Quantifying the effect of fire disturbance on free-living nitrogen

fixation in tropical ecosystems

Barbara D Bomfim1, Lucas C R Silva1, Timothy Doane1, Ben Hur Marimon2, Beatriz S Marimon2, Lais S Neves2, William R Horwath1
1University of California Davis, Department of Land, Air and Water Resources. bdbomfim@ucdavis.edu

(Presenting author); 2 Universidade Estadual do Mato Grosso, Department of Plant Biology.

Introduction Results/Discussion
Biological nitrogen fixation (BNF) is one of the most metabolically costly processes on Earth (Gutschick
1981). Given that not all legume species (N fixers) are actively fixing N (Nardoto et al. 2014), free-living BNF Correlations between free-living BNF rates Biplot of PLSR Components
is expected to be a significant source of N inputs to terrestrial ecosystems (Reed et al. 2011; Cleveland et al. and soil and litter properties
1999), especially in the tropics, where its energy costs can be met. The variability in the rate of free-living N2
fixation rates across different ecosystems could be a function of disturbance regimes, which strongly influence
resource availability (e.g., C, N, P, Fe, Mo) (Vitousek et al. 1979). The effect of fire regimes on free-living BNF
rates in tropical forests is currently uncertain. We measured free-living BNF and quantified rates of N2 fixation
in different substrates of seasonally flooded tropical forests under contrasting fire regimes in the Amazon-
Cerrado transition of central Brazil. We tested the hypothesis that environmental legacies of previous
disturbances influence free-living N2 fixation rates in different layers of the forest floor. Specifically, we
expected to identify a negative relationship between fire frequency and flooding regime, which in turn, should
influence species diversity and nutrient stocks (Silva et al. 2013) and affect free-living BNF rates as a result of
differences in litter and soil carbon and nutrient stoichiometry.

a b

Figure 3. a) Correlations between substrate properties and free-living BNF rates (15N2 and ARA) measured
in the forests assessed at PEA, southern Amazonia, Brazil. b) Partial Least Squares Regression (PLSR)
biplot for components 1 and 2 (x and y-scores) of substrate properties and free-living BNF rates (15N2 and
ARA): arrows are substrate properties and free-living BNF rates.

Variables with the

a a b c highest importance Best model:
Figure 1. a) Photos of seasonally flooded tropical forests in southern Amazonia during dry season, water levels reach up to 1 m for projection (VIP): Log free-living BNF rate ~ N:P + PMN + Fe
height during the wet season; b) Typical soil profile (Histosol – Typic Haplosaprist); c) Fire removing the litter layer on forest floor, - N:P ratio R2adj = 0.85 (p = 0.002)
litter removal is expected to alter biogeochemical cycles (including BNF) at the stand level. - PMN PRESS = 1.16
- Fe
Material And Methods
Sampling design
v 5 ha of seasonally flooded tropical forests were assessed, establishing a chronosequence of fire frequency, 15N
2 and ARA Free-living BNF rates
based on recurrence (0 to 8 events) from 2000 to 2013 (Figure 2 and Table 1);
v We used a cluster sampling design (Wang et al. 2012) at each ha of forest by randomly selecting one point 15N
2 free-living BNF rates ARA free-living BNF rates
within the contiguous plots (100 of 10 x 10 m), previously established for vegetation survey, and
systematically establishing 4 N-S and E-W oriented points; 6

v At each sampling point, soil (0 – 10, 10 – 30 cm) and standing litter samples were collected for free-living µg N fixed gDW-1 h-1
2 6
4 Substrate
BNF rate measurements using two methods: 15N2 (98 atom% 15N) (Hsu & Buckley 2009) and acetylene Soil 0-10 cm
reduction assay (ARA) (Sullivan et al. 2014); Soil 10-30 cm

1 Normal Q-Q plot Litter

2
Table 1. Seasonally flooded tropical forests, respective fire Layer
Litter

frequency and aboveground vegetation data (DBH ≥10 cm).

H’ = Shannon’s Diversity Index; J = Pielou’s Coefficient of Equability; IVI = 0 0
Importance Value. DBH = diameter at breast height. *2008 survey data – 4
2 3 1 4 3 2 3 1 4 3
100 10m x 10m permanent plots per forest.
Fire recurrence Fire recurrence

Fire Yrs Tree basal Figure 4. Free-living N fixation rates (15N2 and ARA) in µg N fixed gDW-1 h-1 measured in litter and soil (0-10
Forest Highest IV
recurrence since H’ J area (m2 and 10-30 cm) in the fire-disturbed seasonally flooded tropical forests studied at PEA, southern Amazonia,
ID species
(yrs) last fire ha-1) Brazil. 2 = 2 years of fire legacy.

Micropholis
F3 1 3 2.49 0.70 40.76
gardneriana
- Free-living BNF rates varied significantly between forests with distinct fire legacies, substrates and in
F1 2 3 2.63 0.75 25.19
Licania the interaction between fire legacy and substrate (p < 0.05);
apetala

Micropholis - The most severely burnt forest (4 yrs) showed lower mean free-living BNF rates, although not
F2 3 3 2.87 0.75 35.39 significantly distinct from rates found in the forest burned once (1 yr) over the 14 years;
gardneriana

Figure 2. Study site: Araguaia State Park (PEA), southern Licania

F7 3 3 1.99 0.76 22.36
Amazonia, Brazil. In detail, map of natural fire frequency (0 to 8 apetala
years) between 2000 and 2013, used to establish a
Micropholis
chronosequence of sampling sites under different disturbance F5 4 3 2.98 0.77 33.75
regimes. Adapted from Neves (2013).
gardneriana Conclusions
Soil and litter chemical analyses o Free-living BNF rates in forest soil and litter were significantly affected by fire in seasonally flooded
vSoil pH in H2O (2:1) and KCl (1M); tropical forests;
vTotal Carbon and Nitrogen by elemental analysis (Costech Elemental Combustion System 4010);
vPotentially Mineralizable Nitrogen (PMN) by anaerobic incubation (Waring & Bremner 1964);
o Nutrient stoichiometry (N:P), PMN and total Iron were the main drivers of variations in free-living BNF
vTotal Phosphorus (P) and Iron (Fe) by sulfuric acid digestion (Claessen et al. 1997) and colorimetric
rates in soil and litter of the fire-disturbed forests;
determination (Murphy and Riley 1962; Stookey 1970);
vSoil available P and Fe by Mehlich-3 extractant (Claessen et al. 1997) followed by colorimetric
determination (Murphy and Riley 1962).
Acknowledgements
Data analyses This work was financially supported by CAPES – Science without Borders and University of California
Partial Least Squares Regression Multiple Linear Regressions Two-way Anova, Tukey HSD Davis. Special thanks to Eder Carvalho and Ben Hur Marimon for field sampling assistance, Tad for
Y = 15N2 and ARA free-living N fixation rates guidance in laboratory analyses, Secretaria Estadual do Meio Ambiente – MT, Parque Estadual do
Best model = lowest PRESS and R2adj. Factors = fire and layer
X = Soil and litter chemical properties Araguaia – MT and Universidade Estadual do Mato Grosso for logistics support.