METHODS•Soil cores (n=30) were collected from the base of alder genets in early (alder-willow), mid (balsam popular) and late (white spruces) successional stands (20m X 20m plots) along the Tanana River floodplain in mid-August 2009•Cores were split into organic and mineral fractions•Cores were stored on ice and alder roots were removed from cores within 2-3 hours•Single healthy mycorrhizal tips (n = 7 tips/core) were excised from root systems under 10-40x magnification

•Enzyme assays were of single root tips, held in PCR strips with mesh bottoms•Acid phosphatase and phosphodiesterase (Fig. 1) were assayed with 4-methylumbelliferyl (MU) linked substrates in acetate buffer (pH 5.5)•Phytase activity was measured by incubating tips in phytic acid and measuring [Pi] fluorometrically

Alder strategies for phosphorus assimilation across a boreal forest successional sequence

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Michaela Swanson1, Roger Ruess1, Karl Olson1, Knut Kielland1, Lee Taylor1 and Jack McFarland2

1 Institute of Arctic Biology, Department of Biology and Wildlife, Fairbanks, AK, 99775, 2 U.S. Geological Survey, Menlo Park, CA 94025

RESULTS•All enzymes activities were positively inter-correlated across stages and horizons (all P< .001); the strongest of these relationships was between acid phosphatase and phosphodiesterase (P<.001, R2 = 0.32) (Fig. 2)•

•Enzyme activities were ~double in late compared to early and mid-succession (all P<.001) (Fig. 3)•Higher acid phosphatase in organic vs. mineral soils (P<.001) (Fig. 4)•Stage by horizon interactions:

•Acid phosphatase (P<.01) activity was higher on tips from organic verses mineral horizons (P<.001) in early succession •Phosphodiesterase (P<.001) activity was higher on tips from organic horizons of early (P<.05) and mid (P<.10) successional stands, but lower in organic horizons of late succession (P<.01)

INTRODUCTION•Rates of carbon cycling and NPP in boreal forests are heavily dependent on inputs of biologically fixed N from Alnus tenuifolia

•N fixing species, including alder have exceptionally high demands for phosphorus (P) and N-fixation rates have been shown to respond positively to P fertilization

•One strategy that alder uses to meet its P requirements is to associate with ectomycorrhizal fungi (EMF), which produce surface bound enzymes that mobilize organic and recalcitrant P forms

ACKNOWLEDGEMENTSThis work was jointly funded by a UAF Global Change Research Grant, NSF DEB-0641033, and the BNZ LTER. Thank you to Ivan Lien and Collin Stackhouse for their hard work and help with field and laboratory portions of this project.

CONCLUSIONS•Functional traits of individual EMF species vary between soil horizons and throughout succession

•High activities of enzymes that mineralize complex forms of organic P in late successional white spruce sites are likely related to the more recalcitrant forms of organic matter found in these soils

FUTURE WORK•Enzyme activities are being matched with digital estimates of root tip surface area•Automated ribosomal intergenic spacer analysis (ARISA) is being used to analyze fungal ITS rDNA sequences isolated from individual alder root tips for community analysis and paired with enzyme activity •Forms and availability of P are being quantified using Hedley fractionation method•Measures of plant N:P balance: foliar nutrient analyses and leaf resorption – from trees above where soil cores were taken

Fig 3. Activities of phosphorous mobilizing enzymes on the surfaces of mycorrhizally infected alder root tips collected from cores in early, mid and late successional sites along the Tanana River floodplain.

Fig 4. Activities of phosphorous mobilizing enzymes on the surfaces of mycorrhizally infected alder root tips in organic and mineral horizons collected from cores across successional stages along the Tanana River floodplain.

Fig. 1. Bis 4-methylumbelliferyl phosphate: structure of fluorogenic enzyme substrate for phosphodiesterase

Fig 2. Relationship between acid phosphatase and phosphodiesterase activity on individual mycorrhizally-infected alder root tips collected from cores in early, mid and late successional sites along the Tanana River floodplain.

acid phosphatase phosphodiesterase phytaseEnz

yme

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HYPOTHESISN-fixation inputs across the landscape are controlled by the ability of alder to assimilate P through associations with EMF species of varying capacities to mobilize P. Because the forms and availability of P are known to change throughout forest succession and between soil horizons, we expected to see parallel shifts in EMF communities and function at these same scales.

Alder/willow site

Balsam popular siteWhite spruce site