GreenGrass soil guidelines for physico-chemical

Version 2 - 14.10.2002
WP1 – Texture & physico-chemical analysis
Sampling method A
Objective
To characterise the soil physico-chemical parameters of a plot on the basis of 3 composite samples per
treatment.
Sampling
3 depths of soil sampling : 0-10, 10-30 & 30-50 cm. These can be altered with distinct soil horizon.
3 (ideally 4) replicates.
A known volume of soil should be sampled and dried at 30°C. The dry fraction less than 2 mm should
be kept for analysis after sieving and grinding. Stones and roots should be weighted and their volume
estimated in order to calculate soil density. 200 g of fine homogenised soil would be used for soil
analysis.
3 replicates = 3 composites samples - 1 composite sample = mixing of a minimum of 10 soil cores on a same area
(10 m²)
number of soil samples per
treatment
Country
Site
Number of
treatments
Ireland
Scotland
Scotland
Denmark
The Netherlands
France
Switzerland
Hungary
Italy
Carlow
Eastern Bush
Poldean
Lille Valby
Lelystad
Laqueuille
Oesingen
Bugacpuszta
Malga Arpaco
1
2
1
1
1
2
2
1
1
9
9
9
9
9
9
9
9
9
9
18
9
9
9
18
18
9
9
Total
-
-
-
108
(reminder :4 soil depths, 3
composites samples)
Laboratory in charge of the analyses
All the samples will be analysed by a common laboratory : INRA Arras - France
Total number of soil
samples per site
Sending
The samples must be sent first to Sloan Salètes in INRA-Clermont-Ferrand, who will be in charge to
centralise them, then forward to the INRAS Arras’laboratory.
The model sheet here below can be used as an “Order for Analysis”.
Analyses to be done :
Parameter
Requirement
Method of analysis
Code for INRA
Arras
Price (€)
Soil Texture
(in each layer) Clay/Sand/Silt
fractions
Mandatory
5 classes without
decarbonation
114
11.80 €
Mandatory
Wet oxidation or
“Carbo-Erba”
178
6.65 €
Mandatory
pH electrode
144
2.35 €
Mandatory
Method Metson
158
5.85 €
16.10
11.90
Soil organic carbon & nitrogen
(in each layer)
pH in water
(in each layer)
CEC Total
Exchangeable
ions Ca2+, Mg2+,
+
+
K , Na , P2O5 (Dyer’s
method)
Mandatory
Ammonium acetate
respectively 161,
162, 163, 164, 151 +
160 (extraction by
ammonium acetate
NH4+ and NO3-
Mandatory
Extrait 1/10 KCl 0.5
mol/l
244, 245 + 382
(extraction 1/10 by
KCL 0.5 mol/l)
Total : 42.75 €
Price
42.75 € / sample
Payment will be done by cheque or bank transfer to INRA Arras whose bank details are below :
• Numero SIRET : 180 070 039 00045
• Order : TRESORERIE GENERALE ARRAS
• VAT number (intra EC): FR 571 800 700 39
• Bank Code : 10071 CAB : 62000 bank account : : 00003001942 KEY : 39
Contact & Address for sending
Sloan Salètes
INRA - Agronomy Unit
234, avenue du Brézet
63039 Clermont-Ferrand Cedex 02
France
Tel : +33 (0)473 62 44 25
Fax : +33 (0)473 62 44 57
e-mail : [email protected]
Protocols
•
Soil Texture - 5 classes without decarbonation
On détermine la proportion des classes de particules suivantes :
•
•
•
•
•
Argile <2 um
Limons fins 2 um à 20 um
Limons grossiers 20 um à 50 um
Sables fins 0.050 mm à 0.200 mm
Sables grossiers 0.200 mm à 2.00 mm
La détermination des fractions fines (<50 um) s'effectue au moyen de 3 prélèvements à la pipette dite
de Robinson. Les fractions de sable sont déterminées par tamisage sous courant d'eau après
élimination d'un maximum de particules fines par lavages successifs.
Prélèvements et tamisages sont réalisés après destruction de la matière organique par l'eau oxygénée
(H2O2). La dispersion finale est réalisée par passage aux ultrasons après addition de dispersant
(NaPO3 + NH4OH) et après avoir au préalable éliminé les sables grossiers (> 0.200 mm) par
tamisage. Cette méthode équivalente en justesse aux méthodes classiques procédant par simple
agitation est cependant plus reproductible.
Les résultats sont exprimés par rapport à la phase minérale (somme des 5 fractions = 1000).
•
Soil organic carbon & nitrogen
CARBONE ET AZOTE TOTAUX PAR ANALYSE ÉLÉMENTAIRE (g/kg)
Détermination de la teneur en carbone et azote par combustion de l'échantillon en présence de
catalyseur. Les gaz de combustion (N2 et CO2) sont séparés sur une colonne de chromatographie et
détectés sur catharomètres.
La méthode utilisable pour les sols et végétaux utilise des prises d'essai de quelques mg à quelques
dizaines de mg.
•
pH in water (pH electrode)
Mesure du pH après mise en suspension de l’échantillon dans l’eau dans un rapport 1/5 (v/v).
•
P2O5 (Dyer’s method)
Méthode d’extraction des phosphates initialement mise au point pour les sols acides. Elle est
inutilisable pour les sols calcaires sans traitement préalable de l’échantillon. La tendance actuelle est
de la mettre en œuvre uniquement quand le pH est inférieur à 7.
Prise d’essai : 10 g de sol broyé et tamisé à 2 mm.
Réactif : 50 ml d’acide citrique à 20 g/l.
Le dosage du phosphore extrait s’effectue par spectrocolorimétrie à 825 nm après développement de
la coloration d’un complexe phosphomolybdique.
La procédure utilisée est décrite dans la norme AFNOR NF X 31-160 (1993)
Bibliographie succincte :
Dyer B., 1894. On the analytical détermination of probably available mineral plant food in soils. J.
Chem. Soc., 65, 115-167.
•
CEC Total - Method Metson
2.5 g d’échantillon sont saturés en ion ammonium (NH4+) par percolation d’environ 75 ml d’une
solution d’acétate d’ammonium à 1 mole/l. Après rinçage à l’alcool éthylique et séchage à l’air,
l’ammonium fixé est échangé en présence de 50 ml d’une solution de chlorure de sodium à 1 mole/l.
Le dosage des ions NH4+ est réalisé par spectrocolorimétrie, les concentrations trouvées sont
converties en cmol+/kg (centimole de charges (+) par kilogramme).
La procédure utilisée est décrite dans la norme AFNOR NF X 31-130 (1993).
Bibliographie succincte :
Metson AJ (1956) Methods of chemical analysis for soil survey samples. NZ Soil Bur Bull n°12
Ciesielski H, Sterckeman T (1997) A comparison between three methods for the détermination of
cation exchange capacity and exchangeable cations in soils. agronomie, 17, 9-15.
•
Exchangeable ions Ca2+, Mg2+, K+, Na+
* 160 - EXTRACTION DES CATIONS à L’ACéTATE D’AMMONIUM à 1 MOL/L
2.5 g d’échantillon sont agités en présence de 50 ml d’une solution d’acétate d’ammonium à 1 mole/l
pendant une heure. Après filtration la solution est récupérée en vue du dosage des cations
échangeables.
La procédure utilisée est décrite dans la norme AFNOR NF X 31-108 (1992).
Bibliographie succincte :
Thomas GW (1982) Exchangeable cations. In : Methods of soil Analysis. (AL Page et al, eds)
Agronomy 9 ; 154-157 (Madison)
Ciesielski H, Sterckeman T (1997) A comparison between three methods for the détermination of
cation exchange capacity and exchangeable cations in soils. agronomie, 17, 9-15
* 161 Ca éCHANGEABLE EN CMOL+/KG
Le dosage est réalisé par absorption atomique de flamme à partir de l’extrait 160. Les codes 165 et
170 correspondent à la même détermination mais exprimée dans des unités différentes :
165 (g/kg de CaO) = 161 x 0.28 170 (g/kg de Ca) = 161 x 0.20
* 162 Mg éCHANGEABLE EN CMOL+/KG
Le dosage est réalisé par absorption atomique de flamme à partir de l’extrait 160. Les codes 166 et
171 correspondent à la même détermination mais exprimée dans des unités différentes :
166 (g/kg de MgO) = 162 x 0.202 171 (g/kg de Mg) = 162 x 0.122
* 163 K éCHANGEABLE EN CMOL+/KG
Le dosage est réalisé par émission atomique de flamme à partir de l’extrait 160. Les codes 172 et 175
correspondent à la même détermination mais exprimée dans des unités différentes :
172 (g/kg de K) = 163 x 0.391 175 (g/kg de K2O) = 163 x 0.47
* 164 Na éCHANGEABLE EN CMOL+/KG
Le dosage est réalisé par émission atomique de flamme à partir de l’extrait 160. Les codes 168 et 173
correspondent à la même détermination mais exprimée dans des unités différentes :
168 (g/kg de Na2O) = 164 x 0.31 173 (g/kg de Na) = 164 x 0.23
Version 2 - 14.10.2002
WP1 - Microbial biomass
& Extractible living Organic Matter
Sampling method B
by Catherine Hénault & Rachida Nouaïm, INRA Dijon
Soil Microbiology Unit France
Objectives
To characterise the microbial biomass & the labile pool of O.M, which is defined as the amount of
organic carbon solubilized by hot water.
After the measurement of the microbial biomass by fumigation-extraction, the extractible part of
the living organic matter is removed (i.e. the supernatant is discarded) and the fumigated soil sample
is re-suspended in K2SO4 25 mM (ratio soil/water = 1/5) and autoclaved for 16h at 121°C. After
centrifugation, the soluble organic C is measured in the supernatant. The amount of “labile” carbon
represents about 12 to 25% of total organic carbon, according to the soil type and management (crop,
“organic status”…)
Sampling
Country
Site
Ireland
Scotland
Scotland
Denmark
The Netherlands
France
Switzerland
Hungary
Italy
Carlow
Eastern Bush
Poldean
Lille Valby
Lelystad
Laqueuille
Oesingen
Bugacpuszta
Malga Arpaco
Number of
treatments
1
2
1
1
1
2
2
1
1
Number of samples
per treatment
1 composite *
ditto
ditto
ditto
ditto
ditto
ditto
ditto
ditto
Total
Total number
of samples
1
2
1
1
1
2
2
1
1
12
* 1 composite sample = mixing of several soil cores (12 to 20) on a same area (10 m²)
Depth of soil sampling
The main part of the biological activity occurs in the surface layers. The soil uptake must therefore
be done between 0-20 cm.
1-1.5 kg of soil must be sent to allow the analysis of all the parameters required
The samples must not me sieved.
Date of soil uptake
The optimal dates of soil sampling are within the spring and autumn, out of periods of hydric or
thermic stress, and preferably before the beginning of the treatments.
In the frame of GREENGRASS, it is suggested to collect :
-
1 sample during autumn 2002
1 sample during autumn 2003
1 sample during autumn 2004
Laboratory in charge of the analyses
All the samples will be analysed by a common laboratory : INRA Dijon - France
Storage and sending
The samples must be stored at 4°C for maximum 2 weeks and sent directly to INRA-Dijon using
packets with ice-blocks inside (see address below). The sending must be done at the beginning of the
week (do not send any packet after Wednesday…) : It would be necessary to establish maximum
times (perhaps 7 days) between soil sampling and analysis in the lab
Price
The price includes :
- Soil preparation (sieving, drying, packaging…)
-
Basic physico-chemical soil analysis to match the microbial biomass analyses with some physicochemical parameters (including NH4+ and NO3- extraction 1/10 by KCL 0.5 mol/l)
-
Measures of Microbial Biomass (sample analysed in triplicate)
-
Measure of the labile pool of O.M (sample analysed in triplicate)
Total price : 230 € per sample
Protocols
•
ALVAREZ, G., CHAUSSOD R., LOISEAU P., and R. DELPY. 1998. Soil indicators of C and N
transformations under pure and mixed grass-clover swards. European J. Agro.. 9:157-172.
•
CHAUSSOD R. et HOUOT S., 1993. La Biomasse Microbienne des sols : perspectives
d'utilisation de cette mesure pour l'estimation de la fourniture d'azote par les sols. In : Matières
organiques et agricultures, Blois (16-18/11/93) ; Decroux & Ignazi, Eds, pp 17-26.
•
CHAUSSOD R., NICOLARDOT B., CATROUX G. et CHRETIEN J. 1986. Relations entre les
caractéristiques physico-chimiques et microbiologiques de quelques sols cultivés. Science du Sol,
24, pp 213-226.
•
WU J. JOERGENSEN R.G., POMMERENING B., CHAUSSOD R. and BROOKES P.C. 1990.
Measurement of microbial biomass C by fumigation-extraction - An automated procedure. Soil
Biology and Biochemistry, 22, pp 1167-1169.
•
LEMAÎTRE A., CHAUSSOD R., TAVANT Y., BRUCKERT S. 1995. An attempt to determine a
pool of labile organic matter associated with the soil microbial biomass. Eur.J.Soil.Biol., 31 (3),
pp 121-125
•
LEMAÎTRE A., TAVANT Y, CHAUSSOD R., ANDREUX F. 1995 ; Characterization of
microbial components and metabolites isolated from humic calcic soil. Eur.J.Soil.Biol., 31 (3), pp
127-133.
Contact
Rachida NOUAÏM
Office :
Université de Bourgogne
C.A.E. / SEMSE
6 Boulevard Gabriel
21000 DIJON
e-mail : [email protected]
Address of sending
Sloan Salètes
INRA - Agronomy Unit
234, avenue du Brézet
63039 Clermont-Ferrand Cedex 02
France
Tel : +33 (0)473 62 44 25
Fax : +33 (0)473 62 44 57
e-mail : [email protected]
Laboratory
Laboratory Soil Microbiology / SEMSE
To the attention of Rachida NOUAÏM
17 rue Sully
BP 86510
21065 DIJON cedex
Version 2 - 14.10.2002
WP1 - Measures of Advanced soil OM
Sampling method C
by L. Dawson, B. Mayes, K. Väisänen
Plant and Soil Interactions - The Macaulay Institute - Craigiebuckler - Aberdeen - AB15 8QH - UK
Objectives
To characterise the advanced soil OM of a plot, on the basis of 4 composite samples from 2 soil
depths/horizons and in relation with roots/rootmat & the vegetation.
Sampling
The samples for the n-alkane analysis should be taken from 2 soil depths/horizons :
- Rootmat, i.e. the O-horizon
- and also the A-horizon
The specific depths of sampling will depend on the soil type and vegetation history. It is important
then to make a note of the soil depths from where the samples were taken and keep the depth
constant for all replicates.
Samples of roots/ rootmat are required, as the concentrations of alkanes tend to differ between
shoot and root tissues of the same species.
Additionally, a sample of vegetation should also be taken to get a reference signature for each of the
species present at each site. For plant samples, it is crucial to obtain a sample that is representative of
the field. Therefore, for the species rich sites, a larger sample size is required. In addition it should be
ensured that the leaf blade and sheath material is collected.
Additionally, it would be useful to get samples of plant litter (e.g. leaves) from any adjacent
vegetation type to the study sites if it is possible to get carry over of that litter to the site by wind. For
example, it was said in the document describing the 9 sites that the Irish site is next to woodland. In
this case, it could be possible that leaf litter is blown over to the study site by wind and this could
potentially have contributed to the n-alkane patterns of the soil. In addition, if any plant species are
currently in flower, then please separate out the flower head parts of the sample and make a note
of which species it is (the flower heads of plants can have very distinctly different alkane spectra).
Additionally, the samples for the n- alkane analysis should be taken as close as possible to the areas
from which the samples for the amino acid analyses were taken. This is to ensure that the two methods
can be most effectively compared, minimising spatial variability, and to ensure the results are not
biased because of differences in vegetation cover
For each sample site, 4 replicate samples should be taken from each of 2 soil depths, roots and
from the vegetation. Sample size of 10 g is adequate for soil and roots. However, larger sample
size may be required for sampling the vegetation, depending on the species richness. It is important
that sample can be deemed as representative of the field. Sampling could be carried out, for example,
using a soil corer. If the soil corer has a smaller diameter than 10cm, 2 soil cores should be taken
adjacent to each other and bulked.
Number of
treatments
Total number of
Total number of
Total number of
soil samples
vegetation
root samples
(reminder :
samples
2 soil depths)
Country
Site
Ireland
Scotland
Scotland
Denmark
The Netherlands
France
Switzerland
Hungary
Italy
Carlow
Eastern Bush
Poldean
Lille Valby
Lelystad
Laqueuille
Oesingen
Bugacpuszta
1
2
1
1
1
2
2
1
1
8
16
8
8
8
16
16
8
8
4
8
4
4
4
8
8
4
4
4
8
4
4
4
8
8
4
4
Total
-
-
96
48
48
Reminder : be ensured that the leaf blade and sheath material is collected - it would be useful to get samples of plant
litter (e.g. leaves) from any adjacent vegetation type to the experimental plots
Storage
The samples can be stored in strong polythene bags or storage jars. Roots should be separated from
soil when it is still fresh using moist sieving with a 5-mm sieve. If no sieves are available, then hand
sorting of roots from the bulk soil will have to be done prior to storage. Stones separated from the soil
by sieving can be discharged. All the samples should be frozen as quickly as possible. Further
instructions will follow on how to treat the samples prior to transportation to Macaulay.
Additionally
•
•
•
•
No waxy material (e.g. storage bags with waxy lining) should be used at any point of sampling,
sample storage or sample preparation. It has been established that this can lead to significant
contamination of the samples.
No hand creams/lotions or equivalent should be used when taking handling the samples for the
same reason.
If samples are stored in jars, no parafilm should be used for sealing them. Parafilm has also been
discovered to affect the alkane concentrations in samples.
If at any point, alterations to this protocol are made, careful notes of the changes should be taken
and send along with the samples to the staff at the Macaulay Institute.
Sending
This point is still under discussion ; the samples will be :
- either sent to Britain through Gary Campbell (University of Aberdeen) and then directly sent to
Katariina Väisänen (Macaulay) ;
- or collected in a place still to define within a next GREENGRASS meeting (avoiding fastidious
customs obligations) and brought back by Gary or Pete to Aberdeen.
Do not forget to join to the sending the description of present vegetation (list of species, estimate of
abundance of each) and management options adopted should also be included. Additionally, a
description of past vegetation cover and land use, including time scale of changes in land use,
would be useful. Future treatments to be applied at each site would also be useful.
Price
Cost of chemicals is approximately £6 per sample. The Macaulay Laboratory will process and analyze
using its own labour. The true cost of analysis is £ 33 per sample, but The Macaulay Laboratory
provides this at a subsidized price (only the chemicals need to be paid for).
Protocols / References
To be provided soon
Contact
Katariina Vaisanen
PhD student
Plant and Soil Interactions
The Macaulay Institute
Craigiebuckler
Aberdeen
AB15 8QH
UK
Tel: (+44)- (0) 1224-498 200 (extn: 2138)
Fax: (+44)- (0) 1224-498 206
e-mail : [email protected]
Version 1 - 4.4.2003
WP1 & 2 – Particulate Organic Matter
Sampling method C
by Bob Rees, SAC Edinburgh
Objectives
To characterise the particulate organic matter pool, which can be considered to represent a relatively
labile fraction of the soil’s organic matter, and as such one that is sensitive to management.
Principle
The method is based on a procedure developed by Besnard et al (1996) in which a soil sample is
dispersed in ZnCl2, and organic matter is separated by flotation. Sonification is used to release the
organic matter contained within soil aggregates.
Sampling procedure
Suggested soil layers for these one-off measurements are: 0-10, 10-20, and 20-50 cm.
Air dry the soil (in an oven at 60 oC), then remove large fragments of organic matter such as roots and
coarse plant material that would not readily pass through a 2 mm sieve. Pass the whole soil (approx
500 g) through a 2 mm sieve and record the weight of stones and any further organic/root material
retained. A 100 g sample of sieved soil is sent to SAC for analysis together with information on stone
and organic residue content of the whole soil.
Date of soil sampling
It has been proposed that samples should be taken at the start and end of the experimental period for
analysis.
Storage and sending
Dried samples can be stored for up to 12 months prior to analysis
Price
The price includes the costs of labour and materials for the above analysis, which is carried out in
triplicate on each soil sample received.
Total price: 138 € per sample
Experimental Protocol
•
•
•
•
•
•
Weigh out 20g of sieved, air dried soil into a centrifuge tube and add 200 cm3 of ZnCl2 with a
density of 1.6g cm-3.
Disperse sample by placing on an end over end shaker for 30 seconds then diperse by sonification
for 5 mins.
Centrifuge for 30 minutes at 2400 rpm.
The light fraction is collected using the suction apparatus described by Strickland and Sollins
(1987) to remove the suspension until about 3 cm remain above the heavy fraction.
The collected suspension is filtered using a Whatman GF/A glass-fibre filter paper with a 1.6um
retention and washed with water to remove the free dispersion solution.
Dry overnight at 60 oC. Record the weight.
Results will be reported as mass of particulate organic matter/unit mass of sieved dry soil and mass of
particulate organic matter/unit mass of whole dry soil with standard errors
References
Besnard E, Chenu C, Balesdent J, Puget P, Arrouays D (1996). Fate of particulate organic matter in
soil aggregates during cultivation. European Journal of Soil Science, 47 (4): 495-503.
Strickland TC, Sollins P (1987). Improved method for separating light-fraction and heavy-fraction
organic material from soil. Soil Science Society of America Journal, 51 (5): 1390-1393.
Postal Address
Dr Bob Rees
SAC Edinburgh
Bush Estate, Penicuik, EH26 0PH
UK
Tel 0131 535 3011
Fax 0131 535 3031
E-mail [email protected]