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Cytokinin-deficient CRISPR/Cas9 mutants of Chlamydomonas reinhardtii show reduced ability to prime resistance of tobacco against bacterial infection
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  • Sopan Ganpatrao Wagh,
  • Roman Sandor,
  • Simon Kelterborn,
  • Ondrej Novak,
  • Niels Olsen,
  • Bichitra Paul,
  • Shujie Wu,
  • Peter Hegemann,
  • Miroslav Strnad,
  • Jan Červený,
  • Thomas Roitsch
Sopan Ganpatrao Wagh
Ustav vyzkumu globalni zmeny Akademie ved Ceske republiky
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Roman Sandor
Ustav vyzkumu globalni zmeny Akademie ved Ceske republiky
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Simon Kelterborn
Humboldt-Universitat zu Berlin Institut fur Biologie
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Ondrej Novak
Palacký University
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Niels Olsen
University of Copenhagen
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Bichitra Paul
University of Copenhagen
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Shujie Wu
University of Copenhagen
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Peter Hegemann
Humboldt-Universitat zu Berlin Institut fur Biologie
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Miroslav Strnad
Palacký University
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Jan Červený
Ustav vyzkumu globalni zmeny Akademie ved Ceske republiky
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Thomas Roitsch
Ustav vyzkumu globalni zmeny Akademie ved Ceske republiky

Corresponding Author:[email protected]

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Abstract

Microalgae play an essential role in agriculture as bio-fertilizers and biostimulants, but some also produce various plant hormones, such as cytokinins (CK). CK are a class of natural plant hormones and there is evidence that exogenous or bacterial CK can modulate plant defense. However, there is a lack of knowledge about the effect of specific microalgae or microalgae-produced cytokinins on this response. In our study, we have evaluated the cytokinins-producing microalgae Chlamydomonas reinhardtii (Cri) on its ability to prime Nicotiana tabacum plant defenses against its natural pathogen Pseudomonas syringae pv. tabaci. Our work functionally verifies that Cri primes plant defense response, with cytokinins being a crucial component. We have used the CRISPR/Cas9 system to generate Cri LOG and IPT gene knockouts, which are major points of the CK biosynthesis pathways. While Cri shows strong protection potential, the CK-deficient mutants have a reduced ability to affect plant defense, where the degree of protection correlates with the CK levels - the IPT mutants show less protection than the LOG mutants. Additionally, by measuring plant defense gene expression by RTq-PCR we have shown that Cri treatment stimulates tobacco defense response by priming.