Abstract
The hyphae and spores of arbuscular mycorrhizal (AM) fungi represent an essential component in the extraradical zone due to their role in nutrients and water uptake and as propagules that allow the perpetuation of the AM symbiosis over time, respectively. However, the attention of scientific literature is usually more focused on root colonization than on the study of the extraradical components of AM fungi, especially their vital, active, or functional fractions. This chapter presents some easy-to-use alternatives for staining vital, active, or functional structures of AM fungi for their subsequent microscopic visualization, such as the application of enzyme-based stains, NADPH formation, and also nucleus staining. Some modified methods for the extraction of mycelium from the soil are also presented.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Phillips JM, Hayman DS (1970) Improved procedures for clearing roots and staining parasitic and vesicular-arbuscular mycorrhizal fungi for rapid assessment of infection. Trans Br Mycol Soc 55:158–161
Giovannetti M, Mosse B (1980) An evaluation of techniques for measuring vesicular arbuscular mycorrhizal infection in roots. New Phytol 84:489–500
McGonigle TP, Miller MH, Evans DG et al (1990) A new method which gives an objective measure of colonization of roots by vesicular-arbuscular mycorrhizal fungi. New Phytol 115:495–501
Trouvelot A, Kough JL, Gianinazzi-Pearson V (1986) Mesure du taux de mycorhization VA d’un systeme radiculaire. Recherche de methodes d’estimation ayant une signification fonctionnelle. In: Gianinazzi-Pearson V, Gianinazzi S (eds) Physiological and genetical aspects of mycorrhizae. Proceedings of the 1st European symposium on Mycorrhizae. Institut National de la Recherche Agronomique, Paris, pp 217–221
Barea JM, Palenzuela J, Cornejo P et al (2011) Ecological and functional roles of mycorrhizas in semi-arid ecosystems of Southeast Spain. J Arid Environ 75:1292–1301
Cornejo P, Seguel A, Aguilera P et al (2017) In: Singh D, Singh H, Prabha R (eds) Plant-microbe interactions in agro-ecological perspectives. Springer, Singapore, pp 55–80
Santander C, Aroca R, Ruiz-Lozano JM et al (2017) Arbuscular mycorrhiza effects on plant performance under osmotic stress. Mycorrhiza 27:639–657
Borie F, Aguilera P, Castillo C et al (2019) Revisiting the nature of phosphorus pools in Chilean volcanic soils as a basis for arbuscular mycorrhizal management in plant P acquisition. J Soil Sci Plant Nutr 19:390–401
Kemper WD, Rosenau RC (1986) Aggregate stability and size distribution. In: Klute A (ed) Methods of soil analysis. Part1 physical and mineralogical methods, Agron .Monogr.9, 2nd edn. American Society of Agronomy, Inc. Soil Science Society of America, Inc, Madison, WI, pp 425–442
Rubio R, Borie F, Schalchli C et al (2003) Occurrence and effect of arbuscular mycorrhizal propagules in wheat as affected by the source and amount of phosphorus fertilizer and fungal inoculation. Appl Soil Ecol 23:245–255
MacDonald RM, Lewis M (1978) The occurrence of some acid phosphatases and dehydrogenases in the vesicular-arbuscular mycorrhizal fungus Glomus mosseae. New Phytol 80:135–141
Abdel-Fattah GM (2011) Measurement of the viability of arbuscular-mycorrhizal fungi using three different stains; relation to growth and metabolic activities of soybean plants. Microbiol Res 156:359–367
Vierheilig H, Schweiger P, Brundrett M (2005) An overview of methods for the detection and observation of arbuscular mycorrhizal fungi in roots. Physiol Plant 125:393–404
Tisserant B, Gianinazzi-Pearson V, Gianinazzi S, Gollotte A (1993) In planta histochemical staining of fungal alkaline phosphatase activity for analysis of efficient arbuscular mycorrhizal infections. Mycol Res 97:245–250
Gerdemann JW, Nicolson TH (1963) Spores of mycorrhizal endogone species extracted from soil by wet sieving and decanting. Trans Br Mycol Soc 46:235–244
Cornejo P, Pérez-Tienda J, Meier S et al (2013) Copper compartmentalization in spores as a survival strategy of arbuscular mycorrhizal fungi in cu-polluted environments. Soil Biol Biochem 57:925–928
Newman EI (1996) A method of estimating the total length of root in a sample. J Appl Ecol 3:139–145
Gavito ME, Olsson P, Rouhier H et al (2005) Temperature constraints on the growth and functioning of root organ cultures with arbuscular mycorrhizal fungi. New Phytol 168:179–188
Acknowledgments
The authors thank Comisión Nacional de Investigación Científica y Tecnológica, CONICYT-Chile, for the Doctoral Scholarship No. 21160049 (H. Aponte), CONICYT/FONDECYT 1170264, CONICYT/FONDAP/15130015, and CONICYT/RING/ACM170002 (P. Cornejo).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Cornejo, P., Aponte, H. (2020). Visualization of Arbuscular Mycorrhizal Fungal Extraradical Hyphae and Spores Vitality and Activity. In: Ferrol, N., Lanfranco, L. (eds) Arbuscular Mycorrhizal Fungi. Methods in Molecular Biology, vol 2146. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0603-2_6
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0603-2_6
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0602-5
Online ISBN: 978-1-0716-0603-2
eBook Packages: Springer Protocols