Transformation of European Ash (Fraxinus excelsior L.) Callus as a Starting Point for Understanding the Molecular Basis of Ash Dieback

The population of European ash (Fraxinus excelsior L.) is currently facing the risk of collapse, mainly due to ash dieback, a disease caused by a pathogenic fungus, Hymenoscyphus fraxineus. To facilitate studies into the molecular basis of ash dieback and design...

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Autores principales:	Anna Hebda, Aleksandra Liszka, Piotr Zgłobicki, Katarzyna Nawrot-Chorabik, Jan J. Lyczakowski
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<i>Agrobacterium tumefaciens</i> ash dieback callus <i>Fraxinus excelsior</i> transformation Botany QK1-989
Acceso en línea:	https://doaj.org/article/1c46780895334fe5afbfd79653886c7d
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spelling	oai:doaj.org-article:1c46780895334fe5afbfd79653886c7d2021-11-25T18:47:23ZTransformation of European Ash (<i>Fraxinus excelsior</i> L.) Callus as a Starting Point for Understanding the Molecular Basis of Ash Dieback10.3390/plants101125242223-7747https://doaj.org/article/1c46780895334fe5afbfd79653886c7d2021-11-01T00:00:00Zhttps://www.mdpi.com/2223-7747/10/11/2524https://doaj.org/toc/2223-7747The population of European ash (<i>Fraxinus excelsior</i> L.) is currently facing the risk of collapse, mainly due to ash dieback, a disease caused by a pathogenic fungus, <i>Hymenoscyphus fraxineus</i>. To facilitate studies into the molecular basis of ash dieback and design breeding strategies for a generation of resistant trees, it is necessary to develop tools enabling the study of gene function in <i>F. excelsior</i>. Despite this, a method for the genetic engineering of <i>F. excelsior</i> is still missing. Here, we report the first successful genetic transformation of <i>F. excelsior</i> callus and a selection process enabling the formation of stable transgenic callus lines. The protocol relies on the use of <i>Agrobacterium tumefaciens</i> to transform callus tissue derived from embryos of <i>F. excelsior</i>. In our experiments, we used the β-glucuronidase (GUS) reporter system to demonstrate the transformation of callus cells and performed RT-PCR experiments to confirm the stable expression of the transgene. Since ash dieback threatens the long-term stability of many native <i>F. excelsior</i> populations, we hope that the transformation techniques described in this manuscript will facilitate rapid progress in uncovering the molecular basis of the disease and the validation of gene targets previously proposed to be linked to the resistance of trees to <i>H. fraxineus</i> pathogenicity.Anna HebdaAleksandra LiszkaPiotr ZgłobickiKatarzyna Nawrot-ChorabikJan J. LyczakowskiMDPI AGarticle<i>Agrobacterium tumefaciens</i>ash diebackcallus<i>Fraxinus excelsior</i>transformationBotanyQK1-989ENPlants, Vol 10, Iss 2524, p 2524 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<i>Agrobacterium tumefaciens</i> ash dieback callus <i>Fraxinus excelsior</i> transformation Botany QK1-989
spellingShingle	<i>Agrobacterium tumefaciens</i> ash dieback callus <i>Fraxinus excelsior</i> transformation Botany QK1-989 Anna Hebda Aleksandra Liszka Piotr Zgłobicki Katarzyna Nawrot-Chorabik Jan J. Lyczakowski Transformation of European Ash (<i>Fraxinus excelsior</i> L.) Callus as a Starting Point for Understanding the Molecular Basis of Ash Dieback
description	The population of European ash (<i>Fraxinus excelsior</i> L.) is currently facing the risk of collapse, mainly due to ash dieback, a disease caused by a pathogenic fungus, <i>Hymenoscyphus fraxineus</i>. To facilitate studies into the molecular basis of ash dieback and design breeding strategies for a generation of resistant trees, it is necessary to develop tools enabling the study of gene function in <i>F. excelsior</i>. Despite this, a method for the genetic engineering of <i>F. excelsior</i> is still missing. Here, we report the first successful genetic transformation of <i>F. excelsior</i> callus and a selection process enabling the formation of stable transgenic callus lines. The protocol relies on the use of <i>Agrobacterium tumefaciens</i> to transform callus tissue derived from embryos of <i>F. excelsior</i>. In our experiments, we used the β-glucuronidase (GUS) reporter system to demonstrate the transformation of callus cells and performed RT-PCR experiments to confirm the stable expression of the transgene. Since ash dieback threatens the long-term stability of many native <i>F. excelsior</i> populations, we hope that the transformation techniques described in this manuscript will facilitate rapid progress in uncovering the molecular basis of the disease and the validation of gene targets previously proposed to be linked to the resistance of trees to <i>H. fraxineus</i> pathogenicity.
format	article
author	Anna Hebda Aleksandra Liszka Piotr Zgłobicki Katarzyna Nawrot-Chorabik Jan J. Lyczakowski
author_facet	Anna Hebda Aleksandra Liszka Piotr Zgłobicki Katarzyna Nawrot-Chorabik Jan J. Lyczakowski
author_sort	Anna Hebda
title	Transformation of European Ash (<i>Fraxinus excelsior</i> L.) Callus as a Starting Point for Understanding the Molecular Basis of Ash Dieback
title_short	Transformation of European Ash (<i>Fraxinus excelsior</i> L.) Callus as a Starting Point for Understanding the Molecular Basis of Ash Dieback
title_full	Transformation of European Ash (<i>Fraxinus excelsior</i> L.) Callus as a Starting Point for Understanding the Molecular Basis of Ash Dieback
title_fullStr	Transformation of European Ash (<i>Fraxinus excelsior</i> L.) Callus as a Starting Point for Understanding the Molecular Basis of Ash Dieback
title_full_unstemmed	Transformation of European Ash (<i>Fraxinus excelsior</i> L.) Callus as a Starting Point for Understanding the Molecular Basis of Ash Dieback
title_sort	transformation of european ash (<i>fraxinus excelsior</i> l.) callus as a starting point for understanding the molecular basis of ash dieback
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/1c46780895334fe5afbfd79653886c7d
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Transformation of European Ash (<i>Fraxinus excelsior</i> L.) Callus as a Starting Point for Understanding the Molecular Basis of Ash Dieback

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