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Genetic Technologies

Junior Research Group

The "Genetic Technologies" Group is an independent junior research group at the Thünen Institute of Forest Genetics within the project "TreeEdit", funded by the Agency of Renewable Resources ("Fachagentur Nachwachsende Rohstoffe", FNR).

We are studying CRISPR/Cas-mediated genome editing of trees, in particular whether and how these methods work in woody plants and what potentials and risks are associated with them. Thus, we are expanding the knowledge regarding these technologies in tree species, which is often based on results from crop research.

In addition to this method development, the Genetic Technologies Group also conducts research in the field of climate adaptation. Poplars serve us as model organisms on which the genetic background of drought stress tolerance is investigated. By experimentally modifying the genes involved, we aim to draw conclusions about their function. The results will be transferred to the most important German deciduous tree species, the European beech Fagus sylvatica.

In a subproject, we are investigating the gene expression of European beech of different provenances under drought stress. With this, we hope to obtain insights into the natural stress response of European beech and which differences exist between Central and Southern European beech. With our partners we combine genetic, molecular and physiological analyses.


People

Dr. Tobias Brügmann (group leader)
Alexander Fendel (PhD student)
Virginia Zahn (PhD student)
Susanne Jelkmann (technical assistant)

Sarah Wodtke (master student, since July 2024)
+ one technician trainee in lab rotation


Stress trials in the climate forest

Project module

European beech trees have a large distribution area. At its southern edge, they are confronted with different climatic conditions than in Germany: the average annual temperature is higher and the amount of precipitation is lower than in Germany. To investigate the different drought stress tolerance of beech provenances, a provenance trial ("climate forest") was planted in Berlin. Here, individual trial members (a group of trees) can be irrigated, while other trial members are affected by the (naturally occurring) drought stress in summer. A drought stress experiment was initiated in the summer of 2022 with one provenance each from Germany, France, Italy and Spain. Ecophysiological, biochemical, and genetic analyses were conducted using different stress intensities. The data will be used to estimate the drought stress tolerance of individual European beech provenances.

Genetic regulations will be investigated by transcriptome analysis. For this purpose, all mRNA molecules produced in the leaves will be decoded. Biochemical analyses include measuring the levels of the drought stress markers proline, malondialdehyde (MDA), and peroxide. Together with physiological data such as photosynthetic activity, stomatal conductance, and water potential, the analytical triad should provide a comprehensive picture of drought stress tolerance.

We expect answers to the questions: What is the response of European beech under drought stress? Do the selected provenances differ in their drought stress response? Which genes are switched on or off and how fast or how strongly is gene activity regulated? Is provenance-dependent differential stress tolerance related to differential gene regulation? The findings from this collaborative research may be helpful in selecting suitable drought-tolerant provenances.

Beech provenances

Cooperation partners

Ecology of Plants, Free University of Berlin (PD Dr. M. Forstreuter)

Plant Biotechnology and Bioinformatics, Braunschweig University of Technology (Prof. Dr. B. Pucker)


Study, Training & Guest Scientists

Bachelor's and master's theses as well as university internships can be carried out in our laboratory. Furthermore, we participate in the training of biological-technical assistants. The trainees are integrated into our research activities in four-month blocks. If you are interested or have any questions, please contact Tobias Brügmann.

Felix Wiedemann (2024) Charakterisierung von PcP5Cs1-Überexpressionslinien in Populus x canescens unter kontrollierten Trockenstressbedingungen
Master thesis, course biotechnology, University of Hannover

Dr. Anke Bollen (2024) Infiltration rekombinanter Ribonukleotidproteinkomplexe (RNP) zur DNA-freien Genomeditierung an Pflanzen als unterstützende Maßnahme zum Erhalt der Biodiversität
Master thesis, course applied environmental sciences, University of Koblenz

Yuna Hartschen (2024)
Student trainee, University of Düsseldorf

Ina Martini (2023) Etablierung der Fluoreszenzmarker-basierten Effizienzbestimmung modifizierter guide-RNAs in Pappelprotoplasten [mit SpCas9 und Lb-ttCas12a]
Bachelor thesis, course Biotechnology, Hochschule Emden-Leer
[Establishment of Fluorescence Marker-Based Efficiency Determination of Modified Guide RNAs in Poplar Protoplasts [with SpCas9 and Lb-ttCas12a]]

Alina Fomin (2023) Biochemical analyses of drought stress-related markers and promoter analysis of P5CS1 in European beech (Fagus sylvatica) of different provenances
Bachelor thesis, course Biochemistry, University of Lübeck

Keelin Meyer (2023) Agrobakterium-vermittelte Transformation der Europäischen Buche (Fagus sylvatica)
Bachelor thesis, course Biotechnology, Hochschule Emden-Leer
[Agrobacterium-mediated transformation of European beech (Fagus sylvatica)]

Franziska Brunozzi (2023)
Student trainee, TU Munich

Prof. Dr. Stefano Biricolti (2022): Guest scientist
University of Florence

Thalia von Nethen (2022) Promotor-Tests in Nicotiana benthamiana zur Optimierung der Effizienz von CRISPR/Cas-vermittelter Genomeditierung
Bachelor thesis, course Biotechnology, Hochschule Emden-Leer

Stefan Zebbedies (2021) Etablierung der Nickase-Genomeditierung in Pappeln.
Bachelor thesis, course Renewable Resources, Hochschule Hannover
[Establishing the nickase-mediated genome editing in poplars]

Oliver Sulkowski (2020) Molekularbiologische Untersuchung auf erhöhte Salz- und Trockenstresstoleranz in SCL4- und SCL7-überexprimierenden Pappelhybriden (Populus × canescens).
Bachelor thesis, course Biotechnology, Hochschule Emden-Leer
[Analysis of salt and drought stress tolerance in
SCL4 and SCL7 overexpression poplar hybrids]


Partners

The Genetic Technologies Group interacts with scientists from several research institutions. These are in particular:

Thünen Institute of Forest Genetics

  • Genome Research, Thünen Institute of Forest Genetics (PD Dr. M. Fladung)
  • Resistance Research, Thünen Institute of Forest Genetics (Dr. B. Bubner)

National Partners

International Partners

  • Instytut Dendrologii Polskiej Akademii Nauk, Kornik, Poland (Prof. P. Chmielarz)
  • Universität Concepción, Chile (Dr. Sofia Valenzuela)
  • Universität Novi Sad, Bosnien-Herzegowina (Dr. V. Galovic)

Former Partners


Publications

  1. 0

    Brügmann T, Zahn V, Fendel A, Fladung M (2024) Biotechnologische Innovationen an Rotbuchen und weiteren Baumarten. In: Liesebach M, Tröber U, Neophytou C (eds) 8. Tagung der Sektion Forstgenetik/Forstpflanzenzüchtung "Wald der Zukunft - Beitrag von Forstgenetik und Forstpflanzenzüchtung" : Freiburg, 11. bis 13. September 2024 ; Abstract-Band und Exkursionsführer. p 66

  2. 1

    Grünhofer P, Herzig L, Zhang Q, Vitt S, Stöcker T, Malkowsky Y, Brügmann T, Fladung M, Schreiber L (2024) Changes in wax composition but not amount enhance cuticular transpiration. Plant Cell Environ 47(1):91-105, DOI:10.1111/pce.14719

    https://literatur.thuenen.de/digbib_extern/dn066889.pdf

  3. 2

    Brügmann T, Fendel A, Zahn V, Fladung M (2024) Enhancing tree resilience to climate change: Biotechnological strategies and genome editing. In: Forests & society towards 2050 : 26th IUFRO World Congress, Stockholm, Sweden, 23-29 June 2024 ; Book of abstracts. p 1032

    https://literatur.thuenen.de/digbib_extern/dn068696.pdf

  4. 3

    Fendel A, Fladung M, Brügmann T (2024) Erforschung der Trockenstresstoleranz in Bäumen durch Einzelgen-Modifikationen in Pappeln (Populus). In: Liesebach M, Tröber U, Neophytou C (eds) 8. Tagung der Sektion Forstgenetik/Forstpflanzenzüchtung "Wald der Zukunft - Beitrag von Forstgenetik und Forstpflanzenzüchtung" : Freiburg, 11. bis 13. September 2024 ; Abstract-Band und Exkursionsführer. p 10

  5. 4

    Brügmann T, Fendel A, Zahn V, Fladung M (2024) Genome editing in forest trees. In: Ricroch A, Eriksson D, Miladinovic D, Sweet JB, Laere K van, Wozniak-Gientka E (eds) A roadmap for plant genome editing. Cham: Springer, pp 347-372, DOI:10.1007/978-3-031-46150-7_20

    https://literatur.thuenen.de/digbib_extern/dn067381.pdf

  6. 5

    Brügmann T, Fladung M (2024) Genscheren in der Forstpflanzenzüchtung? AFZ Der Wald 79(5):27-30

  7. 6

    Rupps A, Thiesen T, Raschke J, Walther M, Ehmke E, Safranek L, Eckardt J, Schmidt A, Beck W, Bubner B, Eisold A-M, Zahn V, Fendel A, Brügmann T (2024) PInK-NET: Resuming the network idea of in vitro tree labs in Germany. In: Sota V, Werbrouck S (eds) In vitro culture of woody crops: problem solving by new approaches : The 2nd Conference of Cost Action CA21157 CopyTree, 22-24 April, 2024, Bulduri Technical School, Jurmala, Latvia ; Book of proceedings. pp 221-230

  8. 7

    Brügmann T, Fendel A, Woith M, Schwarzer F, Fomin A, Haß S, Pucker B, Fladung M, Forstreuter M (2024) Stronger together: genetics and ecophysiology of drought stress in Fagus sylvatica. In: Forests & society towards 2050 : 26th IUFRO World Congress, Stockholm, Sweden, 23-29 June 2024 ; Book of abstracts. p 2125

    https://literatur.thuenen.de/digbib_extern/dn068712.pdf

  9. 8

    Brügmann T, Zahn V, Fendel A, Sievers A-J, Jelkmann S, Fladung M (2024) The triad of tree biotechnology. In: Technische Universität Dresden (ed) FowiTa 2023 : Forstwissenschaftliche Tagung vom 11.-13. September 2023 in Dresden ; Wald- und Holzforschung zwischen Klimawandel, Bioökonomie und gesellschaftlichen Umbrüchen [Abstractband]. p 179, DOI:10.25368/2024.36

  10. 9

    Brügmann T, Fendel A, Woith M, Schwarzer F, Fomin A, Haß S, Pucker B, Fladung M, Forstreuter M (2024) Zusammenbringen, was zusammengehört: Genetik und Ökophysiologie von Trockenstress in Rotbuchen (Fagus sylvatica). In: Technische Universität Dresden (ed) FowiTa 2023 : Forstwissenschaftliche Tagung vom 11.-13. September 2023 in Dresden ; Wald- und Holzforschung zwischen Klimawandel, Bioökonomie und gesellschaftlichen Umbrüchen [Abstractband]. pp 47-48, DOI:10.25368/2024.36

  11. 10

    Cardi T, Murovec J, Bakhsh A, Boniecka J, Brügmann T, Bull SE, Eeckhaut T, Fladung M, Galovic V, Linkiewicz A, Lukan T, Mafra I, Michalski K, Kavas M, Nicolia A, Nowakowska J, Sagi L, Sarmiento C, Yildirim K, Zlatkovic M, et al (2023) CRISPR/Cas-mediated plant genome editing: outstanding challenges a decade after implementation. Trends Plant Sci 28(10):1144-1165, DOI:10.1016/j.tplants.2023.05.012

    https://literatur.thuenen.de/digbib_extern/dn066450.pdf

  12. 11

    Shahzaib M, Brügmann T, Shakeel M, Khan SH, Azhar MT, Atif RM, Fladung M, Rana IA (2023) Development of climate smart fruit plants via CRISPR/Cas genome editing systems: A spatiotemporal review [Preprint]. 2023051887. Basel: Preprints, 22 p, DOI:10.20944/preprints202305.1887.v1

    https://literatur.thuenen.de/digbib_extern/dn067511.pdf

  13. 12

    Zahn V, Sievers A-J, Fladung M, Brügmann T (2023) Etablierung einer stabilen in vitro-Kultur für Fagus sylvatica - Ein erster Schritt hin zur Genomeditierung in Buchen. Thünen Rep 105:9-10

    https://literatur.thuenen.de/digbib_extern/dn066285.pdf

  14. 13

    Fendel A, Brügmann T (2023) Kontrolliert unter Stress. AFZ Der Wald 78(20):12-15

  15. 14

    Brügmann T, Zahn V, Fendel A, Zebbedies S, Sievers A-J, Becker D, Fladung M (2023) Neue biotechnologische Methoden für Gehölze. Thünen Rep 105:6-8

    https://literatur.thuenen.de/digbib_extern/dn066284.pdf

  16. 15

    Fendel A, Fladung M, Brügmann T (2023) Steigerung der Trockenstresstoleranz in Bäumen durch genetische Modifikationen. Thünen Rep 105:88-89

    https://literatur.thuenen.de/digbib_extern/dn066288.pdf

  17. 16

    Riefler M, Brügmann T, Fladung M, Schmülling T (2022) A constitutively active cytokinin receptor variant increases cambial activity and stem growth in poplar. Int J Mol Sci 23(15):8321, DOI:10.3390/ijms23158321

    https://literatur.thuenen.de/digbib_extern/dn065121.pdf

  18. 17

    Zahn V, Sievers A-J, Fladung M, Brügmann T (2022) A reliable in vitro culture system for Fagus sylvatica - first steps to expend the scope of genome editing in forest trees to beech. In: Plant Sciences for a Sustainable Future : Botanik-Tagung, International Conference of the German Society for Plant Sciences, 2022 Bonn, 28 August - 01 September ; Programme. Deutsche Botanische Gesellschaft, p 249

  19. 18

    Zahn V, Brügmann T, Fladung M (2022) Combining bacterial and viral elements for efficient gene targeting in poplar. In: Plant Sciences for a Sustainable Future : Botanik-Tagung, International Conference of the German Society for Plant Sciences, 2022 Bonn, 28 August - 01 September ; Programme. Deutsche Botanische Gesellschaft, p 85

  20. 19

    Zahn V, Fladung M, Brügmann T (2022) Combining bacterial and viral elements for efficient gene targeting in poplar. In: COST Action CA18111 "Genome Editing in Plants" : Book of Abstracts ; 3rd PlantEd Conference, 5 - 7 September 2022, Düsseldorf, Germany. p 24

  21. 20

    Brügmann T, Zebbedies S, Zahn V, Fladung M (2022) Diverse mutation patterns include large deletions in a CRISPR/nCas9 double nicking approach poplar. In: COST Action CA18111 "Genome Editing in Plants" : Book of Abstracts ; 3rd PlantEd Conference, 5 - 7 September 2022, Düsseldorf, Germany. p 48

  22. 21

    Brügmann T, Zebbedies S, Zahn V, Fladung M (2022) Diverse mutation patterns include large deletions in a CRISPR/nCas9 double nicking approach poplar. In: Plant Sciences for a Sustainable Future : Botanik-Tagung, International Conference of the German Society for Plant Sciences, 2022 Bonn, 28 August - 01 September ; Programme. Deutsche Botanische Gesellschaft, p 248

  23. 22

    Brügmann T, Fladung M, Schröder H (2022) Flexible DNA isolation procedure for different tree species as a convenient lab routine. Silvae Genetica 71:20-30, DOI:10.2478/sg-2022-0003

    https://literatur.thuenen.de/digbib_extern/dn065030.pdf

  24. 23

    Fendel A, Fladung M, Brügmann T (2022) Improvement of drought stress tolerance in poplars (Populus) by modification of candidate genes. In: COST Action CA18111 "Genome Editing in Plants" : Book of Abstracts ; 3rd PlantEd Conference, 5 - 7 September 2022, Düsseldorf, Germany. p 35

  25. 24

    Fendel A, Fladung M, Brügmann T (2022) Improvement of drought stress tolerance in poplars (Populus) by modification of candidate genes. In: Plant Sciences for a Sustainable Future : Botanik-Tagung, International Conference of the German Society for Plant Sciences, 2022 Bonn, 28 August - 01 September ; Programme. Deutsche Botanische Gesellschaft, p 104

  26. 25

    Shrestha A, Fendel A, Nguyen TH, Adebabay A, Kullik AS, Benndorf J, Leon J, Naz AA (2022) Natural diversity uncovers P5CS1 regulation and its role in drought stress tolerance and yield sustainability in barley. Plant Cell Environ 45(12):3523-3536, DOI:10.1111/pce.14445

    https://literatur.thuenen.de/digbib_extern/dn065457.pdf

  27. 26

    Brügmann T, Zahn V, Fendel A, Zebbedies S, Sievers A-J, Becker D, Fladung M (2022) Neue biotechnologische Methoden für Gehölze. In: Liesebach M, Tröber U (eds) 7. Tagung der Sektion Forstgenetik/Forstpflanzenzüchtung : "Beiträge von Forstpflanzenzüchtung und Forstgenetik für den Wald von Morgen" ; Ahrensburg, 12. bis 14. September 2022, Abstract-Band und Exkursionsführer. p 11

  28. 27

    Brügmann T, Biricolti S, Becker D, Fladung M (2022) On track towards DNA-free genome editing. In: COST Action CA18111 "Genome Editing in Plants" : Book of Abstracts ; 3rd PlantEd Conference, 5 - 7 September 2022, Düsseldorf, Germany. p 49

  29. 28

    Brügmann T, Deecke K, Fladung M (2019) Evaluating the efficiency of gRNAs in CRISPR/Cas9 mediated genome editing in poplars. Int J Mol Sci 20(15):3623, DOI:10.3390/ijms20153623

    https://literatur.thuenen.de/digbib_extern/dn061163.pdf

  30. 29

    Brügmann T, Fladung M (2019) Genom-Editierung in Bäumen. AFZ Der Wald 74(5):16-18

    https://literatur.thuenen.de/digbib_extern/dn060910.pdf

  31. 30

    Brügmann T, Wetzel H, Hettrich K, Smeds A, Willför S, Kersten B, Fladung M (2019) Knockdown of PCBER1, a gene of neolignan biosynthesis, resulted in increased poplar growth. Planta 249(2):515-525, DOI:10.1007/s00425-018-3021-8

  32. 31

    Brügmann T, Fladung M (2019) Overexpression of both flowering time genes AtSOC1 and SaFUL revealed huge influence onto plant habitus in poplar. Tree Genetics Genomes 15:20, DOI:10.1007/s11295-019-1326-9

  33. 32

    Brügmann T, Polak O, Deecke K, Nietsch J, Fladung M (2019) Poplar transformation. Meth Mol Biol 1864:165-177

  34. 33

    Brügmann T, Fladung M (2016) Flowering time genes influence biomass production in poplars. In: Hensel G (ed) Annual meeting : new breeding technologies in times of politically enforces research prohibition ; German Society of Plant Biotechnology e.V. ; book of abstracts ; Gatersleben, 02.-04.05.2016. Gatersleben: Gesellschaft für Pflanzenbiotechnologie, p 23

  35. 34

    Brügmann T, Polak O, Fladung M (2015) Development and use of novel gene technologies to increase biomass yield in the woody perennial Populus spec. (PopMass) : project A - modification of genes expressed in xylem and flower. In: Conference documents Plant 2040 Status Seminar, March 4 - 6, 2015 in Potsdam. pp 67-68

  36. 35

    Brügmann T, Fladung M (2015) Genetic approaches to increase biomass yield in the woody perennial Populus. In: Vettori C, Vendramin GG, Paffetti D, Travaglini D (eds) IUFRO Tree Biotechnology 2015 Conference : "Forests: the importance to the planet and society" ; 8-12 June 2015, Florence, Italy ; Proceedings. Florenz: IUFRO, pp 507-509

  37. 36

    Schröder H, Brügmann T, Fladung M (2015) Nuclear and chloroplast SNP markers support successful poplar breeding. In: Vettori C, Vendramin GG, Paffetti D, Travaglini D (eds) IUFRO Tree Biotechnology 2015 Conference : "Forests: the importance to the planet and society" ; 8-12 June 2015, Florence, Italy ; Proceedings. Florenz: IUFRO, pp 295-296

  38. 37

    Hönicka H, Fladung M, Lehnhardt D, Brügmann T (2013) Pleiotropic effects of "flowering" genes in transgenic poplar. In: SPP1530 workshop: Pleiotropic effects of flowering time genes and impact on adaptation and speciation, January 21-23,2013, Max Planck Institute for Plant Breeding Research, Köln. p 13

  39. 38

    Brügmann T, Fladung M (2013) Potentials and limitations of the cross-species transfer of nuclear microsatellite marker in six species belonging to three sections of the genus PopulusL.. Tree Genetics Genomes 9(6):1413-1421, DOI:10.1007/s11295-013-0647-3

  40. 39

    Brügmann T (2011) Entwicklung molekularer Marker zur genetischen Charakterisierung von Genotypen verschiedener Arten der Gattung Populus L.. Hamburg: Universität, 134 p, Hamburg, Univ, Fachber Biologie, Masterarbeit, 2011

  41. 40

    Brügmann T (2009) Optimierung von RNA-Extraktion und cDNA-Synthese aus Quercus robur. Hamburg: Univ, 76 p, Hamburg, Univ, Department Biologie, Bachelorarbeit, 2009


    Funding

    The Genetic Technologies Junior Research Group is financially supported by the Federal Ministry of Food and Agriculture through the Agency of Renewable Resources (FNR) (funding code 2219NR359).

    The related project "TreeEdit" is part of the Department of Genome Research.


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