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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 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

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  1. The soil food web under stress by application of ashes from biofuel ASHBACK WP3 Ecotoxicology Jiayi QinInternship: Margarita Ustyuzhanina Supervisor P.H. Krogh

  2. Outline

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

  4. 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

  5. SoilBioStoreCounting webinterface and MySQL database storage Trait database

  6. Metabarcoding — DNA extraction Environmental DNA (eDNA) extraction: (Bienertet al.2012; Taberletet al. 2012, Yu et al. 2012, Yang et al. 2013 )

  7. 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 )

  8. 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)

  9. 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)

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

  11. 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

  12. 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

  13. 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)

  14. Result Mortality Survival

  15. THANK YOU

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