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Methods for Experiment 279 -

affe279 - Root biochemistry

Root biochemistry protocol

In each old field (site numbers assigned from e054), we collected soil cores (3 cm diameter) to a depth of 5 cm from 20 randomly selected locations across two previously established transects. In the forests, we established two parallel 40 m transects and randomly sampled at 10 points along each transect. Within each old field and forest, soil samples were composited and sieved (2 mm) to homogenize soil and separate roots. Roots were collected, dried and sent to University of Wisconsin Soil Laboratories (Verona, WI) for biochemical analysis.

Root biochemistry was characterized by lignin, cellulose, hemicellulose, and total N (Goering and Van Soest 1970, Van Soest et al. 1991). Root lignin was determined by the acid detergent lignin (ADL) procedure, in which ADL is determined gravimetrically as the residue remaining upon ignition after H2SO4 treatment (Goering & Van Soest, 1970). Root cellulose was calculated by subtracting percent acid detergent fiber (ADF) and lignin from root dry mass. ADF was determined gravimetrically as the residue remaining after dissolution and extraction of cell solubles, hemicellulose and soluble minerals with hexadecyltrimethylammonium bromide and sulfuric acid. Hemicellulose was determined by subtracting ADF and neutral detergent fiber (NDF; Van Soest et al. 1991).

Results from this are data published in:
Cline, Lauren C. and Donald R. Zak. Soil Microbial Communities are Shaped by Plant-Driven Changes in Resource Availability During Secondary Succession. Ecology. 96(12), 2015, pp. 3374?3385 http://dx.doi.org/10.1890/15-0184.1 2015


References cited:
Goering, H. K., and P. J. Van Soest. 1970. Forage fiber analysis. Agricultural Research Service Handbook number 379. U.S. Department of Agriculture, Washington, D.C., USA.

Van Soest, P. J., J. B. Robertson, and B. A. Lewis. 1991. Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. Journal of Dairy Science 74:3583?3597.

afge279 - Soil organic matter, total nitrogen and pH

Soil organic matter, total nitrogen and pH - instrumentation

Soil pH was quantified in deionized water using a pH meter (ThermoFischer Scientific).
Soil organic matter was determined using a Leco CNS2000 Analyzer (LECO? St. Joseph, MI).
Total soil N was measured colorimetrically following digestion in concentrated H2SO4 (Lachet Instruments, Loveland, CO)

Soil organic matter, total nitrogen and pH methods

In each old field (site numbers assigned from e054), we collected soil cores (3 cm diameter) to a depth of 5 cm from 20 randomly selected locations across two previously established transects. In the forests, we established two parallel 40 m transects and randomly sampled at 10 points along each transect. Within each old field and forest, soil samples were composited and sieved (2 mm) to homogenize soil and separate roots.

A soil subsample from each old field and forest was sent to University of Wisconsin Soil Laboratories (Verona, WI) to quantity soil pH, organic matter (SOM), and total Nitrogen.

Results from this are data published in:
Cline, Lauren C. and Donald R. Zak. Soil Microbial Communities are Shaped by Plant-Driven Changes in Resource Availability During Secondary Succession. Ecology. 96(12), 2015, pp. 3374?3385 http://dx.doi.org/10.1890/15-0184.1 2015