Soil organic carbon stabilization: Mapping carbon speciation from

Oct 17, 2018 - The clearing of land for agricultural production depletes soil organic carbon (OC) reservoirs, yet despite their importance, the mechan...
0 downloads 0 Views 1MB Size
Subscriber access provided by UNIV OF NEW ENGLAND ARMIDALE

Environmental Processes

Soil organic carbon stabilization: Mapping carbon speciation from intact microaggregates Maria Hernandez-Soriano, Ram Dalal, Frederick J. Warren, Peng Wang, Kathryn Green, Mark James Tobin, Neal W Menzies, and Peter M Kopittke Environ. Sci. Technol., Just Accepted Manuscript • DOI: 10.1021/acs.est.8b03095 • Publication Date (Web): 17 Oct 2018 Downloaded from http://pubs.acs.org on October 19, 2018

Just Accepted “Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted online prior to technical editing, formatting for publication and author proofing. The American Chemical Society provides “Just Accepted” as a service to the research community to expedite the dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been fully peer reviewed, but should not be considered the official version of record. They are citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors or consequences arising from the use of information contained in these “Just Accepted” manuscripts.

is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.

Page 1 of 35

Environmental Science & Technology

1

Soil organic carbon stabilization: Mapping carbon speciation from intact

2

microaggregates

3 4

Maria C. Hernandez-Soriano1, Ram C. Dalal1, Frederick J. Warren2, Peng Wang1,3, Kathryn

5

Green4, Mark J. Tobin5, Neal W. Menzies1, Peter M. Kopittke1,*

6 7

1

8

Queensland, 4072, Australia

9

2

The University of Queensland, School of Agriculture and Food Sciences, St. Lucia,

The University of Queensland, Centre for Nutrition and Food Sciences, Queensland Alliance

10

for Agriculture and Food Innovation, St. Lucia, Queensland, 4072, Australia

11

3

12

Nanjing, Jiangsu, 210009, China

13

4

14

Queensland, 4072, Australia

15

5

Nanjing Agricultural University, College of Resources and Environmental Sciences,

The University of Queensland, Centre for Microscopy and Microanalysis, St. Lucia,

Australian Synchrotron, Clayton, Victoria, 3168, Australia

16 17

*Corresponding author: Peter M. Kopittke, [email protected], +61 7 3346 9149

18

1 ACS Paragon Plus Environment

Environmental Science & Technology

Page 2 of 35

19

Abstract

20

The clearing of land for agricultural production depletes soil organic carbon (OC) reservoirs,

21

yet despite their importance, the mechanisms by which C is stabilized in soils remain unclear.

22

Using synchrotron-based infrared microspectroscopy, we have for the first time obtained in

23

situ, laterally-resolved data regarding the speciation of C within sections taken from intact

24

free microaggregates from two contrasting soils (Vertisol and Oxisol, 0-20 cm depth)

25

impacted upon by long-term (up to 79 y) agricultural production. There was no apparent

26

gradient in the C concentration from the aggregate surface to the interior for any of the three

27

forms of C examined (aliphatic C, aromatic C, and polysaccharide C). Rather, organo-mineral

28

interactions were of critical importance in influencing overall C stability, particularly for

29

aliphatic C, supporting the hypothesis that microaggregates form through organo-mineral

30

interactions. However, long-term cropping substantially decreased the magnitude of the

31

organo-mineral interactions for all three forms of C. Thus, although organo-mineral

32

interactions are important for OC stability, C forms associated with the mineral phases are

33

not entirely resistant to degradation. These results provide important insights into the

34

underlying mechanisms by which microaggregates form, and the factors influencing the

35

persistence of OC in soils.

36

2 ACS Paragon Plus Environment

Page 3 of 35

Environmental Science & Technology

37

TOC

38 39

3 ACS Paragon Plus Environment

Environmental Science & Technology

Page 4 of 35

40

INTRODUCTION

41

Carbon (C) reservoirs in soil (ca. 2,344 Gt in the surface 3 m) exceed those in both the

42

atmosphere (850 Gt) and the biotic pool (560 Gt) combined.1 However, the conversion of

43

land from native vegetation to long-term agricultural cropping often reduces soil organic C

44

(SOC) stocks by 20-60%.2-4 This marked depletion of SOC stocks results in the release of

45

greenhouse gases. Given that the soil represents a large C pool, only comparatively small

46

changes in SOC stocks can have important impacts on greenhouse gas concentrations. Not

47

only does the decrease in SOC in long-term agricultural systems increase greenhouse gas

48

emissions, it also results in the loss of soil quality and fertility, with land use change

49

influencing both the content and composition of soil organic matter (SOM) with concomitant

50

changes in the chemical and biological processes that determine soil functioning.5

51

Accordingly, to both improve soil fertility and decrease release of greenhouse gases, it is

52

important to optimize the management of SOM and to develop strategies for stabilizing C in

53

soils.1, 6, 7 This has recently been recognized in the ‘soil carbon 4 per mille’ initiative which

54

has an aspiration to increase global soil OC stocks by 4 per 1000 (0.4 %) per year as a

55

compensation for global emissions of greenhouse gases.8

56 57

In order to effectively understand and predict the potential sequestration of C in soils, it is

58

first necessary to understand the underlying mechanisms of C stabilization and processing.7, 9,

59

10

60

mechanisms by which organic C is stabilized in soils. It has traditionally been assumed that

61

the stability of SOC is regulated by the (i) preservation of complex (recalcitrant) compounds

62

resistant to microbial degradation6 (ii) formation of organo-mineral associations11 and (iii)

63

physical (spatial) inaccessibility of occluded SOC to microbial communities and enzymes.12-

64

14

Indeed, despite its importance, there remains considerable uncertainty regarding the

The overall relative importance of each of these three underlying mechanisms to the overall

4 ACS Paragon Plus Environment

Page 5 of 35

Environmental Science & Technology

65

stabilization of OC across a broad range of soils remains somewhat uncertain, with

66

investigation of these mechanisms being the focus of the present study (for example, see 15).

67

It is important to note that the currently-available evidence generally does not appear to

68

support the first of these three hypotheses [i.e. that recalcitrant compounds, “humic

69

substances”, form during the degradation process],7, 14, 16, 17 and this hypothesis cannot

70

account for the long-term stabilization of otherwise labile SOC in soils.17 Nevertheless, it is

71

known that the degree of ‘recalcitrance’ is important in the early stages of decomposition,

72

although ‘spatial inaccessibility’ and ‘organo‐mineral interactions’ are important during the

73

later stages of decomposition.18

74 75

Previously, progress in identifying and confirming these underlying processes that regulate

76

the cycling of SOC has been hindered by a lack of suitable approaches, such as those that

77

provide in situ, laterally-resolved analyses of SOC composition within intact soil

78

microaggregates. Previously, scanning electron microscopy coupled with energy-dispersive

79

X-ray spectroscopy (SEM-EDS) has been found to be useful in this regard.19 Similarly, other

80

approaches such as nanoscale secondary ion mass spectrometry (NanoSIMS) and µ-

81

tomography are being used increasingly to investigate the cycling of OC in soils.20, 21 It has

82

been shown comparatively recently that synchrotron-based spectroscopy [such as scanning

83

transmission X-ray microspectroscopy (STXM) and infrared (IR) microspectroscopy] can

84

provide laterally-resolved information on the molecular organization of SOC,13, 22, 23 the

85

physical protection in the soil particles, and the co-localization of C sources with microbial

86

processes.12, 13, 24 However, we are unaware of any studies that have provided laterally-

87

resolved analyses of SOC speciation from entire, intact sections of microaggregates from

88

soils impacted upon by long-term agricultural production. Such information is important in

89

understanding the factors influencing the stability and turnover of organic C in soil systems.

5 ACS Paragon Plus Environment

Environmental Science & Technology

Page 6 of 35

90 91

In the present study, we have utilized two paired-sites (Oxisol and Vertisol) from subtropical

92

Australia to compare undisturbed soils to adjacent soils that have been used for agricultural

93

production for either 60 y (Oxisol) or 79 y (Vertisol). The SOC content of these soils has

94

been reduced markedly by agricultural production, with the decrease in SOC being 85% for

95

the Oxisol (with the corresponding reduction in C stock being from 26 to 7 Mg C/ha) and

96

76% for the Vertisol (with the corresponding reduction in C stock being from 61 to 10 Mg

97

C/ha) (0–20 cm depth). Comparable C losses (up to 75%) have been previously reported for

98

long-term (> 50 y) cultivation in tropical and sub-tropical systems.25-27 For the two soils, we

99

used in situ, laterally-resolved synchrotron-based analyses to examine changes in the

100

speciation and distribution of SOC using thin-sections obtained from free intact

101

microaggregates (250-53 µm). For the first time using in situ, laterally-resolved analyses, our

102

results link long-term changes in organo-mineral associations of various forms of C with the

103

decline of SOC stocks – this information being crucial in understanding the mechanisms

104

which influence SOC stability and cycling. It is important to note that it was not the aim of

105

our study to perform a statistically-based experiment on land use change effects based on

106

replicated samples, but rather to highlight the great potential of synchrotron-based methods to

107

elucidate land-use change effects on important soil properties at the aggregate and sub-

108

aggregate scale. We hypothesized that the sections taken from intact microaggregates would

109

show a distinct gradient from the interior to the exterior of the microaggregates resulting from

110

the occlusion of organic C by the mineral phase during microaggregate formation.

111

Furthermore, we hypothesized that there would be a strong correlation between the clay

112

mineral phase and the various forms of SOC due to the importance of organo-mineral

113

associations.

114

6 ACS Paragon Plus Environment

Page 7 of 35

Environmental Science & Technology

115

MATERIALS AND METHODS

116

Soil collection and general analyses

117

We collected two soils (Oxisol and Vertisol) from land-use pairs at two locations in

118

Queensland (Australia). The two soils were selected as they differ markedly in their

119

properties, including mineralogy (see later). For the Oxisol (26° 42’ S, 151° 48’ E), the native

120

vegetation was ‘vine scrub’, while for the Vertisol (26° 48’ S, 150° 54’ E), the dominant

121

native vegetation was brigalow (Acacia harpophylla). The Oxisol has an average annual

122

rainfall of 770 mm, with a yearly average maximum temperature of 24.7 °C and average

123

minimum of 11.4 °C. The Vertisol has an average annual rainfall of 610 mm, with a yearly

124

average maximum temperature of 26.9 °C and average minimum of 11.9 °C. For both sites,

125

the rainfall is summer-dominant and the climate classified as sub-tropical. The paired sites for

126

both the Oxisol and the Vertisol encompass two land-use types < 100 m apart, being

127

undisturbed soil (native vegetation) and cropped soil. The conversion of native vegetation to

128

agricultural cropping occurred 60 y ago for the Oxisol and 79 y ago for the Vertisol. The

129

Oxisol has mainly been used for cropping of peanut (Arachis hypogaea) and maize (Zea

130

mays), while the Vertisol has mainly been used for winter cropping of wheat (Triticum

131

aestivum). Both soils are low input systems. For the Oxisol, the soils received no fertilizer for

132

the first 55 y of cropping, with fertilizer for the last 5 y applied as 50 kg/ha/y as diammonium

133

phosphate. For the Vertisol, soils received only 30 kg N/ha/y for 51 y, with 100 kg N/ha/y

134

and 10 kg P/ha/y since this time for 28 y. No organic amendments (such as manures) have

135

been applied at either site. For the cropping soils, cultivation consisted of two or three

136

operations per year with a chisel plough (approximately 20 cm depth) to control weeds during

137

fallow.

138

7 ACS Paragon Plus Environment

Environmental Science & Technology

Page 8 of 35

139

Soil samples were collected from both land-uses (undisturbed and cropping) at the two sites

140

from a depth of 0-20 cm. The samples were collected in the October-November period (mid-

141

spring), with those from the cropping soils collected either at post-harvest (Vertisol) or at

142

maturity (Oxisol). Three individual replicate samples were collected, sealed in plastic

143

containers, and stored at 4° C until further analysis. For background analyses, soils were first

144

sieved to 2 mm and air-dried. Soil pH was measured using 1:5 soil:water suspensions with

145

air-dried soil. For organic C, we measured concentrations in the bulk soil plus four different

146

size fractions that were obtained by wet sieving,28 being > 2000 (large macroaggregates),

147

2000-250 (small macroaggregates), 250-53 (free microaggregates), and < 53 µm (silt+clay).

148

The free water-stable microaggregates (250-53 µm) accounted for 5.3-9.6 % of the total

149

aggregates in the undisturbed soils and increased to 8.9-13 % in the cropped soils, although

150

occluded microaggregates are also present within macroaggregates. Organic C concentrations

151

were measured by dry combustion using a CN analyzer (VarioMax) after oven-drying (60

152

°C). Particle size analysis was performed using the pipet method.29 Mineralogy was

153

determined for the undisturbed soils using X-ray diffraction.

154 155

Infrared microspectroscopy

156

For infrared microspectroscopy, subsamples of the three air-dried individual replicates were

157

combined to form a composite sample. A composite sample was used as only limited samples

158

could be examined using synchrotron-based infrared microspectroscopy. Soils were sieved (2

159

mm), and intact microaggregates particles (53–250 µm in diameter) were isolated by wet

160

sieving28 from both land uses of the two soil types. The use of this approach resulted in the

161

collection of free water-stable microaggregates rather than microaggregates occluded within

162

macroaggregates.28 The free water-stable microaggregates (53–250 µm in diameter) were

163

then air-dried until needed. Approximately 20-30 typical example free microaggregates (53–

8 ACS Paragon Plus Environment

Page 9 of 35

Environmental Science & Technology

164

250 µm in diameter) were selected, placed on filter paper, moistened gently through

165

humidification.22 Once moistened, the free microaggregates were frozen at -20 °C and semi-

166

thin sections of microaggregates were cut using a diamond knife without embedding media

167

using a cryo-ultramicrotome. The sections were cut with the ultramicrotome set to a thickness

168

of 200 nm, although the actual thickness of each section was not measured. Sections were

169

then transferred to Cu grids (carbon free, 200 mesh, silicon monoxide coating) in order to

170

allow the sections to dry and be transferred to the Australian Synchrotron (below).

171 172

The sections were removed from the Cu grids and analyzed at the IR Microspectroscopy

173

beamline at the Australian Synchrotron (Melbourne, Australia), using a Bruker Hyperion

174

2000 infrared microscope and V80v Fourier transform infrared (FT-IR) spectrometer. Due to

175

time constraints, only a single replicate section from each of the four treatments could be

176

examined. The microscope was equipped with 36× (0.5 numerical aperture) condenser and

177

objective optics, a narrowband mercury cadmium telluride detector, and a detection aperture

178

was selected to sample an area of 5 µm × 5 µm. The maps (5 µm step size over ca. 150 µm ×

179

150 µm) were obtained from 64 co-added scans (4 cm-1 resolution) and the spectra were

180

acquired in transmission mode. A single map (150 µm × 150 µm) was obtained for each of

181

the four sections collected from the intact microaggregates.

182 183

Spectral maps were processed using the software OPUS 7.2 (Bruker Optik GmbH, Germany).

184

Map profiles were created for absorbance at 3630 cm-1 (O–H groups of clays), 2920 cm-1

185

(aliphatic-C), 1600 cm-1 (aromatic-C), and 1035 cm-1 (polysaccharides-C).22, 30 The peak at

186

3630 cm-1 corresponds to the stretching vibrations of O-H groups of clay minerals, the peak

187

at 2920 cm-1 corresponds to the C-H stretching vibrations of aliphatic biopolymers, the peak

188

at 1600 cm-1 corresponds to C=C stretching of aromatic C or N-H deformations, and the peak

9 ACS Paragon Plus Environment

Environmental Science & Technology

Page 10 of 35

189

at 1035 cm-1 corresponds to C-O stretching vibrations of polysaccharide C.30 In each case the

190

integrated area under the absorbance peak was applied to the map, selecting appropriate

191

baseline points either side of the absorbance peak [3550-3740 cm-1 (O–H groups of clays),

192

2800-3000 cm-1 (aliphatic C), 1500-1750 cm-1 (aromatic C), and 950-1170 cm-1

193

(polysaccharide C)].

194 195

The spectra collected for each map were examined and a linear regression (Genstat v16) used

196

to examine the correlation between the amount of clay (absorbance at 3630 cm-1) and either

197

polysaccharides C (absorbance at 1035 cm-1), aromatic C (absorbance at 1600 cm-1) or

198

aliphatic C (absorbance at 2920 cm-1).22, 30 The R2 coefficients and regression slopes were

199

used as indices for associations, the former indicating the residual variability around the

200

association while the latter the relative strength of association. Based upon the signal to noise

201

ratio, it was estimated that these synchrotron-based analyses were ca. 100-fold more sensitive

202

than conventional (globar source) IR microspectroscopy (also see Miller and Dumas 31 for a

203

comparison of a synchrotron source and a globar source).

204 205

Bulk infrared spectroscopy

206

Soil samples were sieved (2 mm) and finely ground using a mortar and pestle. FTIR-ATR

207

spectra were collected for all soil samples using a PerkinElmer Spectrum 100 FTIR

208

spectrometer equipped with a deuterated triglycine sulfate (DTGS) detector. The ATR

209

accessory (PerkinElmer UATR) was equipped with a diamond crystal. Background spectra

210

were collected every five samples and subtracted from each recorded spectrum. For each

211

spectrum, 16 scans were co-added at a resolution of 4 cm-1 from 4000 to 650 cm-1. The

212

spectra were vector-normalized in order to minimize intensity variations between the spectral

213

data set.

10 ACS Paragon Plus Environment

Page 11 of 35

Environmental Science & Technology

214 215

RESULTS

216

Undisturbed soils

217

Using synchrotron-based IR microspectroscopy, we obtained laterally-resolved information

218

on C speciation [polysaccharide C (1035 cm-1), aromatic C (1600 cm-1), aliphatic C (2920

219

cm-1), and clay minerals (3650 cm-1)]22, 30 and distribution in thin sections taken from entire,

220

intact microaggregates of the Oxisol and Vertisol (Table 1 and Figure 1). Giving

221

consideration firstly to the undisturbed soils, it was observed that the distribution of the

222

various C forms was not homogenous, but rather, they were distributed as discrete areas of C

223

accumulation in the microaggregate section both in the interior and the edges of the

224

microaggregate sections (see top panels of Figure 2 and Figure 3). For these undisturbed

225

soils, it was apparent that there were relationships between the distribution of clay minerals

226

and the various forms of C. Indeed, correlation analysis (which indicates co-localization)

227

identified that aliphatic-C was closely associated with clay minerals in both the undisturbed

228

Oxisol (R2 = 0.97) and Vertisol (R2 = 0.72) (Figure 2 and Figure 3). For the two other forms

229

of C (polysaccharide C and aromatic C), whilst relationships with clay minerals were

230

generally strong in the Oxisol (R2 values 0.83 and 0.77), relationships with clay minerals in

231

the Vertisol were poor (R2 values 0.29 and 0.22) (Figure 2 and Figure 3). Thus, whilst some

232

of the polysaccharide C and aromatic C was associated with clay minerals, a higher

233

proportion of this C was associated with areas spatially unrelated to clay minerals. It is

234

possible that this poorer relationship for the Vertisol was due to its lower C concentration

235

(Table 1), with not all reactive sites being occupied.

236 237

It was also noted that there was no apparent gradient in accumulation of the various C forms

238

from the interior to the exterior of the microaggregate, as might occur if organic C was

11 ACS Paragon Plus Environment

Environmental Science & Technology

Page 12 of 35

239

surrounded (occluded) in a central core by clay minerals, for example. To examine this more

240

closely, changes in the various forms of organic C and clay minerals were examined across a

241

transect through the middle of the microaggregate section. For both the Oxisol (Figure 4) and

242

the Vertisol (Figure 5), there was no clear evidence of a distinct trend in organic C, nor was

243

there a clear core of organic C surrounded by clay minerals.

244

Vertisol

Oxisol 800

Undisturbed Cropped

600

-1

Aggregates (g kg soil)

Undisturbed Cropped

400

200

0 ates ates ilt+clay ates ates ilt+clay ates ates S S greg aggreg aggreg greg aggreg aggreg g g a a icro icro acro l macro acro l macro M M m m e e al al Sm Sm Larg Larg

245 246

Figure 1. Distribution of aggregate size fractions in undisturbed and cropped soils, from an

247

Oxisol (left) and Vertisol (right).

248

12 ACS Paragon Plus Environment

Page 13 of 35

Environmental Science & Technology

249

Table 1. Soil pH, clay content, concentration of organic carbon (g OC kg soil -1) in bulk soil and soil size fractions, being > 2000, 2000-250,

250

250-53, and < 53 µm. pH (1:5

Sand

Silt

Clay

water)

(%)

(%)

(%)

Mineralogy*

C/N

OC (g kg

ratio

soil-1) Bulk soil

Oxisol

Vertisol

251

Undisturbed

5.2

55

8

36

Cropped

5.7

62

8

30

Undisturbed

6.9

31

18

51

Cropped

7.9

34

16

50

Q, K, H

Q, S, K

OC (g kg aggregate-1)

> 2000

2000-250

250-53

µm

µm

µm

< 53 µm

13.8

52

53

47

56

46

11.5

7.9

7.3

6.7

8.1

20

11.4

22

21

21

16

19

12.2

5.3

5.7

5.0

3.4

7.0

* Listed in decreasing order. Abbreviations: Q, quartz; H, hematite; K, kaolinite; S, smectite.

252

13

ACS Paragon Plus Environment

Environmental Science & Technology

Page 14 of 35

253 254

Figure 2. Semi-thin (200 nm) sections of microaggregates (