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Growth of Mixoploid GIBBERELLIC ACID 20 OXIDASE (GA20-OXIDASE) Overexpressing Transgenic Populus

Wachstum von mixoploiden GIBBERELLINSÄURE 20 OXIDASE (GA20-OXIDASE) überexprimierenden transgenen Pappeln

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Abstract

Biomass production is traditionally an important goal in forest tree breeding programs. However, progress in classical tree breeding is limited because of long generation cycles of many forest tree species. Due to this limitation, genetic engineering, and also the quite recently developed genome editing technique, are very attractive tools to improve tree characteristics in a relatively short time. As a proof-of-concept approach, the GIBBERELLIC ACID 20 OXIDASE (GA20-OXIDASE) gene was constitutively overexpressed in poplar leading to significantly higher plants. Unfortunately, deleterious side-effects were also observed, like longer and thinner shoots leading to plants with unstable stems unable to grow without external stabilization. In the literature, polyploidization was reported to increase stem thickness relatively to plant height in several plant species. Therefore, polyploidization of GA20-OXIDASE overexpressing transgenic poplar lines was attempted. Mixoploid (ploidy chimeric: diploid and tetraploid nuclei) plants were obtained and assessed for several years for ploidy status and growth performance (stem height and diameter, bud set) in climate chamber and glasshouse. The results indicate that mixoploid GA20-OXIDASE overexpressing transgenic poplar still reveal increased growth but also showed a lower height-to-stem ratio compared to the diploid GA20-OXIDASE transgenic poplar, enabling the mixoploid transgenic plants to grow without external stabilization.

Zusammenfassung

Die Biomasseproduktion ist traditionell ein wichtiges Ziel in der Forstpflanzenzüchtung. Die Fortschritte in der klassischen Forstpflanzenzüchtung sind jedoch wegen der langen Generationszyklen vieler Waldbaumarten begrenzt. Aufgrund dieser Einschränkung stellen die klassische Gentechnik sowie auch die vor kurzem entwickelte Genom-Editierung-Technik sehr attraktive Werkzeuge dar, um Eigenschaften von langlebigen Bäumen in relativ kurzer Zeit zu verbessern. Als „Proof-of-Concept“-Ansatz wurde das GIBBERELLINSÄURE 20 OXIDASE (GA20-OXIDASE) Gen in der Pappel konstitutiv überexprimiert, was zu signifikant höheren Pflanzen führte. Leider wurden auch unliebsame Nebeneffekte beobachtet, wie längere und dünnere Triebe, die zu Pflanzen mit instabilen Stämmen führen, die ohne externe Stabilisierung nicht wachsen können. In der Literatur wurde berichtet, dass die Polyploidisierung bei verschiedenen Pflanzenarten die Stammdicke relativ zur Pflanzenhöhe erhöht. Daher wurde eine Polyploidisierung von GA20-OXIDASE-überexprimierenden transgenen Pappellinien versucht. Mixoploide (ploidische Chimäre: diploide und tetraploide Kerne) Pflanzen wurden erhalten und über mehrere Jahre auf Ploidiestatus und Wachstumsleistung (Stammhöhe und -durchmesser, Knospensatz) in der Klimakammer und im Gewächshaus untersucht. Die Ergebnisse zeigen, dass die mixoploiden GA20-OXIDASE-überexprimierenden transgenen Pappeln immer noch ein erhöhtes Wachstum aufweisen, aber auch im Vergleich zu diploiden GA20-OXIDASE-transgenen Pappeln ein geringeres Verhältnis von Höhe zu Stängel zeigen, wodurch die mixoploiden transgenen Pflanzen ohne externe Stabilisierung wachsen können.

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Acknowledgements

Thanks are due to Maria E. Eriksson (University Umeå, Sweden) for providing the 35S::GA20-OXIDASE construct. I thank A. Schellhorn and Olaf Polak for excellent technical assistance in the lab, and the glasshouse staff (M. Hunger, G. Wiemann, R. Ebbinghaus, M. Spauszus) for plant cultivation. I also thank anonymous reviewers for their valuable comments.

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  1. Thuenen-Institute of Forest Genetics (TI-FG), Sieker Landstr. 2, 22927, Grosshansdorf, Germany

    Matthias Fladung

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  1. Matthias Fladung
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Correspondence to Matthias Fladung.

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M. Fladung declares that he has no competing interests.

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M. Fladung: experimental design, genetic transformation, molecular analysis, polyploidization, data analyses, and manuscript preparation.

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Fladung, M. Growth of Mixoploid GIBBERELLIC ACID 20 OXIDASE (GA20-OXIDASE) Overexpressing Transgenic Populus . Gesunde Pflanzen 70, 91–98 (2018). https://doi.org/10.1007/s10343-018-0418-z

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  • DOI: https://doi.org/10.1007/s10343-018-0418-z

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