Endophytic colonization of Arabidopsis thaliana by Gluconacetobacter diazotrophicus and its effect on plant growth promotion, plant physiology, and activation of plant defense
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Background and aims
Gluconacetobacter diazotrophicus is a plant growth-promoting bacteria (PGPB) that colonizes several plant species. Here, we studied the internal colonization of Arabidopsis thaliana tissues by G. diazotrophicus and analyzed its effects on physiology, growth, and activation of plant immune system during such association.
A. thaliana seedlings were inoculated with G. diazotrophicus and grown in substrate for 50 days. Effects on plant growth were estimated by quantifying number of leaves, leaf area, and fresh and dry weight. Endophytic bacterial population was determined by colony-forming unit (CFU), and its location in plant tissues was assayed by epifluorescence microscopy of red fluorescent protein-labeled bacterium. Whole canopy gas exchange (photosynthesis and transpiration) was determined using a portable photosynthesis system.
G. diazotrophicus efficiently promoted A. thaliana plant growth at 50 days after inoculation. Inoculated plants showed higher whole canopy photosynthesis, lower whole plant transpiration, and increased water-use efficiency. The bacterium colonized preferentially root xylem. The inoculation of plants defective in systemic acquired resistance (SAR)-associated defense revealed that plant immune system plays an important role during the early association stages.
G. diazotrophicus endophytically colonizes A. thaliana roots, promotes plant growth, and increases whole canopy photosynthesis. Our results indicate that A. thaliana is useful for molecular studies of the mechanisms involved in the interaction between plants and PGPB, especially those involving G. diazotrophicus.
KeywordsPGPB Endophytic bacteria BNF Plant defense Plant immunity G.diazotrophicus
This research was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Financiadora de Estudos e Projetos (FINEP), Fundação de Amparo a Pesquisa do Estado do Rio de Janeiro (FAPERJ), and Instituto Nacional de Ciências e Tecnologia em Fixação Biológica de Nitrogênio (INCT-FBN). First author received fellowship from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior-CAPES.
- Alquéres S, Meneses C, Rouws L, Rothballer M, Baldani I, Schmid M, Hartmann A (2013) The bacterial superoxide dismutase and glutathione reductase are crucial for endophytic colonization of rice roots by Gluconacetobacter diazotrophicus PAL5. Mol Plant-Microbe Interact 26:937–945CrossRefPubMedGoogle Scholar
- Anitha KG, Thangaraju M (2010) Growth promotion of rice seedling by Gluconacetobacter diazotrophicus under in vivo conditions. J Cell Plant Sci 1:6–12Google Scholar
- Galisa PS, da Silva HA, Macedo AV, Reis VM, Vidal MS, Baldani JI, Simões-Araújo JL (2012) Identification and validation of reference genes to study the gene expression in Gluconacetobacter diazotrophicus grown in different carbon sources using RT-qPCR. J Microbiol Methods 91(1):1–7CrossRefPubMedGoogle Scholar
- Hoagland DR, Arnon DI (1950) The water-culture method for growing plants without soil. Circular Calif Agric Exp Sta 347Google Scholar
- Jimenez-Salgado T, Fuentes-Ramirez LE, Tapia-Hernandez A, Mascarua-Esparza MA, Martinez-Romero E, Caballero-Mellado J (1997) Coffea arabica L., a new host plant for Acetobacter diazotrophicus, and isolation of other nitrogen fixing acetobacteria. Appl Environ Microbiol 63(9):3676–3683PubMedPubMedCentralGoogle Scholar
- Kechid M, Desbrosses G, Rokhsi W, Varoquaux F, Djekoun A, Touraine B (2013) The NRT2.5 and NRT2.6 genes are involved in growth promotion of Arabidopsis by the plant growth-promoting rhizobacterium (PGPR) strain Phyllobacterium brassicacearum STM196. New Phytol 198(2):514–524CrossRefPubMedGoogle Scholar
- Lee S, Flores-Encarnación M, Contreras-Zentella M, Garcia-Flores L, Escamilla JE, Kennedy C (2004) Indole-3-acetic acid biosynthesis is deficient in Gluconacetobacter diazotrophicus strains with mutations in cytochrome c biogenesis genes. J Bacteriol 186(16):5384–5391CrossRefPubMedPubMedCentralGoogle Scholar
- Maclean AM, Sugio A, Kingdom HN, Grieve VM, Hogenhout SA (2011) Arabidopsis thaliana as a model plant for understanding phytoplasma interactions with plant and insect hosts. Bull Insectol 64(Supplement):S173–S174Google Scholar
- Matiru V, Thomson J (1998) Can Acetobacter diazotrophicus be used as a growth promoter for coffee, tea, and banana plants? In Proc 8th Cong Afric Assoc Biol Nitro Fixat, F. D. Dakora (Eds.) 129–130 University of Cape TownGoogle Scholar
- Meenakshisundaram M, Santhaguru K (2010) Isolation and nitrogen fixing efficiency of a novel endophytic diazotroph Gluconacetobacter diazotrophicus associated with Saccharum officinarum from southern district of Tamilnadu. Int J Biol Med Res 1:298–300Google Scholar
- Muthukumarasamy R, Revathi G, Vadivelu M (2000) Antagonistic potential of N2-fixing Acetobacter diazotrophicus against Colletotrichum falcatum Went., a causal organism of red-rot of sugarcane. Curr Sci 78:1063–1065Google Scholar
- Muthukumarasamy R, Cleenwerck I, Revathi G, Vadivelu M, Janssens D, Hoste B, Gum KU, Ki-Do P, Son CY, Sa T, Caballero-Mellado J (2005) Natural association of Gluconacetobacter diazotrophicus and diazotrophic Acetobacter peroxydans with wetland rice. Syst Appl Microbiol 28(3):277–286CrossRefPubMedGoogle Scholar
- Niu DD, Liu HX, Jiang CH, Wang YP, Wang QY, Jin HL, Guo JH (2011) The plant growth-promoting rhizobacterium Bacillus cereus AR156 induces systemic resistance in Arabidopsis thaliana by simultaneously activating salicylate-and jasmonate/ethylene-dependent signaling pathways. Mol Plant-Microbe Interact 24(5):533–542CrossRefPubMedGoogle Scholar
- Riggs PJ, Chelius MK, Iniguez AL, Kaeppler SM, Triplett EW (2001) Enhanced maize productivity by inoculation with diazotrophic bacteria. Aust J Plant Physiol 28(9):829–836Google Scholar
- Rodrigues Neto J, Malavolta V Jr, Victor O (1986) Meio simples para o isolamento e cultivo de Xanthomonas campestris pv. citri tipo B. Summa Phytopathol 12(1–2):32Google Scholar
- Rouws LF, Meneses CH, Guedes HV, Vidal MS, Baldani JI, Schwab S (2010) Monitoring the colonization of sugarcane and rice plants by the endophytic diazotrophic bacterium Gluconacetobacter diazotrophicus marked with gfp and gusA reporter genes. Lett Appl Microbiol 51:325–330CrossRefPubMedGoogle Scholar
- Savci S (2012) An agricultural pollutant: chemical fertilizer. Int J Environ Sci Develop 3:77–80Google Scholar
- Shirano Y, Kachroo P, Shah J, Klessig DF (2002) A gain-of-function mutation in an Arabidopsis Toll interleukin1 receptor-nucleotide binding site-leucine-rich repeat type R gene triggers defense responses and results in enhanced disease resistance. Plant Cell 14:3149–3162CrossRefPubMedPubMedCentralGoogle Scholar
- Vargas L, Santa Brígida AB, Mota Filho JP, de Carvalho TG, Rojas CA, Vaneechoutte D, Van Bel M, Farrinelli L, Ferriera PCG, Hemerly AS (2014) Drought tolerance conferred to sugarcane by association with Gluconacetobacter diazotrophicus: a transcriptomic view of hormone pathways. PLoS One 9(12):e114744CrossRefPubMedPubMedCentralGoogle Scholar