African Journal of
Agricultural Research

  • Abbreviation: Afr. J. Agric. Res.
  • Language: English
  • ISSN: 1991-637X
  • DOI: 10.5897/AJAR
  • Start Year: 2006
  • Published Articles: 6865

Full Length Research Paper

Azospirillum brasilense and nitrogen fertilization affecting wheat productivity

Junia Maria Clemente
  • Junia Maria Clemente
  • Pós-Doutora - Faculdade do Noroeste de Minas, FINOM, Brazil
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Aurinelza Batista Teixeira Condé
  • Aurinelza Batista Teixeira Condé
  • Bolsistas BIP da FAPEMIG; Empresa de Pesquisa Agropecuária de Minas Gerais – EMBRAPA, Brazil
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Alex Teixeira Andrade
  • Alex Teixeira Andrade
  • Bolsistas BIP da FAPEMIG; Empresa de Pesquisa Agropecuária de Minas Gerais – EMBRAPA, Brazil
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Carine Rezende Cardoso
  • Carine Rezende Cardoso
  • Laboratório de Biocontrole Farroupilha, Brazil.
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Iara da Mata Flor
  • Iara da Mata Flor
  • Laboratório de Biocontrole Farroupilha, Brazil.
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Fábio Aurélio Dias Martins
  • Fábio Aurélio Dias Martins
  • Laboratório de Biocontrole Farroupilha, Brazil.
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Willian Tadeu de Lima
  • Willian Tadeu de Lima
  • Laboratório de Biocontrole Farroupilha, Brazil.
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Cleiton Burnier de Oliveira
  • Cleiton Burnier de Oliveira
  • Laboratório de Biocontrole Farroupilha, Brazil.
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  •  Received: 15 April 2016
  •  Accepted: 20 May 2016
  •  Published: 16 June 2016

 ABSTRACT

In order to optimize the use of nitrogen (N), the aim of this work was to evaluate the efficiency of foliar application of Azospirillum brasilense Ab-V5 strain, as regards the productivity of wheat plantations combined with different N doses. The experiments were carried out in four municipalities in Minas Gerais (Brazil) under randomized block design with four replications. Forty percent of the N dose was applied at planting time and 60% as topdressing at tillering stage of the crop. Azospirillum brasilense was applied as foliar spray at a dose of 500 ml/ha. The treatments consisted of: (1) Control-without N or Ab-V5; (2) 50% of the N recommended; (3) 100% of the N; (4) Application of Ab-V5 strain; (5) 50% of N and Ab-V5 strain; (6) 100% of N and Ab-V5 strain; (7) 100% of N and seed inoculation with a commercial product (Master Fix). For all the locations, productivity increased with application of 100% of N recommended and foliar spray containing A. brasilense compared to the treatment that had only 100% of N recommended.

 

Key words: Foliar spray, nitrogen fertilization, diazotrophic bacteria, Triticum aestivum L.


 INTRODUCTION

Wheat is the second cereal most produced in the world. In Brazil, it is grown in the South, Southeast and Midwest regions. In the 2015 to 2016 crop season, the cultivated area with wheat in Brazil was 2103 million hectares; with an average yield of 2770 kg ha-1 (Conab, 2016).
 
Nitrogen fertilization is essential, in order to ensure the production and quality of grains. This is the most limiting nutrient for wheat productivity (Rodrigues et al., 2014). Adequate N supplying determines the number of tillers, which may favor the nodes formation and cause the stem elongation. The increase in the number of tillers and greater elongation of the stem allow higher uptake of solar radiation and, therefore, greater productivity (Fornasieri Filho, 2008). In addition, the number of ears per area and the number of spikelets per ears increase the adequate availability and  application  of  this  nutrient (Megda et al., 2009).
 
 
Bacteria that belong to the Azospirillum genus are known to associate symbiotically with grass forming specialized structures in the roots in which there is conversion of N2 to NH3 (Radwan et al., 2004). For the establishment of a symbiotic relationship, germinating seeds and roots exude molecules that chemically attract the N fixing bacteria and stimulate its development in the rhizosphere and the gene expression of the bacteria related to the biological N fixation (Hungria et al., 2001).
 
The use of Azospirillum spp. in the development of grasses has been researched in recent years, not only regarding the yields but also regarding physiological effects. Recent reports states its positive effects on the growth and on the N accumulation in wheat plants (Sala et al., 2005), maintaining the fertile tillers, higher extraction and N accumulation in grains (Jezewski et al., 2010), more efficient translocation of N to the grains, heavier and full grains, better distribution of N in the grains biomass (Didonet et al., 2000) and root development. However, some authors did not find any difference between treatments with or without  inoculation (Campos et al., 1999). Studies on the application of Azospirillum brasilense via foliar spray in wheat plants are inconclusive and scarce. Farther, N fertilization is one of the most expensive management practices for the growers and the application of A. brasilense may increase the profits as well the use efficiency of this nutrient.
 
The objective of this study was to evaluate in field conditions the efficiency of foliar application of A. brasilense, Ab-V5 strain, as regard to the productivity of wheat cultivated with different N doses.


 MATERIALS AND METHODS

The experiments were carried out in four experimental fields located in the following municipalities of Minas Gerais state: Madre de Deus de Minas, Uberaba, Lambari and Patos de Minas. The plots were 6.0 m long and 4.0 m wide and presented 10 rows spaced at 0.17 m with 80 seeds/linear meter at sowing; the four central rows were considered the useful plot (12 m2). The experiment was designed as randomized block with 4 replications. The chemical and physical characteristics of the soil are presented in Table 1.
 
The dose of N indicated for the crop  under  irrigation  was  60 kg per hectare. It was applied as 40% of the N dose at planting time and 60% as topdressing at tillering stage. The A. brasilense strain Ab-V5 was used at a concentration of 1×108 colony forming units per ml. The Ab-V5 bacterium has been widely studied by “Embrapa Soja” and Federal University of Paraná; nowadays it is in the list of inoculant bacteria approved by Ministry of Agriculture, Livestock and Food Supply (MAPA) for corn and wheat. Ab-V5 was applied via foliar spray at the dose of 500 ml/ha at the beginning of tillering stage with a flow of 200 l/ha.
 
 
 
Nitrogen fertilization varied according to the treatments as it follows: (1) Absolute control,  without N or Ab-V5 strain; (2) 50% of the N dose recommended for the crop; (3) 100% of the N dose recommended for the crop; (4) Ab-V5 without N; (5) 50% of the N recommended for the crop with Ab-V5; (6) 100% of the N recommended for the crop with Ab-V5; (7) 100% of the N recommended for the crop and seeds inoculation with Master Fix (100 ml/20 kg of seeds) which is a commercial product containing A. brasilense.
 
Inoculation with the commercial product “Master Fix” composed by A. brasilense (Ab-V5 and Ab-V6 strains) was directly performed in the seed previous to sowing at a dose of 100 ml ha-1 of the inoculant liquid, containing around 2.0 × 108 colony forming units/ ml.
 
The crop management practices were uniform in all the experiments, except for N fertilization and in the experiment of Madre de Deus de Minas city where the wheat was grown in rainfed system. Sowing was made using a seeder for grooves opening, as well as marking the sowing lines and the N fertilizer was manually distributed. The experiments were performed in four different locations, as described as follow:
 
(a) Madre de Deus de Minas municipality in a field of “Fazenda Liberdade” located at ''21°27’04” S and ''44°19’14” W at the altitude of approximately 1026 m. Coodetec 108 wheat cultivar was cultivated. Sowing was performed on February 29th 2012 and the harvest was done on June 08th 2012.
(b) Uberaba municipality in a field of IFTM (Federal Institute of Triângulo Mineiro) located at ''19°43'6.94'' S and ''47°57'28.29'' W at the altitude of approximately 810 m. Coodetec 108 wheat cultivar was cultivated. Sowing was performed on May 24th 2012 and the harvest on September 02nd 2012.
(c) Lambari municipality in a field of EPAMIG (Agricultural Research Company of Minas Gerais) located at ''21°56´40.85'' S and ''45°18´40.91'' W at the altitude of approximately 883 m. Coodetec 207 cultivar was cultivated. Sowing was performed on April 03rd 2013 and harvest on September 15th 2013.
(d) Patos de Minas municipality in a field of EPAMIG (Agricultural Research Company of Minas Gerais) located at ''18°31´01.90'' S and ''46°26´19.08'' W at the altitude of approximately 926 m. Coodetec 207 cultivar was cultivated. Sowing was performed on May 17th 2013 and the harvest on September 17th 2013.
 
In all experiments, the ears were harvested from the useful plots, corresponding to 12 m2 (four central rows). To determine the grain yield, the ears were threshed and weighed in threshing electric machine. The grains had moisture corrected to 13% on wet basis.
 
The experimental data were submitted to variance analysis and the effects of treatments and interactions were evaluated by F test, whereas the treatment means were compared by the Scott-Knott test (Ferreira et al., 2014).
 
 

 


 RESULTS AND DISCUSSION

The individual variance analyses showed significant results for productivity. In this experiment, for most of the treatments, wheat cultivars showed higher yield than the national average which is 47.65 bags/ha (Conab, 2016). The analysis showed significant results for different locations, treatments and their interaction (Table 2).
 
 

Three groups of treatments were performed in Uberaba and Patos de Minas municipalities and two groups in Lambari and Madre de Deus de Minas. For all of them, the use of 100% N + Azospirillum via foliar spray yielded the best results. For the treatments with no application of A. brasilense (0, 50 and 100% of N) the productivity was significantly lower compared to the treatments with Azospirillum, however they belonged to the same group (Table 3). This lack of response may be related to the residual effect of N fertilizers applied to cultivate previous crops in all of these areas. In these regions the adoption of crop rotation system with leguminous and horticulture crops which leaves high amounts of N in the soil is common. It ensures the positive effect of Azospirillum on the N use efficiency. 

 

As this bacterium was applied via foliar spray, probably this increasing productivity might be related to changes in phytohormone metabolism and N metabolism, even in rainfed systems such as Lambari municipality. However, Azospirillum`s action on the plant`s metabolism is found to be controversial.
 
Recent reports states its increasing ability to fix N from atmosphere (Huergo et al., 2008); increasing effects on activity of nitrate reductase when they grow endophytically (Cassán et al., 2008), production of hormones such as auxins, cytokinins (Tien et al., 1979), gibberellins (Bottini et al., 1989); ethylene (Strzelczyk et al., 1994) and a variety of other molecules in the cell (Perrig et al., 2007); phosphate solubilization (Rodriguez  et al., 2004) and biological control of pathogens (Correa et al., 2008). So they are able to promote the development of roots and shoots, increase water and mineral absorption and optimize the tolerance to abiotic stresses such as salinity or drought (Roscoe and Miranda, 2013).
 
It is widely known that nitrate reduction occurs in the cytosol and involves the action of the nitrate reductase producing nitrite; it enters the plastids of roots or chloroplasts of leaves and it is reduced to ammonia by the  action  of  the  enzyme nitrite reductase, which   is attached via glutamate synthase/glutamine synthase (GS/GOGAT) in amino acids such as glutamine and glutamate which, in its turn serve as substrate for transamination reactions that are essential for the production of amino acids and proteins (Donato et al., 2004).
 
 
Nitrate reductase is one of the most sensitive enzymes to any stress in the plants, because it is highly dependent on NADPH derived from photosynthesis. Therefore, factors that enhances the photosynthetic efficiency, probably improves the N use efficiency. Nitrate reductase has been widely studied, because it controls protein synthesis in plants that absorbs nitrate as the main source of N (Marschner, 2011).
 
Possibly A. brasilense applied via foliar spray had a great effect on nitrate reductase and it increased the use efficiency of N applied via fertilization. Besides this, A. brasilense can also work fixing N from the atmosphere, which may help the plants save energy with the N reduction. Adequate nitrate reductase activity is primordial to guarantee high productivity; once N is one of the most limiting nutrients to form organic molecules (Taiz and Zeiger, 2013). This effect was observed in this experiment.
 
According to Sala et al. (2007) some wheat cultivars may present increases around 27 to 45% on grain production with A. brasilense inoculation. Nozaki et al. (2013) observed a significant increase on wheat productivity applying 290 kg/ha with 1.5 ml of Azzospirillum spp. Martins et al. (2012) observed that A. brasilense inoculation as foliar spray was more efficient in different corn hybrids and showed an excellent choice for use on grass, because it coincided with the herbicide application phase. However, Mendes et al. (2011) did not observed any difference between the treatments with reduction of the N fertilization and inoculation of A. brasilense for the number of tillers, number of ears and weight of 1000 grains.
 
According to Kapulnik et al. (1983), wheat plants inoculated with A. brasilense increased the contents of N, P and K. The contents of nitrate in the vacuole are directly related to nitrate reductase activity (Li and Gresshoff, 1990). Panwar (1991) observed that seeds of wheat inoculated with A. brasilense increased intensively the activity of this enzyme. Didonet et al. (2000) inoculated 245 strains with 10 isolates of A. brasilense in wheat plants with different doses of N and concluded that they provide better use of N accumulated in the biomass, translocating N more efficiently.  SwÄ™drzyÅ„ska (2000) concluded that A. brasilense can be a factor to increase vigor and yield of wheat. In water stress conditions the author observed an increase of 27% in wheat productivity.
 
Initially, some authors expected that the benefits with the use of A. brasilense were basically derived from biological N fixation (Dobbelaere et al., 2004). But it seems that the positive effects provided by these microorganisms are mainly derived from the morpho-logical and physiological changes in the roots of inoculated plants, causing an increase in the uptake of water and nutrients (Okon and Vanderleyden, 1997).
 
Probably this is the reason that the productivity had been so high, even in rainfed systems such as in Lambari municipality. Previous studies show an increasing concentration of the following phytohormones when Ab-V5 strain was inoculated in the plants: Kinetin which induces root growth; salicylic acid which may have an acclimatization effect providing increased tolerance to many different kinds of abiotic stresses; jasmonic acid that may induce gene expression regarding stress defense; indolbutyric acid which is a root promoter; indoleacetic acid which is growth promoter and gibberellic acid that stimulates plant growth.
 
Quadros et al. (2014) concluded that the use of Azospirillum stimulated the growth of plants in the vegetative period, which increased the uniformity of plant stand, greater resistance to stress and greater concentration of chlorophyll in leaves. The Ab-V5 strain induces the production of these phytohormones in a balanced way and possibly is capable to be absorbed by leaves, demonstrating the effectiveness of foliar application. Therefore, in the present study the use of A. brasilense may have a combined effect on wheat productivity, by phytohormones and N metabolism.
 
 

 

 

 

 
 


 CONCLUSION

The foliar application of A. brasilense, Ab-V5 strain, promotes an increasing productivity combined with 100% of the N dose. A. brasilense is a complementary technology focused on increasing wheat productivity especially under water stress conditions.


 CONFLICT OF INTERESTS

The authors have not declared any conflict of interests.


 ACKNOWLEDGEMENTS

The authors appreciate the Foundation of the State of Minas Gerais Research (FAPEMIG) for financial support and the grants awarded and Biocontrol Laboratory Farrukhabad for their help in conducting the tests.



 REFERENCES

Bottini R, Fulchieri M, Pearce D, Pharis R (1989). Identification of gibberelins A1, A3, and iso-A3 in cultures of A. lipoferum. Plant Physiol. 90:45-47.
Crossref
 

Campos BHC, Theisen S, Gnatta V (1999). Inoculante "Graminante" nas culturas de trigo e aveia. Cienc. Rural 23(3): 401-407.
Crossref

  
 

Cassán F, Sgroy V, Perrig D, Masciarelli O, Luna V (2008). Producción de fitohormonas por Azospirillum sp. Aspectos fisiológicos y tecnológicos de la promoción del crecimiento vegetal. In: Cassán FD, Garcia de Salamone I (Eds) Azospirillum sp.: cell physiology, plant interactions and agronomic research in Argentina. Argentina: Asoc. Argent. Microbiol. pp. 61-86.

  
 

Conab (2016). Companhia Nacional de Abastecimento : Acompanhamento da safra brasileira de grãos. Available at: 

View acessed date May 2016.

  
 

Correa OS, Romero AM, Soria MA, De Estrada M (2008). Azospirillum brasilense-plant genotype interactions modify tomato response to bacterial diseases, and root and foliar microbial communities. In: Cassán FD, Garcia de Salamone, I. (Eds) Azospirillum sp.: cell physiology, plant interactions and agronomic research in Argentina. Argentina: Asoc. Argent. Microbiol. pp. 87-95.

  
 

Didonet AD, Lima OS, Candaten AA, Rodrigues O (2000). Realocação de nitrogênio e de biomassa para os grãos, em trigo submetido a inoculação de Azospirillum. Pesq. Agropec. Bras. 35:401-411.
Crossref

  
 

Dobbelaere S, Croonenborghs A, Thys A, Ptacek D, Donato VMTS, Andrade AG, Souza ES, França JGE, Maciel GA (2004). Atividade enzimática em variedades de cana-de-açúcar cultivadas in vitro sob diferentes níveis de nitrogênio. Pesq. Agropec. Bras. 39(11):1087-1093.
Crossref

  
 

Donato VMTS, Andrade AG, Souza ES, França JGE, Maciel GA (2004). Atividade enzimática em variedades de cana-de-açúcar cultivadas in vitro sob diferentes níveis de nitrogênio. Pesq. Agropec. Bras. 39(11):1087-1093.
Crossref

  
 

Ferreira DF (2014). Sisvar a Guide for its Bootstrap procedures in multiple comparisons. Cien. Agrotecnologia 38:109-112.
Crossref

  
 

Fornasieri FD (2008). Manual da cultura do trigo. Jaboticabal: Fundação de Apoio à Pesquisa, Ensino e Extensão P 338.

  
 

Huergo LF, Monteiro RA, Bonato AC, Rigo LU, Steffens MBR, Cruz LM, Chubatsu LS, Souza EM, Pedrosa FO (2008). Regulation of nitrogen fixation in Azospirillum brasilense. In: Cassán FD, Garcia de Salamone I. Azospirillum sp.: cell physiology, plant interactions and agronomic research in Argentina. Asociación Argentina de Microbiologia, Ciudad de Buenos Aires, Argentina P 268.

  
 

Hungria M, Campo RJ, Mendes IC (2001). Fixação biológica do nitrogênio na cultura da soja. Londrina: Embrapa Soja, (Circular Técnica 35).

  
 

Jezewski TJ, Silva JAG, Fernandes SBV (2010). Efeito da inoculação de Azospirillum em trigo, isolado e associado a estimulante de crescimento no noroeste do RS. In: Congresso de Iniciação Científica da UFPEL, Pelotas/RS. XIX Congresso de Iniciação Científica, XII Encontro de Pós-Graduação e II Mostra Científica da UFPEL. Anais... Pelotas 1:568-571.

  
 

Kapulnik Y, Sarig S, Nur I, Okon Y (1983). Effect of Azospirillum inoculation on yield of field-grown wheat. Ca. J. Microbiol. 29:895-899.
Crossref

  
 

Li ZZ, Gresshoff PM (1990). Developmental and biochemical regulation of constitutive' nitrate reductase activity in leaves of nodulating soybean. J. Exp. Bot. 41:1231-1238.
Crossref

  
 

Marschner P (2011). Mineral Nutrition of Higher Plants, Third Edition. Acad. Press P 672.

  
 

Martins FAD, Andrade AT, Condé ABT, Godinho DB, Caixeta CG, Costa RL, Pomela AWV, Soares CMS (2012). Avaliação de híbridos de milho inoculados com Azospirillum brasilense. Pesq. Agrop. Gaúcha 18(2):113-128.

  
 

Megda MM, Buzetti S, Andreotti M, Teixeira Filho MMC, Vieira MX (2009). Resposta de cultivares de trigo ao nitorgênio em relação às fontes e épocas de aplicação sob plantio direto e irrigação por aspersão. Cienc. Agrotec. 33:1055-1060.
Crossref

  
 

Mendes MC, Rosário JG do, Faria MV, Zocche JC, Walter AL (2011). Avaliação da eficiência agronômica deAzospirillum brasilense na cultura do trigo e os efeitos na qualidade de farinha. Rev. Bras. Tecnol. Aplicada nas Ciênc. Agrárias 4:95-102.

  
 

Nozaki MH, Lorenzatto R, Mancini M (2013). Efeito do Azospirillum spp. em associação com diferentes doses de adubação mineral na cultura do trigo. Ensaios e Ciência: Ciências Biológicas, Agrárias da Saúde 17(6):27-35.

  
 

Okon Y, Vanderleyden J (1997). Root-associated Azospirillum species can stimulate plants. Appl. Environ. Microbiol. 63(7):364-370.

  
 

Panwar JDS (1991). Effect of VAM and Azospirillum brasilense on photosynthesis nitrogen metabolism and grain yield in wheat. Indian J. Plant Physiol. 34:357-361.

  
 

Perrig D, Boeiro L, Masciarelli O, Penna C, Cassán F, Luna V (2007). Plant growth promoting compounds produced by two agronomically important strains of Azospirillum brasiliense, and their implications for inoculants formulation. Appl. Environ. Microbiol. 75:1143-1150.

  
 

Quadros PD, Roesch LFW, Silva PRF, Vieira VM, Roehrs DD, Camargo FAO (2014). Desempenho agronômico a campo de híbridos de milho inoculados com Azospirillum. Rev. Ceres 61:209-218.
Crossref

  
 

Radwan TEE, Mahamed ZK, Reis VM (2004). Efeito de inoculação de Azzospirillum e Herbaspirillum na produção de compostos indólicos em plântulas de milho e arroz. Pes. Agropec. Bras. 39:987-994.

  
 

Rodrigues LFOS, Guimarães VF, Silva MB, Pinto Junior AS, Klein J, Costa ACPR (2014). Características agronômicas do trigo em função de Azospirillum brasiliense, ácidos húmicos e nitrogênio em casa de vegetação. R. Bras. Eng. Agríc. Amb. 18:31-37.

  
 

Rodriguez H, Gonzalez T, Goire I, Bashan Y (2004). Gluconic acid production and phosphate solubilization by the plant growth-promoting bacterium Azospirillum spp. Naturwissenschaften 91:552-555.
Crossref

  
 

Roscoe R, Miranda RAS (2013). Fixação Biológica de Nitrogênio e Promoção de Crescimento em Milho Safrinha. In: Roscoe R, Lourenção ALF, Grigolli JFJ, Melotto AM, Pitol C, Miranda RAS. Tecnologia e produção: milho safrinha e culturas de inverno. Midiograf, Curitiba, Brasil P 70.

  
 

Sala VMR, Cardoso EJBN, Freitas JG, Silveira APD. (2007). Resposta de genótipos de trigo à inoculação de bactérias diazotróficas em condições de campo. Pesqui. Agropecu. Bras. 42:833-842.
Crossref

  
 

Sala VMR, Freitas SS, Donzeli VP, Freitas JG, Gallo PB, Silveira APD (2005). Ocorrência e efeito de bactérias diazotróficas em genótipos de trigo. Rev. Bras. Cienc. Solo 29(3):345-352.
Crossref

  
 

Strzelczyk E, Kamper M, Li C (1994). Cytocinin-like-substances and ethylene production by Azospirillum in media with different carbon sources. Microbiol. Res. 149:55-60.
Crossref

  
 

Swędrzyńska D (2000). Effect of inoculation with Azospirillum brasilense on development and yielding of winter wheat and oat under different cultivation conditions. Pol. J. Environ. Stud. 9(5):423-428.

  
 

Taiz L, Zeiger E (2013). Fisiologia Vegetal. Porto Alegre: Artmed, P 954.

  
 

Tien TM, Gaskins MH, Hubbell DH (1979) Plant growth substances produced by Azospirillum brasilense and their effect on the growth of pearl millet (Pennisetum americanum L.). Appl. Environ. Microbiol. 37:1016-1024.

  

 




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