DOCSLIB.ORG

Diversity of Free-Living Nitrogen Fixing Bacteria in the Badlands of South Dakota Bibha Dahal South Dakota State University

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Diversity of Free-Living Nitrogen Fixing Bacteria in the Badlands of South Dakota Bibha Dahal South Dakota State University South Dakota State University Open PRAIRIE: Open Public Research Access Institutional Repository and Information Exchange Theses and Dissertations 2016 Diversity of Free-living Nitrogen Fixing Bacteria in the Badlands of South Dakota Bibha Dahal South Dakota State University Follow this and additional works at: http://openprairie.sdstate.edu/etd Part of the Bacteriology Commons, and the Environmental Microbiology and Microbial Ecology Commons Recommended Citation Dahal, Bibha, "Diversity of Free-living Nitrogen Fixing Bacteria in the Badlands of South Dakota" (2016). Theses and Dissertations. 688. http://openprairie.sdstate.edu/etd/688 This Thesis - Open Access is brought to you for free and open access by Open PRAIRIE: Open Public Research Access Institutional Repository and Information Exchange. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of Open PRAIRIE: Open Public Research Access Institutional Repository and Information Exchange. For more information, please contact [email protected]. DIVERSITY OF FREE-LIVING NITROGEN FIXING BACTERIA IN THE BADLANDS OF SOUTH DAKOTA BY BIBHA DAHAL A thesis submitted in partial fulfillment of the requirements for the Master of Science Major in Biological Sciences Specialization in Microbiology South Dakota State University 2016 iii ACKNOWLEDGEMENTS “Always aim for the moon, even if you miss, you’ll land among the stars”.- W. Clement Stone I would like to express my profuse gratitude and heartfelt appreciation to my advisor Dr. Volker Brӧzel for providing me a rewarding place to foster my career as a scientist. I am thankful for his implicit encouragement, guidance, and support throughout my research. This research would not be successful without his guidance and inspiration. I would like to thank all my POS Committee for supporting me in various stages of my research and encouraging me to achieve my goal swiftly. My sincere thanks goes to my committee members Dr. Lora Perkins and Dr. Senthil Subramanian for their assistance, support, and direction when I needed. I would like to thank all the faculties and staffs of Department of Biology and Microbiology who have directly or indirectly supported me to achieve this goal. I am thankful to my lab mate Gitanjali Nanda Kafle and my friend Tyrel Deutscher who have assisted me in my research in various ways. The conversations we had will remain in my memories. I would like to acknowledge the funding source South Dakota Agricultural Experiment Station for its financial support. Last but not the least, I would like to thank my husband Nirmal Paudel for his encouragement and moral support throughout my study period. My acknowledgement goes to my sister Subha Dahal and my parents who have been very supportive and encouraging. iv TABLE OF CONTENTS ABBREVIATIONS………………………………………………………….viii LIST OF FIGURES……………………………………………………….…..xi LIST OF TABLES…………………………………………………………...xiv ABSTRACT………………………………………………………………….xv Chapter 1: Literature Review 1.1 Introduction .................................................................................................. 1 1.1.1 Biological Nitrogen Fixation ..................................................................... 2 1.1.2 Nitrogen cycle ............................................................................................ 2 1.2 The Nitrogenase enzyme .............................................................................. 4 1.2.1. The Molybdenum containing nitrogenase ................................................. 4 1.2.2. The Vanadium nitrogenase ....................................................................... 8 1.2.3. The Iron-only nitrogenase………………………………………………10 1.2.4. Oxygen insensitive Streptomyces thermoautotrophicus nitrogenase ...... 11 1.3 Diversity of Nitrogen fixing bacteria ......................................................... 12 1.3.1. Life styles of nitrogen fixers ................................................................... 13 1.3.1.1. Symbiotic association .......................................................................... 13 1.3.1.2. Associative nitrogen fixing bacteria .................................................... 14 1.3.1.3. Free-living nitrogen fixers .................................................................... 14 v 1.3.2. Origins and evolution of nitrogenases .................................................... 15 1.4. N2 fixation in arid lands ............................................................................ 17 1.5. Methods for Evaluating of Nitrogen Fixation ........................................... 18 1.5.1. Culture Dependent Methods ................................................................... 18 1.5.2. Culture Independent Methods ................................................................. 19 1.5.2.1. The 16S rRNA gene ............................................................................. 20 1.5.2.2. The nifH gene to study the diversity of diazotrophs ............................ 22 1.5.2.3. Denaturing Gradient Gel Electrophoresis (DGGE) ............................. 24 1.5.3. The Acetylene Reduction Assay ............................................................. 25 15 1.5.4. Detecting nitrogen incorporation using stable isotope N2 .................... 26 1.6. Aim of Study ............................................................................................. 28 Chapter 2: Diversity of free-living nitrogen fixing bacteria in the Badlands of South Dakota………………………………………………………………….30 2.1. Introduction ............................................................................................... 30 2.2. Materials and Methods .............................................................................. 33 2.2.1. Study Area ............................................................................................... 33 2.2.2. Sampling.................................................................................................. 33 2.2.3. Isolation of free-living putative diazotrophs from soil. ........................... 35 2.2.4. DNA Extraction and PCR Amplification of 16S rRNA and nifH from pure cultures ...................................................................................................... 36 vi 2.2.5. DNA Extraction and PCR Amplification of nifH gene from soil samples and DGGE analysis: .......................................................................................... 38 2.2.6. Determination of nifH sequences in soil samples ................................... 38 2.2.7. PCR amplification of Cyanobacterial 16S rRNA genes from soil samples ................................................................................................................. 39 2.2.8. Acetylene Reduction Assay…………………………………………..39 15 2.2.9. Incorporation of N2 isotope in pure cultures ........................................ 40 2.2.10. Confirming nitrogen fixation in an ammonia free atmosphere ............. 40 2.2.11 Determination of 16S rRNA gene sequence of putative Streptomyces isolates……………........................................................................................... 41 2.3. Results ....................................................................................................... 42 2.3.1 Diversity of free-living putative nitrogen fixing bacteria ....................... 42 2.3.2 The nifH gene amplification and growth in liquid NFM with clinoptilolite…………………………………………………………………...45 2.3.3 V1-9 region 16S rRNA gene amplification of isolates allocated to Streptomyces ..................................................................................................... 47 2.3.4. DGGE analysis of microbial community and sequencing of cloned DNA ................................................................................................................. 49 2.3.5. Nitrogen fixation by bacterial isolates……………………………...…..49 2.4. Discussion ............................................................................................... 51 Chapter 3: Fungal isolates on Nitrogen Free Medium…...…………………...55 vii 3.1 Introduction……………………………………………………………….55 3.2 Materials and Methods……………………………………………………57 3.2.1. Isolation of microbial communities from rhizosphere, root, and stem...57 3.2.2. DNA Extraction and PCR Amplification of internal transcribed spacer (ITS) region, 16S rRNA gene and nifH gene…………………………………58 3.2.3. Culture of fungi like isolates on various media………...…….………..59 3.2.4. Isolation of bacterial endophytes………………………………………60 3.2.5. Microscopy…………………………………………………………….60 15 3.2.6. Incorporation of N2 isotope in pure cultures…………………………60 3.3. Results…………………………………………………………………...61 3.4. Discussion……………………………………………………………….64 Chapter 4: Conclusions and Question for future study………………………67 4.1.Conclusion………………………………………………………………..67 4.2.Question for future study…………………………………………………68 References…………………………………………………………………….70 Appendix……………………………………………………………………...99 viii ABBREVIATIONS α: Alpha β: Beta °C: Degrees Celsius µ: Micro ADP: Adenosine Di-Phosphate ARA: Acetylene Reduction Assay ATP: Adenosine Tri- Phosphate BLAST: Basic Local Alignment Search Tool BNF: Biological Nitrogen Fixation bp: base pair BSC: Biological Soil Crust δ: Delta Da: Dalton DGGE: Denaturing Gradient Gel Electrophoresis DNA: Deoxyribonucleic acid dNTP: Deoxyribonucleic triphosphate EPR: Electron paramagnetic resonance ix EXAFS: Extended X-ray Absorption Fine Structure HGT: Horizontal Gene Transfer ITS: Internal Transcribed Spacer kDa: Kilo Dalton
Load more

Recommended publications
  • Bacteria Belonging to Pseudomonas Typographi Sp. Nov. from the Bark Beetle Ips Typographus Have Genomic Potential to Aid in the Host Ecology
    insects Article Bacteria Belonging to Pseudomonas typographi sp. nov. from the Bark Beetle Ips typographus Have Genomic Potential to Aid in the Host Ecology Ezequiel Peral-Aranega 1,2 , Zaki Saati-Santamaría 1,2 , Miroslav Kolaˇrik 3,4, Raúl Rivas 1,2,5 and Paula García-Fraile 1,2,4,5,* 1 Microbiology and Genetics Department, University of Salamanca, 37007 Salamanca, Spain; [email protected] (E.P.-A.); [email protected] (Z.S.-S.); [email protected] (R.R.) 2 Spanish-Portuguese Institute for Agricultural Research (CIALE), 37185 Salamanca, Spain 3 Department of Botany, Faculty of Science, Charles University, Benátská 2, 128 01 Prague, Czech Republic; [email protected] 4 Laboratory of Fungal Genetics and Metabolism, Institute of Microbiology of the Academy of Sciences of the Czech Republic, 142 20 Prague, Czech Republic 5 Associated Research Unit of Plant-Microorganism Interaction, University of Salamanca-IRNASA-CSIC, 37008 Salamanca, Spain * Correspondence: [email protected] Received: 4 July 2020; Accepted: 1 September 2020; Published: 3 September 2020 Simple Summary: European Bark Beetle (Ips typographus) is a pest that affects dead and weakened spruce trees. Under certain environmental conditions, it has massive outbreaks, resulting in attacks of healthy trees, becoming a forest pest. It has been proposed that the bark beetle’s microbiome plays a key role in the insect’s ecology, providing nutrients, inhibiting pathogens, and degrading tree defense compounds, among other probable traits. During a study of bacterial associates from I. typographus, we isolated three strains identified as Pseudomonas from different beetle life stages. In this work, we aimed to reveal the taxonomic status of these bacterial strains and to sequence and annotate their genomes to mine possible traits related to a role within the bark beetle holobiont.
    [Show full text]
  • Response of Azotobacter , Pseudomonas and Trichoderma on Growth of Apple Seedling
    2012 International Conference on Biological and Life Sciences IPCBEE vol.40 (2012) © (2012) IACSIT Press, Singapore Response of Azotobacter , Pseudomonas and Trichoderma on Growth of Apple Seedling + Jayant Raman Department of Botany and Microbiology, Gurukul Kangri Vishwavidyalaya, Hardwar-249404, Uttarakhand Abstract. The investigation on the efficacy of biofertilizers (Azotobacter chroococcum, Pseudomonas striata and Trichoderma viride) considering different aspects of seedling growth under apple nursery conditions enabled to reach the positive conclusions. Azotobacter chroococcum and Trichoderma viride increased the maximum germination per cent of seeds while the maximum survival of seedlings was observed on inoculation with Azotobacter chroococcum and Trichoderma viride. Azotobacter chroococcum and Pseudomonas striata was found beneficial in promoting the growth of seedling by increasing the growth rate of length and diameter of seedlings. Trichoderma viride also helped against the pests and diseases at the same time provided sufficient moisture in dry spell. The highest growth rate of length and diameter of seedling was observed on inoculation with Azotobacter chroococcum + Pseudomonas striata + Trichoderma viride. The maximum initiation and increase rate in production of number of leaves was observed on inoculation with Azotobacter chroococcum and Pseudomonas striata. This treatment also produced large sized leaves. The highest content of N,P,K, Zn and Cu in leaves was analyzed on inoculation with Azotobacter chroococcum + Pseudomonas striata + Trichoderma viride. The highest reduction of the pests attack was observed on inoculation with Pseudomonas striata and Trichoderma viride, while the infestation was least in seedling due to antifungal fungistatic organic compounds and toxins on inoculation with Azotobacter chroococcum + Pseudomonas striata + Trichoderma viride. The good growth of root in seedlings was observed on inoculation with Azotobacter chroococcum + Pseudomonas striata + Trichoderma viride.
    [Show full text]
  • Archaea, Bacteria and Termite, Nitrogen Fixation and Sustainable Plants Production
    Sun W et al . (2021) Notulae Botanicae Horti Agrobotanici Cluj-Napoca Volume 49, Issue 2, Article number 12172 Notulae Botanicae Horti AcademicPres DOI:10.15835/nbha49212172 Agrobotanici Cluj-Napoca Re view Article Archaea, bacteria and termite, nitrogen fixation and sustainable plants production Wenli SUN 1a , Mohamad H. SHAHRAJABIAN 1a , Qi CHENG 1,2 * 1Chinese Academy of Agricultural Sciences, Biotechnology Research Institute, Beijing 100081, China; [email protected] ; [email protected] 2Hebei Agricultural University, College of Life Sciences, Baoding, Hebei, 071000, China; Global Alliance of HeBAU-CLS&HeQiS for BioAl-Manufacturing, Baoding, Hebei 071000, China; [email protected] (*corresponding author) a,b These authors contributed equally to the work Abstract Certain bacteria and archaea are responsible for biological nitrogen fixation. Metabolic pathways usually are common between archaea and bacteria. Diazotrophs are categorized into two main groups namely: root- nodule bacteria and plant growth-promoting rhizobacteria. Diazotrophs include free living bacteria, such as Azospirillum , Cupriavidus , and some sulfate reducing bacteria, and symbiotic diazotrophs such Rhizobium and Frankia . Three types of nitrogenase are iron and molybdenum (Fe/Mo), iron and vanadium (Fe/V) or iron only (Fe). The Mo-nitrogenase have a higher specific activity which is expressed better when Molybdenum is available. The best hosts for Rhizobium legumiosarum are Pisum , Vicia , Lathyrus and Lens ; Trifolium for Rhizobium trifolii ; Phaseolus vulgaris , Prunus angustifolia for Rhizobium phaseoli ; Medicago, Melilotus and Trigonella for Rhizobium meliloti ; Lupinus and Ornithopus for Lupini, and Glycine max for Rhizobium japonicum . Termites have significant key role in soil ecology, transporting and mixing soil. Termite gut microbes supply the enzymes required to degrade plant polymers, synthesize amino acids, recycle nitrogenous waste and fix atmospheric nitrogen.
    [Show full text]
  • Host-Secreted Antimicrobial Peptide Enforces Symbiotic Selectivity in Medicago Truncatula
    Host-secreted antimicrobial peptide enforces symbiotic selectivity in Medicago truncatula Qi Wanga, Shengming Yanga, Jinge Liua, Kata Terecskeib, Edit Ábrahámb, Anikó Gombárc, Ágota Domonkosc, Attila Szucs} b, Péter Körmöczib, Ting Wangb, Lili Fodorc, Linyong Maod,e, Zhangjun Feid,e, Éva Kondorosib,1, Péter Kalóc, Attila Keresztb, and Hongyan Zhua,1 aDepartment of Plant and Soil Sciences, University of Kentucky, Lexington, KY 40546; bInstitute of Biochemistry, Biological Research Center, Szeged 6726, Hungary; cNational Agricultural Research and Innovation Centre, Agricultural Biotechnology Institute, Gödöllo} 2100, Hungary; dBoyce Thompson Institute for Plant Research, Cornell University, Ithaca, NY 14853; and eU.S. Department of Agriculture–Agricultural Research Service Robert W. Holley Center for Agriculture and Health, Cornell University, Ithaca, NY 14853 Contributed by Éva Kondorosi, February 14, 2017 (sent for review January 17, 2017; reviewed by Rebecca Dickstein and Julia Frugoli) Legumes engage in root nodule symbioses with nitrogen-fixing effectors or microbe-associated molecular patterns (MAMPs) soil bacteria known as rhizobia. In nodule cells, bacteria are enclosed such as surface polysaccharides to facilitate their invasion of the in membrane-bound vesicles called symbiosomes and differentiate host (7, 8). Therefore, effector- or MAMP-triggered plant im- into bacteroids that are capable of converting atmospheric nitrogen munity mediated by intracellular nucleotide binding/leucine-rich into ammonia. Bacteroid differentiation
    [Show full text]
  • Methanotrophs Are Core Members of the Diazotroph Community in Decaying T Norway Spruce Logs
    Soil Biology and Biochemistry 120 (2018) 230–232 Contents lists available at ScienceDirect Soil Biology and Biochemistry journal homepage: www.elsevier.com/locate/soilbio Short Communication Methanotrophs are core members of the diazotroph community in decaying T Norway spruce logs ∗ Raisa Mäkipääa, , Sanna M. Leppänena, Sonia Sanz Munoza, Aino Smolandera, Marja Tiirolab, Tero Tuomivirtaa, Hannu Fritzea a Natural Resources Institute Finland, Finland b University of Jyväskylä, Finland ARTICLE INFO ABSTRACT Keywords: Dead wood is initially a nitrogen (N) poor substrate, where the N content increases with decay, partly due to fi Asymbiotic nitrogen xation biological N2 fixation, but the drivers of the N accumulation are poorly known. We quantified the rate of N2 Coarse woody debris fixation in decaying Norway spruce logs of different decay stages and studied the potential regulators of the N2- − − Dead wood fixation activity. The average rate for acetylene reduction in the decaying wood was 7.5 nmol ethylene g 1d 1, nifH − − − which corresponds to 52.9 μgN kg 1d 1. The number of nifH copies (g 1 dry matter) was higher at the later Picea abies decay stages, but no correlation between the copy number and the in vitro N2 fixation rate was found. All recovered nifH sequences were assigned to the order Rhizobiales, and therein mostly (60%) to methane oxidizing genera. We confirm that nitrogen fixing methanotrophs are present in all the wood decay phases and suggest that their interaction between methane producing organisms in decaying wood should be further studied. In boreal forests with nitrogen (N) supply deficiency, asymbiotic N2 (Karst)) logs (using four replicates per sample log) followed the fixation occurs in the dead wood (Brunner and Kimmins, 2003; Hicks methods described in Rinne et al.
    [Show full text]
  • Electron Flow and Management in Living Systems: Advancing Understanding of Electron Transfer to Nitrogenase
    Utah State University DigitalCommons@USU All Graduate Theses and Dissertations Graduate Studies 8-2018 Electron Flow and Management in Living Systems: Advancing Understanding of Electron Transfer to Nitrogenase Rhesa N. Ledbetter Utah State University Follow this and additional works at: https://digitalcommons.usu.edu/etd Part of the Biochemistry Commons Recommended Citation Ledbetter, Rhesa N., "Electron Flow and Management in Living Systems: Advancing Understanding of Electron Transfer to Nitrogenase" (2018). All Graduate Theses and Dissertations. 7197. https://digitalcommons.usu.edu/etd/7197 This Dissertation is brought to you for free and open access by the Graduate Studies at DigitalCommons@USU. It has been accepted for inclusion in All Graduate Theses and Dissertations by an authorized administrator of DigitalCommons@USU. For more information, please contact [email protected]. ELECTRON FLOW AND MANAGEMENT IN LIVING SYSTEMS: ADVANCING UNDERSTANDING OF ELECTRON TRANSFER TO NITROGENASE by Rhesa N. Ledbetter A dissertation submitted in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY in Biochemistry Approved: ______________________ ______________________ Lance C. Seefeldt, Ph.D. Scott A. Ensign, Ph.D. Biochemistry Biochemistry Major Professor Committee Member ______________________ ______________________ Bruce Bugbee, Ph.D. Sean J. Johnson, Ph.D. Plant Physiology Biochemistry Committee Member Committee Member ______________________ ______________________ Nicholas E. Dickenson, Ph.D. Mark R. McLellan, Ph.D. Biochemistry Vice President for Research and Committee Member Dean of the School of Graduate Studies UTAH STATE UNIVERSITY Logan, Utah 2018 ii Copyright © Rhesa N. Ledbetter 2018 All Rights Reserved iii ABSTRACT Electron Flow and Management in Living Systems: Advancing Understanding of Electron Transfer to Nitrogenase by Rhesa N.
    [Show full text]
  • 0041085-15082018101610.Pdf
    Cronfa - Swansea University Open Access Repository _____________________________________________________________ This is an author produced version of a paper published in: The Journal of Antibiotics Cronfa URL for this paper: http://cronfa.swan.ac.uk/Record/cronfa41085 _____________________________________________________________ Paper: Zhang, B., Tang, S., Chen, X., Zhang, G., Zhang, W., Chen, T., Liu, G., Li, S., Dos Santos, L., et. al. (2018). Streptomyces qaidamensis sp. nov., isolated from sand in the Qaidam Basin, China. The Journal of Antibiotics http://dx.doi.org/10.1038/s41429-018-0080-9 _____________________________________________________________ This item is brought to you by Swansea University. Any person downloading material is agreeing to abide by the terms of the repository licence. Copies of full text items may be used or reproduced in any format or medium, without prior permission for personal research or study, educational or non-commercial purposes only. The copyright for any work remains with the original author unless otherwise specified. The full-text must not be sold in any format or medium without the formal permission of the copyright holder. Permission for multiple reproductions should be obtained from the original author. Authors are personally responsible for adhering to copyright and publisher restrictions when uploading content to the repository. http://www.swansea.ac.uk/library/researchsupport/ris-support/ Streptomyces qaidamensis sp. nov., isolated from sand in the Qaidam Basin, China Binglin Zhang1,2,3, Shukun Tang4, Ximing Chen1,3, Gaoseng Zhang1,3, Wei Zhang1,3, Tuo Chen2,3, Guangxiu Liu1,3, Shiweng Li3,5, Luciana Terra Dos Santos6, Helena Carla Castro6, Paul Facey7, Matthew Hitchings7 and Paul Dyson7 1 Key Laboratory of Desert and Desertification, Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences, Lanzhou 730000, China.
    [Show full text]
  • Diversity and Activity of Sulfate-Reducing Bacteria In
    Diversity and Activity of Sulfate- reducing bacteria in Sulfidogenic Wastewater Treatment Reactors If we knew what we are doing, it would not be called research would it? Albert Einstien ii Diversity and Activity of Sulfate- reducing bacteria in Sulfidogenic Wastewater Treatment Reactors Proefschrift ter verkrijging van de graad van doctor aan de Technische Universiteit Delft, op gezag van de Rector Magnificus Prof. dr. ir. J. T. Fokkema, voorzitter van het College voor Promoties, in het openbaar te verdedigen op vrijdag 19 oktober 2007 om 10.00 uur door Shabir Ahmad DAR Master in Science of Bioprocess Technology, Asian Institute of Technology (AIT), Thailand geboren te Srinagar, J&K, India iii Dit proefschrift is goedgekeurd door de promotor: Prof. dr. J. G. Kuenen Toegevoegd promotor Dr. G. Muyzer Samenstelling promotie commissie: Rector Magnificus Voorzitter Prof. dr. J.G. Kuenen Delft University of Technology, Promotor Dr. G. Muyzer Delft University of Technology, Toegevoegd promotor Prof. dr. F. Widdel Max-Planck-Institute for Marine Microbiology, Bremen, Germany Prof. dr. ir. M.C.M. van Loosdrecht Delft University of Technology Prof. dr. H.J. Laanbroek Utrecht University Prof. dr. ir.A.J.M. Stams Wageningen University Prof. dr. ir. P.N.L. Lens Wageningen University This study was carried out in the Environmental Biotechnology group of the Department of Biotechnology at Delft University of Technology, Delft, the Netherlands. This research was financially supported by The Netherlands Organization for Scientific Research – (NWO Earth
    [Show full text]
  • Diversity and Characterization of Azotobacter Isolates Obtained from Rice Rhizosphere Soils in Taiwan
    Ann Microbiol (2018) 68:17–26 https://doi.org/10.1007/s13213-017-1312-0 ORIGINAL ARTICLE Diversity and characterization of Azotobacter isolates obtained from rice rhizosphere soils in Taiwan Syuan-Lu Chen 1 & Meng-Ke Tsai1 & Yuh-Ming Huang1 & Cheng-Hua Huang1 Received: 4 March 2017 /Accepted: 7 November 2017 /Published online: 15 December 2017 # Springer-Verlag GmbH Germany, part of Springer Nature and the University of Milan 2017 Abstract Azotobacter species, free-living nitrogen-fixing A. chroococcum CHB 869 may be used to develop bacteria, have been used as biofertilizers to improve the pro- biofertilizers for rice cultivation because they significantly ductivity of non-leguminous crops, including rice, due to their promoted rice growth. This study contributes to the selection various plant growth-promoting traits. The purposes of this of suitable Azotobacter strains for developing biofertilizer for- study were to characterize Azotobacter species isolated from mulations and soil management strategies of Azotobacter for rice rhizospheres in Taiwan and to determine the relationship paddy fields. between the species diversity of Azotobacter and soil proper- ties. A total of 98 Azotobacter isolates were isolated from 27 Keywords Nitrogen-fixing bacteria . Rice . Diazotropic paddy fields, and 16S rRNA gene sequences were used to bacteria . Nitrogenase . Diversity identify Azotobacter species. The characteristics of these Azotobacter strains were analyzed including carbon source utilization and plant growth-promoting traits such as nitrogen Introduction fixation activity, indole acetic acid production, phosphate- solubilizing ability, and siderophore secretion. Of the 98 Rice is one of the main food crops in the world, being con- strains isolated in this study, 12 were selected to evaluate their sumed by about 50% of world and 85% of Asian populations effects on rice growth.
    [Show full text]
  • Human Alteration of the Global Nitrogen Cycle
    What is Nitrogen? Human Alteration of the Nitrogen is the most abundant element in Global Nitrogen Cycle the Earth’s atmosphere. Nitrogen makes up 78% of the troposphere. Nitrogen cannot be absorbed directly by the plants and animals until it is converted into compounds they can use. This process is called the Nitrogen Cycle. Heather McGraw, Mandy Williams, Suzanne Heinzel, and Cristen Whorl, Give SIUE Permission to Put Our Presentation on E-reserve at Lovejoy Library. The Nitrogen Cycle How does the nitrogen cycle work? Step 1- Nitrogen Fixation- Special bacteria convert the nitrogen gas (N2 ) to ammonia (NH3) which the plants can use. Step 2- Nitrification- Nitrification is the process which converts the ammonia into nitrite ions which the plants can take in as nutrients. Step 3- Ammonification- After all of the living organisms have used the nitrogen, decomposer bacteria convert the nitrogen-rich waste compounds into simpler ones. Step 4- Denitrification- Denitrification is the final step in which other bacteria convert the simple nitrogen compounds back into nitrogen gas (N2 ), which is then released back into the atmosphere to begin the cycle again. How does human intervention affect the nitrogen cycle? Nitric Oxide (NO) is released into the atmosphere when any type of fuel is burned. This includes byproducts of internal combustion engines. Production and Use of Nitrous Oxide (N2O) is released into the atmosphere through Nitrogen Fertilizers bacteria in livestock waste and commercial fertilizers applied to the soil. Removing nitrogen from the Earth’s crust and soil when we mine nitrogen-rich mineral deposits. Discharge of municipal sewage adds nitrogen compounds to aquatic ecosystems which disrupts the ecosystem and kills fish.
    [Show full text]
  • Genome-Wide Identification of Azospirillum Brasilense Sp245
    Koul et al. BMC Genomics (2020) 21:821 https://doi.org/10.1186/s12864-020-07212-7 RESEARCH ARTICLE Open Access Genome-wide identification of Azospirillum brasilense Sp245 small RNAs responsive to nitrogen starvation and likely involvement in plant-microbe interactions Vatsala Koul1,2†, Divya Srivastava2†, Pushplata Prasad Singh2* and Mandira Kochar2* Abstract Background: Small RNAs (sRNAs) are non-coding RNAs known to regulate various biological functions such as stress adaptation, metabolism, virulence as well as pathogenicity across a wide range of bacteria, mainly by controlling mRNA stabilization or regulating translation. Identification and functional characterization of sRNAs has been carried out in various plant growth-promoting bacteria and they have been shown to help the cells cope up with environmental stress. No study has been carried out to uncover these regulatory molecules in the diazotrophic alpha-proteobacterium Azospirillum brasilense Sp245 to date. Results: Expression-based sRNA identification (RNA-seq) revealed the first list of ~ 468 sRNA candidate genes in A. brasilense Sp245 that were differentially expressed in nitrogen starvation versus non-starved conditions. In parallel, in silico tools also identified 2 of the above as candidate sRNAs. Altogether, putative candidates were stringently curated from RNA-seq data based on known sRNA parameters (size, location, secondary structure, and abundance). In total, ~ 59 significantly expressed sRNAs were identified in this study of which 53 are potentially novel sRNAs as they have no Rfam and BSRD homologs. Sixteen sRNAs were randomly selected and validated for differential expression, which largely was found to be in congruence with the RNA-seq data. Conclusions: Differential expression of 468 A.
    [Show full text]
  • Respiration in Azotobacter Vinelandii and Its Relationship with the Synthesis of Biopolymers
    Electronic Journal of Biotechnology 48 (2020) 36–45 Contents lists available at ScienceDirect Electronic Journal of Biotechnology Review Respiration in Azotobacter vinelandii and its relationship with the synthesis of biopolymers Tania Castillo a,AndrésGarcíab, Claudio Padilla-Córdova c, Alvaro Díaz-Barrera c,CarlosPeñaa,⁎ a Departamento de Ingeniería Celular y Biocatálisis, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo. Post. 510-3, Cuernavaca 62250, Morelos, Mexico b Laboratorio de Biotecnología Ambiental, Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Av. Universidad 1001, Col. Chamilpa, Cuernavaca, Morelos 62209, Mexico c Escuela de Ingeniería Bioquímica, Facultad de Ingeniería, Pontificia Universidad Católica de Valparaíso, Av. Brasil 2147, Casilla 4059, Valparaíso, Chile article info abstract Article history: Azotobacter vinelandii is a gram-negative soil bacterium that produces two biopolymers of biotechnological Received 20 April 2020 interest, alginate and poly(3-hydroxybutyrate), and it has been widely studied because of its capability to fix Accepted 27 August 2020 nitrogen even in the presence of oxygen. This bacterium is characterized by its high respiration rates, which Available online 4 September 2020 are almost 10-fold higher than those of Escherichia coli and are a disadvantage for fermentation processes. On the other hand, several works have demonstrated that adequate control of the oxygen supply in A. vinelandii Keywords: cultivations determines the yields and physicochemical characteristics of alginate and poly(3-hydroxybutyrate). poly(3-hydroxybutyrate) Azotobacter vinelandii Here, we summarize a review of the characteristics of A. vinelandii related to its respiration systems, as well as Alginate some of the most important findings on the oxygen consumption rates as a function of the cultivation Biopolymers parameters and biopolymer production.
    [Show full text]