Abstract
Quantitative and reproducible information concerning the development of the extraradical mycelium of arbuscular mycorrhizal fungi (AMF) is lacking due to the difficulties in extracting, identifying and estimating hyphal lengths. In this study, using a rhizobox growth system, the lengths of hyphae of AMF estimated using an image analysis system were not significantly different from data obtained by a trained observer using a modified grid-line intersect method. The assessment of lengths of hyphae on membrane filters or slides was, however, much quicker using image analysis, and allowed the complete sample to be quantified, unlike the grid-line method where a limited number of fields of view are assessed. The image analysis procedure is objective, observer-independent and less laborious than the manual method of assessment. Of the four different methods of sample preparation compared, membrane filter methods were found to be the most appropriate for quantitative sampling from three non-soil substrates. Glomus monosporum (UKC M3) produced twice as much extraradical mycelium and hyphal length per centimetre of colonised root than G. geosporum (BEG 11) on both leek and linseed in a durite sand at final harvest (63 days). Both AMF also produced more hyphal length per centimetre of colonised root on linseed than on leek. The spatial distribution of both AMF, however, was similar in durite sand and no correlation with levels of NaHCO3-extractable phosphorus was noted. In a third experiment, with G. manihotis (UKC INDO-1) colonising a tropical forage legume, Pueraria phaseoloides, in two other growth substrates, a different pattern of development of the extra-radical mycelium was observed. Because of a higher content of particulate matter, which collected on the membrane filters, the extraction technique had to be modified to give optimal performance of the image analysis system.
Similar content being viewed by others
References
Abbott LK, Robson AD (1985) Formation of external hyphae in soil by four species of vesicular-arbuscular mycorrhizal fungi. New Phytol 99:245–255
Bethlenfalvay GJ, Ames RN (1987) Comparison of two methods for quantifying extraradical mycelium of vesicular-arbuscular mycorrhizal fungi. Soil Sci Soc Am J 51:834–837
Bolan NS (1991) A critical review on the role of mycorrhizal fungi in the uptake of phosphorus by plants. Plant Soil 134:189–207
Capaccio LCM, Callow JA (1982) The enzymes of polyphosphate metabolism in vesicular-arbuscular mycorrhizas. New Phytol 91:81–91
Cooper KM, Tinker PB (1978) Translocation and transfer of nutrients in vesicular mycorrhizas. II. Uptake and translocation of phosphorus, zinc and sulphur. New Phytol 81:43–52
Dodd JC (1994) Approaches to the study of the extraradical mycelium of arbuscular mycorrhizal fungi. In: Gianinazzi S, Schuepp H (eds) Impact of arbuscular mycorrhizal on sustainable agriculture and natural ecosystems. Birkhäuser, Basel, pp 147–166
Dodd JC, Jeffries P (1989) Effect of fungicides on three vesicular-arbuscular mycorrhizal fungi associated with winter wheat (Triticum aestivum L.). Biol Fertil Soils 7:120–128
Dodd JC, Krikun J, Haas J (1983) Relative effectiveness of indigenous populations of vesicular-arbuscular mycorrhizal fungi from four sites in the Negev. Israel J Bot 32:10–21
Dodd JC, Burton CC, Burns RG, Jeffries P (1987) Phosphatase activity associated with the roots and the rhizosphere of plants infected with vesicular-arbuscular mycorrhizal fungi. New Phytol 107:163–172
Graham JH, Linderman RG, Menge JA (1982) Development of the external hyphae by different isolates of mycorrhizal Glomus spp. in relation to root colonization and growth of Troyer citrange. New Phytol 91:183–189
Hanssen JF, Thingstad TF, Gohsøyr J (1974) Evaluation of hyphal lengths and fungal biomass in soil by a membrane filter method. Oikos 25:102–107
Hardie K, Leyton L (1981) The influence of vesicular-arbuscular mycorrhiza on growth and water relations of red clover. I. In phosphate deficient soil. New Phytol 89:599–608
Jakobsen I, Abbott LK, Robson AD (1992) External hyphae of vesicular-arbuscular mycorrhizal fungi associated with Trifolium subterraneum L. New Phytol 120:371–380
Johansen A, Jakobsen I, Jensen ES (1993) External hyphae of vesicular-arbuscular mycorrhizal fungi associated with Trifolium subterraneum L. 3. Hyphal transport of 32P and 15N. New Phytol 124:61–68
Koske RE, Gemma JN (1989) A modified procedure for staining roots to detect VA mycorrhizas. Mycol Res 92:486–505
Miller RM, Jastrow JD (1990) Hierarchy of root and mycorrhizal fungal interactions with soil aggregation. Soil Biol Biochem 22:579–584
Morgan P, Cooper CJ, Battersby NS, Lee SA, Lewis ST, Machin TM, Graham SC, Watkinson RJ (1991) Automated image analysis method to determine fungal biomass in soils and on solid matrices. Soil Biol Biochem 23:609–616
Newman EI (1966) A method for estimating the total length of root in a sample. J Appl Ecol 3:139–145
Ojala JC, Jarrel WM, Menge JA, Johnson ELV (1983) Comparison of soil phosphorus extradants as predictors of mycorrhizal dependency. Soil Sci Soc Am J 47:958–962
Olsen SR, Sommers LE (1982) Phosphorus. In: Page AL, Miller RH, Keeney DR (eds) Methods of soil analysis, part 2: Chemical and microbiological properties, 2nd edn. American Society of Agronomy, Madison, Wis, pp 403–430
Pearson JN, Jakobsen I (1993a) Symbiotic exchange of carbon and phosphorus between cucumber and three arbuscular mycorrhizal fungi. New Phytol 124:481–488
Pearson JN, Jakobsen I (1993b) The relative contribution of hyphae and roots to phosphorus uptake by arbuscular mycorrhizal plants, measured by dual labelling with 32P and 33P. New Phytol 124:489–494
Schubert A, Marzachí C, Mazzitelli M, Cravero MC, Bonfante-Fasolo P (1987) Development of total and viable extraradical mycelium in the vesicular-arbuscular mycorrhizal fungus Glomus darum Nicol. & Schenck. New Phytol 107:183–190
Smith SE, Dickson S (1991) Quantification of active vesiculararbuscular mycorrhizal infection using image analysis and other techniques. Aust J Plant Physiol 18:637–648
Smith SE, Gianinazzi-Pearson V (1988) Physiological interactions between symbionts in vesicular-arbuscular mycorrhizal plants. Annu Rev Plant Physiol Plant Mol Biol 39:221–244
Sylvia DM (1992) Quantification of external hyphae of vesicular-arbuscular mycorrhizal fungi. In: Norris JR, Read DJ, Varma AK (eds) Techniques for the study of mycorrhiza. (Methods in microbiology, vol 24). Academic Press, London, pp 53–65
Sylvia DM, Burks JN (1988) Selection of a vesicular-arbuscular mycorrhizal fungus for practical inoculation of Uniola paniculata. Mycologia 80:565–568
Vilariño A, Arines J, Schuepp H (1993) Extraction of vesicular-arbuscular mycorrhizal mycelium from sand samples. Soil Biol Biochem 25:99–100
Xiao-Lin L, George E, Marschner H (1991) Extension of the phosphorus depletion zone in VA-mycorrhizal white clover in a calcareous soil. Plant Soil 136:41–48
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Green, D.C., Newsam, R., Jeffries, P. et al. Quantification of mycelial development of arbuscular mycorrhizal fungi using image analysis. Mycorrhiza 5, 105–113 (1994). https://doi.org/10.1007/BF00202341
Issue Date:
DOI: https://doi.org/10.1007/BF00202341