The soil food web under stress by application of ashes from biofuel
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The soil food web under stress by application of ashes from biofuel. ASHBACK WP3 Ecotoxicology. Jiayi Qin Internship : Margarita Ustyuzhanina Supervisor P.H. Krogh. Outline. Develop DNA metabarcoding method used for the investigating changes of the soil mesofauna biodiversity.

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The soil food web under stress by application of ashes from biofuel

The soil food web under stress by application of ashes from biofuel

ASHBACK WP3 Ecotoxicology

Jiayi QinInternship: Margarita Ustyuzhanina

Supervisor P.H. Krogh


Outline

Outline


The soil food web under stress by application of ashes from biofuel

Develop DNA metabarcodingmethodused for the investigating changes of the soil mesofauna biodiversity.


Morphospecies

Morphospecies

  • Take photo of each morphospecies.

  • Sort morphospeciesinto different bottles.

  • Use SoilBioStoreto count the number of animals.

  • Identify species bybarcoding and verify by conventional taxonomic identification of physical voucher subsequently archived


Soil bio store counting webinterface and mysql database storage

SoilBioStoreCounting webinterface and MySQL database storage

Trait database


Metabarcoding dna extraction

Metabarcoding — DNA extraction

Environmental DNA (eDNA) extraction:

(Bienertet al.2012; Taberletet al. 2012, Yu et al. 2012, Yang et al. 2013 )


Metabarcoding dna extraction1

Metabarcoding — DNA extraction

Environmental DNA (eDNA) extraction:

Hypothesis 1:

Total eDNAcould represent the real biodiversity.

(Bienertet al.2012; Taberletet al. 2012, Yu et al. 2012, Yang et al. 2013 )


Metabarcoding pcr strategy

metabarcoding—PCR strategy

Primer:

mt-COI

12S/16S mt-rDNA

18S/28S rDNA

nuclear Internal Transcribed Spacer (ITS)

Advantages and disadvantages of each gene region for metagenetic analyses.

(Machida and Knowlton 2012)


Metabarcoding pcr strategy1

metabarcoding—PCR strategy

Primer:

mt-COI

12S/16S mt-rDNA

18S/28S rDNA

nuclear Internal Transcribed Spacer (ITS)

Hypothesis 2:

Minibarcodefrom mitochondrial DNA could amplify sequence for specie identification.

Advantages and disadvantages of each gene region for metagenetic analyses.

(Machida and Knowlton 2012)


The soil food web under stress by application of ashes from biofuel

Development of DNA metabarcodingfor the investigating of changes in the soil mesofauna biodiversity.


B iodiversity change in spruce forest under the stress of ash

Biodiversity change in spruce forest under the stress of ash

  • Ash treatment

    0, 3, 4.5, 6 t/ha

  • Sampling occasions

    • Pretreatment sampling, April 1, 2014

    • Spring 2014,

    • June 2-3 Autumn 2014,

    • (spring 2015) autumn 2015

    • spring 2016

    • autumn 2016 ?

  • Analysis strategy

    • Conventional method - morphospecies

    • Metabarcoding


Comparison of food web changes when affected by different levels of ash

Comparison of food web changes when affected by different levels of ash

  • Location: Agricultural field, Foulum

  • Ash treatments:

    super phosphate at 30 and 300 kg ha-1 combined with either

    • wood pill ash

    • straw ash

    • Norway spruce, Piceaabies, ash

  • Sampling:

    Time: September, 2014

    Content: soil cores for eDNA analysis

  • Analysis strategy: conventional method/metabarcoding


Ecotoxicology of ash exposed to mesofauna in a spruce forest soil and an agricultural soil

Ecotoxicology of ash exposed to mesofauna in a spruce forest soil and an agricultural soil

Definitive test protocol in spruce forest soil and agricultural soil

(OECD 2009)


Result

Result

Mortality

Survival


Thank you

THANK YOU


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