Biomass of belowground community groups (bacteria, fungi, protozoa, nematodes, rotifers, tardigrades) determined for organic and mineral soils in moist acidic tundra.
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In June and August 2010, soil samples were collected from control, and N+P plots from within a set of treatments in Moist Acidic Tundra plots established in 1989. We also sampled set of N+P plots within the same block array established in 1996 as part of a fertilization and herbivore exclusion factorial experiment. A set of of Control and N+P plots established on an adjacent hillslope in 2006 with both equivalent (high) and half (low) the fertilization rate of the 89/96 plots were also sampled. At the time of sample collection we separated the soil into organic horizon, organic/mineral interface, and the upper 5cm of the mineral soil. We determined percent moisture for all soils and quantified the biomass of members of the belowground community.
Direct count slides for microbial biomass estimation were made using the methods of Bloem (1995) as adapted by Frey et al. (1999). 10ml of a soil solution diluted 1:100 (5g initial soil mass) was added to each of 5 6mm diameter wells on a slide. Bacteria slides were stained with DTAF (5-(4, 6 dichlorotriazin-2-yl) aminofluorescein) and Fungi slides were stained with calcifluor M2R fluorescence brightener. Direct counts were made using epifluoresence microscopy. Bacteria were counted, 10 images per well, 2 wells per sample. Fungal hyphal length (Lodge and Ingham 1991) was estimated using the formula of Newman (1966): R=πNA/2H where N=number of hyphae crossing transect line, A=area of well, H=total length of transects. 2 wells, each with 10 images, 3 transects per image were used.
Protozoa densities were estimated from 10 g soil samples using Most Probable Number MPN (Darbyshire et al. 1974; Ingham 1994). 10 g soil was serially diluted 10-1-10-6. Using a 24-well tissue culture plate, four replicate wells of each of the six dilutions were created by adding 0.5 ml soil solution to the wells (Rusterholz and Mallory 1994). E. coli, a bacterium food source for protozoa was added to each of the wells. Protozoa were separated into the broad categories of Amoebae, Flagellates and Ciliates.
Nematodes were isolated from 20 g of soil samples using Baermann Funnels (Hall 1996). Nematodes were preserved in formalin, counted and identified to functional group based on the morphology of the stoma and stylet (Niles 1994).
Rotifers, Tardigrades, and Enchytraeids were isolated from 5 g of soil using the methods of Peters et al. (1993).
Microarthropods were heat-extracted into 70% ethanol from soils using Tullgren funnels (Moore et al. 2000). The intact sample pairs from each plot were wrapped together in cotton cheesecloth, weighed, placed over a funnel, and heated with a 9-W incandescent lightbulb for 5 days until dry.
Bloem, J. 1995. Fluorescent staining of microbes for total direct counts. In A. D. L. Akkermans, J. D. van Elsas, and F. J. de Bruijn (ed.), Molecular microbial ecology manual. Kluwer Academic Publishers, Dordrecht, The Netherlands. pp. 1-12.
Darbyshire, J. F., R. F. Wheatley, M. P. Greaves, and R. H. E. Inkson. 1974. A rapid micromethod for estimating bacteria and protozoa in soils. Rev. Ecol. Biol. Sol. 11:465-475.
Frey, S.D., E.T. Elliott and K. Paustian. 1999. Bacterial and fungal biomass in conventional and no-tillage agroecosystems along two climatic gradients. Soil Biology and Biochemistry. 31: 573-585.
Hall, G.S. 1996. Methods for the examination of organismal diversity in soils and sediments. CAB International: New York.
Ingham, E.R. 1994. Protozoa. In: R.W. Weaver et al. (ed.), Methods of Soil Analysis. Part 2: Microbiological and Biochemical properties. Soil Science Society of America, Inc., USA. pp. 491-512.
Lodge, D.J. and E.R. Ingham. 1991. A comparison of agar film techniques for estimating fungal biovolumes in litter and soil. Agriculture, Ecosystems and Environment 34: 131-144.
Moore, J. C., B. B. Tripp, R. T. Simpson, and D. C. Coleman. 2000. Springtails in the classroom: Collembola as model
organisms for inquiry-based laboratories. American Biology Teacher 62:512–519.
Newman, E.L. 1966. A method of estimating the total length of root in a sample. J. Appl. Ecol., 3: 139-145.
Niles, R. 1994. Identification of nematode feeding groups. Unpublished manuscript.
Peters, U., Koste, W., and Westheide W. 1993. A quantitative method to extract moss-dwelling rotifers. Hydrobiologia 255-256: 330-341.
Rusterholz, K.J. and L.M. Mallory. 1994. Density, activity, and diversity of bacteria indigenous to a karstic aquifer. Microbial Ecology 28: 79-99.
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