Associative diazotrophic bacteria from forage grasses in the Brazilian semi-arid region are effective plant growth promoters
Gabiane dos Reis Antunes A , Sheilla Rios Assis Santana A , Indra Elena Costa Escobar B , Marivaine da Silva Brasil C , Gherman Garcia Leal de Araújo D , Tadeu Vinhas Voltolini D and Paulo Ivan Fernandes-Júnior D E
+ Author Affiliations
- Author Affiliations
A Colegiado de Zootecnia, Universidade Federal do Vale do São Francisco, Petrolina, PE, 56300-000, Brazil.
B Colegiado de Farmácia, Universidade Federal do Vale do São Francisco, Petrolina, PE, 56304-917, Brazil.
C Campus Pantanal, Universidade Federal do Mato Grosso do Sul, Corumbá, MS, 79304-902, Brazil.
D Embrapa Semiárido, Petrolina, PE, 56302-970, Brazil.
E Corresponding author. Email: paulo.ivan@embrapa.br
Crop and Pasture Science 70(10) 899-907 https://doi.org/10.1071/CP19076
Submitted: 21 February 2019 Accepted: 14 August 2019 Published: 22 October 2019
Abstract
The study of plant growth-promoting bacteria (PGPB) can identify outstanding bacteria for crops. For forage grasses adapted to drylands, the selection of PGPB can increase the field performance of pastures. The aim of this study was to isolate, and characterise at molecular, biochemical and symbiotic levels, diazotrophic bacteria obtained from buffel grass (Cenchrus ciliaris), sorghum (Sorghum bicolor) and Tifton 85 (Cynodon spp.) from Brazilian semi-arid region fields. Field-grown plants were collected, and the roots were surface-disinfected, crushed and inoculated in a semi-solid medium. After the formation and confirmation of microaerophilic pellicles, the bacteria were isolated and purified. All bacterial isolates were subjected to nifH gene amplification and identified by their partial 16S rRNA gene sequences. The bacteria were evaluated for the production of auxins and siderophores, calcium phosphate solubilisation, and diazotrophic ability as ‘in vitro’ plant growth-promotion traits. A plant inoculation assay was conducted to assess the plant growth-promotion abilities of the bacterial isolates. Twenty-one bacterial isolates harboured the nifH gene (nifH+), among which nine were obtained from sorghum, eight from buffel grass, and four from Tifton 85. The bacterial isolates were classified as Bacillus (8), Stenotrophomonas (7), Agrobacterium (4), Cellulomonas (1) and Paenibacillus (1). All were shown to be auxin producers, with 14 isolates showing diazotrophic capacity ‘in vitro’. Fourteen isolates increased plant N content, but the bacterial strains ESA 392 and ESA 398 (Bacillus), ESA 397 and ESA 407 (Stenotrophomonas), and ESA 401 (Agrobacterium) were shown to promote both plant growth and N nutrition. These strains are candidates for further assays to evaluate their agronomic performance under field conditions, aiming inoculant production for forage grasses in drylands.
Additional keywords: biological nitrogen fixation, Caatinga, plant growth promotion traits, strain selection.
References
Ambrosini A, Stefanski T, Lisboa BB, Beneduzi A, Vargas LK, Passaglia LMP (2016) Diazotrophic bacilli isolated from the sunflower rhizosphere and the potential of Bacillus mycoides B38V as biofertiliser. Annals of Applied Biology 168, 93–110.| Diazotrophic bacilli isolated from the sunflower rhizosphere and the potential of Bacillus mycoides B38V as biofertiliser.Crossref | GoogleScholarGoogle Scholar |
Antunes GR, Carvalho BR, Silva TR, Neiva JNM, Araújo GGL, Fernandes-Júnior PI (2017) Quantification and characterization of putative diazotrophic bacteria from forage palm under saline water irrigation. Geama 3, 261–268.
Baldani JI, Reis VM, Videira SS, Boddey LH, Baldani VLD (2014) The art of isolating nitrogen-fixing bacteria from non-leguminous plants using N-free semi-solid media: a practical guide for microbiologists. Plant and Soil 384, 413–431.
| The art of isolating nitrogen-fixing bacteria from non-leguminous plants using N-free semi-solid media: a practical guide for microbiologists.Crossref | GoogleScholarGoogle Scholar |
Barazani O, Friedman J (1999) Is IAA the major root growth factor secreted from plant-growth-mediating bacteria? Journal of Chemical Ecology 25, 2397–2406.
| Is IAA the major root growth factor secreted from plant-growth-mediating bacteria?Crossref | GoogleScholarGoogle Scholar |
Berraquero FR, Baya B, Cormenzana AR (1976) Establecimiento de índices para el estudio de la solubilización de fosfatos por bacterias del suelo. Ars Pharmaceutica 17, 399–406.
Bertrand H, Nalin R, Bally R, Cleyet-Marel J-C (2001) Isolation and identification of the most efficient plant growth-promoting bacteria associated with canola (Brassica napus). Biology and Fertility of Soils 33, 152–156.
| Isolation and identification of the most efficient plant growth-promoting bacteria associated with canola (Brassica napus).Crossref | GoogleScholarGoogle Scholar |
Boddey RM, De Moraes Sá JC, Alves BJR, Urquiaga S (1997) The contribution of biological nitrogen fixation for sustainable agricultural systems in the tropics. Soil Biology & Biochemistry 29, 787–799.
| The contribution of biological nitrogen fixation for sustainable agricultural systems in the tropics.Crossref | GoogleScholarGoogle Scholar |
Brasil M da S, Baldani JI, Baldani VLD (2005) Ocorrência e diversidade de bactérias diazotróficas associadas a gramíneas forrageiras do Pantanal Sul Matogrossense. Revista Brasileira de Ciência do Solo 29, 179–190.
| Ocorrência e diversidade de bactérias diazotróficas associadas a gramíneas forrageiras do Pantanal Sul Matogrossense.Crossref | GoogleScholarGoogle Scholar |
Brígido C, Oliveira S (2013) Most acid-tolerant chickpea mesorhizobia show induction of major chaperone genes upon acid shock. Microbial Ecology
| Most acid-tolerant chickpea mesorhizobia show induction of major chaperone genes upon acid shock.Crossref | GoogleScholarGoogle Scholar | 22890730PubMed |
da Silva JF, da Silva TR, Escobar IEC, Fraiz ACR, dos Santos JWM, do Nascimento TR, dos Santos JMR, Peters SJW, de Melo RF, Signor D, Fernandes-Júnior PI (2018) Screening of plant growth promotion ability among bacteria isolated from field-grown sorghum under different managements in Brazilian drylands. World Journal of Microbiology & Biotechnology 34, 186
| Screening of plant growth promotion ability among bacteria isolated from field-grown sorghum under different managements in Brazilian drylands.Crossref | GoogleScholarGoogle Scholar |
de Oliveira DM, de Lima ALA, Diniz NB, Santos CERS, da Silva SLF, Simões AN (2018) Inoculation of plant-growth-promoting rhizobacteria in Myracrodruon urundeuva Allemão supports in tolerance to drought stress. Journal of Plant Interactions 13, 91–99.
| Inoculation of plant-growth-promoting rhizobacteria in Myracrodruon urundeuva Allemão supports in tolerance to drought stress.Crossref | GoogleScholarGoogle Scholar |
de Souza R, Ambrosini A, Passaglia LMP (2015) Plant growth-promoting bacteria as inoculants in agricultural soils. Genetics and Molecular Biology 38, 401–419.
| Plant growth-promoting bacteria as inoculants in agricultural soils.Crossref | GoogleScholarGoogle Scholar |
dos Santos MCM, dos Santos DR, Bakke A, Bakke IA (2013) Ocorrência e atividade de bactérias diazotróficas em forrageiras cultivadas na rregião semiárida do Brasil. Caatinga 26, 27–34.
dos Santos CLR, Alves GC, de Matos Macedo AV, Giori FG, Pereira W, Urquiaga S, Reis VM (2017) Contribution of a mixed inoculant containing strains of Burkholderia spp. and Herbaspirillum ssp. to the growth of three sorghum genotypes under increased nitrogen fertilization levels. Applied Soil Ecology 113, 96–106.
| Contribution of a mixed inoculant containing strains of Burkholderia spp. and Herbaspirillum ssp. to the growth of three sorghum genotypes under increased nitrogen fertilization levels.Crossref | GoogleScholarGoogle Scholar |
Duca D, Lorv J, Patten CL, Rose D, Glick BR (2014) Indole-3-acetic acid in plant–microbe interactions. Antonie van Leeuwenhoek 106, 85–125.
| Indole-3-acetic acid in plant–microbe interactions.Crossref | GoogleScholarGoogle Scholar | 24445491PubMed |
Egamberdieva D (2008) Plant growth promoting properties of rhizobacteria isolated from wheat and pea grown in loamy sand soil. Turkish Journal of Biology 32, 9–15.
Egamberdiyeva D, Höflich G (2002) Root colonization and growth promotion of winter wheat and pea by Cellulomonas spp. at different temperatures. Plant Growth Regulation 38, 219–224.
| Root colonization and growth promotion of winter wheat and pea by Cellulomonas spp. at different temperatures.Crossref | GoogleScholarGoogle Scholar |
Estrada-de Los Santos P, Bustillos-Cristales R, Caballero-Mellado J (2001) Burkholderia, a genus rich in plant-associated nitrogen fixers with wide environmental and geographic distribution. Applied and Environmental Microbiology 67, 2790–2798.
| Burkholderia, a genus rich in plant-associated nitrogen fixers with wide environmental and geographic distribution.Crossref | GoogleScholarGoogle Scholar | 11375196PubMed |
Fernandes Júnior PI, Pereira GMD, Perin L, da Silva LM, Baraúna AC, Alvess FM, Passos SR, Zilli JE (2013) Diazotrophic bacteria isolated from wild rice Oryza glumaepatula (Poaceae) in the Brazilian Amazon. Revista de Biología Tropical 61, 991–999.
| Diazotrophic bacteria isolated from wild rice Oryza glumaepatula (Poaceae) in the Brazilian Amazon.Crossref | GoogleScholarGoogle Scholar |
Fernandes-Júnior PI, Aidar S de T, Morgante CV, Gava CAT, Zilli JÉ, de Souza LSB, Marinho R de CN, Nóbrega RSA, Brasil M da S, Seido SL, Martins LMV (2015) The resurrection plant Tripogon spicatus (Poaceae) harbors a diversity of plant growth promoting bacteria in northeastern Brazilian Caatinga. Revista Brasileira de Ciência do Solo 39, 993–1002.
| The resurrection plant Tripogon spicatus (Poaceae) harbors a diversity of plant growth promoting bacteria in northeastern Brazilian Caatinga.Crossref | GoogleScholarGoogle Scholar |
Ferreira DF (2011) Sisvar: a computer statistical analysis system. Ciência e Agrotecnologia 35, 1039–1042.
| Sisvar: a computer statistical analysis system.Crossref | GoogleScholarGoogle Scholar |
Glick BR (2005) Modulation of plant ethylene levels by the bacterial enzyme ACC deaminase. FEMS Microbiology Letters 251, 1–7.
| Modulation of plant ethylene levels by the bacterial enzyme ACC deaminase.Crossref | GoogleScholarGoogle Scholar | 16099604PubMed |
Gontijo JB, Andrade GVS, Baldotto MA, Baldotto LEB (2018) Bioprospecting and selection of growth-promoting bacteria for Cymbidium sp. orchids. Scientia Agrícola 75, 368–374.
| Bioprospecting and selection of growth-promoting bacteria for Cymbidium sp. orchids.Crossref | GoogleScholarGoogle Scholar |
Haiyambo DH, Chimwamurombe PM, Reinhold-Hurek B (2015a) Isolation and screening of rhizosphere bacteria from grasses in East Kavango region of Namibia for plant growth promoting characteristics. Current Microbiology 71, 566–571.
| Isolation and screening of rhizosphere bacteria from grasses in East Kavango region of Namibia for plant growth promoting characteristics.Crossref | GoogleScholarGoogle Scholar | 26254764PubMed |
Haiyambo DH, Reinhold-Hurek B, Chimwamurombe PM (2015b) Effects of plant growth promoting bacterial isolates from Kavango on the vegetative growth of Sorghum bicolor. African Journal of Microbiological Research 9, 725–729.
| Effects of plant growth promoting bacterial isolates from Kavango on the vegetative growth of Sorghum bicolor.Crossref | GoogleScholarGoogle Scholar |
Hungria M, Campo RJ, Souza EM, Pedrosa FO (2010) Inoculation with selected strains of Azospirillum brasilense and A. lipoferum improves yields of maize and wheat in Brazil. Plant and Soil 331, 413–425.
| Inoculation with selected strains of Azospirillum brasilense and A. lipoferum improves yields of maize and wheat in Brazil.Crossref | GoogleScholarGoogle Scholar |
Hungria M, Nogueira MA, Araujo RS (2016) Inoculation of Brachiaria spp. with the plant growth-promoting bacterium Azospirillum brasilense: an environment-friendly component in the reclamation of degraded pastures in the tropics. Agriculture, Ecosystems & Environment 221, 125–131.
| Inoculation of Brachiaria spp. with the plant growth-promoting bacterium Azospirillum brasilense: an environment-friendly component in the reclamation of degraded pastures in the tropics.Crossref | GoogleScholarGoogle Scholar |
IBGE (2017) Censo Agropecuário 2017. IBGE, Rio de Janeiro, Brazil. Available at: https://censoagro2017.ibge.gov.br/templates/censo_agro/resultadosagro/index.html
Kavamura VN, Santos SN, da Silva JL, Parma MM, Ávila LA, Visconti A, Zucchi TD, Taketani RG, Andreote FD, de Melo IS (2013) Screening of Brazilian cacti rhizobacteria for plant growth promotion under drought. Microbiological Research 168, 183–191.
| Screening of Brazilian cacti rhizobacteria for plant growth promotion under drought.Crossref | GoogleScholarGoogle Scholar | 23279812PubMed |
Kuss AV, Kuss VV, Lovato T, Flôres L (2007) Fixação de nitrogênio e produção de ácido indolacético in vitro por bactérias diazotróficas endofíticas. Pesquisa Agropecuária Brasileira 42, 1459–1465.
| Fixação de nitrogênio e produção de ácido indolacético in vitro por bactérias diazotróficas endofíticas.Crossref | GoogleScholarGoogle Scholar |
Leite R da C, Santos JGD dos, Silva EL, Alves CRCR, Hungria M, Leite R da C, Santos AC dos (2019a) Productivity increase, reduction of nitrogen fertiliser use and drought-stress mitigation by inoculation of Marandu grass (Urochloa brizantha) with Azospirillum brasilense. Crop & Pasture Science 70, 61–67.
| Productivity increase, reduction of nitrogen fertiliser use and drought-stress mitigation by inoculation of Marandu grass (Urochloa brizantha) with Azospirillum brasilense.Crossref | GoogleScholarGoogle Scholar |
Leite R da C, Santos AC dos, Santos JGD dos, Leite R da C, Oliveira LBT de, Hungria M (2019b) Mitigation of Mombasa grass (Megathyrsus maximus) dependence on nitrogen fertilization as a function of inoculation with Azospirillum brasilense. Revista Brasileira de Ciência do Solo 43, 1–14.
| Mitigation of Mombasa grass (Megathyrsus maximus) dependence on nitrogen fertilization as a function of inoculation with Azospirillum brasilense.Crossref | GoogleScholarGoogle Scholar |
Liao CFH (1981) Devarda’s alloy method for total nitrogen determination. Soil Science Society of America Journal 45, 852–855.
| Devarda’s alloy method for total nitrogen determination.Crossref | GoogleScholarGoogle Scholar |
Lima JVL, Weber OB, Correia D, Soares MA, Senabio JA (2015) Endophytic bacteria in cacti native to a Brazilian semi-arid region. Plant and Soil 389, 25–33.
| Endophytic bacteria in cacti native to a Brazilian semi-arid region.Crossref | GoogleScholarGoogle Scholar |
Mazzetto AM, Barneze AS (2016) Nitrogen fertilizer effects on nitrous oxide emission from southwest Brazilian Amazon pastures. Journal of Fertilizers & Pesticides 07, 1–5.
| Nitrogen fertilizer effects on nitrous oxide emission from southwest Brazilian Amazon pastures.Crossref | GoogleScholarGoogle Scholar |
Moreira JN, Guimarães Filho C (2011) Sistemas tradicionais para produção de caprinos e ovinos. ‘Produção caprinos e ovinos no Semiárido’. (Ed. TV Voltolini) pp. 49–67. (Embrapa Semiárido: Petrolina, Brazil)
Moreira JN, Lira M de A, dos Santos MVF, Ferreira M de A, de Araújo GGL, Ferreira RLC, da Silva GC (2006) Caracterização da vegetação de Caatinga e da dieta de novilhos no Sertão de Pernambuco. Pesquisa Agropecuária Brasileira 41, 1643–1651.
| Caracterização da vegetação de Caatinga e da dieta de novilhos no Sertão de Pernambuco.Crossref | GoogleScholarGoogle Scholar |
Moreira FTA, Santos DR, Silva GH, Alencar LS (2013) Ocorrência de bactérias do gênero Azospirillum spp. associadas a gramíneas forrageiras no semiárido nordestino. Holos 3, 205–212.
| Ocorrência de bactérias do gênero Azospirillum spp. associadas a gramíneas forrageiras no semiárido nordestino.Crossref | GoogleScholarGoogle Scholar |
Poly F, Monrozier LJ, Bally R (2001) Improvement in the RFLP procedure for studying the diversity of nifH genes in communities of nitrogen fixers in soil. Research in Microbiology 152, 95–103.
| Improvement in the RFLP procedure for studying the diversity of nifH genes in communities of nitrogen fixers in soil.Crossref | GoogleScholarGoogle Scholar | 11281330PubMed |
Reis Junior, FB, Silva MF, Teixeira KRS, Urquiaga S, Reis VM (2004) Identificação de isolados de Azospirillum amazonense associados a Brachiaria spp., em diferentes épocas e condições de cultivo e produção de fitormônio pela bactéria. Revista Brasileira de Ciência do Solo 28, 103–113.
| Identificação de isolados de Azospirillum amazonense associados a Brachiaria spp., em diferentes épocas e condições de cultivo e produção de fitormônio pela bactéria.Crossref | GoogleScholarGoogle Scholar |
Ribeiro CM, Cardoso EJBN (2012) Isolation, selection and characterization of root-associated growth promoting bacteria in Brazil pine (Araucaria angustifolia). Microbiological Research 167, 69–78.
| Isolation, selection and characterization of root-associated growth promoting bacteria in Brazil pine (Araucaria angustifolia).Crossref | GoogleScholarGoogle Scholar | 21596540PubMed |
Rodrigues Neto J, Malavolta Jr VA, Victor O (1986) Meio simples para o isolamento e cultivo de Xanthomonas campestris pv. citri tipo B. Summa Phytopathologica 12, 32
Sarwar M, Kremer RJ (1995) Determination of bacterially derived auxins using a microplate method. Letters in Applied Microbiology 20, 282–285.
| Determination of bacterially derived auxins using a microplate method.Crossref | GoogleScholarGoogle Scholar |
Schwyn B, Neilands JB (1987) Universal chemical assay for the detection and determination of siderophores. Analytical Biochemistry 160, 47–56.
| Universal chemical assay for the detection and determination of siderophores.Crossref | GoogleScholarGoogle Scholar | 2952030PubMed |
Signor D, Cerri CEP (2013) Nitrous oxide emissions in agricultural soils: a review. Pesquisa Agropecuária Tropical 43, 322–338.
| Nitrous oxide emissions in agricultural soils: a review.Crossref | GoogleScholarGoogle Scholar |
Silva MCP, Figueiredo AF, Andreote FD, Cardoso EJBN (2013) Plant growth promoting bacteria in Brachiaria brizantha. World Journal of Microbiology & Biotechnology 29, 163–171.
| Plant growth promoting bacteria in Brachiaria brizantha.Crossref | GoogleScholarGoogle Scholar |
Singh RP, Jha PN (2017) The PGPR Stenotrophomonas maltophilia SBP-9 augments resistance against biotic and abiotic stress in wheat plants. Frontiers in Microbiology 8,
| The PGPR Stenotrophomonas maltophilia SBP-9 augments resistance against biotic and abiotic stress in wheat plants.Crossref | GoogleScholarGoogle Scholar | 29062306PubMed |
Soares RA, Roesch LFW, Zanatta G, de Oliveira Camargo FA, Passaglia LMP (2006) Occurrence and distribution of nitrogen fixing bacterial community associated with oat (Avena sativa) assessed by molecular and microbiological techniques. Applied Soil Ecology 33, 221–234.
| Occurrence and distribution of nitrogen fixing bacterial community associated with oat (Avena sativa) assessed by molecular and microbiological techniques.Crossref | GoogleScholarGoogle Scholar |
Souza MST, Baura VA, Santos SA, Fernandes-Júnior PI, Reis Junior, FB, Marques RM, Paggi GM, Brasil MS (2017) Azospirillum spp. from native forage grasses in Brazilian Pantanal floodplain: biodiversity and plant growth promotion potential. World Journal of Microbiology & Biotechnology 33, 81
| Azospirillum spp. from native forage grasses in Brazilian Pantanal floodplain: biodiversity and plant growth promotion potential.Crossref | GoogleScholarGoogle Scholar |
Sylvester-Bradley R, Asakawa N, La Torraca S, Magalhães FM, Oliveira LA, Pereira RM (1982) Levantamento quantitativo de microrganismos solubilizadores de fosfatos na rizosfera de gramíneas e leguminosas forrageiras na Amazônia. Acta Amazonica 12, 15–22.
| Levantamento quantitativo de microrganismos solubilizadores de fosfatos na rizosfera de gramíneas e leguminosas forrageiras na Amazônia.Crossref | GoogleScholarGoogle Scholar |
Vendan RT, Yu YJ, Lee SH, Rhee YH (2010) Diversity of endophytic bacteria in ginseng and their potential for plant growth promotion. Journal of Microbiology 48, 559–565.
| Diversity of endophytic bacteria in ginseng and their potential for plant growth promotion.Crossref | GoogleScholarGoogle Scholar |
Voltolini TV (Ed.) (2011) ‘Produção de caprinos e ovinos no Semiárido.’ (Embrapa Semiárido: Petrolina, Brazil)
Wang H, Qi M, Cutler AJ (1993) A simple method of preparing plant samples for PCR. Nucleic Acids Research 21, 4153–4154.
| A simple method of preparing plant samples for PCR.Crossref | GoogleScholarGoogle Scholar | 8371994PubMed |
Weisburg WG, Barns SM, Pelletier DA, Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. Journal of Bacteriology 173, 697–703.
| 16S ribosomal DNA amplification for phylogenetic study.Crossref | GoogleScholarGoogle Scholar | 1987160PubMed |
Yoon S-H, Ha S-M, Kwon S, Lim J, Kim Y, Seo H, Chun J (2017) Introducing EzBioCloud: a taxonomically united database of 16S rRNA and whole genome assemblies. International Journal of Systematic and Evolutionary Microbiology 67, 1613–1617.
| Introducing EzBioCloud: a taxonomically united database of 16S rRNA and whole genome assemblies.Crossref | GoogleScholarGoogle Scholar | 28005526PubMed |
Zinniel DK, Lambrecht P, Harris NB, Feng Z, Kuczmarski D, Higley P, Ishimaru CA, Arunakumari A, Barletta RG, Vidaver AK (2002) Isolation and characterization of endophytic colonizing bacteria from agronomic crops and prairie plants. Applied and Environmental Microbiology 68, 2198–2208.
| Isolation and characterization of endophytic colonizing bacteria from agronomic crops and prairie plants.Crossref | GoogleScholarGoogle Scholar | 11976089PubMed |
