovoid to amorphous parasporal crystals and was designated Bt_UNVM-84. Its genome encodes genes for the production of two pairs of binary Vpb1/Vpa2 proteins and three Cry-like proteins showing similarity with different Cry8 proteins. Interestingly, this gene content was found to be conserved in a previously characterized Argentine isolate of B. thuringiensis designated INTA Fr7-4. SDS-PAGE analysis revealed a major band of 130 kDa that is proteolytically processed to an approximately 66-kDa protein fragment by trypsin. Bioassays performed with spore-crystal mixtures demonstrated an interesting insecticidal activity against the cotton boll weevil A. grandis neonate larvae, resulting in 91% mortality. Strain Bt_UNVM-84 is, therefore, an interesting candidate for the efficient biological control of this species, causing significant economic losses in the cotton industry in the Americas.
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- 2023
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3. Induced-feeding bioassays for detection of Bacillus thuringiensis insecticidal activity against Epilachna paenulata (Coleoptera) Bioensaios de alimentação induzida para determinar a atividade inseticida de Bacillus thuringiensis contra Epilachna paenulata (Coleoptera)
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Diego Herman Sauka, Rodrigo Horacio Monella, and Graciela Beatriz Benintende
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controle biológico ,Coccinellidae ,Cucurbitaceae ,joaninha ,toxicidade esporo-cristal ,biological control ,ladybird beetle ,spore-crystal toxicity ,Agriculture (General) ,S1-972 - Abstract
The objective of this work was to establish and test the induced-feeding bioassay in order to detect Bacillus thuringiensis insecticidal activity against Epilachna paenulata (Coleoptera: Coccinellidae). Larvae were induced to swallow high concentrations of spore-crystal suspensions of seven exotic and 30 Argentine B. thuringiensis strains. The great majority of strains showed no toxicity to E. paenulata larvae, and observed mortality was lower than 30%. Induced-feeding bioassay is feasible, and should be used for prospecting strains that produce right combinations of Cry proteins needed to an efficient pest control.O objetivo deste trabalho foi estabelecer e testar o bioensaio de alimentação induzida para a determinação da atividade inseticida de Bacillus thuringiensis contra larvas de Epilachna paenulata (Coleoptera: Coccinellidae). As larvas foram induzidas a ingerir concentrações elevadas de suspensões esporo-cristal de 7 cepas exóticas e 30 argentinas de B. thuringiensis. A maioria das cepas não apresentou toxicidade a larvas de E. paenulata, com mortalidade inferior a 30%. O bioensaio de alimentação induzida se mostrou efetivo, e pode ser usado para identificar cepas que produzam combinações adequadas de proteínas Cry, necessárias para o controle de pragas.
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- 2010
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4. Paenibacillus xylanivorans sp. nov., a xylan-degrading bacterium isolated from decaying forest soil
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Eleonora Campos, Diego Herman Sauka, Silvina Ghio, Florencia E Piccini, Ornella Mailén Ontañon, and Alejandro E. Ferrari
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0106 biological sciences ,0301 basic medicine ,chemistry.chemical_classification ,biology ,Strain (chemistry) ,Nucleic acid sequence ,Fatty acid ,General Medicine ,16S ribosomal RNA ,biology.organism_classification ,010603 evolutionary biology ,01 natural sciences ,Microbiology ,DNA sequencing ,03 medical and health sciences ,Paenibacillus ,030104 developmental biology ,chemistry ,Botany ,Xylanase ,Ecology, Evolution, Behavior and Systematics ,Bacteria - Abstract
A xylanolytic bacterial strain, named A59T, was isolated from a forest soil consortium in southern Argentina. Strain A59T is a Gram-stain-positive, facultative anaerobic, endospore-forming and rod-shaped bacterium. Its optimal growth conditions are 30 °C (range, 28–37 °C), pH 7 (range, pH 5–10) and it tolerates up to 7 % of NaCl (range, 2–7 %). Chemotaxonomic analysis revealed that strain A59Tpossesses meso-diaminopimelic acid in the cell wall. It contains menaquinone MK-7 as the predominant isoprenoid quinone and the major fatty acid is anteiso-C15 : 0 (35.1 %), with a moderate amount of C16 : 0 (6.9 %). According to 16S RNA gene sequence analysis, the isolate is phylogenetically placed in the same cluster as Paenibacillus taichungensis BCRC 17757T (99.7 % nucleotide sequence identity) and Paenibacillus pabuli NBRC 13638T (99.1 %) and is closely related to Paenibacillus tundrae A10bT (98.8 %). However, phylogenetic studies based on the housekeeping gyrB gene placed A59T in a separate branch from all other related type strains. Furthermore, the results of whole genome average nucleotide identity analysis (gANI) with related type strains was lower than 91.10 % and the digital DNA–DNA hybridization value was lower than 44.30 %, which are below the threshold values for separating two species. The DNA G+C content was estimated as 46.09 mol%, based on genome sequencing. On the basis of these results, A59T represents a new species of the genus Paenibacillus , and we propose the name Paenibacillus xylanivorans sp. nov. The type strain is A59T (=DSM 107920T=NCIMB 15123T).
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- 2019
5. Cepas argentinas de Bacillus thuringiensis con distinta morfología en sus cristales paraesporales
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Antonela Alejandra Marozzi, Leopoldo Palma, Diego Herman Sauka, Eleodoro Eduardo del Valle, and Cecilia Peralta
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Microbiology (medical) ,Spores, Bacterial ,Morphology ,Bacteria ,Biopesticidas ,Bacillus thuringiensis ,Argentina ,Morphology (biology) ,General Medicine ,Biology ,biology.organism_classification ,Microbiology ,purl.org/becyt/ford/1 [https] ,Endotoxins ,Hemolysin Proteins ,Bacterial Proteins ,Morfología ,Cristales parasporales ,purl.org/becyt/ford/1.6 [https] - Abstract
Bacillus thuringiensis is a Gram-positive and sporulated bacterium exhibiting insecticidal activity against a wide range of insects.3 During sporulation, this bacterium produces a number of different proteins forming crystalline inclusions adjacent to the spores (parasporal crystals). Among these insecticidal proteins, the most abundant are those commonly known as Cry (Crystal) proteins, which are responsible for exerting a toxic activity (upon ingestion) against insects of different species.5 For this reason, B. thuringiensis has proved to be the most efficient and used bioinsecticide to date.2 However, Spodoptera cosmioides, Spodoptera eridania and Agrotis sp. (Lepidoptera) are species that are not yet controlled by some transgenic crops (e.g. Intacta RR2Pro soybean). Thus, in an attempt to enlarge the host spectrum of this bacterium it is necessary to search for novel strains. In this work we show a sporulated B. thuringiensis Bt-UNVM_84 strain exhibiting a number of rare amorphous to spherical crystal combinations, whereas sporulated B. thuringiensis strain Bt-UNVM-94 showed quasi symmetric bipyramidal parasporal crystals, by using scanning electron microscopy (SEM) (Fig. 1). Strains Bt-UNVM_84 and Bt-UNVM_94 were isolated from Oncativo (Córdoba, Argentina) and Cululú (Santa Fe, Argentina), respectively. The insecticidal activity of these different B. thuringiensis strains is currently under investigation. Each strain was grown in liquid CCY sporulation medium6 for ∼48 h (150 rpm) until no vegetative cells were observed under a light microscope. The presence of parasporal crystals was first determined using Coomassie blue stained slides1 (1000×) under a Nikon E100 light microscope and confirmed later by a Nikon Ti-Eclipse phase contrast microscope (1000×) (data not shown). For the SEM analysis, aliquots of 1 ml were centrifuged for 5 minutes (16,000 g) at room temperature. Each pellet was washed three times with sterile distilled water and fixed with 100 μl 4% formaldehyde. Each fixed preparation was then sent to Centro Integral de Microscopía Electrónica (CIME ? CONICET ? UNT) for SEM examination. Fil: Peralta, Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones y Transferencia de Villa María. Universidad Nacional de Villa María. Centro de Investigaciones y Transferencia de Villa María; Argentina Fil: Sauka, Diego Herman. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Marozzi, Antonela Alejandra. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Patagonia Norte. Instituto de Investigaciones En Biodiversidad y Medioambiente. Subsede San Martín de Los Andes-inibioma | Universidad Nacional del Comahue. Centro Regional Universitario Bariloche. Instituto de Investigaciones En Biodiversidad y Medioambiente. Subsede San Martín de Los Andes-inibioma.; Argentina Fil: del Valle, Eleodoro Eduardo. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Instituto de Ciencias Agropecuarias del Litoral. - Universidad Nacional del Litoral. Instituto de Ciencias Agropecuarias del Litoral.; Argentina Fil: Palma, Leopoldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones y Transferencia de Villa María. Universidad Nacional de Villa María. Centro de Investigaciones y Transferencia de Villa María; Argentina
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- 2021
6. Bacillus toyonensis biovar Thuringiensis: A novel entomopathogen with insecticidal activity against lepidopteran and coleopteran pests
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Diego Herman Sauka, Cecilia Peralta, Melisa Paula Pérez, María Inés Onco, Angelika Fiodor, Javier Caballero, Primitivo Caballero, Colin Berry, Eleodoro Eduardo Del Valle, and Leopoldo Palma
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Insect Science ,Agronomy and Crop Science - Abstract
The Bacillus cereus group includes eight species: Bacillus anthracis, B. cereus, Bacillus cytotoxicus, Bacillus mycoides, Bacillus pseudomycoides, Bacillus thuringiensis, Bacillus weihenstephanensis and Bacillus toyonensis, which are highly related at the phylogenetic level. In this work, we performed the isolation and characterization of a novel Bacillus sp. strain exhibiting parasporal crystals with insecticidal activity that was initially classified as a Bacillus thuringiensis strain. Its genome encoded three genes showing homology to known pesticidal proteins from B. thuringiensis and Lysinibacillus sphaericus proteins: Cry7Ga1 (a crystal protein), a Mpp2Aa3 (ETX/Mtx2 family) homolog and a mosquitocidal-like protein (NPP1). However, since its genome sequence shared > 98% ANI with several Bacillus toyonensis genomes, the strain has subsequently been renamed as Bacillus toyonensis biovar Thuringiensis and designated Bto-UNVM_94. Bioassays demonstrated that this novel strain exhibited toxicity against Cydia pomonella (Lepidoptera: Tortricidae) and Anthonomus grandis (Coleoptera: Curculionidae), a low toxicity against Aedes aegypti (Diptera: Culicidae) and Alphitobius diaperinus (Coleoptera: Tenebrionidae) whereas no toxicity was shown to the free-living nematode Panagrellus redivivus (Rhabditida: Panagrolaimidae).
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- 2022
7. Genome sequence analysis and insecticidal characterization of Bacillus thuringiensis Bt-UNVM_94, a strain showing dual insecticidal activity against lepidopteran and coleopteran pests
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Leopoldo Palma, Javier Caballero, Cecilia Peralta, Angelika Fiodor, Primitivo Caballero, Colin Berry, Eleodoro Eduardo del Valle, Melisa Paula Perez, Diego Herman Sauka, and Maria Ines Onco
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Lepidoptera genitalia ,Rhabditidae ,biology ,Strain (chemistry) ,Muscidae ,Bacillus thuringiensis ,Panagrellus redivivus ,fungi ,Biological pest control ,Bioassay ,biology.organism_classification ,Microbiology - Abstract
Bacillus thuringiensis is a gram-positive and spore-forming bacterium that synthesizes a wide diversity of proteins with insecticidal activity and which has demonstrated its potential and safety as a biocontrol agent for more than four decades. However, several susceptible insect species have been reported for evolving resistance, which demands screening for strains exhibiting novel insecticidal properties. In this work, we performed the genome sequence analysis and the insecticidal characterization of B. thuringiensis strain Bt-UNVM_94 isolated from Argentina. This strain produced quasi symmetric bipyramidal parasporal crystals as shown using Scanning Electron Microscopy. Its genomic sequence harbours one coding sequence showing homology to the crystal toxin Cry7Ga2 and another, with similarity to the Mpp2Aa3 (Mtx2) toxin. Known Cry7A and Cry7B are known to be active against some coleopteran and lepidopteran larvae, respectively. However, bioassays performed with spore-crystal mixtures of strain Bt-UNVM_94 exhibited dual toxicity with 50% and 91% mortality against Cydia pomonella (Lepidoptera: Tortricidae) and Anthonomus grandis (Coleoptera: Curculionidae), respectively. No toxicity was detected against the free-living nematode Panagrellus redivivus (Rhabditidae: Panagrolaimidae). This strain also showed no PCR amplification of the type I b-exotoxin thuE gene, consistent with the absence of mortality in b-exotoxin bioassays with Musca domestica (Diptera: Muscidae). Screenings of novel B. thuringiensis strains may provide toxins with novel insecticidal properties that can be used to suppress insect resistance to the most used B. thuringiensis-crops in the field.
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- 2021
8. Efecto nematicida de una cepa argentina de Photorabdus laumondi laumondi (Enterobacteriaceae) sobre el nematodo de vida libre Panagrellus redivivus (Rhabditidae: Panagrolaimidae)
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Augusto Salas, Diego Herman Sauka, and María Fernanda Achinelly
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Microbiology (medical) ,Rhabditidae ,Photorhabdus laumondi laumondi ,NACOBBUS ,Microbiology ,Panagrolaimidae ,purl.org/becyt/ford/1 [https] ,Rhabditida ,Enterobacteriaceae ,Nematicides ,NEMATICIDA ,Nematocidas ,Animals ,Panagrellus redivivus ,purl.org/becyt/ford/1.6 [https] ,ARGENTINA ,biology ,Panagrellus ,Strain (biology) ,General Medicine ,biology.organism_classification ,PHOTORHABDUS ,Nematode ,Photorhabdus - Abstract
Bacteria with nematicidal effects have attracted interest as biocontrol agents of pests and diseases due to their relatively low environmental impact compared to chemical management strategies1. The damage they cause to target nematodes is diverse and imperfectly understood4.Photorhabdus spp. (Enterobacteriaceae) lives symbiotically within the nematode Heterorhabditis spp. (Rhabditida: Heterorhabditidae). It is known that secondary metabolites of this gram-negative bacterium exhibit insecticidal activity, mainly in Lepidopteran and Coleopteran insects2. Studies have also shown that cell-free culture filtrates of Photorhabdus spp. have nematicidal activity on free-living nematodes and pest phytonematodes; however, the causes of death in these targets are still being studied. Fil: Salas, Augusto. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Achinelly, Maria Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; Argentina Fil: Sauka, Diego Herman. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
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- 2021
9. New entomopathogenic strain of Bacillus thuringiensis is able to solubilize different sources of inorganic phosphates
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Carlos Fabian Piccinetti, Daniela Adriana Vallejo, Melisa Paula Perez, Diego Herman Sauka, Maria Ines Onco, and Graciela Beatriz Benintende
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0106 biological sciences ,Ecology ,biology ,Strain (chemistry) ,Chemistry ,fungi ,PH reduction ,Soil Science ,04 agricultural and veterinary sciences ,biology.organism_classification ,Phosphate ,01 natural sciences ,Agricultural and Biological Sciences (miscellaneous) ,Spore ,Lepidoptera genitalia ,chemistry.chemical_compound ,Cry1Ac ,Phosphorite ,Bacillus thuringiensis ,040103 agronomy & agriculture ,0401 agriculture, forestry, and fisheries ,Food science ,010606 plant biology & botany - Abstract
Phosphate-solubilizing strain INTA 293-1 from a soil sample of Corrientes province (Argentina) was identified as Bacillus thuringiensis based on its faculty to produce crystalline inclusions during sporulation and 16S rDNA sequence similarity. INTA 293-1 was found to produce bipyramidal and cuboidal crystals, containing major proteins of ca. 130 and 65 kDa respectively, resembling those present in Lepidoptera- and Diptera-active strains. Accordingly, INTA 293-1 showed insecticidal activity against lepidopteran and dipteran larvae. No insecticidal activity nor evident effects on the normal development of coleopteran larvae or nematodes were observed. Screening of cry, cyt and vip genes showed that INTA 293-1 contains cry1Ac, cry1Ia, cry2Aa, cry2Ab and vip3Aa genes. Furthermore, this new entomopathogenic strain of B. thuringiensis was able to solubilize different sources of inorganic phosphates in liquid media following similar solubilization kinetics. INTA 293-1 was effective in solubilizing hydroxyapatite, patagonic phosphate rock, and to a lesser extent the aluminium phosphate. The differential behaviour between them may be explained by differences in INTA 293-1 survival. Phosphate solubilization was associated with pH reduction. This study reveals a little-explored aspect of B. thuringiensis and paves the way for the development of new agrobiological products with multiple modes of action.
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- 2021
10. Complete Sequence and Organization of pFR260, the Bacillus thuringiensis INTA Fr7-4 Plasmid Harboring Insecticidal Genes
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Diego Herman Sauka, Rubén O. Zandomeni, Elio Maximiliano Ortiz, Marcelo Facundo Berretta, Ariel Fernando Amadio, Laura Emilce Navas, and Graciela Beatriz Benintende
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0301 basic medicine ,Genetics ,Physiology ,Cell Biology ,Biology ,biology.organism_classification ,Applied Microbiology and Biotechnology ,Biochemistry ,Microbiology ,03 medical and health sciences ,Complete sequence ,030104 developmental biology ,Plasmid ,Bacillus thuringiensis ,Gene ,Biotechnology - Abstract
We report the complete sequence and analysis of pFR260, a novel megaplasmid of 260,595 bp from the Bacillus thuringiensis strain INTA Fr7-4 isolated in Argentina. It carries 7 insecticidal genes: 3 cry8 copies previously reported, 2 vip1, and 2 vip2. Also, it carries a gene encoding a putative atypical Cry protein. These genes are arranged in a region of approximately 105 kbp in size with characteristics of a pathogenicity island with a potential coleopteran-specific insecticide profile. DNA strand composition asymmetry, as determined by GC skew analysis, and the presence of a Rep protein involved in the initiation of replication suggest a bidirectional theta mechanism of replication. In addition, many genes involved in conjugation and a CRISPR-Cas system were detected. The pFR260 sequence was deposited in GenBank under accession number KX258624.
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- 2017
11. Secretome profile of Cellulomonas sp. B6 growing on lignocellulosic substrates
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Paola Talia, Ornella Mailén Ontañon, Maria Pia Valacco, Eleonora Campos, Diego Herman Sauka, Silvina Ghio, Maximo Rivarola, and Florencia Elizabeth Piccinni
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chemistry.chemical_classification ,biology ,020209 energy ,Lignocellulosic biomass ,02 engineering and technology ,General Medicine ,Straw ,biology.organism_classification ,Lignin ,Applied Microbiology and Biotechnology ,Cellulomonas sp ,Enzyme ,Bacterial Proteins ,Biochemistry ,chemistry ,Biofuel ,Metabolome ,0202 electrical engineering, electronic engineering, information engineering ,Glycoside hydrolase ,Biomass ,Cellulomonas ,Bioprocess ,Biotechnology - Abstract
AIMS Lignocellulosic biomass deconstruction is a bottleneck for obtaining biofuels and value-added products. Our main goal was to characterize the secretome of a novel isolate, Cellulomonas sp. B6, when grown on residual biomass for the formulation of cost-efficient enzymatic cocktails. METHODS AND RESULTS We identified 205 potential CAZymes in the genome of Cellulomonas sp. B6, 91 of which were glycoside hydrolases (GH). By secretome analysis of supernatants from cultures in either extruded wheat straw (EWS), grinded sugar cane straw (SCR) or carboxymethylcellulose (CMC), we identified which proteins played a role in lignocellulose deconstruction. Growth on CMC resulted in the secretion of two exoglucanases (GH6 and GH48) and two GH10 xylanases, while growth on SCR or EWS resulted in the identification of a diversity of CAZymes. From the 32 GHs predicted to be secreted, 22 were identified in supernatants from EWS and/or SCR cultures, including endo- and exoglucanases, xylanases, a xyloglucanase, an arabinofuranosidase/β-xylosidase, a β-glucosidase and an AA10. Surprisingly, among the xylanases, seven were GH10. CONCLUSIONS Growth of Cellulomonas sp. B6 on lignocellulosic biomass induced the secretion of a diverse repertoire of CAZymes. SIGNIFICANCE AND IMPACT OF THE STUDY Cellulomonas sp. B6 could serve as a source of lignocellulose-degrading enzymes applicable to bioprocessing and biotechnological industries.
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- 2018
12. Predatory journals: An evil of this time?
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Diego Herman Sauka
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0301 basic medicine ,Microbiology (medical) ,030106 microbiology ,05 social sciences ,Publishing ethics ,050801 communication & media studies ,General Medicine ,Microbiology ,MICROBIOLOGÍA ,Ciencias Biológicas ,purl.org/becyt/ford/1 [https] ,03 medical and health sciences ,0508 media and communications ,Biología Celular, Microbiología ,Open access publishing ,Political science ,REVISTAS DEPREDADORAS ,purl.org/becyt/ford/1.6 [https] ,Humanities ,CIENCIAS NATURALES Y EXACTAS - Abstract
¿Qué investigador no ha recibido en este último tiempo alguna invitación electrónica para publicar un artículo en una revista desconocida para él hasta ese momento? Hay que estar atento, porque podría estar frente al acecho de una revista depredadora. Establecido por Jeffrey Beall -bibliotecario y académico de la Universidad de Colorado-, Este término se refiere un Aquellas revistas Científicas Que explotan, de forma poco ética, el Valioso modelo de acceso abierto ( Open Access ) para su propio beneficio. Fil: Sauka, Diego Herman. Asociación Argentina de Microbiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
- Published
- 2018
13. Sequence and Expression of Two cry8 Genes from Bacillus thuringiensis INTA Fr7-4, a Native Strain from Argentina
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Graciela Beatriz Benintende, Rubén O. Zandomeni, Ariel Fernando Amadio, Melisa Paula Perez, Marcelo Facundo Berretta, Laura Emilce Navas, Elio Maximiliano Ortiz, and Diego Herman Sauka
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Cloning ,Strain (chemistry) ,Physiology ,Sequence analysis ,fungi ,Cell Biology ,Biology ,biology.organism_classification ,Applied Microbiology and Biotechnology ,Biochemistry ,Microbiology ,Molecular biology ,law.invention ,chemistry.chemical_compound ,chemistry ,law ,Bacillus thuringiensis ,Gene expression ,Recombinant DNA ,Gene ,DNA ,Biotechnology - Abstract
We found and characterized two cry8 genes from the Bacillus thuringiensis strain INTA Fr7-4 isolated in Argentina. These genes, cry8Kb3 and cry8Pa3, are located in a tandem array within a 13,200-bp DNA segment sequenced from a preparation of total DNA. They encode 1,169- and 1,176-amino-acid proteins, respectively. Both genes were cloned with their promoter sequences and the proteins were expressed separately in an acrystalliferous strain of B. thuringiensis leading to the formation of ovoid crystals in the recombinant strains. The toxicity against larvae of Anthonomus grandis Bh. (Coleoptera: Curculionidae) of a spore-crystal suspension from the recombinant strain containing cry8Pa3 was similar to that of the parent strain INTA Fr7-4.
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- 2014
14. Identification, Cloning and Expression of an Insecticide cry8 Gene from Bacillusthuringiensis INTA Fr7-4
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Marcelo Facundo Berretta, Rubén O. Zandomeni, Diego Herman Sauka, Laura Emilce Navas, Graciela Beatriz Benintende, and Ariel Fernando Amadio
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Cloning ,biology ,Strain (chemistry) ,Physiology ,fungi ,Cell Biology ,biology.organism_classification ,Applied Microbiology and Biotechnology ,Biochemistry ,Microbiology ,law.invention ,Lepidoptera genitalia ,Anticarsia gemmatalis ,law ,Bacillus thuringiensis ,Recombinant DNA ,Noctuidae ,Gene ,Biotechnology - Abstract
Insecticidal activity of Bacillus thuringiensis is attributed largely to the crystal proteins. These were found with specific toxic activity against insects in different orders. A novel cry8 gene from B. thuringiensis strain INTA Fr7-4 from Argentina was characterized. The encoded protein, Cry8Qa2, is 1184 amino acids long and its sequence is more similar to those of Cry8F class. We cloned and expressed the protein in an acrystalliferous strain of B. thuringiensis using two differentially regulated promoters. The recombinant strains produced ovoid crystals with low toxicity against larvae of Anticarsia gemmatalis (Lepidoptera: Noctuidae). The morphology and insecticidal properties of these crystals resembled those produced by the native strain INTA Fr7-4.
- Published
- 2013
15. PCR-based prediction of type I β-exotoxin production in Bacillus thuringiensis strains
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Diego Herman Sauka, Marcelo Facundo Berretta, Graciela Beatriz Benintende, Nanci N. Lopez, Maria Ines Onco, and Melisa Paula Perez
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chemistry.chemical_classification ,β-exotoxin ,Adenosine ,biology ,Otras Ciencias Biológicas ,Bacillus thuringiensis ,Sugar Acids ,Adenine nucleoside ,Oligosaccharide ,medicine.disease_cause ,biology.organism_classification ,Polymerase Chain Reaction ,Microbiology ,Ciencias Biológicas ,thuE gene ,PCR ,chemistry ,medicine ,Bioassay ,Ecology, Evolution, Behavior and Systematics ,Exotoxin ,CIENCIAS NATURALES Y EXACTAS - Abstract
Some Bacillus thuringiensis strains secrete type I β-exotoxin, which is a non-specific insecticidal and thermostable adenine nucleoside oligosaccharide. Toxicity bioassays and HPLC are traditional methods for detecting β-exotoxin. With the aim of establish a first rapid approach for prediction of type I β-exotoxin production, two PCR-based methods were successfully evaluated in B. thuringiensis strains and native isolates. In order to validate a reliable technology, results obtained by this method were correlated with that obtained from Musca domestica bioassays. Fil: Sauka, Diego Herman. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Perez, Melisa Paula. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Lopez, Nanci N.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Onco, María Inés. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Berretta, Marcelo Facundo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Benintende, Graciela Beatriz. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina
- Published
- 2014
16. Induced-feeding bioassays for detection of Bacillus thuringiensis insecticidal activity against Epilachna paenulata (Coleoptera)
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Graciela Beatriz Benintende, Diego Herman Sauka, and Rodrigo Horacio Monella
- Subjects
Veterinary medicine ,Larva ,biology ,business.industry ,fungi ,Biological pest control ,Pest control ,biological control ,biology.organism_classification ,Cucurbitaceae ,spore-crystal toxicity ,ladybird beetle ,Coccinellidae ,Bacillus thuringiensis ,Botany ,Bioassay ,Animal Science and Zoology ,Natural enemies ,business ,Agronomy and Crop Science - Abstract
The objective of this work was to establish and test the induced-feeding bioassay in order to detect Bacillus thuringiensis insecticidal activity against Epilachna paenulata (Coleoptera: Coccinellidae). Larvae were induced to swallow high concentrations of spore-crystal suspensions of seven exotic and 30 Argentine B. thuringiensis strains. The great majority of strains showed no toxicity to E. paenulata larvae, and observed mortality was lower than 30%. Induced-feeding bioassay is feasible, and should be used for prospecting strains that produce right combinations of Cry proteins needed to an efficient pest control.
- Published
- 2010
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