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  • Bacterial chemotaxis coupling protein: Structure, function and diversity
    Microbiol. Res. (IF 2.777) Pub Date : 2018-11-06
    Zhiwei Huang, Xiaoyue Pan, Nan Xu, Minliang Guo

    In most signal transduction systems, coupling or scaffold proteins establish crucial connections between receptors and histidine kinases. These connections are important for signal transduction. The bacterial chemotaxis system is a canonical signal transduction system that relies on coupling proteins. The coupling proteins in the chemotaxis system have two architectures: CheW or CheV. In a typical chemotaxis signal transduction system, two CheW coupling protein molecules bridge a histidine kinase CheA dimer and two chemoreceptor (also called as methyl-accepting chemotaxis protein, MCP) trimers of dimers to form a core signaling complex and couple CheA activity to chemoreceptor control. Although CheW is a small cytoplasmic protein, it plays multiple functions in chemotaxis. CheW also builds connections between core signaling complexes, which leads to the formation of large chemosensory arrays that are responsible for collecting and amplifying signals from various chemoreceptors. Another coupling protein, CheV, shares a largely redundant ability with CheW; however, the function of CheV is not identical to that of CheW in chemotaxis. In this article, we summarize the molecular mechanism of chemotaxis in Escherichia coli and review the recent advances in the structural details and functions of CheW and CheV. Furthermore, we focus on the diversity of coupling proteins and discuss the relationship among multiple coupling proteins in one organism.

  • Physiological response of Lactuca sativa exposed to 2-nonanone emitted by Bacillus sp. BCT9
    Microbiol. Res. (IF 2.777) Pub Date : 2018-11-03
    Paola Fincheira, Andrés Quiroz

    Volatile organic compounds (VOCs) released from bacterial species have been reported as plant growth inducers. In this sense, Lactuca sativa was used as model vegetable to prospect the effects of 2-nonanone released by Bacillus sp. BCT9 at cellular and organ structure level, so we present preliminary results about the physiological effects. In this study, 2-day-old L. sativa were exposed to 2-nonanone for 10 days under two delivery systems: 1) 2-nonanone (abrupt delivery) and 2) 2-nonanone + lanolin (controlled delivery). The X-ray elemental microanalysis, scanning electron and confocal laser microscopies techniques were used to evaluate physiological changes “in vivo” conditions. The results indicated that 2-nonanone increased root and shoot length independently of 2-nonanone delivery system after 7 days of exposition. Additionally, 2-nonanone elicited the increase of anthocyanin and not affects chlorophyll content and electrolyte leakage percentage. The abrupt delivery elicited the increase of both length and density of root hair without causing changes in size of cell epidermis, while controlled delivery induced stomatal opening. Besides, 2-nonanone exposition did not modify the composition and distribution of carbon, nitrogen, phosphorus, potassium, and chlorine in the surface of plant tissue. The results suggested that 2-nonanone acts as a bacterial signal molecule to elicit changes related to root development without damaging the external morphology while epidermal cells at leaf level are not affected, suggesting that 2-nonanone can be an important tool to apply to vegetables.

  • The Potential of Bacilli Rhizobacteria for Sustainable Crop Production and Environmental Sustainability
    Microbiol. Res. (IF 2.777) Pub Date : 2018-11-02
    B.N. Aloo, B.A. Makumba, E.R. Mbega

    Conventional agricultural practices often rely on synthetic fertilizers and pesticides which have immense and adverse effects on humans, animals and environments. To minimize these effects, scientists world over are now deeply engaged in finding alternative approached for crop production which are less dependent on chemical inputs. One such approach is the use of rhizospheric bacteria as vital components of soil fertility and plant growth promotion (PGP) through their direct and indirect processes in plant rhizospheres. Among the most studied rhizobacteria are the Bacilli, particularly for production of antibiotics, enzymes and siderophores all of which are important aspects of PGP. Despite this, little information is available especially on their potentiality in crop production and their direct application only involves a few species, leaving a majority of these important rhizobacteria exploited. This paper gives an overview of the unique properties of Bacilli rhizobacteria as well as their different PGP mechanisms that if mined can lead to their successful application and agricultural sustainability. It further points out the missing aspects with regards to these important rhizobacteria that should be considered for future research. This information will be useful in analyzing the PGP abilities of Bacilli rhizobacteria with an aim of fully mining their potential for crop production and environmental sustainability.

  • Current situation of biofuel production and its enhancement by CRISPR/Cas9-mediated genome engineering of microbial cells
    Microbiol. Res. (IF 2.777) Pub Date : 2018-11-02
    Muhammad Rizwan Javed, Muhammad Noman, Muhammad Shahid, Temoor Ahmed, Mohsin Khurshid, Muhammad Hamid Rashid, Muhammad Ismail, Maria Sadaf, Fahad Khan

    Geopolitical and economic factors have motivated the scientific community to utilize renewable energy resources. In addition to the modifications in major steps and processes of biofuel production, manipulation of microbial genome engineering tools is essential in order to find sustainable solution of continuous depletion of fossil-fuels. Recently, the CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 (CRISPR-associated nuclease 9), a prokaryotic molecular immunity system, has emerged as a novel technology for targeted genomic engineering. This genetic machinery seems to be a groundbreaking discovery to engineer the microbial genomes for desired traits such as enhancing the biofuel tolerance, inhibitor tolerance and thermotolerance as well as modifying the cellulases and hemicelluloses enzymes. In this review, a summary of different generations of biofuels, integrated processes of bioconversion of raw materials into biofuels and role of microbes in biofuel production has been presented. However, the ultimate focus of the review is on major discoveries of CRISPR/Cas9-mediated genome editing in microorganisms and exploitation of these discoveries for enhanced biofuel production.

  • Development of low-cost formulations of plant growth-promoting bacteria to be used as inoculants in beneficial agricultural technologies
    Microbiol. Res. (IF 2.777) Pub Date : 2018-11-02
    Constanza Belén Lobo, María Silvina Juárez Tomás, Emilce Viruel, Marcela Alejandra Ferrero, María Ester Lucca
  • Far-reaching cellular consequences of tat deletion in Escherichia coli revealed by comprehensive proteome analyses
    Microbiol. Res. (IF 2.777) Pub Date : 2018-11-01
    Katarzyna M. Dolata, Isabel Guerrero Montero, Wayne Miller, Susanne Sievers, Thomas Sura, Christian Wolff, Rabea Schlüter, Katharina Riedel, Colin Robinson
  • Transcriptome analysis of hopanoid deficient mutant of Rhodopseuodomonas palustris TIE-1
    Microbiol. Res. (IF 2.777) Pub Date : 2018-10-29
    Tushar D. Lodha, Indu B., Sasikala Ch., Ramana Ch.V.

    All three domains of life have an ordered plasma membrane which is pivotal in the selective fitness of primitive life. Like cholesterol in eukaryotes, hopanoids are important in bacteria to modulate membrane order. Hopanoids are pentacyclic triterpenoid lipids biosynthesised in many eubacteria, few ferns and lichens. Hopanoid modulates outer membrane order and hopanoid deficiency results in the weakened structural integrity of the membrane which may intern affect the other structures within or spanning the cell envelope and contributing to various membrane functions. Hence, to decipher the role of hopanoid, genome-wide transcriptome of wild-type and Δshc mutant of Rhodopseudomonas palustris TIE-1 was studied which indicated 299 genes were upregulated and 306 genes were downregulated in hopanoid deficient mutant, representing ~11.5% of the genome. Thirty-eight genes involved in chemotaxis, response to stimuli and signal transduction were differentially regulated and impaired motility in hopanoid deficient mutant showed that hopanoid plays a crucial role in chemotaxis. The docking study demonstrated that diguanylate cyclase which catalyses the synthesis of secondary messenger exhibited the capability to interact with hopanoids and might be confederating in chemotaxis and signal transduction. Seventy-four genes involved in membrane transport were differentially expressed and cell assays have explicit that the multidrug transport is compromised in Δshc mutant. Membrane transport is reliant on hopanoids which may explain the basis for previous observations linking hopanoids to antibiotic resistance. Disturbing the membrane order by targeting lipid synthesis can be a possible novel approach in developing new antimicrobials and hopanoid biosynthesis could be a potential target.

  • The histidine kinase slnCl1 of Colletotrichum lindemuthianum as a pathogenicity factor against Phaseolus vulgaris L
    Microbiol. Res. (IF 2.777) Pub Date : 2018-10-28
    Guilherme Bicalho Nogueira, Leandro Vieira dos Santos, Casley Borges de Queiroz, Thamy Lívia Ribeiro Corrêa, Renato Pedrozo Menicucci, Denise Mara Soares Bazzolli, Elza Fernandes de Araújo, Marisa Vieira de Queiroz

    Colletotrichum lindemuthianum, the causal agent of anthracnose, is responsible for significant damage in the common bean (Phaseolus vulgaris L.). Unraveling the genetic mechanisms involved in the plant/pathogen interaction is a powerful approach for devising efficient methods to control this disease. In the present study, we employed the Restriction Enzyme-Mediated Integration (REMI) methodology to identify the gene slnCl1, encoding a histidine kinase protein, as involved in pathogenicity. The mutant strain, MutCl1, generated by REMI, showed an insertion in the slnCl1 gene, deficiency of the production and melanization of appressoria, as well as the absence of pathogenicity on bean leaves when compared with the wild-type strain. The slnCl1 gene encodes a histidine kinase class IV called SlnCl1 showing identity of 97% and 83% with histidine kinases from Colletotrichum orbiculare and Colletotrichum gloesporioides, respectively. RNA interference was used for silencing the histidine kinase gene and confirm slnCl1 as a pathogenicity factor. Furthermore, we identified four major genes involved in the RNA interference-mediated gene silencing in Colletotrichum spp. and demonstrated the functionality of this process in C. lindemuthianum. Silencing of the EGFP reporter gene and slnCl1 were demonstrated using qPCR. This work reports for the first time the isolation and characterization of a HK in C. lindemuthianum and the occurrence of gene silencing mediated by RNA interference in this organism, demonstrating its potential use in the functional characterization of pathogenicity genes.

  • Plant-microbe interactions endorse growth by uplifting microbial community structure of Bacopa monnieri rhizosphere under nematode stress
    Microbiol. Res. (IF 2.777) Pub Date : 2018-10-26
    Rupali Gupta, Akanksha Singh, Madhumita Srivastava, Karuna Shanker, Rakesh Pandey

    The modification of rhizosphere microbial diversity and ecological processes are of rising interest as shifting in microbial community structure impacts the mutual role of host–microbe interactions. Nevertheless, the connection between host-microbial community diversity, their function under biotic stress in addition to their impact on plant performances is poorly understood. The study was designed with the aim to analyze the tripartite interactions among Chitiniphilus sp., Streptomyces sp. and their combination with indigenous rhizospheric microbial population of Bacopa monnieri for enhancing the plant growth and bacoside A content under Meloidogyne incognita stress. Overall, plants treated with the microbial combination recorded enhanced growth as illustrated by significantly higher biomass (2.0 fold), nitrogen uptake (1.8 fold) and bacoside A content (1.3 fold) along with biocontrol efficacy (58.5 %) under nematode infected field. The denaturing gradient gel electrophoresis (DGGE) fingerprints of 16S-rDNA revealed that microbial inoculations are major initiators of bacterial community structure in the plant rhizosphere. Additionally, the plants treated with microbial combination showed maximum diversity viz., Shannon’s (3.29), Margalef’s (4.21), and Simpson’s (0.96) indices. Likewise the metabolic profiling data also showed a significant variation among the diversity and evenness indices upon microbial application on the native microflora. We surmise that the application of beneficial microbes in combinational mode not only helped in improving the microbial community structure but also successfully enhanced plant and soil health under biotic stress.

  • Deletion of the epigenetic regulator GcnE in Aspergillus niger FGSC A1279 activates the production of multiple polyketide metabolites
    Microbiol. Res. (IF 2.777) Pub Date : 2018-10-15
    Bin Wang, Xuejie Li, Dou Yu, Xiaoyi Chen, Jioji Tabudravu, Hai Deng, Pan Li

    Epigenetic modification is an important regulatory mechanism in the biosynthesis of secondary metabolites in Aspergillus species, which have been considered to be the treasure trove of new bioactive secondary metabolites. In this study, we reported that deletion of the epigenetic regulator gcnE, a histone acetyltransferase in the SAGA/ADA complex, resulted in the production of 12 polyketide secondary metabolites in A. niger FGSC A1279, which was previously not known to produce toxins or secondary metabolites. Chemical workup and structural elucidation by 1D/2D NMR and high resolution electrospray ionization mass (HR-ESIMS) yielded the novel compound nigerpyrone (1) and five known compounds: carbonarone A (2), pestalamide A (3), funalenone (4), aurasperone E (5), and aurasperone A (6). Based on chemical information and the literature, the biosynthetic gene clusters of funalenone (4), aurasperone E (5), and aurasperone A (6) were located on chromosomes of A. niger FGSC A1279. This study found that inactivation of GcnE activated the production of secondary metabolites in A. niger. The biosynthetic pathway for nigerpyrone and its derivatives was identified and characterized via gene knockout and complementation experiments. A biosynthetic model of this group of pyran-based fungal metabolites was proposed.

  • Partner-triggered proteome changes in the cell wall of Bacillus sonorensis and roots of groundnut benefit each other
    Microbiol. Res. (IF 2.777) Pub Date : 2018-10-09
    Sravani Ankati, T. Swaroopa Rani, Appa Rao Podile

    Plant growth promoting rhizobacteria (PGPR) promote plant growth and activate defense response against phytopathogens. At the subcellular level plant-PGPR interaction is less understood, which would be essential for future improvement(s) of PGPR formulations. In a rigorous screening process, that also involved efficient PGPR strains, Bacillus sonorensis RS4 was selected to study partner-triggered interactions. The potential of B. sonorensis RS4 to improve growth of groundnut, efficiency to colonize roots, and influence on root topology was assessed. Twenty four cell wall proteins of B. sonorensis RS4 [in presence of groundnut root exudates (REs)], and 22 groundnut root proteins (in RS4-bacterized plants) were differentially expressed. The alterations in cell wall proteins of B. sonorensis RS4 were primarily related to the amino acids synthesis, chemotaxis, antioxidant-metabolism, carbohydrate metabolism, transporters, and antibiosis-related secondary metabolites. Root proteins that were differentially expressed during the interaction may be involved in plant growth, defense responses, and in transportation. The changes in B. sonorensis RS4 cell wall proteome and groundnut root proteome, suggest that at least a part of the proteome changes triggered by each of the partners appear to play a significant role in helping each other akin to symbiosis.

  • Antimicrobial, plant growth-promoting and genomic properties of the peanut endophyte Bacillus velezensis LDO2
    Microbiol. Res. (IF 2.777) Pub Date : 2018-10-09
    Liang Chen, Hao Shi, Junying Heng, Dianxuan Wang, Ke Bian

    Peanut suffer from a number of fungal and bacterial pathogens, while plant endophytes were considered excellent candidates as biocontrol agents. In this study, the peanut endophytic bacterium LDO2 was evaluated for the potential of peanut pathogens inhibition and growth-promotion, and the genetic mechanisms were explored by genome mining. Strain LDO2 significantly inhibited the growth of peanut pathogenic fungi and pathogenic bacteria, and specifically, it showed pronounced inhibition on mycelia growth of Aspergillus flavus mycelia and caused mycelial deformity. Gene clusters responsible for antifungal metabolites (fengycin, surfactin, bacilysin) and antibacterial metabolites (butirosin, bacillaene, difficidin, macrolactin, surfactin, bacilysin) were identified. Strain LDO2 also exhibited several growth-promoting related features including phosphate solubilization, siderophore production and growth promotion of peanut root. Genes associated with plant growth promotion were also identified and analyzed, as well as genes related to secreted proteins. These findings suggested that this peanut endophyte could be a potential biocontrol agent in peanut production and a source of antimicrobial compounds for further exploitation.

  • Recombineering Pseudomonas protegens CHA0: An innovative approach that improves nitrogen fixation with impressive bactericidal potency
    Microbiol. Res. (IF 2.777) Pub Date : 2018-10-06
    Fangnan Yu, Xiaoshu Jing, Xiaochen Li, Hailong Wang, Hanna Chen, Lin Zhong, Jia Yin, Deng Pan, Yulong Yin, Jun Fu, Liqiu Xia, Xiaoying Bian, Qiang Tu, Youming Zhang
  • Environmental conditions shape the biofilm of the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125
    Microbiol. Res. (IF 2.777) Pub Date : 2018-10-06
    Annarita Ricciardelli, Angela Casillo, Alessandro Vergara, Nicole Balasco, Maria Michela Corsaro, Maria Luisa Tutino, Ermenegilda Parrilli

    Biofilms are the most widely distributed and successful microbial modes of life. The capacity of bacteria to colonize surfaces provides stability in the growth environment, allows the capturing of nutrients and affords protection from a range of environmental challenges and stress. Bacteria living in cold environments, like Antarctica, can be found as biofilms, even though the mechanisms of how this lifestyle is related to their environmental adaptation have been poorly investigated. In this paper, the biofilm of Pseudoalteromonas haloplanktis TAC125, one of the model organisms of cold-adapted bacteria, has been characterized in terms of biofilm typology and matrix composition. The characterization was performed on biofilms produced by the bacterium in response to different nutrient abundance and temperatures; in particular, this is the first report describing the structure of a biofilm formed at 0 °C. The results reported demonstrate that PhTAC125 produces biofilms in different amount and endowed with different physico-chemical properties, like hydrophobicity and roughness, by modulating the relative amount of the different macromolecules present in the biofilm matrix. The capability of PhTAC125 to adopt different biofilm structures in response to environment changes appears to be an interesting adaptation strategy and gives the first hints about the biofilm formation in cold environments.

  • Isolation and characterization of endophytes from nodules of Mimosa pudica with biotechnological potential
    Microbiol. Res. (IF 2.777) Pub Date : 2018-10-06
    Ricardo Sánchez-Cruz, Irán Tapia Vázquez, Ramón Alberto Batista-García, Erick Williams Méndez Santiago, María del Rayo Sánchez Carbente, Alfonso Leija, Verónica Lira-Ruan, Gerogina Hernández, Arnoldo Wong-Villarreal, Jorge Luis Folch-Mallol

    Legumes establish symbiotic relationships with different microorganisms, which could function as plant growth promotion microorganisms (PGPM). The finding of new PGPM strains is important to increase plant production avoiding or diminishing the use of industrial fertilizers. Thus, in this work we evaluated the plant growth promotion traits of ten strains isolated from Mimosa pudica root nodules. According to the 16S rDNA sequence, the microorganisms were identified as Enterobacter sp. and Serratia sp.. To the best of our knowledge this is the first report describing and endophytic interaction between Mimosa pudica and Enterobacter sp. These strains have some plant growth promoting traits such as phosphate solubilization, auxin production and cellulase and chitinase activity. Strains identified as Serratia sp. inhibited the growth of the phytopathogenic fungi Fusarium sp., and Alternaria solani and the oomycete Phytophthora capsici. According to their biochemical characteristics, three strains were selected to test their plant growth promoting activity in a medium with an insoluble phosphate source. These bacteria show low specificity for their hosts as endophytes, since they were able to colonize two very different legumes: Phaseolus vulgaris and M. pudica. Seedlings of P. vulgaris were inoculated and grown for fifteen days. Enterobacter sp. NOD1 and NOD10, promoted growth as reflected by an increase in shoot height as well as an increase in the size and emergence of the first two trifolia. We could localize NOD5 as an endophyte in roots in P. vulgaris by transforming the strain with a Green Fluorescent Protein carrying plasmid. Experiments of co-inoculation with different Rhizobium etli strains allowed us to discard that NOD5 can fix nitrogen in the nodules formed by a R. etli Fix- strain. The isolates described in this work show biotechnological potential for plant growth promoting activity and production of indoleacetic acid and siderophores.

  • Helicobacter pylori evasion strategies of the host innate and adaptive immune responses to survive and develop gastrointestinal disease
    Microbiol. Res. (IF 2.777) Pub Date : 2018-10-06
    Ahmad Karkhah, Soheil Ebrahimpour, Maryam Rostamtabar, Veerendra Koppolu, Sorena Darvish, Veneela Krishna Rekha Vasigala, Majid Validi, Hamid Reza Nouri

    Helicobacter pylori (H. pylori) is a bacterial pathogen that resides in more than half of the human population and has co-evolved with humans for more than 58000 years. This bacterium is orally transmitted during childhood and is a key cause of chronic gastritis, peptic ulcers and two malignant cancers including MALT (mucosa-associated lymphoid tissue) lymphoma and adenocarcinoma. Despite the strong innate and adaptive immune responses, H. pylori has a long-term survival in the gastric mucosa. In addition to the virulence factors, survival of H. pylori is strongly influenced by the ability of bacteria to escape, disrupt and manipulate the host immune system. This bacterium can escape from recognition by innate immune receptors via altering its surface molecules. Moreover, H. pylori subverts adaptive immune response by modulation of effector T cell. In this review, we discuss the immune-pathogenicity of H. pylori by focusing on its ability to manipulate the innate and acquired immune responses to increase its survival in the gastric mucosa, leading up to gastrointestinal disorders. We also highlight the mechanisms that resulted to the persistence of H. pylori in gastric mucosa.

  • Organization of multi-binding to host proteins: the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of Mycoplasma pneumoniae
    Microbiol. Res. (IF 2.777) Pub Date : 2018-09-26
    Juliane Grimmer, Roger Dumke

    Mycoplasma pneumoniae is a frequent cause of community-acquired infections of the human respiratory tract. During the evolutionary adaptation of the bacteria to the host, the genome of the pathogen is strongly reduced resulting in the loss of cell wall, limited metabolic pathways and a relatively small number of virulence factors. As interacting with host proteins, surface-exposed proteins with a primary function in cytosol-located processes of metabolism and regulation such as glycolytic enzymes, heat-shock proteins and chaperones have been considered as contributing to pathogenesis. Among these moonlighting proteins, some members are confirmed as binding to several host components. The glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of M. pneumoniae is a typical example of such multi-binding proteins. To investigate the organization of these interactions, GAPDH was divided into four parts. Recombinant proteins were successfully expressed in Escherichia coli and polyclonal antisera were produced. Binding of full length and parts of GAPDH to human A549 cells was proven. Furthermore, interactions with human plasminogen, vitronectin, fibronectin and fibrinogen were demonstrated for nearly all recombinant GAPDH proteins. In the presence of these proteins, plasminogen can be activated to the protease plasmin. In contrast, the localization on the surface of bacterial cell was confirmed for the C-terminal part of GAPDH only. By using overlapping peptides covering this region, binding of the investigated host components to the sequence 326QLVRVVNYCAKL337 was found. The results of the study suggest a prominent role of the surface-localized C-terminal part of GAPDH in associations with different human proteins indicating its importance for host-pathogen-interactions.

  • Genome sequence and comparative genomics of Rhizobium sp. Td3, a novel plant growth promoting phosphate solubilizing Cajanus cajan symbiont
    Microbiol. Res. (IF 2.777) Pub Date : 2018-09-24
    Bhagya Iyer, Shalini Rajkumar

    Rhizobium sp. Td3 is a Sesbania plant growth promoting, Cajanus cajan nodulating rhizobia. Studying its whole genome was important as it is a potent phosphate solubilizer with constitutive gluconic acid production ability through operation of the periplasmic glucose oxidation pathway even under conditions of catabolite repression. This is in contrast to the other explored phosphate solubilizers. Rhizobial isolates sequenced so far are known to lack components of the direct glucose oxidation pathway and cannot produce gluconic acid on its own. Here, we present the genome sequence of Rhizobium sp. Td3. Genome comprises of a single chromosome of size 5,606,547 bp (5.6 Mb) with no symbiotic plasmid. Rhizobium leguminosarum bv. viciae USDA2370 was the closest whole genome known. 109 genes responsible for diverse plant growth promoting activities like P solubilization, synthesis of acetoin, nitric oxide, indole-3 acetic acid, exopolysaccharide, siderophore and trehalose have been identified. Flagellar proteins, genes encoding antibiotic and metal resistance, enzymes required for combating oxidative stress as well as attachment and colonization in the plant rhizosphere are also present. Availability of genome sequence of such a versatile plant growth promoting agent will help in exploiting all the phyto-beneficial traits of Td3 for its use as a biofertilizer.

  • Cordycepin and Pentostatin Biosynthesis Gene Identified through Transcriptome and Proteomics Analysis of Cordyceps kyushuensis Kob
    Microbiol. Res. (IF 2.777) Pub Date : 2018-09-17
    Xuan Zhao, Guoying Zhang, Caiyi Li, Jianya Ling

    Cordyceps kyushuensis is the only species of cordyceps growing on the larvae of Clanis bilineata Walker, and has been demonstrated that there are lots of pharmacological components including cordycepin. Cordycepin shows lots of pharmacological action but it could be converted to 3’-deoxyinosine by adenosine deaminase in vivo, which weakens the efficiency of cordycepin. That pentostatin, which has been reported to inhibit adenosine deaminase, combining cordycepin could enhance the efficiency of cordycepin in vivo. During transcriptome and proteomics analysis of Cordyceps kyushuensis, a single gene cluster including four genes we named ck1-ck4 which can synthesis both cordycepin and pentostatin has been identified using BLAST. Meanwhile, KEGG, KOG, GO analysis and differentially expressed genes were analyzed in transcriptome and proteomics. This study first sequenced transcriptome and proteomics of C. kyushuensis, and demonstrated that there is a single gene cluster related to biosynthesis of cordycepin and pentostatin, which can be employed to improve the yield of cordycepin and find more functional proteins.

  • Enhanced molecular visualization of root colonization and growth promotion by Bacillus subtilis EA-CB0575 in different growth systems
    Microbiol. Res. (IF 2.777) Pub Date : 2018-09-11
    Luisa F. Posada, J.C. Álvarez, Magally Romero-Tabarez, Luz de-Bashan, Valeska Villegas-Escobar

    Bacillus subtilis EA-CB0575 is a plant growth-promoting bacterium (PGPB) associated with banana and tomato crops. Root colonization is an important trait for PGPB microorganisms and potentiates the bacterial effect related to the mechanisms of plant growth promotion. Therefore, detection of bacterial colonization of roots in different culture systems is important in the study of plant–microorganism interactions. In this study, fluorescent in situ hybridization (FISH) and catalyzed reporter deposition–FISH (CARD–FISH) were evaluated to determine the colonization ability of B. subtilis EA-CB0575 on banana and tomato roots planted on solid and liquid Murashige and Skoog medium (MS(S) and MS(L), respectively) and in soil for tomato plants. Results showed B. subtilis colonization 0–30 days post inoculation for banana and tomato plants in different culture systems with differential distribution of bacterial cells along tomato and banana roots. FISH and CARD–FISH methodologies were both successful in detecting B. subtilis colonies, but CARD–FISH proved to be superior due to its enhanced fluorescence signal. The presence of bacteria correlated with the promotion of plant growth in both plant species, providing clues to relate rhizospheric colonization with improvement in plant growth. FISH and CARD–FISH analysis results suggested the presence of native microbiota on the roots of in vitro banana plants, but not on those of tomato plants.

  • Application of laser microdissection to study phytoplasma site-specific gene expression in the model plant Arabidopsis thaliana
    Microbiol. Res. (IF 2.777) Pub Date : 2018-09-10
    Marika Rossi, Massimo Pesando, Marta Vallino, Luciana Galetto, Cristina Marzachì, Raffaella Balestrini

    Many aspects of plant diseases caused by phytoplasmas are still unknown, as these pathogens are phloem restricted, uncultivable wall-less bacteria and must be studied always in association with their host. Phytoplasma transcripts are strongly underrepresented within host tissues and this poses problems for gene expression analyses. In this study, a procedure was established to infect the model plant Arabidopsis thaliana with the phytoplasma Flavescence dorée, a serious threat to European viticulture. Rates of phytoplasma infective insects and transmission efficiency to A. thaliana as well as pathogen loads were measured in different tissues of infected A. thaliana plants, and modification of phloem cell ultrastructure was observed in infected plant tissues at microscopic level. Moreover, a protocol for the application of laser microdissection to analyze plant and phytoplasma gene expression profiles in the specific colonized tissue was designed. The procedure allowed a good preservation of the plant tissue anatomy. Results showed that the extracted RNA was suitable for qualitative and quantitative RT-PCR, since both plant and pathogen transcripts, either abundant or rare ones, could be detected without any pre-amplification step. The combined use of laser microdissection approach and A. thaliana to study phytoplasmas opens the way to exploit biological, molecular and bioinformatic tools available for the model plant and to elucidate key pathways of the infection mechanisms of these important plant pathogen.

  • The exopolysaccharide gene cluster pea is transcriptionally controlled by RpoS and repressed by AmrZ in Pseudomonas putida KT2440
    Microbiol. Res. (IF 2.777) Pub Date : 2018-09-10
    Huizhong Liu, Huaduo Yan, Yujie Xiao, Hailing Nie, Qiaoyun Huang, Wenli Chen

    In Pseudomonas putida KT2440, the exopolysaccharide Pea is associated with biofilm stability and pellicle formation; however, little is known about its regulatory pathway. In this study, we identified that the gene cluster pea was transcribed from 25 bp upstream of the operon and the stationary phase alternative sigma factor RpoS regulated the transcription of pea. When RpoS was absent, another sigma factor, likely the housekeeping sigma factor RpoD, could also mediate pea transcription but at a low level. The function of Pea polysaccharide was further confirmed to be necessary for full production of biofilm, formation of pellicle and c-di-GMP-dependent wrinkly colony morphology. Additionally, evidences were provided to demonstrate that the transcriptional regulator AmrZ was a negative regulator for pea expression. DNase I footprinting studies verified that AmrZ bound directly to the site overlapping the pea promoter, which might interfere with the binding of RNA polymerase to the promoter and resulted in inhibition of transcription initiation.

  • Blue light exposure and nutrient conditions influence the expression of genes involved in simultaneous hyphal knot formation in Coprinopsis cinerea
    Microbiol. Res. (IF 2.777) Pub Date : 2018-09-08
    Yuichi Sakamoto, Shiho Sato, Miyuki Ito, Yuki Ando, Kiyoshi Nakahori, Hajime Muraguchi
  • Isolation, characterization, and selection of heavy metal-resistant and plant growth-promoting endophytic bacteria from root nodules of Robinia pseudoacacia in a Pb/Zn mining area
    Microbiol. Res. (IF 2.777) Pub Date : 2018-09-07
    Miaochun Fan, Zhenshan Liu, Lijun Nan, Entao Wang, Weimin Chen, Yanbing Lin, Gehong Wei

    Multiple heavy metals (HMs) commonly coexist in mining areas, which highlights the necessity to select multiple HM-resistant plant growth-promoting bacteria for improving phytoremediation efficiency. In this study, we isolated and characterized 82 endophytic bacteria from the root nodules of black locust (Robinia pseudoacacia) grown in a Pb-Zn mining area. There were 80 isolates showing resistance to four HMs, 0.01–18.0 mM/L for Cd, 0.2–40.0 mM/L for Zn, 0.3–2.2 mM/L for Pb, and 0.2–1.4 mM/L for Cu. Indole-3-acetic acid production, siderophore production, and 1-aminocyclopropane-1-carboxylate deaminase activity were detected in 43, 50, and 17 isolates, respectively. Two symbiotic isolates selected with the highest potential for HM resistance and PGP traits, designated Mesorhizobium loti HZ76 and Agrobacterium radiobacter HZ6, were evaluated for promotion of plant growth and metal uptake by R. pseudoacacia seedlings grown in pots containing different levels of Cd, Zn, Pb, or Cu. HZ76 significantly increased plant shoot biomass, while HZ6 did not, compared with non-inoculated controls. The results indicate that inoculation with HZ76 or HZ6 relieved HM stress in the plants, depending on the type and concentration of HM in the treatment. Mesorhizobium loti HZ76 may be a better candidate for application in phytoremediation than A. radiobacter HZ6. The microsymbiosis between HM-resistant rhizobia and R. pseudoacacia is an interesting mutualistic system for phytoremediation in mining areas contaminated with multiple HMs.

  • Biological control of the soft rot bacterium Pectobacterium carotovorum by Bacillus amyloliquefaciens strain Ar10 producing glycolipid-like compounds
    Microbiol. Res. (IF 2.777) Pub Date : 2018-08-31
    Sana Azaiez, Imen Ben Slimene, Ines Karkouch, Rym Essid, Selim Jallouli, Naceur Djebali, Salem Elkahoui, Ferid Limam, Olfa Tabbene

    Four hundred and fifty bacteria were evaluated for antagonistic activity against bacterial soft rot of potato caused by Pectobacterium carotovorum sp strain II16. A strain Ar10 exhibiting potent antagonist activity has been identified as Bacillus amyloliquefaciens on the basis of biochemical and molecular characterization. Cell free supernatant showed a broad spectrum of antibacterial activity against human and phytopathogenic bacteria in the range of 10-60 AU / mL. Incubation of P. carotovorum cells with increasing concentrations of the antibacterial compound showed a killing rate of 94.8 and 96% at MIC and 2xMIC respectively. In addition, the antibacterial agent did not exert haemolytic activity at the active concentration and has been preliminary characterized by TLC and GC-MS as a glycolipid compound. Treatment of potato tubers with strain Ar10 for 72 h significantly reduced the severity of disease symptoms (100 and 85.05% reduction of necrosis deep / area and weight loss respectively). The same levels in disease symptoms severity was also recorded following treatment of potato tubers with cell free supernatant for 1 h. Data suggest that protection from potato soft rot disease may be related to glycolipid production by strain Ar10. The present study affords new alternatives for anti-Pectobacterium carotovorum bioactive compounds against the soft rot disease of potato.

  • Genome-wide transcriptome response of Streptomyces tsukubaensis to N-acetylglucosamine: effect on tacrolimus biosynthesis
    Microbiol. Res. (IF 2.777) Pub Date : 2018-08-31
    María Ordóñez-Robles, Antonio Rodríguez-García, Juan F. Martín

    Chitin is the second most abundant carbohydrate biopolymer present in soils and is utilized by antibiotic–producing Streptomyces species. Its monomer, N-acetylglucosamine (GlcNAc), regulates the developmental program of the model organism Streptomyces coelicolor. GlcNAc blocks differentiation when growing on rich medium whilst it promotes development on poor culture media. However, it is unclear if the same GlcNAc regulatory profile observed in S. coelicolor applies also to other industrially important Streptomyces species. We report here the negative effect of GlcNAc on differentiation and tacrolimus (FK506) production by Streptomyces tsukubaensis NRRL 18488. Using microarrays technology, we found that GlcNAc represses the transcription of fkbN, encoding the main transcriptional activator of the tacrolimus biosynthetic cluster, and of ppt1, encoding a phosphopantheteinyltransferase involved in tacrolimus biosynthesis. On the contrary, GlcNAc stimulated transcription of genes related to amino acid and nucleotide biosynthesis, DNA replication, RNA translation, glycolysis and pyruvate metabolism. The results obtained support those previously reported for S. coelicolor, but some important differences were observed; for example genes involved in GlcNAc transport and metabolism and genes encoding transcriptional regulators such as crr, ptsI, nagE1, nagE2, nagB, chiA, chiJ, ngcE, dasR or atrA are not significantly induced in S. tsukubaensis by GlcNAc addition. Differences in the GlcNAc transport systems in the physiology of S. tsukubaensis and S. coelicolor and/or the different composition of the culture media used are likely to be responsible for the discrepancies observed between these species.

  • Prospects of endophytic fungal entomopathogens as biocontrol and plant growth promoting agents: an insight on how artificial inoculation methods affect endophytic colonization of host plants
    Microbiol. Res. (IF 2.777) Pub Date : 2018-08-31
    Bamisope Steve Bamisile, Chandra Kanta Dash, Komivi Senyo Akutse, Ravindran Keppanan, Oluwatoyin Grace Afolabi, Mubasher Hussain, Muhammad Qasim, Liande Wang

    Entomopathogenic fungi (EPF) can be established as endophytes in the host plants to offer a long-term preventive measure for pests and diseases. This practice serves as a better alternative to the common practice of periodic direct application of EPF on plants or the target pests as a short-term defense strategy against pests and diseases. These fungal endophytes, aside from their role in pests and diseases prevention, also act as plant growth promoters. Several fungal endophytes have been associated with improvement in plant height, dry and wet weight and other growth parameters. However, many limiting factors have been identified as mitigating the successful colonization of the host plants by EPF. The inoculation methods used have been identified as one, but sadly, this has received little or less attention. Some previous studies carried out comparison between various artificial inoculation methods; foliar application, seedling dipping, soil drenching, seed inoculation, direct injection and others. In separate studies, some authors had suggested different application methods that are best suitable for certain fungal entomopathogens. For instance, leaf inoculation with conidial suspensions was suggested to be the best inoculation method for Beauveria bassiana in sorghum, stem injection was suggested as the most suitable for coffee, while, root dipping method proved the most successful for B. bassiana colonization of tomato plants for the management of Helicoverpa armigera. Here, we discussed entomopathogenic fungal endophytes as bio-control agents, plant growth promoters and highlighted the effect of various artificial inoculation methods on their endophytic colonization of the host plants.

  • Profiles of quorum sensing (QS)-related sequences in phycospheric microorganisms during a marine dinoflagellate bloom, as determined by a metagenomic approach
    Microbiol. Res. (IF 2.777) Pub Date : 2018-08-30
    Xinqing Huang, Jianming Zhu, Zhonghua Cai, Yongmin Lao, Hui Jin, Ke Yu, Boya Zhang, Jin Zhou

    The complicated relationships among environmental microorganisms are regulated by quorum sensing (QS). Understanding QS-based signals could shed light on the interactions between microbial communities in certain environments. Although QS characteristics have been widely discussed, few studies have been conducted on the role of QS in phycospheric microorganisms. Here, we used metagenomics to examine the profile of AI-1 (AinS, HdtS, LuxI) and AI-2 (LuxS) autoinducers from a deeply sequenced microbial database, obtained from a complete dinoflagellate bloom. A total of 3001 putative AI-1 homologs and 130 AI-2 homologs were identified. The predominant member among the AI groups was HdtS. The abundance of HdtS, AinS, and LuxS increased as the bloom developed, whereas the abundance of LuxI showed the opposite trend. Phylogenetic analysis suggested that HdtS and LuxI synthase originated mainly from alpha-, beta-, and gamma-Proteobacteria, whereas AinS synthase originated solely from Vibrionales. In comparison to AI-1, the sequences related to AI-2 (LuxS) demonstrated a much wider taxonomic coverage. Some significant correlations were found between dominant species and QS signals. In addition to the QS, we also performed parallel analysis of the quorum quenching (QQ) sequences. In comparison to QS, the relative abundance of QQ signals was lower; however, an obvious frequency correlation was observed. These results suggested that QS and QQ signals co-participate in regulating microbial communities during an algal bloom. These data helped to reveal the characteristic behavior of algal symbiotic bacteria, and facilitated a better understanding of microbial dynamics during an algal bloom event from a chemical ecological perspective.

  • Antifungal activity of spider venom-derived peptide lycosin-I against Candida tropicalis
    Microbiol. Res. (IF 2.777) Pub Date : 2018-08-27
    Li Tan, Le Bai, Ling Wang, Lagu He, Guangdi Li, Wenhan Du, Ting Shen, Zheyi Xiang, Jiali Wu, Zhonghua Liu, Min Hu

    Candida species are a major cause of human mucosal and deep tissue fungal infections, but few antifungal treatments are available. Here, we showed that lycosin-I, a peptide isolated from venom of the spider Lycosa singoriensis, acted as a potent antifungal inhibitor against Candida species. The MIC50 values of lycosin-I reached 8 µg/mL to treat fluconazole-susceptible and fluconazole-resistant C. tropicalis isolates. Time-kill kinetics assays revealed that after a 2-hour exposure, lycosin-I reduced colony-forming units/mL in fluconazole-susceptible and fluconazole-resistant C. tropicalis isolates approximately 70%. Furthermore, salinity tolerance assays suggested that even in the presence of Mg2+, lycosin-I maintained its potent antifungal ability at a high concentration. When the concentration of lycosin-I was increased from 1 × MIC to 8 × MIC, a significant decrease of the biofilm metabolic activity was observed in both fluconazole-susceptible and fluconazole-resistant C. tropicalis isolates. Moreover, the biofilm inhibitory concentration 50 (BIC50) and the biofilm eradicating concentration 50 (BEC50) were approximately 32 µg/mL and 128 µg/mL, respectively. Confocal laser scanning microscopy showed the localization of CY5-labeled lycosin-I mainly in the cell cytoplasm, and lycosin-I was likely to be localized in the cytoplasm after its transportation across the cell wall and membrane. Overall, our work shows that lycosin-I is a potent antifungal agent with a high efficacy, a high salinity tolerance, and potent anti-biofilm properties.

  • Clioquinol induces G2/M cell cycle arrest through the up-regulation of TDH3 in Saccharomyces cerevisiae
    Microbiol. Res. (IF 2.777) Pub Date : 2018-05-04
    Chongjia Yan, Song Wang, Jian Wang, Hui Li, Zhiwei Huang, Jing Sun, Min Peng, Wenbin Liu, Ping Shi

    Clioquinol (CQ) has been used as a classical antimicrobial agent for many years. However, its mode of action is still unclear. In our study, the growth of Candida albicans and Saccharomyces cerevisiae was inhibited by CQ. It did not kill yeast cells, but shortened G1 phase and arrested cell cycle at G2/M phase. By using two-dimensional electrophoresis based proteomic approach, six proteins were found to be significantly affected by CQ. Among them, four (PDC1, ADH1, TDH3, IPP1) were up-regulated and the other two (TDH1 and PGK1) were down-regulated. According to the Saccharomyces Genome Database (SGD), these proteins were involved in various biological processes including glycolytic fermentation, gluconeogenesis, glycolytic process, amino acid catabolism, redox reaction and reactive oxygen species metabolic process. It was noted that there was a link between TDH3 and cell cycle. The overexpression of TDH3 phenocopied CQ treatment and arrested the cell cycle at G2/M phase. RT-PCR analysis showed that the mRNA levels of CLN3 and CDC28, critical genes for passage through G1 phase, were up-regulated after the treatment of CQ as well as the overexpression of TDH3. It demonstrates that CQ inhibits the growth of yeast by up-regulating the expression of TDH3 to influence the cell cycle. It is expected to provide new insights for the antimicrobial mechanism of CQ.

  • Effectiveness of multi-trait Burkholderia contaminans KNU17BI1 in growth promotion and management of banded leaf and sheath blight in maize seedling
    Microbiol. Res. (IF 2.777) Pub Date : 2018-05-06
    Setu Bazie Tagele, Sang Woo Kim, Hyun Gu Lee, Hyun Seung Kim, Youn Su Lee

    Plant growth promoting (PGP) bacteria enhance plant growth and are a green alternative to chemical fertilizers. In our study, an effective plant growth promoting rhizobacteria (PGPR) strain, KNU17BI1, was isolated from rhizospheric soil of maize, South Korea. The strain was tested in vitro for specific PGP and antifungal traits, such as phosphate solubilization, zinc solubilization, indole acetic acid (IAA) production, ammonia production, nitrogen fixation, 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity, siderophore hydrogen cyanide production (HCN) and hydrolytic enzyme activity. Furthermore, in viro antifungal activity was done in a laboratory and in vivo effect of KNU17BI1 on banded leaf and sheath blight intensity as well as plant growth promotion on maize seedling were conducted under greenhouse conditions. The strain was found to be highly effective toward all the parameters except HCN production. The strain KNU17BI1 was identified on the basis of 16S RNA and multilocus sequence analysis (MLSA) and confirmed as Burkholderia contaminans. This study for the first time demonstrated potent in vitro antifungal activity of B. contaminans against Rhizoctonia solani AG-1(IA), Pythium graminicola, Fusarium moniliforme, Alternaria alternata, Alternaria solani, Fusarium graminearum, Stemphylium botryosum Wallr, Colletotrichum dematium, Stemphylium lycopersici and Fusarium oxysporum f.sp. melonis. Furthermore, in this study, for the first time, the potential of B. contaminans stain KNU17BI1 in controlling banded leaf and sheath blight of maize caused by R. solani AG-1(IA) was reported. Therefore, further studies are warranted on the structural identification of actual compounds behind such activities that would be exploited further for biocontrol as well as plant growth promotion.

  • Signature-tagged mutagenesis screening revealed the role of lipopolysaccharide biosynthesis gene rfbH in smooth-to-rough transition in Salmonella Enteritidis
    Microbiol. Res. (IF 2.777) Pub Date : 2018-05-05
    Yang Jiao, Zemiao Xia, Xiaohui Zhou, Yaxin Guo, Rongxian Guo, Xilong Kang, Kaiyue Wu, Jun Sun, Xiulong Xu, Xinan Jiao, Zhiming Pan, Xiufan Liu

    Salmonella enterica serovar Enteritidis (S. Enteritidis, SE) is a major cause of foodborne diseases for humans. The completeness of the O-chain antigen of Lipopolysaccharide (LPS) determines whether a S. Enteritidis strain is smooth or rough. However, genes that are involved in the synthesis of LPS and rough-smooth variation are not completely understood. In this study, we used monoclonal antibody against O-antigens (O9 mAb) to identify novel factors that are involved in LPS synthesis and rough variation in S. Enteritidis by using signature-tagged mutagenesis (STM) technique. Our results showed that transposon insertion in the gene rfbH led to different LPS phenotype, auto-aggregation characteristic, motility and resistance to environmental stress compared with SE wild-type strain C50041. In addition, sera tests showed that rfbH mutant does not elicit specific antibodies against O-antigens in vaccinated animals. Taken together, the S. Enteritidis rfbH gene is implicated in LPS biosynthesis, rough variation, sera distinguishable reaction, motility and stress resistance. The rfbH mutant strain could be potentially used as a distinguishable vaccine or a live vector to deliver drugs and antibodies in vivo.

  • The developmental regulator Pcz1 affects the production of secondary metabolites in the filamentous fungus Penicillium roqueforti
    Microbiol. Res. (IF 2.777) Pub Date : 2018-05-04
    Juan F. Rojas-Aedo, Carlos Gil-Durán, Alejandra Goity, Inmaculada Vaca, Gloria Levicán, Luis F. Larrondo, Renato Chávez

    Penicillium roqueforti is used in the production of several kinds of ripened blue-veined cheeses. In addition, this fungus produces interesting secondary metabolites such as roquefortine C, andrastin A and mycophenolic acid. To date, there is scarce information concerning the regulation of the production of these secondary metabolites. Recently, the gene named pcz1 (Penicillium C6 zinc domain protein 1) was described in P. roqueforti, which encodes for a Zn(II)2Cys6 protein that controls growth and developmental processes in this fungus. However, its effect on secondary metabolism is currently unknown. In this work, we have analyzed how the overexpression and down-regulation of pcz1 affect the production of roquefortine C, andrastin A and mycophenolic acid in P. roqueforti. The three metabolites were drastically reduced in the pcz1 down-regulated strains. However, when pcz1 was overexpressed, only mycophenolic acid was overproduced while, on the contrary, levels of roquefortine C and andrastin A were diminished. Importantly, these results match the expression pattern of key genes involved in the biosynthesis of these metabolites. Taken together, our results suggest that Pcz1 plays a key role in regulating secondary metabolism in the fungus Penicillium roqueforti.

  • Brazilian red propolis: Chemical composition and antibacterial activity determined using bioguided fractionation
    Microbiol. Res. (IF 2.777) Pub Date : 2018-05-04
    Luciane Corbellini Rufatto, Paola Luchtenberg, Charlene Garcia, Christine Thomassigny, Sylvie Bouttier, Françoise Dumas, João Antonio Pêgas Henriques, Mariana Roesch-Ely, Sidnei Moura

    The indiscriminate use of antibiotics is causing an increase in bacterial resistance, complicating therapeutic planning. In this context, natural products have emerged as major providers of bioactive compounds. This work performs a bioguided study of Brazilian red propolis to identify compounds with antibacterial potential and to evaluate their cytotoxicity against non-tumour cells. Using bioguided fractionation performed with the hydroalcoholic extract of red propolis from Alagoas, it was possible to obtain subfractions with remarkable bacteriostatic activity compared with the precursor fractions. The SC2 subfraction was highlighted and showed the best results with minimal inhibitory concentrations (MICs) of 56.75, 28.37, 454.00, and 227.00 μg.mL−1 against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa, respectively. However, this study also revealed a cytotoxic effect against the non-tumour Vero cell line. Furthermore, through chemical analyses using high resolution mass spectrometry, high performance liquid chromatography with UV detection, and gas chromatography coupled to mass spectrometry, we verified the presence of important marker compounds in the fractions and extracts, including formononetin (m/z 267.0663), biochanin A (m/z 283.0601), and liquiritigenin (m/z 255.0655). The results obtained in this study suggest an important antibacterial potential of red propolis subfractions. In this context, the bioguided fractionation has been a useful process, due to its ability to isolate and concentrate active compounds in a logical and rational way.

  • Master mechanisms of Staphylococcus aureus: consider its excellent protective mechanisms hindering vaccine development
    Microbiol. Res. (IF 2.777) Pub Date : 2018-05-04
    Xu Zhang, Manukumar H. Marichannegowda, Kadalipura P. Rakesh, Hua-Li Qin

    Lack of known mechanisms of protection against Staphylococcus aureus in humans represents an important risk factor for skin infections and bacteremia in patients, intern hindering the development of efficacious vaccines. However, development of effective humoral response may be dampened by converging immune-evasion mechanisms of S. aureus. To develop a promising vaccine against S. aureus, it is pre-requisite to clear understanding of cutaneous, innate and adaptive immune response. The S. aureus dampening the humoral response, T cell help, blocking complement factors, and killing immune players by its toxins are the important factors need to understand clearly. We hypothesized that the master mechanism of S. aureus counteracts may hindering the immune action which may result in failure of target-oriented vaccine development. Developing immunological interventions that can effectively block the S. aureus counteracting mechanisms are the key success for a developing vaccine for the future was warranted.

  • Biotechnological applications of bacteriophages: State of the art
    Microbiol. Res. (IF 2.777) Pub Date : 2018-04-30
    Liliam K. Harada, Erica C. Silva, Welida F. Campos, Fernando S. Del Fiol, Marta Vila, Krystyna Dąbrowska, Victor N. Krylov, Victor M. Balcão

    Bacteriophage particles are the most abundant biological entities on our planet, infecting specific bacterial hosts in every known environment and being major drivers of bacterial adaptive evolution. The study of bacteriophage particles potentially sheds light on the development of new biotechnology products. Bacteriophage therapy, although not new, makes use of strictly lytic phage particles as an alternative in the antimicrobial treatment of resistant bacterial infections and is being rediscovered as a safe method due to the fact that these biological entities devoid of any metabolic machinery do not have affinity to eukaryotic cells. Furthermore, bacteriophage-based vaccination is emerging as one of the most promising preventive strategies. This review paper discusses the biological nature of bacteriophage particles, their mode(s) of action and potential exploitation in modern biotechnology. Topics covered in detail include the potential of bacteriophage particles in human infections (bacteriophage therapy), nanocages for gene delivery, food biopreservation and safety, biocontrol of plant pathogens, phage display, bacterial biosensing devices, vaccines and vaccine carriers, biofilm and bacterial growth control, surface disinfection, corrosion control, together with structural and functional stabilization issues.

  • Evidence for Up and Down Regulation of 450 genes by rpoB12 (rif) Mutation and their Implications in Complexity of Transcription Modulation in Escherichia coli
    Microbiol. Res. (IF 2.777) Pub Date : 2018-04-30
    Shanmugaraja Meenakshi, M. Hussain Munavar

    Analyses of mutations in rpoB subunit of Escherichia coli that lead to resistance to rifampicin have been invaluable in providing insight into events during transcription continue to be discovered. Earlier we reported that rpoB12 suppresses over-expression of cps genes in Δlon mutant of E.coli, by interfering with the transcription of rcsA. Here we report Microarray based Transcriptome profile of Δlon and Δlon rpoB12 strains. The data analyses clearly reveal that rpoB12 mutation results in the differential expression of ∼450 genes. The transcription profiles of some of the genes namely, rcsA, gadE, csgD, bolA, ypdI, dnaJ, clpP, csrA and hdeA are significantly altered, particularly the genes implicated in virulence. Some of the phenotypic traits namely, biofilm formation, motility, curli synthesis and ability to withstand acidic stress in a lon+rpoB12 strain were assessed. The results clearly indicate that rpoB12 up-regulates biofilm formation and curli synthesis while it makes the cells sensitive for growth in acidic medium and inhibits motility almost completely. Furthermore, rpoB12 modulates the expression profile of a significant number of genes involved in stress responses, genes encoding small RNAs. Thus, this study reveals the versatile role of the rpoB12 mutation, especially its impact on the regulation of genes related to virulence and highlights its medical importance.

  • Pathogenesis-Related Proteins and Peptides as Promising Tools for Engineering Plants with Multiple Stress Tolerance
    Microbiol. Res. (IF 2.777) Pub Date : 2018-04-30
    Sajad Ali, Bashir Ahmad Ganai, Azra N Kamili, Ajaz Ali Bhat, Zahoor Ahmad Mir, Javaid Akhter Bhat, Anshika Tyagi, Sheikh Tajamul Islam, Muntazir Mushtaq, Prashant Yadav, Sandhya Rawat, Anita Grover

    Pathogenesis-related (PR) proteins and antimicrobial peptides (AMPs) are a group of diverse molecules that are induced by phytopathogens as well as defense related signaling molecules. They are the key components of plant innate immune system especially systemic acquired resistance (SAR), and are widely used as diagnostic molecular markers of defense signaling pathways. Although, PR proteins and peptides have been isolated much before but their biological function remains largely enigmatic despite the availability of new scientific tools. The earlier studies have demonstrated that PR genes provide enhanced resistance against both biotic and abiotic stresses, which make them one of the most promising candidates for developing multiple stress tolerant crop varieties. In this regard, plant genetic engineering technology is widely accepted as one of the most fascinating approach to develop the disease resistant transgenic crops using different antimicrobial genes like PR genes. Overexpression of PR genes (chitinase, glucanase, thaumatin, defensin and thionin) individually or in combination have greatly uplifted the level of defense response in plants against a wide range of pathogens. However, the detailed knowledge of signalling pathways that regulates the expression of these versatile proteins is critical for improving crop plants to multiple stresses, which is the future theme of plant stress biology. Hence, this review provides an overall overview on the PR proteins like their classification, role in multiple stresses (biotic and abiotic) as well as in various plant defense signaling cascades. We also highlight the success and snags of transgenic plants expressing PR proteins and peptides.

  • 更新日期:2018-04-25
  • Bacterial endophytes modulates the withanolide biosynthetic pathway and physiological performance in Withania somnifera under biotic stress
    Microbiol. Res. (IF 2.777) Pub Date : 2018-04-22
    Aradhana Mishra, Satyendra Pratap Singh, Sahil Mahfooz, Arpita Bhattacharya, Nishtha Mishra, Pramod Arvind Shirke, C.S. Nautiyal

    Despite the vast exploration of endophytic microbes for growth enhancement in various crops, knowledge about their impact on the production of therapeutically important secondary metabolites is scarce. In the current investigation, chitinolytic bacterial endophytes were isolated from selected medicinal plants and assessed for their mycolytic as well as plant growth promoting potentials. Among them the two most efficient bacterial endophytes namely Bacillus amyloliquefaciens (MPE20) and Pseudomonas fluorescens (MPE115) individually as well as in combination were able to modulate withanolide biosynthetic pathway and tolerance against Alternaria alternata in Withania somnifera. Interestingly, the expression level of withanolide biosynthetic pathway genes (3-hydroxy-3-methylglutaryl co-enzyme A reductase, 1-deoxy-D-xylulose-5-phosphate reductase, farnesyl di-phosphate synthase, squalene synthase, cytochrome p450, sterol desaturase, sterol Δ-7 reductase and sterol glycosyl transferases) were upregulated in plants treated with the microbial consortium under A. alternata stress. In addition, application of microbes not only augmented withaferin A, withanolide A and withanolide B content (1.52-1.96, 3.32-5.96 and 12.49-21.47 fold, respectively) during A. alternata pathogenicity but also strengthened host resistance via improvement in the photochemical efficiency, normalizing the oxidized and non-oxidized fraction, accelerating photochemical and non-photochemical quantum yield, and electron transport rate. Moreover, reduction in the passively dissipated energy of PSI and PSII in microbial combination treated plants corroborate well with the above findings. Altogether, the above finding highlights novel insights into the underlying mechanisms in application of endophytes and emphasizes their capability to accelerate biosynthesis of withanolides in W. somnifera under biotic stress caused by A. alternata.

  • 更新日期:2018-04-25
  • Comparison of genomic islands in cyanobacteria: evidence of bacteriophage-mediated horizontal gene transfer from eukaryotes
    Microbiol. Res. (IF 2.777) Pub Date : 2018-04-11
    James S. Godde, Shakuntala Baichoo, Zahra Mungloo-Dilmohamud, Yasmina Jaufeerally-Fakim

    A number of examples of putative eukaryote-to-prokaryote horizontal gene transfer (HGT) have been proposed in the past using phylogenetic analysis in support of these claims but none have attempted to map these gene transfers to the presence of genomic islands (GIs) in the host. Two of these cases have been examined in detail, including an ATP sulfurylase (ATPS) gene and a class I fructose bisphosphate aldolase (FBA I) gene that were putatively transferred to cyanobacteria of the genus Prochlorococcus from either green or red algae, respectively. Unlike previous investigations of HGT, parametric methods were initially used to detect genomic islands, then more traditional phylogenomic and phylogenetic methods were used to confirm or deny the HGT status of these genes. The combination of these three methods of analysis- detection of GIs, the determination of genomic neighborhoods, as well as traditional phylogeny, lends strong support to the claim that trans-domain HGT has occurred in only one of these cases and further suggests a new insight into the method of transmission of FBA I, namely that cyanophage-mediated transfer may have been responsible for the HGT event in question. The described methods were then applied to a range of prochlorococcal genomes in order to characterize a candidate for eukaryote-to-prokaryote HGT that had not been previously studied by others. Application of the same methodology used to confirm or deny HGT for ATPS and FBA I identified a ⊗12 fatty acid desaturase (FAD) gene that was likely transferred to Prochlorococcus from either green or red algae.

  • Tissue age and plant genotype affect the microbiota of apple and pear bark
    Microbiol. Res. (IF 2.777) Pub Date : 2018-04-10
    Elena Arrigoni, Livio Antonielli, Massimo Pindo, Ilaria Pertot, Michele Perazzolli

    Plant tissues host complex fungal and bacterial communities, and their composition is determined by host traits such as tissue age, plant genotype and environmental conditions. Despite the importance of bark as a possible reservoir of plant pathogenic microorganisms, little is known about the associated microbial communities. In this work, we evaluated the composition of fungal and bacterial communities in the pear (Abate and Williams cultivars) and apple (Golden Delicious and Gala cultivars) bark of three/four-year-old shoots (old bark) or one-year-old shoots (young bark), using a meta-barcoding approach. The results showed that both fungal and bacterial communities are dominated by genera with ubiquitous attitudes, such as Aureobasidium, Cryptococcus, Deinococcus and Hymenobacter, indicating intense microbial migration to surrounding environments. The shoot age, plant species and plant cultivar influenced the composition of bark fungal and bacterial communities. In particular, bark communities included potential biocontrol agents that could maintain an equilibrium with potential plant pathogens. The abundance of fungal (e.g. Alternaria, Penicillium, Rosellinia, Stemphylium and Taphrina) and bacterial (e.g. Curtobacterium and Pseudomonas) plant pathogens was affected by bark age and host genotype, as well as those of fungal genera (e.g. Arthrinium, Aureobasidium, Rhodotorula, Sporobolomyces) and bacterial genera (e.g. Bacillus, Brevibacillus, Methylobacterium, Sphingomonas and Stenotrophomonas) with possible biocontrol and plant growth promotion properties.

  • Biocontrol of Sclerotinia sclerotiorum (Lib.) de Bary on common bean by native lipopeptide-producer Bacillus strains
    Microbiol. Res. (IF 2.777) Pub Date : 2018-04-07
    Daniela C. Sabaté, Carolina Pérez Brandan, Gabriela Petroselli, Rosa Erra-Balsells, M.Carina Audisio

    Bacillus sp. B19, Bacillus sp. P12 and B. amyloliquefaciens B14 were isolated from soils of Salta province, and PGPR properties on the common bean (Phaseolus vulgaris L.) cv. Alubia and antagonistic activity against Sclerotinia sclerotiorum were studied.It was determined that B19 and P12 increased crop germination potential (GP) from the common bean by 14.5% compared to control seeds; these strains also increased root length (10.4 and 15%, respectively) and stem length (20.2 and 30%, respectively) compared to the control; however, as for the B14 strain, no increases in growth parameters were detected. In addition, all the treatments that combined two bacilli: B14 + B19, B14 + P12 and B19 + P12, generated beneficial effects on GP and seedling growth compared to control seeds, but not compared to a single inoculant. B19 and P12 strains synthesized auxins at concentrations of 5.71 and 4.90 mg/mL, respectively, and it was qualitatively determined that they synthesize siderophores. In addition, previous studies have determined that B14 produces auxins in a concentration of 10.10 mg/ml, and qualitatively synthesizes siderophores.The phytosanitary state of the white bean cv. Alubia control seeds revealed bacterial contamination in 87% of all the evaluated seeds and different fungi such as Cladosporium sp., Fusarium sp., and Rhizopus sp. Bean seeds treated with B14, B19 or P12 showed no growth of contaminating bacteria or of pathogenic fungi; in fact, bacilli inoculum development was observed in all seeds. Additionally, B19, P12 and B14 strains inhibited in vitro the development of 9 native S. sclerotiorum strains isolated from the Salta region, with FI ranging between 60 and 100%. The three Bacillus strains synthesized different isoforms of the lipopeptides: surfactin, iturin, and fengycin in the presence of S. sclerotiorum, as determined by MALDI-TOF.In the in vivo trials, when common bean seeds were grown in soils contaminated with S. sclerotiorum, an incidence of 100% was determined when the seeds were not treated with any Bacillus. Seeds treated with the chemical fungicide and sown in S. sclerotiorum-infested soil did not produce seed emergence, while the inoculation of the seeds with B14 + P12, B14 + B19 or B19 + P12 reduced the effect of the pathogen by 46, 43 and 25%, respectively. Disease progression in B14 + P12 and B14 + B19 treatments was significantly lower than in the remaining treatments, with an AUDPC of 873.75 and 1071, respectively.

Some contents have been Reproduced with permission of the American Chemical Society.
Some contents have been Reproduced by permission of The Royal Society of Chemistry.
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