Skip to main content
SpringerLink
Log in

Microarray analysis of humic acid effects on Brassica napus growth: Involvement of N, C and S metabolisms

  • Regular Article
  • Published:
Plant and Soil Aims and scope Submit manuscript

Abstract

Background & aims

Winter rapeseed (Brassica napus) is characterized by a low N recovery in seeds and requires high rates of fertilization to maintain yield. Its nutrient use efficiency could be improved by addition of a biostimulant such as humic acids whose physiological effects have been described previously in some plant species. However, to our knowledge, no study has focused on transcriptomic analyses to determine metabolic targets of this extract.

Methods

A preliminary screening of ten humic acids revealed a significant effect of one of them (HA7) on rapeseed root growth. Microarray analysis was then used on HA7-treated or non-treated plants to characterize changes in gene expression that were further supported by physiological evidence.

Results

Stimulation of nitrogen uptake (+15% in shoots and +108% in roots) and assimilation was found to be increased in a similar manner to growth while sulfate content (+76% in shoots and +137% in roots) was more strongly stimulated leading to higher sulfate accumulation. In parallel, microscopic analysis showed an enhancement of chloroplast number per cell.

Conclusion

It is therefore suggested that HA7, which promotes plant growth and nutrient uptake, could be used as a supplementary tool to improve rapeseed nitrogen use efficiency.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Abdallah M, Etienne P, Ourry A, Meuriot F (2011) Do initial S reserve and mineral S availability alter leaf S-N mobilization and leaf senescence in oilseed rape? Plant Sci 180:511–520

    Article  PubMed  CAS  Google Scholar 

  • Abiven S, Menasseri S, Chenu C (2009) The effect of organic input over time on soil aggregate stability—a literature analysis. Soil Biol Biochem 41:1–12

    Article  CAS  Google Scholar 

  • Aguirre E, Leménager D, Bacaicoa E, Fuentes M, Baigorri R, Zamarreño AM, García-Mina JM (2009) The root application of a purified leonardite humic acid modifies the transcriptional regulation of the main physiological root responses to Fe deficiency in Fe-sufficient cucumber plants. Plant Physiol Biochem 47:215–223

    Article  PubMed  CAS  Google Scholar 

  • Andersson S, Nilsson SI, Saetre P (2000) Leaching of dissolved organic carbon (DOC) and dissolved organic nitrogen (DON) in mor humus as affected by temperature and pH. Soil Biol Biochem 32:1–10

    Article  CAS  Google Scholar 

  • Arancon NQ, Edwards CA, Bierman P, Metzger JD, Lucht C (2005) Effects of vermicomposts produced from cattle manure, food waste and paper waste on the growth and yield of peppers field. Pedobiologia 49:297–306

    Article  CAS  Google Scholar 

  • Arancon NQ, Adwards CA, Bierman P (2006) Influence of vermicomposts on field strawberries: Part 2. Effects on soil microbiological and chemical properties. Bioresource Technology 97:831–840

    Article  PubMed  CAS  Google Scholar 

  • Atiyeh RM, Lee S, Edwards CA, Arancon NQ, Metzger JD (2002) The influence of humic acids derived from earthworm-processed organic wastes on plant growth. Bioresource Technology 84:7–14

    Article  PubMed  CAS  Google Scholar 

  • Ayuso M, Hernandez T, Garcia C, Pascual JA (1996) Stimulation of barley growth and nutrient absorption by humic substances originating from various organic materials. Bioresource Technology 57:251–257

    Article  CAS  Google Scholar 

  • Bossuyt H, Six J, Hendrix PF (2007) Protection of soil carbon by microaggregates within earthworm casts. Soil Biol Biochem 37:251–258

    Article  Google Scholar 

  • Bungard R, Wingler A, Morton J, Andrews M, Press M, Scholes J (1999) Ammonium can stimulate nitrate and nitrite reductase in the absence of nitrate in Clematis vitalba. Plant Cell Environ 22:859–866

    Article  CAS  Google Scholar 

  • Canellas LP, Olivares FL, Okorokova-Façanha AL, Façanha AR (2002) Humic acids isolated from earthworm compost enhance root elongation, lateral root emergence, and plasma membrane H+-ATPase activity in maize roots. Plant Physiol 130:1951–1957

    Article  PubMed  CAS  Google Scholar 

  • Canellas LP, Piccolo A, Dobbss LB, Spaccini R, Olivares FL, Zandonadi DB, Façanha AR (2010) Chemical composition and bioactivity properties of size-fractions separated from a vermicompost humic acid. Chemosphere 78:457–466

    Article  PubMed  CAS  Google Scholar 

  • Carletti P, Masi A, Spolaore B, Polverino De Laureto P, De Zorzi M, Turetta L, Ferretti M, Nardi S (2008) protein expression changes in maize roots in response to humic substances. J Chem Ecol 34:804–818

    Article  PubMed  CAS  Google Scholar 

  • Castaings L, Marchive C, Meyer C, Krapp A (2010) Nitrogen signaling in Arabidopsis: how to obtain insights into a complex signaling network. J Exp Bot 62:1391–1347

    Article  PubMed  Google Scholar 

  • Cesco S, Nikolic M, Römheld V, Varanini Z, Pinton R (2002) Uptake of 59Fe from 59Fe-humate complexes by cucumber and barley plants. Plant Soil 241:121–128

    Article  CAS  Google Scholar 

  • Christl I, Kretzschmar R (2001) Relating ion binding by fulvic and humic acids to chemical composition and molecular size. 1. Proton binding. Environ Sci Technol 35:2505–2511

    Article  PubMed  CAS  Google Scholar 

  • Conte P, Piccolo A (1999) Conformational arrangement of dissolved humic substances. Influence of solution composition on association of humic molecules. Environ Sci Technol 33:1682–1690

    Article  CAS  Google Scholar 

  • Cookson WR, Abaye DA, Marschner P, Murphy DV, Stockdale EA, Goulding KWT (2005) The contribution of soil organic matter fractions to carbon and nitrogen mineralization and microbial community size and structure. Soil Biol Biochem 37:1726–1737

    Article  CAS  Google Scholar 

  • Daniel-Vedele F, Filleur S, Caboche M (1998) Nitrate transport: a key step in nitrate assimilation. Curr Opin Plant Biol 1:235–239

    Article  PubMed  CAS  Google Scholar 

  • Dejoux JF, Recous S, Meynard JM, Trinsoutrot I, Leterme P (2000) The fate of nitrogen from winter-frozen rapeseed leaves: mineralization, fluxes to the environment and uptake by rapeseed crop in spring. Plant Soil 218:257–272

    Article  CAS  Google Scholar 

  • Dell’Agnola G, Nardi S (1987) Hormone-like effect and enhanced nitrate uptake induced by depolycondensed humic fractions obtained from Allolobophora rosea and A. caliginosa faeces. Biol Fertil Soils 4:115–118

    Google Scholar 

  • Desclos M, Dubousset L, Etienne P, Bonnefoy J, Lecahérec F, Satoh H, Ourry A, Avice JC (2008) A proteomic profiling approach to reveal a novel role of BnD22 (Brassica napus drought 22)/water soluble chlorophyll binding protein in young leaves during nitrogen remobilization induced by stressful condition. Plant Physiol 147:1830–1844

    Article  PubMed  CAS  Google Scholar 

  • Dobrev P, Kamiek M (2002) Fast and efficient separation of cytokinins from auxin and abscisic acid and their purification using mixed-mode solid-phase extraction. J Chromatogr A 950:21–29

    Article  PubMed  Google Scholar 

  • Dreccer MF, Schapendonk AHM, Slafer GA, Rabbinge R (2000) Comparative response of wheat and oilseed rape to nitrogen supply: absorption and utilisation efficiency of radiation and nitrogen during reproductive stage determining yield. Plant Soil 220:189–205

    Article  CAS  Google Scholar 

  • Esparza I, Salinas I, Santamaria C, Garcia-Mina JM, Fernandez JM (2005) Electrochemical and theoretical complexation studies for Zn and Cu with individual polyphenols. Analytica Chimica Acta 543:267–274

    Article  CAS  Google Scholar 

  • Etienne P, Desclos M, Le Gou L, Gombert J, Bonnefoy J, Maurel K, Le Dily F, Ourry A, Avice JC (2007) N-protein mobilization associated with the leaf senescence process in oilseed rape in concomitant with the disappearance of trypsin inhibitor activity. Funct Plant Biol 34:895–906

    Article  CAS  Google Scholar 

  • Eyheraguibel B, Silvestre J, Morard P (2008) Effects of humic substances derived from organic waste enhancement on the growth and mineral nutrition of maize. Bioresource Technology 99:4206–4212

    Article  PubMed  CAS  Google Scholar 

  • Garcia-Mina JM, Antolin MC, Sanchez-Diaz M (2004) Metal-humic complexes and plant micronutrient uptake: a study based on different plant species cultivated in diverse soil types. Plant Soil 258:57–68

    Article  CAS  Google Scholar 

  • Garcia-Mina JM (2006) Stability, solubility and maximum metal-binding capacity in metal-humic complexes involving humic substances extracted from peat and organic compost. Org Geochem 37:1960–1972

    Article  CAS  Google Scholar 

  • Imbufe AU, Patti AF, Burrow D, Surapaneni A, Jackson WR, Milner AD (2005) effects of potassium humate on aggregate stability of two soils from Victoria, Australia. Geoderma 125:321–330

    Article  CAS  Google Scholar 

  • Itoh R, Fujiwara M, Nagata N, Yoshida S (2001) A chloroplast protein homologous to the eubacterial topotogical specificity factor MinE plays a role in chloroplast division. Plant Physiol 127:1644–1655

    Article  PubMed  CAS  Google Scholar 

  • Keeling AA, McCallum KR, Beckwith CP (2003) Mature green waste compost enhances growth and nitrogen uptake in wheat (Triticum aestivum L.) and oilseed rape (Brassica napus L.) through the action of water-extractable factors. Bioresource Technology 90:127–132

    Article  PubMed  CAS  Google Scholar 

  • Kojo KH, Fujiwara MT, Itoh RD (2009) Involvement of AtMinE in plastid morphogenesis in various tissues of Arabidopsis thaliana. Biosci Biotechnol Biochem 73:2632–2639

    Article  PubMed  CAS  Google Scholar 

  • Kozuch J, Pempkowiak J (1996) Molecular weight of humic acids as a major property of the substances influencing the accumulation rate of cadmium by a blue mussel (Mytilus edulis). Environ Int 22:585–589

    Article  CAS  Google Scholar 

  • Krouk G, Lacombe B, Bielach A, Perrine-Walker F, Malinska K, Mounier E, Hoyerova K, Tillard P, Leon S, Ljung K, Zazimalova E, Benkova E, Nacry P, Gojon A (2010) Nitrate-regulated auxin transport by NRT1.1 defines a mechanism for nutrient sensing in plants. Dev Cell 18:927–937

    Article  PubMed  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of gene expression data using real-time quantitative PCR and the 2(-DDc(t)) method. Methods 25:402–408

    Article  PubMed  CAS  Google Scholar 

  • Malagoli P, Laine P, Rossato L, Ourry A (2005) Dynamics of nitrogen uptake and mobilization in field-grown winter oilseed rape (Brassica napus L.) from stem extension to harvest. II A 15N-labelling based simulation model of N partitioning between vegetative and reproductive tissues. Ann of Bot 95:1187–1198

    Article  CAS  Google Scholar 

  • Mora V, Bacaicoa E, Zamarreno AM, Aguirre E, Garnica M, Fuentes M, Garcia-Mina JM (2010) Action of humic acid on promotion of cucumber shoot growth involves nitrate-related changes associated with the root-to-shoot distribution of cytokinins, polyamines and mineral nutrients. J Plant Physiol 167:633–642

    Article  PubMed  CAS  Google Scholar 

  • Moroney JV, Bartlett SG, Samuelsson G (2001) Carbonic anhydrase in plants and algae. Plant Cell Envir 24:141–153

    Article  CAS  Google Scholar 

  • Muscolo A, Panuccio MR, Abenavoli MR, Concheri G, Nardi S (1996) Effect of molecular complexity and acidity of earthworm faeces humic fractions on glutamate dehydrogenase, glutamine synthetase, and phosphoenolpyruvate carboxylase in Daucus carota άII cells. Biol Fertil Soils 22:83–88

    Article  CAS  Google Scholar 

  • Muscolo A, Cutrupi S, Nardi S (1998) IAA detection in humic substances. Soil Biol Biochem 30:1199–1201

    Article  CAS  Google Scholar 

  • Muscolo A, Bovalo F, Gionfriddo F, Nardi S (1999) Earthworm humic matter produces auxin-like effects on Daucus carota cell growth and nitrate metabolism. Soil Biol Biochem 31:1303–1311

    Article  CAS  Google Scholar 

  • Muscolo A, Sidari M, Attina M, Francioso O, Tugnoli V, Nardi S (2007a) Biological activity of humic substances is related to their chemical structure. Soil Sci Soc Am J 71:75–85

    Article  CAS  Google Scholar 

  • Muscolo A, Sidari M, Francioso O, Tugnoli V, Nardi S (2007b) The auxin-like activity of humic substances is related to membrane interactions in carrot cell culture. J Chem Ecol 33:115–129

    Article  PubMed  CAS  Google Scholar 

  • Nardi S, Concheri G, Pizzeghello D, Sturaro A, Rella R, Parvoli G (2000a) Soil organic matter mobilization by root exudates. Chemosphere 41:653–658

    Article  PubMed  CAS  Google Scholar 

  • Nardi S, Pizzeghello D, Gessa C, Ferrarese L, Trainotti L, Casadoro G (2000b) A low molecular weight humic fraction on nitrate uptake and protein synthesis in maize seedlings. Soil Biol Biochem 32:415–419

    Article  CAS  Google Scholar 

  • Nardi S, Pizzeghello D, Reniero F, Rascio N (2000c) Chemical and biochemical properties of humic substances isolated from forest soils and plant growth. Soil Sci Soc Am J 64:639–645

    Article  CAS  Google Scholar 

  • Nardi S, Pizzeghello D, Muscolo A, Vianello A (2002a) Physiological effects of humic substances on higher plants. Soil Biol Biochem 34:1527–1536

    Article  CAS  Google Scholar 

  • Nardi S, Sessi E, Pizzeghello D, Sturaro A, Rella R, Parvoli G (2002b) Biological activity of soil organic matter mobilized by root exudates. Chemosphere 46:1075–1081

    Article  PubMed  CAS  Google Scholar 

  • Nardi S, Tosoni M, Pizzeghello D, Provenzano MR, Cilenti A, Sturaro A, Rella R, Vianello A (2005) Chemical characteristics and biological activity of organic substances extracted from soil by root exudates. Soil Sci Soc Am J 69:2012–2019

    Article  CAS  Google Scholar 

  • Nardi S, Muscolo A, Vaccaro S, Baiano S, Spaccini R, Piccolo A (2007) Relationship between molecular characteristics of soil humic fractions and glycolytic pathway and krebs cycle in maize seedlings. Soil Biol Biochem 39:3138–3146

    Article  CAS  Google Scholar 

  • Okazaki K, Kabeya Y, Suzuki K, Mori T, Ichikawa T, Matsui M, Nakanishi H, Miyagishima S (2009) The PLASTID DIVISION1 and 2 component of the chloroplast division machinery determine the rate of chloroplast division in land plant cell differentiation. Plant Cell 21:1769–1780

    Article  PubMed  CAS  Google Scholar 

  • Park SY, Yu JW, Li J, Yoo SC, Lee NY, Lee SK, Jeong SW, Seo HS, Koh HJ (2007) The senescence-induced staygreen protein regulates chlorophyll degradation. Plant Cell 19:1649–1664

    Article  PubMed  CAS  Google Scholar 

  • Piccolo A, Pietramellara G, Mbagwu JSC (1997a) Reduction in soil loss from erosion-susceptible soils amended with humic substances from oxidized coal. Soil Technology 10:235–245

    Article  Google Scholar 

  • Piccolo A, Pietramellara G, Mbagwu JSC (1997b) Use of humic substances as soil conditioners to increase aggregate stability. Geoderma 75:267–277

    Article  CAS  Google Scholar 

  • Pinton R, Cesco S, Iacolettig G, Astolfi S, Varanini Z (1999) Modulation of NO -3 uptake by water-extractable humic substances: involvement of root plasma membrane H+ATPase. Plant Soil 215:155–161

    Article  CAS  Google Scholar 

  • Puglisi E, Fragoulis G, Ricciuti P, Cappa F, Spaccini R, Piccolo A, Trevisan M, Grecchio C (2009) Effects of humic acid and its size-fractions on the bacterial community of soil rhizosphere under maize (Zea mays L.). Chemosphere 77:829–837

    Article  PubMed  CAS  Google Scholar 

  • Quaggiotti S, Ruperti B, Pizzeghello D, Francioso O, Tugnoli V, Nardi S (2004) Effect of low molecular size humic substances on nitrate uptake and expression of genes involved in nitrate transport in maize (Zea mays L.). J Exp Bot 55:803–813

    Article  PubMed  CAS  Google Scholar 

  • Ramanan R, Kannan K, Vinayagamoorthy N, Ramkumar KM, Sivanesan SD, Chakrabarti T (2009) Purification and characterization of a novel plant-type carbonic anhydrase from Bacillus subtilis. Biotechnol Bioproc Eng 14:32–37

    Article  CAS  Google Scholar 

  • Schmidt W, Santi S, Pinton R, Varanini Z (2007) Water-extractable humic substances alter root development and epidermal cell pattern in Arabidopsis. Plant Soil 300:259–267

    Article  CAS  Google Scholar 

  • Smejkalova D, Piccolo A (2008) Aggregation and disaggregation of humic supramolecular assemblies by NMR diffusion ordered spectrometry (DOSY-NMR). Environ Sci Technol 42:699–706

    Article  PubMed  CAS  Google Scholar 

  • Sutton R, Sposito G (2005) Molecular structure in soil humic substances: the new view. Environ Sci Technol 39:9009–9015

    Article  PubMed  CAS  Google Scholar 

  • Swift RS (1996) Organic matter characterization. In: Sparks DL (ed) Methods of soil analysis Part 3 Chemical methods SSSA Book Ser5. SSSA, Madison, pp 1011–1069

    Google Scholar 

  • Tejada M, Gonzalez JM (2004) Effects of foliar application of a byproduct of the two-step olive oil mill process on rice yield. Eur J Agron 21:31–40

    Article  Google Scholar 

  • Trevisan S, Botton A, Vaccaro S, Vezzaro A, Quaggiotti S, Nardi S (2011) Humic substances affect Arabidopsis thaliana physiology by altering the expression of genes involved in primary metabolism, growth and development. Environ. Exp Bot doi:10.1016/j.envexpbot.2011.04.017

  • Varga B, Kiss G, Galambos I, Gelencser A, Hlavay J, Krivacsy Z (2000) Secondary structure of humic acids. Can micelle-like conformation be proved by aqueous size exclusion chromatography? Environ Sci Technol 34:3303–3306

    Article  CAS  Google Scholar 

  • Vaughan DA, Balazs E, Heslop-Harrison JS (2007) From crop domestication to super-domestication. Annals of Botany 100:893–901

    Article  PubMed  CAS  Google Scholar 

  • Visser SA (1985) Viscometric studies on molecular weight fractions of fulvic and humic acids of aquatic, terrestrial and microbial origin. Plant Soil 87:209–221

    Article  Google Scholar 

  • Zandonadi DB, Canellas LP, Façanha AR (2007) Indolacetic and humic acids induce lateral root development through a concerted plasmalemma and tonoplast H+ pumps activation. Planta 225:1583–1595

    Article  PubMed  CAS  Google Scholar 

  • Zhang X, Schmidt RE (1997) The impact of growth regulators on the α-tocopherol status in water-stressed Poa pratensis. Int Turfgrass Soc Res J 8:1364–1371

    Google Scholar 

Download references

Acknowledgements

This study was a part of AZOSTIMER project selected and supported by the Pôle de compétitivité Mer-Bretagne and funded by the French FUI (Fond Unique Interministériel), Brittany Region and Saint-Malo Agglomeration. We thank Marie-Paule Bataillé and Raphaël Ségura for IRMS analyses. We acknowledge Patrick Beauclair for LICOR measurements, Julie Levallois for technical assistance in RNA extractions and q-PCR analyses, Xavier Sarda and Anne-Françoise Ameline for helping with plant culture and harvest and finally Nicolas Elie from GRECAN (Groupe Régional d’Etude sur le CANcer, Histo-imagerie quantitative, Caen, France) for microscopic analysis. We thank Laurence Cantrill for improving the English of the manuscript.

Author information

Authors and Affiliations

  1. Université de Caen Basse-Normandie, UMR 950 Ecophysiologie Végétale, Agronomie et nutritions N, C, S. Esplanade de la paix, 14032, Caen Cedex, France

    Laëtitia Jannin, Mustapha Arkoun, Alain Ourry, Philippe Laîné & Philippe Etienne

  2. INRA, UMR 950 Ecophysiologie Végétale, Agronomie et nutritions N, C, S. Esplanade de la paix, 14032, Caen Cedex, France

    Laëtitia Jannin, Mustapha Arkoun, Alain Ourry, Philippe Laîné & Philippe Etienne

  3. Université de Caen Basse-Normandie, Centre de Microscopie Appliquée à la Biologie (CMABio), Esplanade de la Paix, 14032, Caen Cedex, France

    Didier Goux

  4. TIMAC Agro Spain, Poligono de Arazuri-Orcoyen, Calle C, n°32, E-31160, Orcoyen, Spain

    Maria Garnica, Marta Fuentes, Sara San Francisco, Roberto Baigorri & José-Maria Garcia-Mina

  5. Centre de Recherche International en Agroscience, CRIAS-TAI, Groupe Roullier, 55 boulevard Jules Verger, 35800, Dinard, France

    Florence Cruz, Fabrice Houdusse & Jean-Claude Yvin

Authors
  1. Laëtitia Jannin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mustapha Arkoun
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Alain Ourry
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Philippe Laîné
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Didier Goux
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Maria Garnica
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Marta Fuentes
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Sara San Francisco
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Roberto Baigorri
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Florence Cruz
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Fabrice Houdusse
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. José-Maria Garcia-Mina
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Jean-Claude Yvin
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Philippe Etienne
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Laëtitia Jannin.

Additional information

Responsible Editor: A.C. Borstlap.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplemental Table S1

List of the differentially expressed genes in shoots and roots of rapeseed after 3 days of HA7 supply to the roots. (XLS 295 kb)

Supplemental Table S2

List of the differentially expressed genes in shoots and roots of rapeseed after 30 days of HA7 supply to the roots. (XLS 35 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jannin, L., Arkoun, M., Ourry, A. et al. Microarray analysis of humic acid effects on Brassica napus growth: Involvement of N, C and S metabolisms. Plant Soil 359, 297–319 (2012). https://doi.org/10.1007/s11104-012-1191-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11104-012-1191-x

Keywords

Search

Navigation