Academic literature on the topic 'Cellulose colonization'

When the attachment of cellulolytic rumen fungi to cellulose is blocked by the addition of methylcellulose, cellulose digestion is entirely inhibited. Even after these fungi have colonized and penetrated the cellulosic fibers of filter paper, the addition of methylcellulose effectively halts cellulose digestion. This effect of methylcellulose is accompanied by the complete inhibition of fungal attachment to cellulose fibers; the addition of methylcellulose does not affect the growth of these organisms on soluble substrates. We conclude that fungal cellulose digestion, like bacterial cellulose digestion, requires the spatial juxtaposition of the cellulolytic organism and its insoluble substrate. The simultaneous inhibition of both attachment and digestion by the same inhibitor suggests that these two processes are functionally linked in the fungi. Key words: cellulolysis, anaerobic rumen fungi, attachment inhibition, digestion inhibition.

Alternatives to synthetic nitrogen fertilizer are needed to reduce the costs of crop production and offset environmental damage. Nitrogen-fixing bacterium Gluconacetobacter diazotrophicus has been proposed as a possible biofertilizer for monocot crop production. However, the colonization of G. diazotrophicus in most monocot crops is limited and deep understanding of the response of host plants to G. diazotrophicus colonization is still lacking. In this study, the molecular response of the monocot plant model Brachypodium distachyon was studied during G. diazotrophicus root colonization. The gene expression profiles of B. distachyon root tissues colonized by G. diazotrophicus were generated via next-generation RNA sequencing, and investigated through gene ontology and metabolic pathway analysis. The RNA sequencing results indicated that Brachypodium is actively involved in G. diazotrophicus colonization via cell wall synthesis. Jasmonic acid, ethylene, gibberellin biosynthesis. nitrogen assimilation, and primary and secondary metabolite pathways are also modulated to accommodate and control the extent of G. diazotrophicus colonization. Cellulose synthesis is significantly downregulated during colonization. The loss of function mutant for Brachypodium cellulose synthase 8 (BdCESA8) showed decreased cellulose content in xylem and increased resistance to G. diazotrophicus colonization. This result suggested that the cellulose synthesis of the secondary cell wall is involved in G. diazotrophicus colonization. The results of this study provide insights for future research in regard to gene manipulation for efficient colonization of nitrogen-fixing bacteria in Brachypodium and monocot crops. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Agrobacterium tumefaciens growing in liquid attaches to the surface of tomato and Arabidopsis thaliana roots, forming a biofilm. The bacteria also colonize roots grown in sterile quartz sand. Attachment, root colonization, and biofilm formation all were markedly reduced in celA and chvB mutants, deficient in production of cellulose and cyclic β-(1,2)-D-glucans, respectively. We have identified two genes (celG and celI) in which mutations result in the overproduction of cellulose as judged by chemical fractionation and methylation analysis. Wild-type and chvB mutant strains carrying a cDNA clone of a cellulose synthase gene from the marine urochordate Ciona savignyi also overproduced cellulose. The overproduction in a wild-type strain resulted in increased biofilm formation on roots, as evaluated by light microscopy, and levels of root colonization intermediate between those of cellulose-minus mutants and the wild type. Overproduction of cellulose by a nonattaching chvB mutant restored biofilm formation and bacterial attachment in microscopic and viable cell count assays and partially restored root colonization. Although attachment to plant surfaces was restored, overproduction of cellulose did not restore virulence in the chvB mutant strain, suggesting that simple bacterial binding to plant surfaces is not sufficient for pathogenesis.

In this study, newly developed polymeric composites based on poly(3-hydroxybutyrate) (PHB), cellulose fibres (CF) and plasticizer (bis[2-(2-butoxyethoxy)ethyl] adipate) (DBEEA) were subjected both to microorganism�s action in controlled conditions, and to soil�s action by sample burial. The weight loss of PHB based composites at 60 days exposure were determined in comparison with neat PHB. The weight variation for PHB based composites buried in a natural soil at 45 days and 90 days was monitored. DSC-differential scanning calorimetry and SEM -scanning electron microscopy analysis were performed on the tested composites after 90 days of soil burial. It was found that the introduction of cellulose fibres into PHB leads to the increase of biodegradability of composites.

Numerous salmonellosis outbreaks have been associated with vegetables, in particular sprouted seed. Thin aggregative fimbriae (Tafi), a component of the extracellular matrix responsible for multicellular behavior, are important for Salmonella enterica attachment and colonization of plants. Here, we demonstrate that the other surface polymers composing the extracellular matrix, cellulose, and O-antigen capsule also play a role in colonization of plants. Mutations in bacterial cellulose synthesis (bcsA) and O-antigen capsule assembly and translocation (yihO) reduced the ability to attach to and colonize alfalfa sprouts. A colanic acid mutant was unaffected in plant attachment or colonization. Tafi, cellulose synthesis, and O-antigen capsule, all of which contribute to attachment and colonization of plants, are regulated by AgfD, suggesting that AgfD is a key regulator for survival outside of hosts of Salmonella spp. The cellulose biosynthesis regulator adrA mutant was not affected in the ability to attach to or colonize plants; however, promoter probe assays revealed expression by cells attached to alfalfa sprouts. Furthermore, quantitative reverse-transcriptase polymerase chain reaction revealed differential expression of agfD and adrA between planktonic and plant-attached cells. In addition, there was no correlation among mutants between biofilm formation in culture and attachment to plants. Outside of animal hosts, S. enterica appears to rely on an arsenal of adhesins to persist on plants, which can act as vectors and perpetuate public health concerns.

This paper provides details on the infection processes at the ultrastructural level in Pinus resinosa Ait. seedlings during early stages of colonization by Gremmeniella abietina (Lagerb.) Morelot. Different gold-conjugated enzymes and antibodies were used to cytochemically localize cellulose, pectin, fungal laccase, and the pathogen cells in host tissues. Gremmeniella abietina penetrated into the host through stomata of the short shoot bracts and sparsely colonized both intercellular and intracellular areas of the bract tissues. The colonizing hyphae usually had a thick wall surrounded by an extracellular sheath composed of fibrillar material. Microhyphaelike cells were observed as having penetrated host cell walls. The fungal cells (except the extracellular sheath), even when embedded in cellulosic or pectic material of host tissues, did not appear to contain cellulose or pectin. We suggest that G. abietina is able to degrade cellulose and pectin and that phenoloxidases secreted by the pathogen could be involved in host cell wall degradation. The results indicate that the extracellular sheath of G. abietina is implicated in host–pathogen interactions such as attachment of hyphae to the host surface and cell wall degradation during colonization of host tissues. Key words: Gremmeniella, Pinus, infection processes, cell wall degradation, extracellular fungal sheath, gold labelling.

Infection and subsequent nodulation of legume host plants by the root nodule symbiote Rhizobium leguminosarum usually require attachment of the bacteria to root-hair tips. Bacterial cellulose fibrils have been shown to be involved in this attachment process but appeared not to be essential for successful nodulation. Detailed analysis of Vicia sativa root-hair infection by wild-type Rhizobium leguminosarum RBL5523 and its cellulose fibril-deficient celE mutant showed that wild-type bacteria infected elongated growing root hairs, whereas cellulose-deficient bacteria infected young emerging root hairs. Exopolysaccharide-deficient strains that retained the ability to produce cellulose fibrils could also infect elongated root hairs but infection thread colonization was defective. Cellulose-mediated agglutination of these bacteria in the root-hair curl appeared to prevent entry into the induced infection thread. Infection experiments with V. sativa roots and an extracellular polysaccharide (EPS)- and cellulose-deficient double mutant showed that cellulose-mediated agglutination of the EPS-deficient bacteria in the infection thread was now abolished and that infection thread colonization was partially restored. Interestingly, in this case, infection threads were initiated in root hairs that originated from the cortical cell layers of the root and not in epidermal root hairs. Apparently, surface polysaccharides of R. leguminosarum, such as cellulose fibrils, are determining factors for infection of different developmental stages of root hairs.

Biocontrol activity of Candida saitoana and its interaction with Botrytis cinerea in apple wounds were investigated. When cultured together, yeast attached to Botrytis sp. hyphal walls. In wounded apple tissue, C. saitoana restricted the proliferation of B. cinerea, multiplied, and suppressed disease caused by either B. cinerea or Penicillium expansum. In inoculated apple tissue without the yeast, fungal colonization caused an extensive degradation of host walls and altered cellulose labeling patterns. Hyphae in close proximity to the antagonistic yeast exhibited severe cytological injury, such as cell wall swelling and protoplasm degeneration. Colonization of the wound site by C. saitoana did not cause degradation of host cell walls. Host cell walls in close contact with C. saitoana cells and B. cinerea hyphae were well preserved and displayed an intense and regular cellulose labeling pattern. In addition to restricting fungal colonization, C. saitoana induced the formation of structural defense responses in apple tissue. The ability of C. saitoana to prevent the necrotrophic growth of the pathogen and stimulate structural defense responses may be the basis of its biocontrol activity.

ABSTRACTThe importance of bacterial adherence has been acknowledged in microbial lignocellulose conversion studies; however, few reports have described the function and structure of biofilms supported by cellulosic substrates. We investigated the organization, dynamic formation, and carbon flow associated with biofilms of the obligately anaerobic cellulolytic bacteriumClostridium thermocellum27405. Using noninvasive,in situfluorescence imaging, we showed biofilms capable of near complete substrate conversion with a characteristic monolayered cell structure without an extracellular polymeric matrix typically seen in biofilms. Cell division at the interface and terminal endospores appeared throughout all stages of biofilm growth. Using continuous-flow reactors with a rate of dilution (2 h−1) 12-fold higher than the bacterium's maximum growth rate, we compared biofilm activity under low (44 g/liter) and high (202 g/liter) initial cellulose loading. The average hydrolysis rate was over 3-fold higher in the latter case, while the proportions of oligomeric cellulose hydrolysis products lost from the biofilm were 13.7% and 29.1% of the total substrate carbon hydrolyzed, respectively. Fermentative catabolism was comparable between the two cellulose loadings, with ca. 4% of metabolized sugar carbon being utilized for cell production, while 75.4% and 66.7% of the two cellulose loadings, respectively, were converted to primary carbon metabolites (ethanol, acetic acid, lactic acid, carbon dioxide). However, there was a notable difference in the ethanol-to-acetic acid ratio (g/g), measured to be 0.91 for the low cellulose loading and 0.41 for the high cellulose loading. The results suggest that substrate availability for cell attachment rather than biofilm colonization rates govern the efficiency of cellulose conversion.

La méthanisation des composés lignocellulosiques présente un fort intérêt en raison de leur haut potentiel énergétique et de leur abondance, notamment dans les ordures ménagères résiduelles. Toutefois, leur complexité de structure et de composition rend ces matériaux difficilement dégradables en conditions anaérobies et l’utilisation de prétraitements est généralement requise afin d’améliorer leurs rendements de biodégradation. Outre l’effet de ces prétraitements sur la biodégradation de ces composés, la colonisation des lignocelluloses par les micro-organismes cellulolytiques est une étape clé pour l’efficacité de sa dégradation. Dans ce cadre, le travail de thèse a pour objectifs de mieux comprendre le déterminisme de la colonisation de déchets, d’établir le lien entre la colonisation des déchets lignocellulosiques et l'efficacité de leur dégradation et enfin de caractériser plus finement les mécanismes et interactions mises en jeu au sein de la biomasse. Afin de répondre à ces questions, une approche transversale a été développée, combinant des modèles de cultures de souches pures et des systèmes de méthanisation en laboratoire par des communautés complexes. Des approches intégratives ont été appliquées à l’étude de ces systèmes, couplant des analyses haut-débit (métagénomique/(méta)protéomique), un suivi physico-chimique de la biodégradation et des caractérisations physico-chimiques des composés lignocellulosiques étudiés. L’ensemble des résultats met en évidence le rôle des propriétés chimiques, micro-et macro¬structurales des composés lignocellulosiques dans leur récalcitrance, leur performances de dégradation et la réponse du compartiment microbien. La réalisation de la première étude de protéomique totale et quantitative sur la souche pure cellulolytique Clostridium cellulolyticum, modèle des Clostridia cellulolytiques mésophiles, a permis de mettre en évidence que la vitesse maximale de biodégradation du mouchoir en papier est supérieure à celle du coton et que cette dégradation est associée à un profil métabolique particulier, à une colonisation plus rapide et plus étendue et à une modulation quantitative du système cellulasique. D’autre part, une étude sur un système plus réaliste pour l’étude de la méthanisation des déchets lignocellulosiques a confirmé la bonne concordance entre ce système et le système modèle utilisé et a également permis de mettre en évidence les effets substrats sur la structure des communautés microbienne avec la dominance de la classe Bacteroidia en présence de mouchoir en papier et la forte proportion de la classe Spirochaetes en présence de coton. Enfin l’étude des effets de broyages très fins de la paille de blé et du carton plat ont mis en évidence les limites de ces prétraitements sur les performances de leur dégradation, avec l’effet positif modéré du broyage fin de la paille. Ils ont également montré la sensibilité des communautés microbiennes aux changements de surface du substrat, qui se manifeste par l'émergence de communautés parfois différentes en fonction du prétraitement mécanique appliqué. En conclusion, ce travail a permis de traiter sous un angle nouveau les questions liées à la récalcitrance des déchets lignocellulosiques en abordant à la fois les aspects structuraux, écologiques et fonctionnels. Ces résultats alimentent le corps de connaissances fondamentales sur les bioprocédés. Ils confirment que les matériaux lignocellulosiques sont particuliers parmi les déchets non-dangereux et qu’une exploitation plus large de leur potentiel énergétique nécessiterait la mise en œuvre de procédés spécifiquement adaptés<br>Lignocellulosic materials have a high energy potential and are abundant, especially in municipal solid waste and their methanization is a promising waste-to-energy bioprocess. However, owing to their highly complex and heterogeneous structure, they are recalcitrant to anaerobic conditions and the use of pre-treatments is usually required to improve their biodegradation yields. Besides, lignocellulose colonization by cellulolytic microorganisms is a key step for an efficient biodegradation. In this context, the PhD work aimed to better understand the factors affecting waste colonization, to establish the link between lignocellulosic waste colonization and its biodegradation efficiency and to characterize more precisely the mechanisms and interactions within the biomass. A transversal approach was developed, combining cultures of model pure strains and lab-scale methanization microcosms with a complex biomass. Integrated approaches were applied to these studies, combining high-throughput analyses (metagenomics/(meta) proteomics), physico-chemical monitoring of bioconversion and finally physico-chemical characterization of substrates. The main results highlight the important role of lignocellulosic materials chemical and micro-and macro -structural features for their recalcitrance, their biodegradation efficiency and the response of the microbial compartment. The first global quantitative proteomic study on the cellulolytic model Clostridium cellulolyticum was conducted. Results showed an increased biodegradation rate of the facial tissue compared to cotton. This enhanced biodegradation was associated to a particular metabolic profile, a faster and more extensive colonization and finally a quantitative modulation of the cellulasic system. On the other hand, study of lignocellulosic waste methanization confirmed the good agreement between this more realistic system and the above-described model system. It also provided new information about the effects of substrate on microbial community structure. Noticeably, Bacteroidia members predominated in the presence of tissue and a high proportion of Spirochaetes members was observed in the presence of cotton. Finally, study of the effects of wheat straw and cardboard dry grinding revealed the limitations of these pretreatments on biodegradation efficiency. Main key points were a moderate positive effect of wheat straw fine grinding, and the sensitivity of the microbial communities to substrate surface characteristics, as evidenced by the emergence of different microbial communities according to the applied mechanical pretreatment. In conclusion, this work brings new perspectives to the study of lignocellulosic waste recalcitrance by addressing both the structural, functional and ecological aspects. These results contribute to the core fundamental knowledge on bioprocesses. They confirm that the lignocellulosic materials are specific among non-hazardous waste and require the implementation of adapted specific processes

Les procédés biotechnologiques mettant en œuvre les micro-organismes ou les cellules en général ont souvent recours à l'observation microscopique pour acquérir des informations concernant la morphologie ou le nombre de cellules. C'est notamment le cas des micro-organismes filamenteux dont la morphologie est très complexe et des cellules animales qui croissent sur des microporteurs. Un moyen sur de quantifier les observations microscopiques est l'analyse quantitative d'images. Nous avons utilisé l'analyse d'images pour caractériser les différentes populations morphologiques de micro-organismes filamenteux. Un critère permettant de déterminer leur degré d'enchevêtrement a été mis au point. Deux grandes familles morphologiques sont reconnues: les pelotes et les mycelia libres appelés non-pelotes. Une classification basée sur le critère d'enchevêtrement des non-pelotes permet d'obtenir trois classes d'individus: les filaments, les entrecroisements et les enchevêtrements. Nous avons réalisé le suivi de l'évolution des différentes formes citées ci-dessus au cours de fermentations de streptomyces ambofaciens produisant ou non un antibiotique: la spiramycine. Nous avons remarqué que la phase de production de ce produit est contemporaine de modifications morphologiques certaines. D'autre part, l'analyse d'images a permis de quantifier le processus de colonisation de microporteurs servant de support à la croissance de cellules animales en culture en réacteurs agités. Nous avons montré que les microporteurs de taille moyenne (163 et 183 um) étaient les plus représentatifs de la croissance globale. L'analyse d'images est un outil performant qui a montré ses possibilités dans ces deux applications. Des extensions pour son application en ligne sont proposées

Scientists have been knocking the wood to ascertain the symbiotic relationships of tiny living creatures, that is, microorganisms with other beings such as plants, animals, insects, and humans. The concept of “symbiosis” got its existence in 1879, which means “living together.” Microorganisms show a great deal of diverse interactions such as commensalism (moochers), mutualism (both benefitted), and parasitism (one benefitted and other unharmed) with other living beings and mutualism being the most common of all, thus forming a range of antagonistic to cooperative symbiotic relationships. These tiny creatures interact with plants by forming lichens (fungi and algae), mycorrhizae (plants and roots of higher plants), root noodles (Rhizobium) and acting as keyworkers in plant’s rhizosphere promoting growth and development. Microbial community also extends itself to kingdom Animalia establishing relationships with phylum Mammalia including humans, animals, and the most abundant species of phylum Arthropoda, that is, insects such as termites, which have colonization of bacteria in gut to digest wood cellulose. Scientists have discovered that most studied organisms—mussels found in deep-sea hydrothermal vents too live in a mutualistic association whereby bacteria get protection and mussels get nutrition as bacteria use chemicals from hydrothermal fluid producing organic compounds.

Clavibacter michiganensis subsp. michiganensis (Cmm), the causal agent of bacterial wilt and canker of tomato, is the most destructive bacterial disease of tomato causing substantial economic losses in Israel, the U.S.A. and worldwide. The molecular strategies that allow Cmm, a Gram-positive bacterium, to develop a successful infection in tomato plants are largely unknown. The goal of the project was to elucidate the molecular interactions between Cmmand tomato. The first objective was to analyze gene expression profiles of susceptible tomato plants infected with pathogenic and endophytic Cmmstrains. Microarray analysis identified 122 genes that were differentially expressed during early stages of infection. Cmm activated typical basal defense responses in the host including induction of defense-related genes, production of scavenging of free oxygen radicals, enhanced protein turnover and hormone synthesis. Proteomic investigation of the Cmm-tomato interaction was performed with Multi-Dimensional Protein Identification Technology (MudPIT) and mass spectroscopy. A wide range of enzymes secreted by Cmm382, including cell-wall degrading enzymes and a large group of serine proteases from different families were identified in the xylem sap of infected tomato. Based on proteomic results, the expression pattern of selected bacterial virulence genes and plant defense genes were examined by qRT-PCR. Expression of the plasmid-borne cellulase (celA), serine protease (pat-1) and serine proteases residing on the chp/tomA pathogenicity island (chpCandppaA), were significantly induced within 96 hr after inoculation. Transcription of chromosomal genes involved in cell wall degradation (i.e., pelA1, celB, xysA and xysB) was also induced in early infection stages. The second objective was to identify by VIGS technology host genes affecting Cmm multiplication and appearance of disease symptoms in plant. VIGS screening showed that out of 160 tomato genes, which could be involved in defense-related signaling, suppression of 14 genes led to increase host susceptibility. Noteworthy are the genes Snakin-2 (inhibitor of Cmm growth) and extensin-like protein (ELP) involved in cell wall fortification. To further test the significance of Snakin -2 and ELP in resistance towards Cmm, transgenic tomato plants over-expressing the two genes were generated. These plants showed partial resistance to Cmm resulting in a significant delay of the wilt symptoms and reduction in size of canker lesion compared to control. Furthermore, colonization of the transgenic plants was significantly lower. The third objective was to assess the involvement of ethylene (ET), jasmonate (JA) and salicylic acid (SA) in Cmm infection. Microarray and proteomic studies showed the induction of enzymes involved in ET and JA biosynthesis. Cmm promoted ET production 8 days after inoculation and SIACO, a key enzyme of ET biosynthesis, was upregulated. Inoculation of the tomato mutants Never ripe (Nr) impaired in ET perception and transgenic plants with reduced ET synthesis significantly delayed wilt symptoms as compared to the wild-type plants. The retarded wilting in Nr plants was shown to be a specific effect of ET insensitivity and was not due to altered expression of defense related genes, reduced bacterial population or decrease in ethylene biosynthesis . In contrast, infection of various tomato mutants impaired in JA biosynthesis (e.g., def1, acx1) and JA insensitive mutant (jai1) yielded unequivocal results. The fourth objective was to determine the role of cell wall degrading enzymes produced by Cmm in xylem colonization and symptoms development. A significance increase (2 to 7 fold) in expression of cellulases (CelA, CelB), pectate lyases (PelA1, PelA2), polygalacturonase and xylanases (XylA, XylB) was detected by qRT-PCR and by proteomic analysis of the xylem sap. However, with the exception of CelA, whose inactivation led to reduced wilt symptoms, inactivation of any of the other cell wall degrading enzymes did not lead to reduced virulence. Results achieved emphasized the complexity involved in Cmm-tomato interactions. Nevertheless they provide the basis for additional research which will unravel the mechanism of Cmm pathogenicity and formulating disease control measures.