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Journal of Integrative Agriculture  2014, Vol. 13 Issue (3): 483-490    DOI: 10.1016/S2095-3119(13)60703-0
Section 1: Biochar Characters and Impacts Advanced Online Publication | Current Issue | Archive | Adv Search |
Microscopy Observations of Habitable Space in Biochar for Colonization by Fungal Hyphae From Soil
 Noraini M Jaafar, Peta L Clode , Lynette K Abbott
1、Soil Biology and Molecular Ecology Group, School of Earth and Environment and UWA Institute of Agriculture, The University of Western
Australia, Crawley 6009, Australia
2、Centre for Microscopy, Characterisation and Analysis, The University of Western Australia, Crawley 6009, Australia
3、Department of Land Management, Faculty of Agriculture, Universiti Putra Malaysia, Serdang, Selangor 43400, Malaysia
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摘要  Biochar is a potential micro-environment for soil microorganisms but evidence to support this suggestion is limited. We explored imaging techniques to visualize and quantify fungal colonization of habitable spaces in a biochar made from a woody feedstock. In addition to characterization of the biochar, it was necessary to optimize preparation and observation methodologies for examining fungal colonization of the biochar. Biochar surfaces and pores were investigated using several microscopy techniques. Biochar particles were compared in soilless media and after deposition in soil. Scanning electron microscopy (SEM) observations and characterization of the biochar demonstrated structural heterogeneity within and among biochar particles. Fungal colonization in and on biochar particles was observed using light, fluorescence and electron microscopy. Fluorescent brightener RR 2200 was more effective than Calcofluor White as a hyphal stain. Biochar retrieved from soil and observed using fluorescence microscopy exhibited distinct hyphal networks on external biochar surfaces. The extent of hyphal colonization of biochar incubated in soil was much less than for biochar artificially inoculated with fungi in a soilless medium. The location of fungal hyphae was more clearly visible using SEM than with fluorescence microscopy. Observations of biochar particles colonized by hyphae from soil posed a range of difficulties including obstruction by the presence of soil particles on biochar surfaces and inside pores. Extensive hyphal colonization of the surface of the biochar in the soilless medium contrasted with limited hyphal colonization of pores within the biochar. Both visualization and quantification of hyphal colonization of surfaces and pores of biochar were restricted by two-dimensional imaging associated with uneven biochar surfaces and variable biochar pore structure. There was very little colonization of biochar from hyphae in the agricultural soil used in this study.

Abstract  Biochar is a potential micro-environment for soil microorganisms but evidence to support this suggestion is limited. We explored imaging techniques to visualize and quantify fungal colonization of habitable spaces in a biochar made from a woody feedstock. In addition to characterization of the biochar, it was necessary to optimize preparation and observation methodologies for examining fungal colonization of the biochar. Biochar surfaces and pores were investigated using several microscopy techniques. Biochar particles were compared in soilless media and after deposition in soil. Scanning electron microscopy (SEM) observations and characterization of the biochar demonstrated structural heterogeneity within and among biochar particles. Fungal colonization in and on biochar particles was observed using light, fluorescence and electron microscopy. Fluorescent brightener RR 2200 was more effective than Calcofluor White as a hyphal stain. Biochar retrieved from soil and observed using fluorescence microscopy exhibited distinct hyphal networks on external biochar surfaces. The extent of hyphal colonization of biochar incubated in soil was much less than for biochar artificially inoculated with fungi in a soilless medium. The location of fungal hyphae was more clearly visible using SEM than with fluorescence microscopy. Observations of biochar particles colonized by hyphae from soil posed a range of difficulties including obstruction by the presence of soil particles on biochar surfaces and inside pores. Extensive hyphal colonization of the surface of the biochar in the soilless medium contrasted with limited hyphal colonization of pores within the biochar. Both visualization and quantification of hyphal colonization of surfaces and pores of biochar were restricted by two-dimensional imaging associated with uneven biochar surfaces and variable biochar pore structure. There was very little colonization of biochar from hyphae in the agricultural soil used in this study.
Keywords:  biochar       agriculture       fungi       soil       habitable pore space       microscopy  
Received: 09 October 2013   Accepted:
Fund: 

The Universiti Putra Malaysia and Government of Malaysia are acknowledged for providing a postgraduate scholarship and study leave to Noraini M Jaafar. The University of Western Australia provided access to facilities, research funds and postgraduate student support.

Corresponding Authors:  Noraini M Jaafar, Tel: +61-603-89474953, Fax : +61-603-89408316, E-mail: noraini_aj@yahoo.com   
About author:  Noraini M Jaafar, Tel: +61-603-89474953, Fax : +61-603-89408316, E-mail: noraini_aj@yahoo.com

Cite this article: 

Noraini M Jaafar, Peta L Clode , Lynette K Abbott. 2014. Microscopy Observations of Habitable Space in Biochar for Colonization by Fungal Hyphae From Soil. Journal of Integrative Agriculture, 13(3): 483-490.

Abdullah H, Wu H. 2009. Biochar as a Fuel: 1. Propertiesand grindability of biochars produced from the pyrolysisof mallee wood under slow-heating conditions. Energyand Fuels, 23, 4174-4181

Ascough P L, Bird M I, Scott A C, Collinson M E, Cohen-Ofri I. 2010a. Charcoal reflectance measurements: implications for structural characterization and assessmentof diagenetic alteration. Journal of Archaeological Science, 37, 1590-1599

Ascough P L, Sturrock C J, Bird M I. 2010b. Investigationof growth responses in saprophytic fungi to charred biomass. Isotopes in Environmental and Health Studies,46, 64-77

Bird M I, Ascough P L, Young I M, Wood C V, Scott A C. 2008. X-ray microtomographic imaging of charcoal.Journal of Archaeological Science, 35, 2698-2706

Blackwell P, Krull E, Butler G, Herbert A, SolaimanZ. 2010. Effect of banded biochar on dryland wheat production and fertiliser use in south-western Australia:an agronomic and economic perspective. Soil Research,48, 531-545

Chia C H, Munroe P, Joseph S, Lin Y. 2010. Microscopiccharacterisation of synthetic Terra Preta. Soil Research,48, 593-605

Downie A, Crosky A, Munroe P. 2009. Physical propertiesof biochar. In: Lehmann J, Joseph S, eds., Biochar for Environmental Management: Science and Technology.Earthscan, London. pp. 13-32

Harris K, Crabb D, Young I M, Weaver H, Gilligan C A, Otten W, Ritz K. 2002. In situ visualisation of fungi insoil thin sections: problems with crystallisation of the fluorochrome FB 28 (Calcofluor M2R) and improvedstaining by SCRI Renaissance 2200. Mycological Research, 106, 293-297

Hockaday W C, Grannas A M, Kim S, Hatcher P G. 2007.The transformation and mobility of charcoal in a fire-impacted watershed. Geochimica et Cosmochimica Acta,71, 3432-3445

Jindo K, Sánchez-Monedero M A, Hernández T, García C,Furukawa T, Matsumoto K, Sonoki T, Bastida F. 2012a.Biochar influences the microbial community structure during manure composting with agricultural wastes.Science of The Total Environment, 416, 476-481

Jindo K, Suto K, Matsumoto K, García C, Sonoki T,Sanchez-Monedero M A. 2012b. Chemical andbiochemical characterisation of biochar-blendedcomposts prepared from poultry manure. BioresourceTechnology, 110, 396-404

Joseph S D, Camps-Arbestain M, Lin Y, Munroe P, Chia CH. 2010. An investigation into the reactions of biochar insoil. Soil Research, 48, 501-515

Lehmann J, Rillig M C, Thies J, Masiello C A, HockadayW C, Crowley D. 2011. Biochar effects on soil biota -Areview. Soil Biology and Biochemistry, 43, 1812-1836

Moskal-del Hoyo M, Wachowiak M, Blanchette R A. 2010.Preservation of fungi in archaeological charcoal. Journalof Archaeological Science, 37, 2106-2116

Pietikäinen J, Kiikkilä O, Fritze H. 2000. Charcoal as a habitat for microbes and its effect on the microbial community of the underlying humus. Oikos, 89, 231-242

Solaiman Z M, Blackwell P, Abbott L K, Storer P. 2010.Direct and residual effect of biochar application onmycorrhizal root colonisation, growth and nutrition of wheat. Soil Research, 48, 546-554

Thies J E, Rillig M. 2009. Characteristics of biochar: Biological properties. In: Lehmann J, Joseph S, eds.,Biochar for Environmental Management. Science andTechnology Earthscan, London. pp. 85-105

Verheijen F, Jeffery S, Bastos A C, van der Velde M, Diafas I.2009. Biochar Application to Soils - A Critical ScientificReview of Effects on Soil Properties, Processes andFunctions. Office for the Official Publications of the European Communities, Luxembourg.

Warnock D, Lehmann J, Kuyper T, Rillig M. 2007.Mycorrhizal responses to biochar in soil-Concepts andmechanisms. Plant and Soil, 300, 9-20

Zackrisson O, Nilsson C, Wardle D A. 1996. Key ecologicalfunction of charcoal from wildfire in the Boreal forest.Oikos, 77, 10-19
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