Abstract
The 15N natural abundance (δ15N) technique offers good prospects for the quantification of the contribution of biological nitrogen fixation (BNF) to crop nutrition. The objectives of this study were to determine which plant parts best represented the whole plant with respect to δ15N and to quantify the BNF contribution to sugarcane. The experiment was carried out in pots of soil (100 kg pot−1) from the field. The sugarcane varieties used were RB867515 and RB92579 with the reference plants Brachiaria decumbens, millet (Pennisetum americanum) and sorghum (Sorghum bicolor). The experimental design was randomised blocks with four replications. The treatments were inoculation of the sugarcane with or without diazotrophic bacteria and the different parts of the sugarcane plant (root, green leaf, senescent leaf and different stem sections). The δ15N of different cane parts and varieties ranged from + 2.68 to + 5.85‰ after 365 days, while the δ15N of reference plants were from + 7.8 to + 8.3‰. The mean contribution of BNF to the sugarcane crop N supply was estimated to be 47% with no effect of variety or inoculation. The results indicate that the δ15N value of the entire shoot can be considered to represent the entire sugarcane plant, but no single organ could be relied on for this purpose.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Code Availability
Not applicable.
References
Aquino GS, Medina CC, Shahab M, Santiago AD, Cunha ACB, Kussaba DAO, Carvalho JB, Moreira A (2018) Does straw mulch partial-removal from soil interfere in yield and industrial quality sugarcane? A long-term study. Ind Crops Prod 111:573–578. https://doi.org/10.1016/j.indcrop.2017.11.026
Araújo KEC, Vergara C, Guimarães AP, Rouws JRC, Jantalia CP, Uquiaga S, Alves BJR, Boddey RM (2018) Changes in 15N natural abundance of biologically fixed N2 in soybean due to shading, rhizobium strain and plant growth stage. Plant Soil 426:413–428. https://doi.org/10.1007/s11104-018-3627-4
Baldani JI, Baldani VLD, Seldin L, Döbereiner J (1986) Characterization of Herbaspirillum seropedicae gen. nov. sp. nov. a root associated nitrogen fixing bacterium. Int J of Syst Evol Bacteriol 36:86–93. https://doi.org/10.1099/00207713-36-1-86
Baldani JI, Pot B, Kirchhof G, Falsen E, Baldani VLD, Olivares FL, Hoste B, Kersters K, Hartmann A, Gillis M, Döbereiner J (1996) Emended description of Herbaspirillum; inclusion of [Pseudomonas] rubrisubalbicans, a mild plant pathogen, as Herbaspirillum rubrisubalbicans comb. nov.; and classification of a group of clinical isolates EF group 1 as Herbaspirillum species 3. Int J Syst Evol Bacteriol 46:802–810. https://doi.org/10.1099/00207713-46-3-802
Baptista RB, Morais RF, Leite JM, Schultz N, Alves BJR, Boddey RM, Urquiaga S (2014) Variations in the 15N natural abundance of plant-available N with soil depth: their influence on estimates of contributions of biological N2 fixation to sugarcane. Appl Soil Ecol 73:124–129. https://doi.org/10.1016/j.apsoil.2013.08.008
Bloch SE, Ryu MH, Ozaydin B, Broglie R (2020) Harnessing atmospheric nitrogen for cereal crop production. Curr Opin Biotechnol 62:181–188. https://doi.org/10.1016/j.copbio.2019.09.024
Boddey RM, Peoples MB, Palmer B, Dart PJ (2000) Use of the 15N natural abundance technique to quantify biological nitrogen fixation by woody perennials. Nut Cycl Agroecosystems 57:235–270. https://doi.org/10.1023/A:1009890514844
Boddey RM, Polidoro JC, Resende AS, Alves BJR, Urquiaga S (2001) Use of the 15N natural abundance technique for the quantification of the contribution of N2 fixation to grasses and cereals. A J Plant Physiology 2:889–895. https://doi.org/10.1071/PP01058
Bordonal RO, Carvalho JLN, Lal R, de Figueiredo EB, Oliveira BG, La Scala N (2018) Sustainability of sugarcane production in Brazil. A Review Agron Sustain Dev 38:1–23. https://doi.org/10.1007/s13593-018-0490-x
Caldeira-Pires A, Benoist A, Luz SM, Silverio VC, Silveira CM, Machado FS (2018) Implications of removing straw from soil for bioenergy: an LCA of ethanol production using total sugarcane biomass. J Clean Prod 181:249–259. https://doi.org/10.1016/j.jclepro.2018.01.119
Carvalho EX, Menezes RSC, Freitas ADS, Sampaio EVS, Neto DES, Tabosa JN, Primo DC, Queiroz RO (2017) The 15N natural abundance technique to assess the potential of biological nitrogen fixation (BNF) in some important C4 grasses. Austr J of Crop Sci 11:1559–1564. https://doi.org/10.21475/ajcs.17.11.12.pne729
Castro-González R, Martínez-Aguilar L, Ramírez-Trujillo A, Estrada-de los Santos P, Caballero-Mellado J, (2011) High diversity of culturable Burkholderia species associated with sugarcane. Plant Soil 345:155–169. https://doi.org/10.1007/s11104-011-0768-0
Castro Gava GJ, Scarpare FV, Cantarella H, Kölln OT, Ruiz-Corrêa ST, Arlanch AB, Trivelin PCO (2019) Nitrogen source contribution in sugarcane-inoculated plants with diazotrophic bacterias under urea-N fertigation management. Sugar Tech 21:462–470. https://doi.org/10.1007/s12355-018-0614-2
Cavalcante VA, Dobereiner J (1988) A new acid tolerant nitrogen-fixing bacterium associated with sugarcane. Plant Soil 108:23–31. https://doi.org/10.1007/BF02370096
Chaves VA, Santos SG, Schultz N, Pereira W, Sousa JS, Monteiro RC, Reis VM (2015) Desenvolvimento inicial de duas variedades de cana-de-açúcar inoculadas com bactérias diazotróficas. Rev Bras Ciênc Solo 39:1595–1602. https://doi.org/10.1590/01000683rbcs20151144
Dos Santos RL, Freire FJ, Oliveira ECA, Trivelin PCO, Freire MBGS, Bezerra PC, Oliveira RI, Santos MBC (2019) Changes in biological nitrogen fixation and natural-abundance N isotopes of sugarcane under molybdenum fertilization. Sugar Tech 21:925–935. https://doi.org/10.1007/s12355-019-00717-w
Fischer D, Pfitzner B, Schmid M et al (2012) Molecular characterisation of the diazotrophic bacterial community in uninoculated and inoculated field-grown sugarcane (Saccharum sp). Plant Soil 356:83–99. https://doi.org/10.1007/s11104-011-0812-0
Landell MGA, Campana MP, Figueiredo P et al (2012) Sistema de multiplicação de cana-de-açúcar com uso de mudas pré-brotadas (MPB), oriundas de gemas individualizadas, Fundação IAC Boletim 109. Campinas, São Paulo, Brazil.
Lima E, Boddey RM, Dobereiner J (1987) Quantification of biological nitrogen fixation associated with sugar cane using a 15N aided nitrogen balance. Soil Biol Biochem 19:165–170. https://doi.org/10.1016/0038-0717(87)90077-0
Lin SY, Hameed A, Shen FT, Liu YC, Hsu YH, Shahina M, Lai WA, Young CC (2014) Description of Niveispirillum fermenti gen. nov., sp. nov., isolated from a fermentor in Taiwan, transfer of Azospirillum irakense 1989 as Niveispirillum irakense comb. nov., and reclassification of Azospirillum amazonense 1983 as Nitrospirillum amazonense gen. nov. Antonie Van Leeuwenhoek 105:1149–1162. https://doi.org/10.1007/s10482-014-0176-6
LSPA-IBGE. Instituto Brasileiro de Geografia e Estatística (2020) Levantamento Sistemático da Produção Agrícola, Rio de Janeiro, Brasil. Disponível em: https://sidra.ibge.gov.br/home/lspa/brasil. Aceessed 23 Nov 2020.
Magalhães FM, Baldani JI, Souto SM, Kuykendall JR, Döbereiner J (1983) A new acid-tolerant Azospirillum species. An Acad Bras Ciênc 55:417–430
Makarov MI, Buzin IS, Tiunov AV, Malysheva TI, Kadulin MS (2019) nitrogen isotopes in soils and plants of tundra ecosystems in the khibiny mountains. Eurasian Soil Sci 52:119–1206. https://doi.org/10.1134/S1064229319100077
Martins DS, Reis VM, Schultz N, Alves BJR, Urquiaga S, Pereira W, Sousa JS, Boddey RM (2020) Both the contribution of soil nitrogen and of biological N2 fixation to sugarcane can increase with the inoculation of diazotrophic bacteria. Plant Soil 452:1–15. https://doi.org/10.1007/s11104-020-04621-1
Morais RF, Quesada DM, Reis VM, Urquiaga S, Alves BJR, Boddey RM (2012) Contribution of biological nitrogen fixation to Elephant grass (Pennisetum purpureum Schum.). Plant Soil 356:23–34. https://doi.org/10.1007/s11104-011-0944-2
Nogueira ARA, Souza GB (2005) Manual de laboratório: solo, água, nutrição vegetal, nutrição animal e alimentos. Embrapa Pecuária Sudeste, São Carlos, SP, Brazil.
Oliveira ALM, Urquiaga S, Döbereiner J, Baldani JI (2002) The effect of inoculating endophytic N2-fixing bacteria on micropropagated sugarcane plants. Plant Soil 242:205–215. https://doi.org/10.1023/A:1016249704336
Oliveira ALM, Canuto EL, Reis VM, Baldani JI (2003) Response of micropropagated sugarcane varieties to inoculation with endophytic diazotrophic bacteria. Braz J Microbiol 34:59–61. https://doi.org/10.1590/S1517-83822003000500020
Oliveira ALM, Canuto EL, Urquiaga S, Reis VM, Baldani JI (2006) Yield of micropropagated sugarcane varieties in different soil types following inoculation with diazotrophic bacteria. Plant Soil 284:23–32. https://doi.org/10.1007/s11104-006-0025-0
Oren A, Garrity GM (2015) List of new names and new combinations previously effective, but not validly, published. Int Syst Evol Microbiol 65:2017–2025. https://doi.org/10.1099/ijs.0.000317
Pardo LH, Semaoune P, Schaberg PG, Eagar C, Sebilo M (2013) Patterns in δ15N in roots, stems, and leaves of sugar maple and American beech seedlings, saplings, and mature trees. Biogeochemistry 112:275–291. https://doi.org/10.1007/s10533-012-9724-1
Pereira W, Sousa JS, Schultz N, Reis VM (2019) Sugarcane productivity as a function of nitrogen fertilization and inoculation with diazotrophic plant growth-promoting bacteria. Sugar Tech 21:71–82. https://doi.org/10.1007/s12355-018-0638-7
Perin L, Martínez-Aguilar L, Castro-González R, Estrada-de los Santos P, Cabellos-Avelar T, Guedes HV, Reis VM, Caballero-Mellado J, (2006) Diazotrophic Burkholderia species associated with field-grown maize and sugarcane. Appl Environ Microbiol 72:3103–3110. https://doi.org/10.1128/AEM.72.5.3103-3110.2006
Reis VM, Olivares FL, Dobereiner J (1994) Improved methodology for isolation of Acetobacter diazotrophicus and confirmation of its endophytic habitat. World J of Microbiol and Biotechnol 10:101–104. https://doi.org/10.1007/BF00144460
Reis VM, Estrada-de los Santos P, Tenorio-Salgado S, Vogel J, Stoffels M, Guyon S, Mavingui P, Baldani VL, Schmid M, Baldani JI, Balandreau J, Hartmann A, Caballero-Mellado J (2004) Burkholderia tropica sp nov, a novel nitrogen-fixing, plant-associated bacterium Int. J. Syst. Evol. Microbiol 4:2155–2162. https://doi.org/10.1099/ijs.0.02879-0
Rouws LFM, Leite J, Mattos GF, Zilli JE, Coelho MRR, Xavier GR, Fischer D, Hartmann A, Reis VM, Baldani JI (2014) Endophytic Bradyrhizobium spp. isolates from sugarcane obtained through different culture strategies. Environ Microbiol Rep 6:354–363. https://doi.org/10.1111/1758-2229.12122
Sanguino A, Moraes VA, Casagrande MV (2006) Training course and management of sugarcane seedling nurseries, Campinas, São Paulo, Brazil.
Santos SG, Ribeiro FS, Fonseca CS, Pereira W, Santos LA, Reis VM (2017) Development and nitrate reductase activity of sugarcane inoculated with five diazotrophic strains. Arch Microbiol 199:863–873. https://doi.org/10.1007/s00203-017-1357-2
Schultz N, Morais RF, Silva JA, Baptista RB, Oliveira RP, Leite JM, Pereira W, Carneiro Júnior JB, Alves BJR, Baldani JI, Boddey RM, Urquiaga S, Reis VM (2012) Agronomic evaluation of sugarcane varieties inoculated with diazotrophic bacteria and fertilized with nitrogen. Pesqui Agropecu Bras 47:261–268. https://doi.org/10.1590/S0100-204X2012000200015
Schultz N, Pereira W, Reis VM, Urquiaga S (2016) Productivity and 15N isotope dilution of sugarcane inoculated with diazotrophic bacteria. Pesqui Agropecu Bras 51:1594–1601. https://doi.org/10.1590/s0100-204x2016000900059
Schultz N, Pereira W, da Silva PA, Baldani JI, Boddey RM, Alves BJR, Urquiaga S, Reis VM (2017) Yield of sugarcane varieties and their sugar quality grown in different soil types and inoculated with diazotrophic bacteria consortium. Plant Prod Sci 20:366–374. https://doi.org/10.1080/1343943X.2017.1374869
Shearer G, Kohl DH, Virginia RA, Bryan BA, Skeeters JL, Nilsen ET, Sharifi MR, Rundel PW (1983) Estimates of N2-fixation from variation in the natural abundance of 15N in sonoran desert ecosystems. Oecologia 56:365–373. https://doi.org/10.1007/BF00379714
Shearer G, Kohl DH (1986) N2-fixation in field settings: estimations based on natural 15N abundance. Func Plant Biol 13:699–756. https://doi.org/10.1071/PP9860699
Silva FAS, Azevedo CAV (2016) The Assistat Software Version 7.7 and its use in the analysis of experimental data. Afr J Agric Res 11:3733–3740. https://doi.org/10.5897/AJAR2016.11522
Souza RBS, Okura VK, Armanhi JSL, Jorrín B, Lozano N, Silva MJ, González-Guerreiro M, Araújo LM, Verza NC, Bagheri HC, Imperial J, Arruda P (2016) Unlocking the bacterial and fungal community’s assemblages of sugarcane microbiome. Sci Rep 6:28774. https://doi.org/10.1038/srep28774
Spaepen S, Bossuyt S, Engelen K, Marchal K, Vanderleyden J (2014) Phenotypical and molecular responses of Arabidopsis thaliana roots as a result of inoculation with the auxin-producing bacterium Azospirillum brasilense. New Phytol 201:850–886. https://doi.org/10.1111/nph.12590
Taulé C, Mareque C, Barlocco C, Hackembruch F, Reis VM, Sicardi M, Battistoni F (2012) The contribution of nitrogen fixation to sugarcane (Saccharum officinarum L.), and the identification and characterization of part of the associated diazotrophic bacterial community. Plant Soil 356:35–49. https://doi.org/10.1007/s11104-011-1023-4
Thaweenut N, Hachisuka Y, Ando S, Yanagisawa, S, Yoneyama T (2011) Estudo de duas temporadas da expressão do gene nifH e fixação de nitrogênio por endófitos diazotróficos em cana-de-açúcar (Saccharum spp. Híbridos): expressão de genes nifH semelhantes aos de rizóbio. Plant Soil 338, 435-449. https://doi.org/10.1007/s11104-010-0557-1
Unkovich M, Herridge D, Peoples M, Cadisch G, Boddey R, Giller K, Alves B, Chalk P (2008) Measuring plant-associated nitrogen fixation in agricultural systems. ACIAR monograph, Australia.
Urquiaga S, Cruz KHS, Boddey RM (1992) Contribution of nitrogen fixation to sugar cane: nitrogen-15 and nitrogen balance estimates. Soil Sci Soc Am J 56:105–114. https://doi.org/10.2136/sssaj1992.03615995005600010017x
Urquiaga S, Xavier R, Morais RF, Batista R, Schultz N, Leite JM, Resende A, Alves BJR, Boddey RM (2012) Evidence from field nitrogen balance and 15N natural abundance data of the contribution of biological N2 fixation to Brazilian sugarcane varieties. Plant Soil 356:5–21. https://doi.org/10.1007/s11104-011-1016-3
Zotarelli L, Zatorre N, Alves BJR, Jantalia CP, Franchini J, Urquiaga S, Boddey RM (2012) Influence of no-tillage and frequency of a green manure legume in crop rotations on balancing n outputs and preserving soil organic c stocks. Field Crop Res 32:185–195. https://doi.org/10.1016/j.fcr.2011.12.013
Acknowledgements
The authors thank Alderi F. da Silva, Ernani Meirelles, and their diligent team of field workers for managing the experiment and Altiberto M. Baêta, Roberto G. de Souza, and Renato M. da Rocha for the nitrogen and 15N abundance measurements.
Funding
The work was funded by Embrapa, the National Council for Scientific and Technological Development (CNPq). The first author, ECM gratefully acknowledges a PhD fellowship from CAPES (Ministry of Education), in agreement with Embrapa Agrobiologia (CAPES/EMBRAPA). The authors VMR, BJRA, SU, and RMB gratefully acknowledge grants from the programme Cientista de Nosso Estado from the Carlos Chagas Rio State Research Foundation (FAPERJ) and Research Productivity Fellowships from CNPq.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of Interest
The authors declare no competing interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Monteiro, E.d., da Silva, C.G.N., dos reis Martins, M. et al. Strategy for the Sampling of Sugarcane Plants for the Reliable Quantification of N2 Fixation Using 15N Natural Abundance. J Soil Sci Plant Nutr 21, 2741–2752 (2021). https://doi.org/10.1007/s42729-021-00561-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s42729-021-00561-6