Journal of Microbiology

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Different distribution patterns of microorganisms between aquaculture pond sediment and water: Lili Dai , Chengqing Liu , Liang Peng , Chaofeng Song , Xiaoli Li , Ling Tao; J. Microbiol. 2021;59(4):376-388. Published online February 25, 2021; DOI: https://doi.org/10.1007/s12275-021-0635-5

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Aquatic microorganisms in the sediment and water column are closely related; however, their distribution patterns between these two habitats still remain largely unknown. In this study, we compared sediment and water microeukaryotic and bacterial microorganisms in aquaculture ponds from different areas in China, and analyzed the influencing environmental factors as well as the inter-taxa relationships. We found that bacteria were significantly more abundant than fungi in both sediment and water, and the bacterial richness and diversity in sediment were higher than in water in all the sampling areas, but no significant differences were found between the two habitats for microeukaryotes. Bacterial taxa could be clearly separated through cluster analysis between the sediment and water, while eukaryotic taxa at all classification levels could not. Spirochaetea, Deltaproteobacteria, Nitrospirae, Ignavibacteriae, Firmicutes, Chloroflexi, and Lentimicrobiaceae were more abundantly distributed in sediment, while Betaproteobacteria, Alphaproteobacter, Cyanobacteria, Roseiflexaceae, Dinghuibacter, Cryomorphaceae, and Actinobacteria were more abundant in water samples. For eukaryotes, only Cryptomonadales were found to be distributed differently between the two habitats. Microorganisms in sediment were mainly correlated with enzymes related to organic matter decomposition, while water temperature, pH, dissolved oxygen, and nutrient levels all showed significant correlation with the microbial communities in pond water. Intensive interspecific relationships were also found among eukaryotes and bacteria. Together, our results indicated that eukaryotic microorganisms are distributed less differently between sediment and water in aquaculture ponds compared to bacteria. This study provides valuable data for evaluating microbial distributions in aquatic environments, which may also be of practical use in aquaculture pond management.

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International Journal of Environmental Studies.2024; 81(4): 1836. CrossRef
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Qi Mang, Jun Gao, Quanjie Li, Yi Sun, Gangchun Xu, Pao Xu
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Yan Zhang, Tiejun Li, Guangzhi Li, Tao Yuan, Yao Zhang, Lei Jin
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Minhai Liu, Zhongneng Yan, Chengwei Huang, Zhihua Lin, Zhilan Peng, Chenxi Zhao, Xiafei Zheng
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Ngoc Minh Ngan Bui, Jasmine Heyse, Jérôme Delamare-Deboutteville, Tom Defoirdt, Ruben Props, Colin Shelley
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Saurabh Chatterjee, Madhura More
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Chaohua Wang, Zhiyuan Yao, Pingping Zhan, Xianghua Yi, Jiong Chen, Jinbo Xiong
Journal of Environmental Sciences.2023; 124: 429. CrossRef
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Antonia Bruno, Alessandra Cafiso, Anna Sandionigi, Andrea Galimberti, Davide Magnani, Amedeo Manfrin, Giulio Petroni, Maurizio Casiraghi, Chiara Bazzocchi
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Zebing Zhu, Siyang Xu, Xiajun Bao, Lili Shan, Yunyan Pei, Wanjun Zheng, Yixing Yuan
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Tianheng Gao, Nannan Li, Wenlei Xue, Yuning Hu, Hai Lin
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Antibiotic resistance gene-free probiont administration to tilapia for growth performance and Streptococcus agalactiae resistance
Mira Mawardi, Agustin Indrawati, Angela Mariana Lusiastuti, I Wayan Teguh Wibawan
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Analysis of the structure and function of microbial community in late-stage of grass carp (Ctenopharyngodon idella) farming ponds
Shuhui Niu, Kai Zhang, Zhifei Li, Jun Xie, Guangjun Wang, Hongyan Li, Ermeng Yu, Yun Xia, Jingjing Tian, Wangbao Gong
Aquaculture Reports.2023; 30: 101556. CrossRef
Case study on the aptness of in-pond raceways for pikeperch, Sander lucioperca, grow-out
Zoltán Nagy, László Ardó, Ferenc Demény, Dénes Gál, Zsuzsanna J. Sándor, Uroš Ljubobratović
Aquaculture Reports.2022; 27: 101356. CrossRef
Structural and functional comparisons of the environmental microbiota of pond and tank environments at different locations for the commercial aquaculture of American shad
J. Du, Q.H. Liu, J.S. Liu, K.-Q. Zhang, W.F. Huang
Letters in Applied Microbiology.2022; 75(1): 51. CrossRef
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Hongyu Feng, Yajun Qiao, Lu Xia, Wen Yang, Yongqiang Zhao, Nasreen Jeelani, Shuqing An
Plant and Soil.2022; 477(1-2): 439. CrossRef
Improvement of fish production and water quality in a recirculating aquaculture pond enhanced with bacteria-microalgae association
Chu Wang, Cancan Jiang, Tianming Gao, Xiawei Peng, Shuanglong Ma, Qian Sun, Bing Xia, Xiangming Xie, Zhihui Bai, Shengjun Xu, Xuliang Zhuang
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Microbial Community Structure and Its Driving Environmental Factors in Black Carp (Mylopharyngodon piceus) Aquaculture Pond
Xuemei Li, Lu Liu, Yongjiu Zhu, Tingbing Zhu, Xingbing Wu, Deguo Yang
Water.2021; 13(21): 3089. CrossRef

Highly diverse endophytes in roots of Cycas bifida (Cycadaceae), an ancient but endangered gymnosperm: Ying Zheng , Tzen-Yuh Chiang , Chao-Li Huang , Xun Gong; J. Microbiol. 2018;56(5):337-345. Published online May 2, 2018; DOI: https://doi.org/10.1007/s12275-018-7438-3

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Abstract

As an ancient seed plant, cycads are one of the few gymnosperms that develop a root symbiosis with cyanobacteria, which has allowed cycads to cope with harsh geologic and climatic conditions during the evolutionary process. However, the endophytic microbes in cycad roots remain poorly identified. In this study, using next-generation sequencing techniques, we investigated the microbial diversity and composition of both the coralloid and regular roots of Cycas bifida (Dyer) K.D. Hill. Highly diverse endophytic communities were observed in both the coralloid and regular roots. Of the associated bacteria, the top five families were the Nostocaceae, Sinobacteraceae, Bradyrhizobiaceae, Bacillaceae, and Hyphomicrobiaceae. The Nectriaceae, Trichocomaceae, and Incertae sedis were the predominant fungal families in all root samples. A significant difference in the endophytic bacterial community was detected between coralloid roots and regular roots, but no difference was observed between the fungal communities in the two root types. Cyanobacteria were more dominant in coralloid roots than in regular roots. The divergence of cycad root structures and the modified physiological processes may have contributed to the abundance of cyanobionts in coralloid roots. Consequently, the colonization of cyanobacteria inhibits the assemblage of other endophytes. Our results contribute to an understanding of the species diversity and composition of the cycad-endophyte microbiome and provide an abbreviated list of potential ecological roles of the core microbes present.

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Colonization study of gfp-tagged Achromobacter marplatensis strain in sugar beet: YingWu Shi , Chun Li Li , HongMei Yang , Tao Zhang , Yan Gao , Min Chu , Jun Zeng , Qing Lin , OuTiKu Er , YuGuo Li , Xiangdong Huo , Kai Lou; J. Microbiol. 2017;55(4):267-272. Published online January 26, 2017; DOI: https://doi.org/10.1007/s12275-017-6371-1

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Abstract

This study details the introduction of a gfp marker into an endophytic bacterial strain (Achromobacter marplatensis strain 17, isolated from sugar beet) to monitor its coloniza-tion of sugar beet (Beta. vulgaris L.). Stability of the plasmid encoding the gfp was confirmed in vitro for at least 72 h of bacterial growth and after the colonization of tissues, under nonselective conditions. The colonization was observed us-ing fluorescence microscopy and enumeration of culturable endophytes in inoculated sugar beet plants that grew for 10 or 20 days. gfp-Expressing strains were re-isolated from the inner tissues of surface-sterilized roots and stems of inocu-lated plants, and the survival of the Achromobacter marpla-tensis 17:gfp strain in plants 20 days after inoculation, even in the absence of selective pressure, suggests that it is good colonizer. These results also suggest that this strain could be a useful tool for the delivery of enzymes or other proteins into plants. In addition, the study highlights that sugar beet plants can be used effectively for detailed in vitro studies on the interactions between A. marplatensis strain 17 and its host, particularly if a gfp-tagged strain of the pathogen is used.

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Research Support, Non-U.S. Gov'ts

An endophytic Coniochaeta velutina producing broad spectrum antimycotics: Jie Xie , Gary A. Strobel , Tao Feng , Huishuang Ren , Morgan T. Mends , Zeyang Zhou , Brad Geary; J. Microbiol. 2015;53(6):390-397. Published online May 30, 2015; DOI: https://doi.org/10.1007/s12275-015-5105-5

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Abstract

An endophyte (PC27-5) was isolated from stem tissue of Western hemlock (Tsuga heterophylla) in a Pacific Northwest temperate rainforest. Phylogenetic analyses, based on ITS- 5.8S rDNA and 18S rDNA sequence data, combined with cultural and morphological analysis showed that endophyte PC27-5 exhibited all characteristics of a fungus identical to Coniochaeta velutina. Furthermore, wide spectrum antimycotics were produced by this endophyte that were active against such plant pathogens as Sclerotinia sclerotiorum, Pythium ultimum, and Verticillium dahliae and lethal to Phythophthora cinnamomi, Pythium ultimum, and Phytophthora palmivora in plate tests. The bioactive components were purified through organic solvent extraction, followed by silica column chromatography, and finally preparative HPLC. The minimum inhibitory concentration of the active fraction to Pythium ultimum, which was gained from preparative HPLC, was 11 ?/ml. UPLC-HRMS analysis showed there were two similar components in the antimycotic fraction. Their molecular formulae were established as C30H22O11 (compound I) and C30H22O10 (compound II) respectively, and preliminary spectral results indicate that they are anthroquinone glycosides. Other non ?biologically active compounds were identified in culture fluids of this fungus by spectral means as emodin and chrysophanol - anthroquinone derivatives. This is the first report that Coniochaeta velutina as an endophyte produces bioactive antifungal components.

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Novel Collophorina and Coniochaeta species from Euphorbia polycaulis, an endemic plant in Iran
Shaghayegh Nasr, Steffen Bien, Mohammad Reza Soudi, Nayyereh Alimadadi, Seyed Abolhassan Shahzadeh Fazeli, Ulrike Damm
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Evaluation of the Antimicrobial Potency of Silver Nanoparticles Biosynthesized by Using an Endophytic Fungus, Cryptosporiopsis ericae PS4: Lamabam Sophiya Devi , Santa Ram Joshi; J. Microbiol. 2014;52(8):667-674. Published online July 4, 2014; DOI: https://doi.org/10.1007/s12275-014-4113-1

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Abstract

In the present study, silver nanoparticles (AgNPs) with an average particle size of 5.5 ± 3.1 nm were biosynthesized using an endophytic fungus Cryptosporiopsis ericae PS4 isolated from the ethno-medicinal plant Potentilla fulgens L. The nanoparticles were characterized using UV-visible spectrophotometer, transmission electron microscopy (TEM), scanning electron microscopy (SEM), selective area electron diffraction (SAED), and energy dispersive X-ray (EDX) spectroscopy analysis. Antimicrobial efficacy of the AgNPs was analyzed singly and in combination with the antibiotic/antifungal agent chloramphenicol/fluconazole, against five pathogenic microorganisms-Staphylococcus aureus MTCC96, Salmonella enteric MTCC735, Escherichia coli MTCC730, Enterococcus faecalis MTCC2729, and Candida albicans MTCC 183. The activity of AgNPs on the growth and morphology of the microorganisms was studied in solid and liquid growth media employing various susceptibility assays. These studies demonstrated that concentrations of AgNPs alone between 10 and 25 μM reduced the growth rates of the tested bacteria and fungus and revealed bactericidal/fungicidal activity of the AgNPs by delaying the exponential and stationary phases. Examination using SEM showed pits and ruptures in bacterial cells indicating fragmented cell membrane and severe cell damage in those cultures treated with AgNPs. These experimental findings suggest that the biosynthesized AgNPs may be a potential antimicrobial agent.

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Characterization of Cellulases of Fungal Endophytes Isolated from Espeletia spp.: Luisa Cabezas , Carolina Calderon , Luis Miguel Medina , Isabela Bahamon , Martha Cardenas , Adriana Jimena Bernal , Andrés Gonzalez , Silvia Restrepo; J. Microbiol. 2012;50(6):1009-1013. Published online December 30, 2012; DOI: https://doi.org/10.1007/s12275-012-2130-5

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Abstract: Endophytes are microorganisms that asymptomatically invade plant tissues. They can stimulate plant growth and/or provide defense against pathogen attacks through the production of secondary metabolites. Most endophyte species are still unknown, and because they may have several applications, the study of their metabolic capabilities is essential. We characterized 100 endophytes isolated from Espeletia spp., a genus unique to the paramo ecosystem, an extreme environment in the Andean mountain range. We evaluated the cellulolytic potential of these endophytes on the saccharification of the oil palm empty fruit bunch (OPEFB). The total cellulolytic activity was measured for each endophyte on filter paper (FPA). In addition, the specific carboxymethyl cellulase (CMCase), exoglucanase, and β-glucosidase activities were determined. We found four fungi positive for cellulases. Of these fungi, Penicillium glabrum had the highest cellulolytic activity after partial purification, with maximal CMCase, exoglucanase and β-glucosidase enzyme activities of 44.5, 48.3, and 0.45 U/ml, respectively. Our data showed that the bioprospection of fungi and the characterization of their enzymes may facilitate the process of biofuel production.

Enhancement of Anti-candidal Activity of Endophytic Fungus Phomopsis sp. ED2, Isolated from Orthosiphon stamineus Benth, by Incorporation of Host Plant Extract in Culture Medium: Tong Woei Yenn , Chong Chai Lee , Darah Ibrahim , Latiffah Zakaria; J. Microbiol. 2012;50(4):581-585. Published online July 21, 2012; DOI: https://doi.org/10.1007/s12275-012-2083-8

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Abstract: This study examined the effect of host extract in the culture medium on anti-candidal activity of Phomopsis sp. ED2, previously isolated from the medicinal herb Orthosiphon stamineus Benth. Interestingly, upon addition of aqueous host extract to the culture medium, the ethyl acetate extract prepared from fermentative broth exhibited moderate anticandidal activity in a disc diffusion assay. The minimal inhibitory concentration of this extract was 62.5 μg/ml and it only exhibited fungistatic activity against C. albicans. In the time-kill study, a 50% growth reduction of C. albicans was observed at 31.4 h for extract from the culture incorporating host extract. In the bioautography assay, only one single spot (Rf 0.59) developed from the extract exhibited anti-candidal activity. A spot with the a similar Rf was not detected for the crude extract from YES broth without host extract. This indicated that the terpenoid anti-candidal compound was only produced when the host extract was introduced into the medium. The study concluded that the incorporation of aqueous extract of the host plant into the culture medium significantly enhanced the anti-candidal activity of Phomopsis sp. ED2.

Effects of a Dark-Septate Endophytic Isolate LBF-2 on the Medicinal Plant Lycium barbarum L.: Hai-han Zhang , Ming Tang , Hui Chen , Ya-jun Wang; J. Microbiol. 2012;50(1):91-96. Published online February 27, 2012; DOI: https://doi.org/10.1007/s12275-012-1159-9

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Abstract: Dark septate endophytes (DSE) are ubiquitous root associated fungi; however, our understanding of their ecological function remains unclear. Here, we investigated the positive effect of a DSE fungus on its host plant Lycium barbarum L. A DSE isolate, LBF-2, isolated from the roots of L. barbarum, was inoculated onto the roots of plants, which were grown under greenhouse conditions for five weeks. The result of molecular analyses of internal transcribed spacer regions indicated that LBF-2 was 96% similar to Paraphoma chrysanthemicola. Melanized septate hyphae were observed in the root cortical cells of L. barbarum using a light microscope. Inoculation with LBF-2 increased the total biomass by 39.2% and also enhanced chlorophyll fluorescence. Inoculation increased the concentration of total chlorophyll by 22.8% and of chlorophyll a by 21.3%, relative to uninoculated controls. These data indicate that the LBF-2 isolate might be used to facilitate the cultivation of L. barbarum, which has medicinal applications.

Endophytic Fungus Trichothecium roseum LZ93 Antagonizing Pathogenic Fungi In Vitro and Its Secondary Metabolites: XiaoMei Zhang , GuoHong Li , Juan Ma , Ying Zeng , WeiGuang Ma , PeiJi Zhao; J. Microbiol. 2010;48(6):784-790. Published online January 9, 2011; DOI: https://doi.org/10.1007/s12275-010-0173-z

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Abstract: The endophytic fungus Trichothecium roseum LZ93 from Maytenus hookeri was found to antagonize other pathogenic fungi in vitro. To identify which compound contributed substantially to the antagonism, we fermented the strain and purified its fermentation products. Eleven compounds were obtained, including two trichothecenes, five rosenonolactones, two cardiotonic cyclodepsipeptides, and two sterols. Compound 11β-hydroxyrosenonolactone (1) was assigned according to 1D and 2D-NMR data for the first time. At the same time, the 1H and 13C-NMR assignments for 6β-hydroxyrosenonolactone (2) were revised. Of all of them, only trichothecin (6) showed strong antifungal activity. Based on our observations of the antagonistic activity and the other experimental results, we suggest that the antifungal compound trichothecin was the main contributor to the antagonistic action of T. roseum LZ93.

Endophytic Fungi Diversity of Aquatic/Riparian Plants and Their Antifungal Activity In Vitro: Hai-Yan Li , Chun-An Zhao , Chen-Jian Liu , Xiao-Fei Xu; J. Microbiol. 2010;48(1):1-6. Published online March 11, 2010; DOI: https://doi.org/10.1007/s12275-009-0163-1

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Abstract

Two hundred and fourteen endophytic fungi were isolated from 500 segments of aquatic/riparian plants Ottelia acuminata, Myriophyllum verticillatum, Equisetum arvense, Cardamine multijuga, and Impatiens chinensis. They were identified to 31 taxa in which Cladosporium, Fusarium, and Geotrichum were the dominant genera. Among all isolates, 169 (79%) were anamorphic fungi, 1 (0.5%) was an teleomorphic ascomycete and 44 (21%) were sterile mycelia. There were significant differences in the colonization frequency of endophytes between the five plant species (X~2=51.128, P<0.001, Chi-square test). The riparian plants harboured more endophytes than the submerged plants. The antifungal activity of these isolates against Fusarium solani and Phytophthora nicotianae in vitro were tested and 28 (13.1%) isolates showed antifungal activities with more than 30% growth inhibition rate against the two pathogens.

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Endophytic Bacterial Diversity in Grapevine (Vitis vinifera L.) Leaves Described by 16S rRNA Gene Sequence Analysis and Length Heterogeneity-PCR: Daniela Bulgari , Paola Casati , Lorenzo Brusetti , Fabio Quaglino , Milena Brasca , Daniele Daffonchio , Piero Attilio Bianco; J. Microbiol. 2009;47(4):393-401. Published online September 9, 2009; DOI: https://doi.org/10.1007/s12275-009-0082-1

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Abstract

Diversity of bacterial endophytes associated with grapevine leaf tissues was analyzed by cultivation and cultivation-independent methods. In order to identify bacterial endophytes directly from metagenome, a protocol for bacteria enrichment and DNA extraction was optimized. Sequence analysis of 16S rRNA gene libraries underscored five diverse Operational Taxonomic Units (OTUs), showing best sequence matches with gamma-Proteobacteria, family Enterobacteriaceae, with a dominance of the genus Pantoea. Bacteria isolation through cultivation revealed the presence of six OTUs, showing best sequence matches with Actinobacteria, genus Curtobacterium, and with Firmicutes genera Bacillus and Enterococcus. Length Heterogeneity-PCR (LH-PCR) electrophoretic peaks from single bacterial clones were used to setup a database representing the bacterial endophytes identified in association with grapevine tissues. Analysis of healthy and phytoplasma- infected grapevine plants showed that LH-PCR could be a useful complementary tool for examining the diversity of bacterial endophytes especially for diversity survey on a large number of samples.

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Microbial detoxification of toxin and antifungal activity of several endophytic bacterial strains against Colletotrichum nymphaeae causal agent of strawberry anthracnose
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Diversity Profiling of Grapevine Microbial Endosphere and Antagonistic Potential of Endophytic Pseudomonas Against Grapevine Trunk Diseases
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Rare Species Shift the Structure of Bacterial Communities Across Sphagnum Compartments in a Subalpine Peatland
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Phenotypic and genotypic characterization of endophytic bacteria associated with transgenic and non-transgenic soybean plants
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The Evolution, Ecology, and Mechanisms of Infection by Gram-Positive, Plant-Associated Bacteria
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Green Technology: Bacteria-Based Approach Could Lead to Unsuspected Microbe–Plant–Animal Interactions
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Insight Into the Microbial Co-occurrence and Diversity of 73 Grapevine (Vitis vinifera) Crown Galls Collected Across the Northern Hemisphere
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Isolation of fruit colonizer yeasts and screening against mango decay caused by multiple pathogens
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Diversity and Applications of Endophytic Actinobacteria of Plants in Special and Other Ecological Niches
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Grape microbiome as a reliable and persistent signature of field origin and environmental conditions in Cannonau wine production
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Microbial diversity in leaves, trunk and rhizosphere of coconut palms (Cocos nucifera L.) associated with the coconut lethal yellowing phytoplasma in Grand-Lahou, Cte dIvoire
P. Morales-Lizcano Nadia, Hasan Ahmed, S. To Henry, T. Lekadou Tacra, Copeland Julia, Wang Pauline, A. Diallo Hortense, L. Konan Konan Jean, Yoshioka Keiko, Moeder Wolfgang, Scott James, Arocha Rosete Yaima
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Identification and Characterizations of Potential Indigenous Endophytic Bacteria which Had Ability to Promote Growth Rate of Tomato and Biocontrol Agents of Ralstonia solanacearum and Fusarium oxysporum fsp. solani
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Leaf Treatments with a Protein-Based Resistance Inducer Partially Modify Phyllosphere Microbial Communities of Grapevine
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Diversity of bacterial endophytes in 3 and 15 year-old grapevines of Vitis vinifera cv. Corvina and their potential for plant growth promotion and phytopathogen control
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Endophytic bacteria with antagonistic traits inhabit the wood tissues of grapevines from Tunisian vineyards
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Whole-Metagenome-Sequencing-Based Community Profiles of Vitis vinifera L. cv. Corvina Berries Withered in Two Post-harvest Conditions
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Frontiers in Microbiology.2016;[Epub] CrossRef
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Enrico Baldan, Sebastiano Nigris, Chiara Romualdi, Stefano D’Alessandro, Anna Clocchiatti, Michela Zottini, Piergiorgio Stevanato, Andrea Squartini, Barbara Baldan, Peter Mergaert
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Pyrosequencing detects human and animal pathogenic taxa in the grapevine endosphere
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Dark Septate Endophyte (DSE) Fungi Isolated from Metal Polluted Soils: Their Taxonomic Position, Tolerance, and Accumulation of Heavy Metals In Vitro: Yujie Zhang , Yan Zhang , Maojun Liu , Xiaodong Shi , Zhiwei Zhao; J. Microbiol. 2008;46(6):624-632. Published online December 24, 2008; DOI: https://doi.org/10.1007/s12275-008-0163-6

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Abstract: To understand the possible role of the plant root associated fungi on metal tolerance, their role in the uptake of heavy metals and the potential transfer of these metal ions to the plant, three strains of dark septate endophytic (DSE) fungi were isolated from a waste smelter site in southwest China, and one strain was isolated from a non-contaminated site. According to molecular phylogenetic analysis of the ITS 1-5.8S rDNA-ITS 2 gene regions and morphological characteristics, one is identified as Exophiala pisciphila, and the other three are non-sporulating fungi under the experiment condition with the nearest phylogenetic affinities to the Thysanorea papuana strain EU041814. Tolerance and accumulation abilities of the three DSE strains for metals were investigated in liquid culture. Minimum inhibitory concentrations (MIC) of Pb, Zn, and Cd were determined. It was demonstrated that the tolerance of the DSE strains varied between metal species and strains. The E. pisciphila strain is able to accumulate lead and cadmium over 20% and 5% of dry weight of biomass, respectively. Partial of the sequestrated metals can be washed with CaCl2. Morphological and enzyme activity changes taking place in the presence of excessive Pb, Cd, and/or Zn also indicate that the mechanism of heavy metal tolerance and accumulation of the DSE strains would be a complex process. The findings indicated promising tolerance and accumulation of the DSE strains with potential values in metal cycling and restoration of soil and water system.

Diversity of Endophytic Enterobacteria Associated with Different Host Plants: Adalgisa Ribeiro Torres , Welington Luiz Araujo , Luciana Cursino , Mariangela Hungria , Fabio Plotegher , Fabio Luis Mostasso , Joao Lucio Azevedo; J. Microbiol. 2008;46(4):373-379. Published online August 31, 2008; DOI: https://doi.org/10.1007/s12275-007-0165-9

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Abstract: Fifty-three endophytic enterobacteria isolates from citrus, cocoa, eucalyptus, soybean, and sugar cane were evaluated for susceptibility to the antibiotics ampicillin and kanamycin, and cellulase production. Susceptibility was found on both tested antibiotics. However, in the case of ampicillin susceptibility changed according to the host plant, while all isolates were susceptible to kanamycin. Cellulase production also changed according to host plants. The diversity of these isolates was estimated by employing BOX-PCR genomic fingerprints and 16S rDNA sequencing. In total, twenty-three distinct operational taxonomic units (OTUs) were identified by employing a criterion of 60% fingerprint similarity as a surrogate for an OTU. The 23 OTUs belong to the Pantoea and Enterobacter genera, while their high diversity could be an indication of paraphyletic classification. Isolates representing nine different OTUs belong to Pantoea agglomerans, P. ananatis, P. stewartii, Enterobacter sp., and E. homaechei. The results of this study suggest that plant species may select endophytic bacterial genotypes. It has also become apparent that a review of the Pantoea/Enterobacter genera may be necessary.

Evaluation of Endophytic Colonization of Citrus sinensis and Catharanthus roseus Seedlings by Endophytic Bacteria: Paulo Teixeira Lacava , Welington Luiz Araujo , Joao Lucio Azevedo; J. Microbiol. 2007;45(1):11-14.; DOI: https://doi.org/2498 [pii]

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Abstract: Over the last few years, the endophytic bacterial community associated with citrus has been studied as an important component interacting with Xylella fastidiosa, the causal agent of citrus variegated chlorosis (CVC). This bacterium may also colonize some model plants, such as Catharanthus roseus and Nicotiana clevelandii. In the present study, we compared the endophytic colonization of Citrus sinensis and Catharanthus roseus using the endophytic bacteria Klebsiella pneumoniae. We chose an appropriate strain, K. pneumoniae 342 (Kp342), labeled with the GFP gene. This strain was inoculated onto seedlings of C. sinensis and C. roseus. The isolation frequency was determined one week after the inoculation and the endophytic colonization of K. pneumoniae was observed using fluorescence microscopy. Although the endophytic bacterium was more frequently isolated from C. roseus than from C. sinensis, the colonization profiles for both host plants were similar, suggesting that C. roseus could be used as a model plant to study the interaction between endophytic bacteria and X. fastidiosa.

Impact of Genetically Modified Enterobacter cloacae on Indigenous Endophytic Community of Citrus sinensis Seedlings: Fernando Dini Andreote , Marcelo Jose Mortatti Gullo , Andre Oliveira de Souza Lima , Walter Maccheroni Junior , Joao Lucio Azevedo , Welington Luiz Araujo; J. Microbiol. 2004;42(3):169-173.

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Abstract: Enterobacter cloacae (strain PR2/7), a genetically modified endophyte (GME) in citrus plants, carrying different plasmids (pEC3.0/18, pCelE, pEglA and pGFP), was inoculated into Citrus sinensis seedlings under greenhouse conditions. The impact of this on the indigenous bacterial endophytic community was studied by analyses of 2 different morphologic groups. The germination rates of inoculated seeds were evaluated in greenhouse, and plasmid stability under in vitro conditions. Results demonstrated a great and diverse endophytic community inside plants, and specialization in tissue colonization by some bacterial groups, in different treatments. Shifts in seed germination rate were observed among treatments: in general, the PR2/7 harboring pEglA bacterial clone significantly reduced seed germination, compared to the PR2/7 harboring pEC3.0/18 clone. This suggests that the presence of the pEglA plasmid changes bacteria-seed interactions. The endophytic community of citrus seedlings changed according to treatment. In seedlings treated with the PR2/7 with pEglA clone, the population of group II decreased significantly, within the context of the total endophytic community. These results indicate that the application of GMEs induces shifts in the endophytic bacterial community of citrus seedlings.

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