Enzymatic activities in crop and grassland soils and the impact of copper addition

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Enzymatic activities in crop and grassland soils and the impact of copper addition

Isabelle Trinsoutrot-Gattin, Marthe Vinceslas-Akpa, Caroline Bailleul, Christian Mougin, Karine Laval

To cite this version:

Isabelle Trinsoutrot-Gattin, Marthe Vinceslas-Akpa, Caroline Bailleul, Christian Mougin, Karine Laval. Enzymatic activities in crop and grassland soils and the impact of copper addition. Third International Conference on Enzymes in the Environment, Jul 2007, Viterbo, Italy. 2007. �hal- 01601168�

Enzymatic activities in crop and grassland soils - Impact of copper addition

Isabelle Trinsoutrot-Gattin, Marthe Vinceslas-Akpa, Caroline Bailleul, Agathe Brault, Christian Mougin and

Karine Laval

Introduction

• Soil : non-renewable resource essential for life

• Several functions :

• Physical and cultural environment for humankind

• Food and biomass production

• Storing, filtering and tranformation of organic matter

• Habitat and gene pool of living organisms...

• Pressures : agricultural production, urban and industrial development, transport networks, leisure activities…

• Necessity to guaranty the protection and sustainable use of the soil

!

!

!

! !

!

! ! P P roject of European policy 2006. roject of European policy 2006.

Context

• Current tools in agronomic practices : physical and chemical properties

• Soil organisms responsible for biogeochemical cycles, ...

• This study is a part of a vast project intending to elaborate an index of the biological state of soil.

! ! !

! !

!

!

! microbial, fungal and bacterial biomass

!

! !

! !

!

!

! Enzymatic activities Enzymatic activities

• Enzymatic activities :

• Soil functioning

• Scientifically recognized

• Integrative tool : pedological characteristics, climate,

soil management

" Impact of copper on soil enzymatic activites

" Considering the natural variations of the

enzymatic activities in our referenced agricultural systems (grassland and crop plots)

Objectives

Material and Methods

Enzymatic activities assessed

Acid Phosphatase

Substrate pH

p-nitrophenyl phosphate (PNP)

p-nitrophenyl b-glucoside (PNP)

p-nitrophenyl acetyl glucosaminide (PNP) Alkaline Phosphatase

ββββ -glucosidase

N-acetyl glucosaminidase 6

6 6,5

11

Enzymes

p-nitrophenyl phosphate (PNP)

Dehydrogenase 1,3,5-triphenyltetrazolium chloride (TTC) 7

urea

Urease 7

Dick et al. (1996)

Material and Methods - Field plots

Saint Georges sur Saint Georges sur

Fontaine Fontaine

Two sites, located in the North West of France

18.0%

18.0% clayclay 67.2% silt 67.2% silt 14.8%

14.8% sansan

Yvetot Yvetot

18.2% clay 18.2% clay 62.1% silt 62.1% silt 19.7 sand 19.7 sand

April, June, October

1 m

0-10cm, 4-kg

20 samples per plot

Crop pH 6,8 Crop pH 6,8

Crop pH 6,8 Crop pH 6,8 Meadow pH 5,8

Meadow pH 5,8

Meadow pH 6,5 Meadow pH 6,5

Undisturbed

Undisturbed cosmes cosmes sampled from sampled from Yvetot grassland and crop plots.

Yvetot grassland and crop plots.

2 ppm Cu 200 ppm Cu Control

Models :

5 5 cosmes cosmes for for each each model model 4 dates: 0, 7, 35

4 dates: 0, 7, 35 and and 70 70 days days Incubation in

Incubation in field field conditions conditions

Material

Material and and Methods Methods Cosme

Cosme experimentation experimentation

15 cm 15 cm

12cm 12cm

Results

2508.8

3022.4

1959.8

1842.5

1291.4

640.8

263.0 551.7

The level of activities is higher in grassland than in crop plots

Temporal and spatial variations for enzymatic

activities

Acidic, alkaline phosphatases and b-glucosidase activities exhibit a great variability

Spatio temporal variability in Yvetot meadow soils

0 500 1000 1500 2000 2500 3000 3500

enzymatic activites in nmol hydrolysed product g-1 h-1Enzymatic activity in nmol hydrolysed product g-1h-1

PAC PAL B-glu NAG DH URE Spatio temporal variability in Yvetot meadow soils

0 500 1000 1500 2000 2500 3000 3500

enzymatic activites in nmol hydrolysed product g-1 h-1Enzymatic activity in nmol hydrolysed product g-1h-1

PAC PAL B-glu NAG DH URE

Spatio temporal variability in Yvetot crop soils

0 500 1000 1500 2000 2500 3000 3500

enzymatic activity in nmol hydrolysed product g-1 h-1

PAC PAL B-glu NAG DH URE Enzymatic activity in nmol hydrolysed product g-1 h-1

Spatio temporal variability in Yvetot crop soils

0 500 1000 1500 2000 2500 3000 3500

enzymatic activity in nmol hydrolysed product g-1 h-1

PAC PAL B-glu NAG DH URE Enzymatic activity in nmol hydrolysed product g-1 h-1

Yvetot site

Enzymatic activities in Yvetot plots and in cosmes

Enzymatic activities are similar in plots and control cosmes

!

! !

! !

!

!

! Validation of Validation of our our undisturbed undisturbed cosmes cosmes approach approach

cosmes versus plots - Yvetot Meadow

0 100 200 300 400 500 600 700 800 900

acid-P alkaline-P b-glu NAG

nmol PNP g-1 h-1

cosme J0 Field plot J0 cosme J70 Field plot J70

day 0

day 70

Alkaline phosphatase - crop

0 200 400 600 800

0 7 35 70

days after Cu inoculation

nmol PNP g-1 h-1

control 2 ppm Cu 200 ppm Cu

N Acetyl Glucosaminidase - crop

0 50 100 150 200 250 300

0 7 35 70

days after Cu inoculation

nmol PNP g-1 h-1

control 2 ppm Cu 200 ppm Cu

Urease - crop

0,00 5,00 10,00 15,00

0 7 35 70

days after Cu inoculation

nmol N-NH4+ g-1 h-

control 2 ppm Cu 200 ppm Cu

Impact of the two copper concentrations on enzymatic activities in crop cosmes

Acid phosphatase - crop

0 100 200 300 400 500 600 700 800

0 7 35 70

days after Cu inoculation

nmol PNP g-1 h-1

b-glucosidase - crop

0,00 100,00 200,00 300,00 400,00 500,00 600,00

0 7 35 70

days after Cu inoculation

dehydrogenase - crop

0 2 4 6 8 10 12

0 7 35 70

days after Cu inoculation

Impact of Cu on enzymatic activities : Impact of Cu on enzymatic activities :

Literature controversy Literature controversy

"

" Differences in experimental conditions Differences in experimental conditions

!

!

!

! !

! !

! Sieved soil Sieved soil vs vs undisturbed undisturbed cosmes cosmes . microbial physiological status . microbial physiological status

. physical protection on soil microorganisms . physical protection on soil microorganisms

. availability of copper . availability of copper

! !

! !

!

! !

! High Cu concentration High Cu concentration vs vs agronomic reality agronomic reality . Status of copper

. Status of copper

. Distribution of copper

. Distribution of copper

Distribution of inoculated copper in microcosms

Cu 330mg/25mL

3535

3232

3535 1919

1313

1313

3030

66

1616 Content (mg) after equilibration

12cm

12cm diameterdiameter

15cm 15cm

Cu distribution in cosmes

Copper distribution according height

0,00 20,00 40,00 60,00 80,00 100,00 120,00 140,00 160,00

1 2 3 4 5 6 7

amount of Cu (ppm) mg

CP Cu 200ppm

0 - 5 cm 5 - 10 cm 10 - 15 cm

ADN Fong.

Ergostérol

ADN total

ADN Bact.

Bactéries Totales Phosphatase Acide

β-D-Glucosidase

N-acétyl-β-D-Glucosaminidase

Evaluation of the copper impact on the descriptors of the microbial communities

Meadow (reference) Meadow (reference) Meadow (200ppm Cu) Meadow (200ppm Cu) Crop

Crop

Crop (200ppm Cu) Crop (200ppm Cu)

Results at T

0 1

C16:1w5

Microbial DNA Fungal DNA

Bacterial DNA Cultivable R2A

Conclusion

"

"Spatiotemporal Spatiotemporal variability variability of of enzymatic enzymatic activities activities ranged ranged from from 6 to 95 % in 6 to 95 % in referenced referenced systems systems

"

" Total Total enzymatic enzymatic activities activities are are higher higher in in meadow meadow related related to to soil soil biomass biomass increase increase

"

" Anova Anova analysis analysis shows : shows :

! ! Agricultural practices Agricultural practices

! ! Sampling Sampling date (including date ( including season season and and plant cover plant cover) )

! ! No No copper copper effect effect

Conclusion

"

" Enzymatic activities as indicator of soil Enzymatic activities as indicator of soil « « quality quality » » ? ?

"

" Links between microbial communities structure and Links between microbial communities structure and functional diversity?

functional diversity?

"

"Enzymatic activity of interest ? Enzymatic activity of interest ?

!stage of organic matter decomposition ? ! stage of organic matter decomposition ?

"

" Methodological strategies for Soil Science : Methodological strategies for Soil Science :

!pH? ! pH?

!Substrate? ! Substrate?

!spectrophotometry ! spectrophotometry versus fluorimetry versus fluorimetry? ?

!Units? ! Units?

"

" Agronomic decision tools? Agronomic decision tools?

Thank

Thank you you for for your your attention attention