DOCSLIB.ORG

Evaluating the Diversity and Phylogeny of Plant Growth Promoting Bacteria Associated with Wheat (Triticum Aestivum) Growing in Central Zone of India

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Evaluating the Diversity and Phylogeny of Plant Growth Promoting Bacteria Associated with Wheat (Triticum Aestivum) Growing in Central Zone of India Int.J.Curr.Microbiol.App.Sci (2014) 3(5): 432-447 ISSN: 2319-7706 Volume 3 Number 5 (2014) pp. 432-447 http://www.ijcmas.com Original Research Article Evaluating the diversity and phylogeny of plant growth promoting bacteria associated with wheat (Triticum aestivum) growing in central zone of India Priyanka Verma1,2, Ajar Nath Yadav1, Sufia Khannam Kazy2, Anil Kumar Saxena1 and Archna Suman1* 1Division of Microbiology, Indian Agricultural Research Institute, New Delhi- 110012, India 2Department of Biotechnology, National Institute of Technology, Durgapur-713209, India *Corresponding author A B S T R A C T The diversity of plant growth promoting bacteria was investigated from wheat growing in different sites in central zone of India. Epiphytic, endophytic and rhizospheric bacteria were isolated using different growth medium. Bacterial diversity was analysed through amplified ribosomal DNA restriction analysis K e y w o r d s (ARDRA) using three restriction enzymes Alu I, Hae III, and Msp I which led to the grouping of 348 isolates into 24-29 clusters at >75% similarity index. 16S Epiphytic; rRNA gene based phylogenetic analysis, revealed that 134 strains belonged to three Endophytic; phyla namely actinobacteria, firmicutes and proteobacteria with 38 distinct species of 17 genera. Bacillus and Pseudomonas were dominant in rhizosphere while Rhizospheric; Methylobacterium were in phyllosphere. Endophytic niche specific bacteria were PGPB; identified as Delftia and Micrococcus. Sampling of different sites showed variation Drought in diversity indices. In vitro plant growth promoting activities of bacteria exposed stress; more than three beneficial traits which may act independently or concurrently. Biocontrol Phosphate solubilization and siderophores production are the predominant traits exhibited by these microbes. The many species of genera Bacillus, Exiguobacterium, Micrococcus, Pseudomonas and Psychrobacter showed antagonistic properties against fungal pathogens Fusarium graminerum, Rhizoctonia solani and Macrophomina phaseoli. These promising isolates showing a range of useful plant growth promoting attributes insist to be explored for agricultural applications. Introduction Wheat (Triticum durum L.) is one of the The central zone of India (Madhaya most important cereals world-wide and it Pradesh, Kota region of Rajasthan and is grown in different environments. Jhansi region of Uttar Pradesh) were Drought is one of the major constraints characterised for water stress ecosystem. which hamper wheat production in India. Plant growth promoting bacteria (PGPB) 432 Int.J.Curr.Microbiol.App.Sci (2014) 3(5): 432-447 are free-living soil, rhizospheric, epiphytic without causing substantive harm to the and endophytic bacteria that can either host. Bacterial endophytes such as directly or indirectly have an impact on Achromobacter, Microbiospora, plant growth (Glick et al. 1999; Verma et Micrococcus, Micromomospora, Pantoea, al. 2013). PGPB stimulate plant growth in Planomonospora, Pseudomonas, multiple ways viz N2 fixation, synthesize Stenotrophomonoas, Streptomyces and phytohormones (auxin and cytokinin), Thermomonospora have been reported production of siderophores and suppress from wheat (Zinniel et al. 2002; Coombs pathogenic organisms. PGPB has been and Franco 2003; Verma et al. 2013). A reported not only to improve plant growth number of bacterial species associated but also to suppress the plant pathogens, of with the wheat rhizosphere were recovered which Pseudomonas and Bacillus were belonging to genera Azospirillum, well characterised. Pink-pigmented Arthrobacter, Acinetobacter, Bacillus, facultative methylotrophs synthesize a Burkholderia, Enterobacter, Erwinia, variety of metabolites useful for the plants Flavobacterium, Methylobacterium, including phytohormones (Ivanova et al. Pseudomonas, Rhizobium and Serratia 2001; Verma et al. 2013) that promote (Xie et al. 1996; Lavania et al. 2006; plant growth and yield. PGPB are used as Chaiharn and Lumyong 2011; Meena et al. biological control agents to reduce the 2012; Verma et al. 2013). development of plant diseases caused by plant pathogenic fungi, bacteria, viruses The present study attempted to elucidate and nematodes. the bacterial diversity associated with wheat growing in central zone of India, In the last decade, a number of PGPB employing different growth media. associated with wheat and different cereals ARDRA analysis was done for crops have been identified including phylogenetic clustering of the moderately Acinetobacter, Arthrobacter, Azospirillum, drought tolerant isolates. Sequencing the Azotobacter, Bacillus, Burkholderia, 16S rRNA gene of representative strains Citricoccus, Kocuria, Lysinibacillus, was undertaken for identification. Methylobacterium, Paenibacillus, Representative strain from each cluster Providencia, Pseudomonas and Serratia was screen in vitro for plant growth (Coombs and Franco 2003; Conn and promotion in drought stress condition. Franco 2004; Streptomyces 2005; Jha and PGPB inoculants are inexpensive, simple Kumar 2009; Wellner et al. 2011; Meena to use and have no unpleasant effects to et al. 2012; Verma et al. 2013). The land. The use of PGPB may prove useful phyllosphere is common niche for in developing strategies to facilitate wheat synergism between bacteria and plant. and other cereals crops growth in drought Many bacteria such as Pseudomonas and area. Methylobacterium have been reported in the wheat phyllosphere (Wellner et al. Materials and Methods 2011; Meena et al. 2012; Verma et al. 2013). Rhizospheric bacteria have the Samples collection ability to attach to the root surfaces (rhizoplane) allowing these to derive Wheat plants of two var. IWP5007 and maximum benefit from root exudates. HD2987 with rhizospheric soil were Endophytic bacteria live in plant tissues collected from five different sites in 433 Int.J.Curr.Microbiol.App.Sci (2014) 3(5): 432-447 central zone of India, which included PCR amplification of 16S rDNA and Gwalior, Sagar and Indore region of amplified rDNA restriction analysis Madhya Pradesh, Jhansi region of Uttar (ARDRA) Pradesh and Kota region of Rajasthan (Table 1). A total forty samples, eight Genomic DNA was extracted by the from each site were collected in sterile method as earlier described by Kumar et polythene bags labelled transported on ice al. (2013). The amount of DNA extracted and processes immediately. was electrophoresed on 0.8% agarose gel. Amplification of 16S rDNA was done by Physico-chemical properties of samples using the universal primers pA (5 - AGAGTTTGATCCTGGCTCAG-3 ) and The pH and conductivity of the soil pH (5 -AAGGAGGTGATCCAGCCGCA- samples were recorded on sampling site. 3 ). The amplification was carried out in a Soil samples analyzed for soil organic 100 l volume and amplification carbon, total nitrogen (%), soil organic conditions were used as described earlier matter, exchangeable cations and available (Pandey et al. 2013). The PCR amplified phosphorus was determined as described 16S rDNA were purified by QIA quick earlier Verma et al. (2013). Soil analysis PCR product purification kit (Qiagen). 100 was done at Division of Soil Sciences, ng purified PCR products were digested Indian Agricultural Research Institute, separately with three restriction New Delhi, India. endonucleases Alu I, Hae III and Msp I (GeNei) in 25 l reaction volumes, using Enumeration of wheat associated the manufacturer s recommended buffer bacteria and temperature. The clustering analysis was undertaken using the software, The culturable bacteria from soil and NTSYS-2.02e package (Numerical rhizosphere soil were isolated through taxonomy analysis program package, enrichment using the standard serial Exeter software, USA). Similarity among dilution plating technique employing nine the isolates was calculated by Jaccard s different growth media as described earlier and dendrogram was constructed using the (Verma et al. 2013). Endophytic and UPGMA method (Nei and Li 1979). epiphytic bacteria were isolated using methods described by Conn and Franco 16S rDNA Sequencing and phylogenetic (2004) and Holland and Polacco (1994) analysis respectively. The plates were incubated at 30 °C and the population was counted PCR amplified 16S rRNA genes were after 3-7 days. Colonies that appeared purified and sequenced using both pA and were purified by repeated re-streaking to pH primers for forward and reverse obtain pure colonies using respective reactions respectively. Sequencing medium plates. The pure cultures were employed a dideoxy cycle with fluorescent maintained at 4 °C as slant and glycerol terminators and was run in a 3130xl stock (25 %) at -80 °C for further use. All Applied Biosystems ABI prism automated the isolates were screened in triplicates for DNA sequencer (Applied Biosystems, tolerance to pH and drought [Low water Foster City, CA) at SCI Genome Chennai, potential on polyethylene glycol (PEG- India. 16S rRNA gene sequences were 8000) - infused plates] as described earlier analysed using codon code aligner v.4.0.4. Yadav et al. (2014). 434 Int.J.Curr.Microbiol.App.Sci (2014) 3(5): 432-447 The 16S rRNA gene sequences were phaseoli according to the method aligned to those of closely related bacterial described by Verma et al. (2013). species available at GenBank database using BLASTn program. Bacterial isolates Statistical Analysis were identified based on percentage of sequence similarity ( 97%) with that of a In order to compare the bacterial diversity prototype strain sequence in the GenBank. among
Load more

Recommended publications
  • Pseudomonas Helmanticensis Sp. Nov., Isolated from Forest Soil
    International Journal of Systematic and Evolutionary Microbiology (2014), 64, 2338–2345 DOI 10.1099/ijs.0.063560-0 Pseudomonas helmanticensis sp. nov., isolated from forest soil Martha-Helena Ramı´rez-Bahena,1,2 Maria Jose´ Cuesta,1 Jose´ David Flores-Fe´lix,3 Rebeca Mulas,4 Rau´l Rivas,2,3 Joao Castro-Pinto,5 Javier Bran˜as,5 Daniel Mulas,5 Fernando Gonza´lez-Andre´s,4 Encarna Vela´zquez2,3 and A´ lvaro Peix1,2 Correspondence 1Instituto de Recursos Naturales y Agrobiologı´a, IRNASA-CSIC, Salamanca, Spain A´ lvaro Peix 2Unidad Asociada Grupo de Interaccio´n Planta-Microorganismo, [email protected] Universidad de Salamanca-IRNASA (CSIC), Salamanca, Spain 3Departamento de Microbiologı´a y Gene´tica, Universidad de Salamanca, Salamanca, Spain 4Instituto de Medio Ambiente, Recursos Naturales y Biodiversidad, Universidad de Leo´n, Leo´n, Spain 5Fertiberia S. A., Madrid, Spain A bacterial strain, OHA11T, was isolated during the course of a study of phosphate-solubilizing bacteria occurring in a forest soil from Salamanca, Spain. The 16S rRNA gene sequence of strain OHA11T shared 99.1 % similarity with respect to Pseudomonas baetica a390T, and 98.9 % similarity with the type strains of Pseudomonas jessenii, Pseudomonas moorei, Pseudomonas umsongensis, Pseudomonas mohnii and Pseudomonas koreensis. The analysis of housekeeping genes rpoB, rpoD and gyrB confirmed its phylogenetic affiliation to the genus Pseudomonas and showed similarities lower than 95 % in almost all cases with respect to the above species. Cells possessed two polar flagella. The respiratory quinone was Q9. The major fatty acids were C16 : 0, C18 : 1v7c and summed feature 3 (C16 : 1v7c/iso-C15 : 0 2-OH).
    [Show full text]
  • 4.2. Biodegradation of Soil Pollutants by Microorganisms
    UNIVERSITY OF PANNONIA GEORGIKON FACULTY FESTETICS DOCTORAL SCHOOL School leader: Dr. habil. Angéla Anda, DSc THE BASICS OF RESEARCH IN MICROBIOLOGICAL SOIL REMEDIATION PRACTICES DOCTORAL (PHD) THESIS Written by: Nikoletta Horváth Thesis supervisors: Dr. Borbála Biró, DSc Dr. Péter Budai, PhD 2017 KESZTHELY HUNGARY CONTENTS 1. ABSTRACTS............................................................................................................................................... 1 1.1. Abstract (English) ................................................................................................................................. 1 1.2. Kivonat (Hungarian) ............................................................................................................................. 2 1.3. Abstract (German) ................................................................................................................................ 3 2. INTRODUCTION AND IMPORTANCE OF THE TOPIC ................................................................... 4 2.1. Research background ............................................................................................................................ 4 2.2. Basis of the research ............................................................................................................................. 4 3. OBJECTIVES ............................................................................................................................................. 6 4. LITERATURE REVIEW..........................................................................................................................
    [Show full text]
  • Isolation of Rhizobacteria in Southwestern Québec, Canada: An
    Isolation of rhizobacteria in Southwestern Québec, Canada: An investigation of their impact on the growth and salinity stress alleviation in Arabidopsis thaliana and crop plants Di Fan Department of Plant Science Faculty of Agricultural and Environmental Sciences Macdonald Campus of McGill University 21111 Lakeshore Road, Sainte-Anne-de-Bellevue, Québec H9X 3V9 December 2017 A thesis submitted to McGill University in partial fulfillment of the requirements of the degree of DOCTOR OF PHILOSOPHY © Di Fan, Canada, 2017 Table of contents Abstract ................................................................................................. x Résumé ................................................................................................ xiii Acknowledegments ............................................................................. xv Preface ................................................................................................ xviii Contribution of authors ................................................................................ xviii Chapter 1................................................................................................. 1 Introduction ............................................................................................ 1 Chapter 2................................................................................................. 5 Literature Review ................................................................................... 5 2.1 What are root exudates? .........................................................................
    [Show full text]
  • Aquatic Microbial Ecology 80:15
    The following supplement accompanies the article Isolates as models to study bacterial ecophysiology and biogeochemistry Åke Hagström*, Farooq Azam, Carlo Berg, Ulla Li Zweifel *Corresponding author: [email protected] Aquatic Microbial Ecology 80: 15–27 (2017) Supplementary Materials & Methods The bacteria characterized in this study were collected from sites at three different sea areas; the Northern Baltic Sea (63°30’N, 19°48’E), Northwest Mediterranean Sea (43°41'N, 7°19'E) and Southern California Bight (32°53'N, 117°15'W). Seawater was spread onto Zobell agar plates or marine agar plates (DIFCO) and incubated at in situ temperature. Colonies were picked and plate- purified before being frozen in liquid medium with 20% glycerol. The collection represents aerobic heterotrophic bacteria from pelagic waters. Bacteria were grown in media according to their physiological needs of salinity. Isolates from the Baltic Sea were grown on Zobell media (ZoBELL, 1941) (800 ml filtered seawater from the Baltic, 200 ml Milli-Q water, 5g Bacto-peptone, 1g Bacto-yeast extract). Isolates from the Mediterranean Sea and the Southern California Bight were grown on marine agar or marine broth (DIFCO laboratories). The optimal temperature for growth was determined by growing each isolate in 4ml of appropriate media at 5, 10, 15, 20, 25, 30, 35, 40, 45 and 50o C with gentle shaking. Growth was measured by an increase in absorbance at 550nm. Statistical analyses The influence of temperature, geographical origin and taxonomic affiliation on growth rates was assessed by a two-way analysis of variance (ANOVA) in R (http://www.r-project.org/) and the “car” package.
    [Show full text]
  • Control of Phytopathogenic Microorganisms with Pseudomonas Sp. and Substances and Compositions Derived Therefrom
    (19) TZZ Z_Z_T (11) EP 2 820 140 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: A01N 63/02 (2006.01) A01N 37/06 (2006.01) 10.01.2018 Bulletin 2018/02 A01N 37/36 (2006.01) A01N 43/08 (2006.01) C12P 1/04 (2006.01) (21) Application number: 13754767.5 (86) International application number: (22) Date of filing: 27.02.2013 PCT/US2013/028112 (87) International publication number: WO 2013/130680 (06.09.2013 Gazette 2013/36) (54) CONTROL OF PHYTOPATHOGENIC MICROORGANISMS WITH PSEUDOMONAS SP. AND SUBSTANCES AND COMPOSITIONS DERIVED THEREFROM BEKÄMPFUNG VON PHYTOPATHOGENEN MIKROORGANISMEN MIT PSEUDOMONAS SP. SOWIE DARAUS HERGESTELLTE SUBSTANZEN UND ZUSAMMENSETZUNGEN RÉGULATION DE MICRO-ORGANISMES PHYTOPATHOGÈNES PAR PSEUDOMONAS SP. ET DES SUBSTANCES ET DES COMPOSITIONS OBTENUES À PARTIR DE CELLE-CI (84) Designated Contracting States: • O. COUILLEROT ET AL: "Pseudomonas AL AT BE BG CH CY CZ DE DK EE ES FI FR GB fluorescens and closely-related fluorescent GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO pseudomonads as biocontrol agents of PL PT RO RS SE SI SK SM TR soil-borne phytopathogens", LETTERS IN APPLIED MICROBIOLOGY, vol. 48, no. 5, 1 May (30) Priority: 28.02.2012 US 201261604507 P 2009 (2009-05-01), pages 505-512, XP55202836, 30.07.2012 US 201261670624 P ISSN: 0266-8254, DOI: 10.1111/j.1472-765X.2009.02566.x (43) Date of publication of application: • GUANPENG GAO ET AL: "Effect of Biocontrol 07.01.2015 Bulletin 2015/02 Agent Pseudomonas fluorescens 2P24 on Soil Fungal Community in Cucumber Rhizosphere (73) Proprietor: Marrone Bio Innovations, Inc.
    [Show full text]
  • Table S8. Species Identified by Random Forests Analysis of Shotgun Sequencing Data That Exhibit Significant Differences In
    Table S8. Species identified by random forests analysis of shotgun sequencing data that exhibit significant differences in their representation in the fecal microbiomes between each two groups of mice. (a) Species discriminating fecal microbiota of the Soil and Control mice. Mean importance of species identified by random forest are shown in the 5th column. Random forests assigns an importance score to each species by estimating the increase in error caused by removing that species from the set of predictors. In our analysis, we considered a species to be “highly predictive” if its importance score was at least 0.001. T-test was performed for the relative abundances of each species between the two groups of mice. P-values were at least 0.05 to be considered statistically significant. Microbiological Taxonomy Random Forests Mean of relative abundance P-Value Species Microbiological Function (T-Test) Classification Bacterial Order Importance Score Soil Control Rhodococcus sp. 2G Engineered strain Bacteria Corynebacteriales 0.002 5.73791E-05 1.9325E-05 9.3737E-06 Herminiimonas arsenitoxidans Engineered strain Bacteria Burkholderiales 0.002 0.005112829 7.1580E-05 1.3995E-05 Aspergillus ibericus Engineered strain Fungi 0.002 0.001061181 9.2368E-05 7.3057E-05 Dichomitus squalens Engineered strain Fungi 0.002 0.018887472 8.0887E-05 4.1254E-05 Acinetobacter sp. TTH0-4 Engineered strain Bacteria Pseudomonadales 0.001333333 0.025523638 2.2311E-05 8.2612E-06 Rhizobium tropici Engineered strain Bacteria Rhizobiales 0.001333333 0.02079554 7.0081E-05 4.2000E-05 Methylocystis bryophila Engineered strain Bacteria Rhizobiales 0.001333333 0.006513543 3.5401E-05 2.2044E-05 Alteromonas naphthalenivorans Engineered strain Bacteria Alteromonadales 0.001 0.000660472 2.0747E-05 4.6463E-05 Saccharomyces cerevisiae Engineered strain Fungi 0.001 0.002980726 3.9901E-05 7.3043E-05 Bacillus phage Belinda Antibiotic Phage 0.002 0.016409765 6.8789E-07 6.0681E-08 Streptomyces sp.
    [Show full text]
  • Pseudomonas Versuta Sp. Nov., Isolated from Antarctic Soil 1 Wah
    *Manuscript 1 Pseudomonas versuta sp. nov., isolated from Antarctic soil 1 2 3 1,2 3 1 2,4 1,5 4 2 Wah Seng See-Too , Sergio Salazar , Robson Ee , Peter Convey , Kok-Gan Chan , 5 6 3 Álvaro Peix 3,6* 7 8 4 1Division of Genetics and Molecular Biology, Institute of Biological Sciences, Faculty of 9 10 11 5 Science University of Malaya, 50603 Kuala Lumpur, Malaysia 12 13 6 2National Antarctic Research Centre (NARC), Institute of Postgraduate Studies, University of 14 15 16 7 Malaya, 50603 Kuala Lumpur, Malaysia 17 18 8 3Instituto de Recursos Naturales y Agrobiología. IRNASA -CSIC, Salamanca, Spain 19 20 4 21 9 British Antarctic Survey, NERC, High Cross, Madingley Road, Cambridge CB3 OET, UK 22 23 10 5UM Omics Centre, University of Malaya, Kuala Lumpur, Malaysia 24 25 11 6Unidad Asociada Grupo de Interacción Planta-Microorganismo Universidad de Salamanca- 26 27 28 12 IRNASA ( CSIC) 29 30 13 , IRNASA-CSIC, 31 32 33 14 c/Cordel de Merinas 40 -52, 37008 Salamanca, Spain. Tel.: +34 923219606. 34 35 15 E-mail address: [email protected] (A. Peix) 36 37 38 39 16 Abstract: 40 41 42 43 17 In this study w e used a polyphas ic taxonomy approach to analyse three bacterial strains 44 45 18 coded L10.10 T, A4R1.5 and A4R1.12 , isolated in the course of a study of quorum -quenching 46 47 19 bacteria occurring Antarctic soil . The 16S rRNA gene sequence was identical in the three 48 49 50 20 strains and showed 99.7% pairwise similarity with respect to the closest related species 51 52 21 Pseudomonas weihenstephanensis WS4993 T, and the next closest related species were P.
    [Show full text]
  • Entomopathogenic Nematode-Associated Microbiota: from Monoxenic Paradigm to Pathobiome Jean-Claude Ogier†, Sylvie Pagès†, Marie Frayssinet and Sophie Gaudriault*
    Ogier et al. Microbiome (2020) 8:25 https://doi.org/10.1186/s40168-020-00800-5 RESEARCH Open Access Entomopathogenic nematode-associated microbiota: from monoxenic paradigm to pathobiome Jean-Claude Ogier†, Sylvie Pagès†, Marie Frayssinet and Sophie Gaudriault* Abstract Background: The holistic view of bacterial symbiosis, incorporating both host and microbial environment, constitutes a major conceptual shift in studies deciphering host-microbe interactions. Interactions between Steinernema entomopathogenic nematodes and their bacterial symbionts, Xenorhabdus, have long been considered monoxenic two partner associations responsible for the killing of the insects and therefore widely used in insect pest biocontrol. We investigated this “monoxenic paradigm” by profiling the microbiota of infective juveniles (IJs), the soil-dwelling form responsible for transmitting Steinernema-Xenorhabdus between insect hosts in the parasitic lifecycle. Results: Multigenic metabarcoding (16S and rpoB markers) showed that the bacterial community associated with laboratory-reared IJs from Steinernema carpocapsae, S. feltiae, S. glaseri and S. weiseri species consisted of several Proteobacteria. The association with Xenorhabdus was never monoxenic. We showed that the laboratory-reared IJs of S. carpocapsae bore a bacterial community composed of the core symbiont (Xenorhabdus nematophila) together with a frequently associated microbiota (FAM) consisting of about a dozen of Proteobacteria (Pseudomonas, Stenotrophomonas, Alcaligenes, Achromobacter, Pseudochrobactrum, Ochrobactrum, Brevundimonas, Deftia, etc.). We validated this set of bacteria by metabarcoding analysis on freshly sampled IJs from natural conditions. We isolated diverse bacterial taxa, validating the profile of the Steinernema FAM. We explored the functions of the FAM members potentially involved in the parasitic lifecycle of Steinernema. Two species, Pseudomonas protegens and P. chlororaphis, displayed entomopathogenic properties suggestive of a role in Steinernema virulence and membership of the Steinernema pathobiome.
    [Show full text]
  • Molecular Identification and Characterization of Phosphate Solubilizing Pseudomonas Sp
    Archive of SID J. Agr. Sci. Tech. (2016) Vol. 18: 775-788 Molecular Identification and Characterization of Phosphate Solubilizing Pseudomonas sp. Isolated from Rhizosphere of Mash Bean ( Vigna Mungo L.) for Growth Promotion in Wheat M. Ehsan 1, 2, 3, I. Ahmed 1, *, R. Hayat 2, M. Iqbal 1, N. Bibi 1, 2, and N. Khalid 4 ABSTRACT Bio-inoculants have potential role in plant growth promotion. The present study evaluated the potential of Pseudomonas strains as bio-inoculants in wheat on the basis of plant growth promotion and physiological characterization. The 16S rRNA gene sequencing and phylogenetic analysis revealed that four isolated strains belonged to genus Pseudomonas . These strains were positive for phosphorus solubilization and indole acetic acid production, whereas only two strains were positive candidate for their nitrogen fixing ability as determined by presence or absence of nifH gene through amplification from polymerase chain reaction. The pot experiment showed that the integrated use of Pseudomonas strains as co-inoculant and 50% applied mineral fertilizers enhanced the maximum wheat growth and development from 58 to 140% for different shoot and root growth parameters. The strain NCCP-45 and NCCP-237 were closely related to Pseudomonas beteli and Pseudomonas lini, respectively. These isolated strains can be used to increase crop productivity by using as a bio-fertilizer inoculum. Keywords: Bio-inoculant, PCR, PGPR, Phylogenetic analysis, 16S rRNA gene. INTRODUCTION different crops for enhancing plant growth, seed emergence, crop yield, and as bio- Microorganisms are abundant in various control agents against certain plant ecologies. For instance, at least 10 4 bacterial pathogens and pests (Dey et al., 2004; taxa and 10 10-10 11 different bacterial copy Herman et al., 2008).
    [Show full text]
  • Bioprospecting Surfactants Produced by Pseudomonas Spp. Isolated from Soil for Potential Application in Biotechnology
    Bioprospecting Surfactants Produced by Pseudomonas spp. Isolated from Soil for Potential Application in Biotechnology A thesis submitted for the degree of Doctor of Philosophy (PhD) by Kamaluddeen Kabir BSc, MSc. School of Science, Engineering and Technology, Abertay University. April, 2017 Preliminary pages K. Kabir Page ii Preliminary pages Declarations I, Kamaluddeen Kabir, hereby certify that this thesis submitted in partial fulfilment of the requirements for the award of Doctor of Philosophy (PhD), Abertay University, is wholly my own work unless otherwise referenced or acknowledged. This work has not been submitted for any other qualification at any other academic institution. 02/08/17 …………………………………………………. ………………………………………………….. Candidate’s Signature Date K. Kabir Page iii Preliminary pages Acknowledgements I would like to express my sincere gratitude to my supervisor, Dr Andrew Spiers. He has continuously supported me during my PhD programme at Abertay University, and his patience, motivation and advice on ideas have been a source of inspiration to me. This thesis would not have been accomplished without his intellectual input. His supervision has guided me throughout my research and writing of this thesis, and I cannot imagine having a better supervisor or mentor. My immense gratitude also goes to my second supervisor, Dr Yusuf Deeni, for his intuitive comments and support. His questions and input truly helped me to widen my research and look at it from different angles. Even with his tight schedule, he took the time to go through my entire thesis and for this I have nothing to say but, ‘JazakAllah khair’. My sincere gratitude also goes to Zuzana Spiers for her help during the Sample collection and for making my stay in Dundee a memorable one.
    [Show full text]
  • Plant Growth Promoting Bacteria: Biodiversity and Multifunctional Attributes for Sustainable Agriculture
    Review Article Adv Biotech & Micro - August Volume 5 Issue 5 2017 Copyright © All rights are reserved by Ajar Nath Yadav DOI: 10.19080/AIBM.2017.05.555671 Plant Growth Promoting Bacteria: Biodiversity and Multifunctional Attributes for Sustainable Agriculture Ajar Nath Yadav1*, Priyanka Verma2, Bhanumati Singh3, Vinay Singh Chauahan3, Archna Suman4 and Anil Kumar Saxena5 1Department of Biotechnology, Akal College of Agriculture, Eternal University, India 2Department of Microbiology, Akal College of Basic Science, Eternal University, India 3Department of Biotechnology, Bundelkhand University, India 4Division of Microbiology, Indian Agricultural Research Institute, India 5ICAR-National Bureau of Agriculturally Important Microorganisms, India Submission: May 24, 2017; Published: August 24, 2017 *Corresponding author: Ajar Nath Yadav, Department of Biotechnology, Akal College of Agriculture, Eternal University, India, Tel: +91- 9882545085; Email: Ajar Nath Yadav and Priyanka Verma contributed equally to the present work. Abstract The use of plant growth promoting bacteria may prove useful in developing strategies to facilitate plant growth under normal as well as diverse abiotic stress conditions. The application of microbes with the aim of improving nutrients availability for plants is an important practice and necessary for sustainable agriculture. During the past couple of decades, the use of microbial inoculants for sustainable agriculture has to inoculation with plant growth promoting (PGP) microbes have been repeatedly reported. The actual biodiversity of PGP microbes belong to differentincreased groups tremendously including in Actinobacteria, various parts of Bacteroidetes, the world. Significant Balneolaeota increases Firmicutes, in growth Proteobacteria and yield of and agronomically Spirochaetes. important PGP bacteria crops are in responsenaturally productionoccurring soil of bacteriaammonia, that hydrogen aggressively cyanide, colonize siderophore plant roots and and biocontrol benefit plants against by different providing plant growth pathogens.
    [Show full text]
  • Uptake of Carbamazepine by Rhizomes and Endophytic Bacteria of Phragmites Australis
    ORIGINAL RESEARCH ARTICLE published: 20 February 2015 doi: 10.3389/fpls.2015.00083 Uptake of carbamazepine by rhizomes and endophytic bacteria of Phragmites australis Andrés Sauvêtre and Peter Schröder* Plant Endophyte Physiology, Research Unit Microbe–Plant Interactions, Helmholtz Zentrum München GmbH, German Research Center for Environmental Health, Munich, Germany Edited by: Carbamazepine is an antiepileptic and mood-stabilizing drug which is used widely in Antonella Furini, University of Verona, Europe and North America. In the environment, it is found as a persistent and recalcitrant Italy contaminant, being one of the most prominent hazardous pharmaceuticals and personal Reviewed by: care products in effluents of wastewater treatment plants. Phragmites australis is one Muhammad Afzal, National Institute of Biotechnology and Genetic of the species with both, the highest potential of detoxification and phytoremediation. Engineering, Pakistan It has been used successfully in the treatment of industrial and municipal wastewater. Josep Maria Bayona, Consejo Recently, the identification of endophytic microorganisms from different plant species Superior de Investigaciones Científicas, Spain growing in contaminated sites has provided a list of candidates which could be used as bio-inoculants for bioremediation of difficult compounds. In this study, Phragmites australis *Correspondence: Peter Schröder, Plant Endophyte plants were exposed to 5 mg/L of carbamazepine. After 9 days the plants had removed Physiology, Research Unit 90% of the initial concentration. Endophytic bacteria were isolated from these plants Microbe–Plant Interactions, Helmholtz and further characterized. Phylogenetic analysis based on 16S rDNA sequencing revealed Zentrum München GmbH, German Research Center for Environmental that the majority of these isolates belong to three groups: Proteobacteria, Actinobacteria, Health, Ingolstädter Landstraße 1, and Bacteroidetes.
    [Show full text]