Introduction of antifungal genes in sunflower via Agrobacterium

There is evidence that overexpression of transgenes codifying antifungal proteins may confer protection to pathogen attack, and that this protection is increased due to the synergic effect of the expression of two or more genes. On the other hand it is well known that sunflower is a recalcitrant spe...

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Autores principales:	Radonic,Laura Mabel, Zimmermann,Julián Marcos, Zavallo,Diego, López,Nilda, López Bilbao,Marisa
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2008
Materias:	antifungal genes double expression cassettes polyprotein cassettes sunflower transgenic plants
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000500008
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spelling	oai:scielo:S0717-345820080005000082009-06-16Introduction of antifungal genes in sunflower via AgrobacteriumRadonic,Laura MabelZimmermann,Julián MarcosZavallo,DiegoLópez,NildaLópez Bilbao,Marisa antifungal genes double expression cassettes polyprotein cassettes sunflower transgenic plants There is evidence that overexpression of transgenes codifying antifungal proteins may confer protection to pathogen attack, and that this protection is increased due to the synergic effect of the expression of two or more genes. On the other hand it is well known that sunflower is a recalcitrant specie, highly difficult to be genetically transformed. In this context, the final aim of this project was to obtain sunflower plants expressing at least two antifungal genes, avoiding sequential transformation. The antifungal genes used encode for two enzymes that degrade the fungal wall (glucanase and chitinase), an osmotin and a ribosome inhibitor protein. Two types of transformation vectors were used: a more traditional system with a double cassette and a novel system producing a unique polyprotein with antifungal proteins released in equimolecular quantities. The polyprotein vector system generated hyperhydric shoots with necrotic areas and abnormal growth at the end of the tissue culture procedure, making impossible the use of this interesting vector in sunflower. Transformation assays carried out with the pHGC39 vector (including glucanase and chitinase genes) vector produced 0.83% efficiency, corresponding to 13 rooted shoots in kanamycin (Km) from a total of 1568 agroinfected shoots. T0 rooted shoots resulted positive by PCR analysis and were transferred to greenhouse to obtain their offspring. In addition, we corroborate the transformation protocol using Km as selective marker, previously described (Radonic et al. 2006) with a reporter gene, but in this opportunity with antifungal genes.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.11 n.5 20082008-12-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000500008en10.4067/S0717-34582008000500008
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	antifungal genes double expression cassettes polyprotein cassettes sunflower transgenic plants
spellingShingle	antifungal genes double expression cassettes polyprotein cassettes sunflower transgenic plants Radonic,Laura Mabel Zimmermann,Julián Marcos Zavallo,Diego López,Nilda López Bilbao,Marisa Introduction of antifungal genes in sunflower via Agrobacterium
description	There is evidence that overexpression of transgenes codifying antifungal proteins may confer protection to pathogen attack, and that this protection is increased due to the synergic effect of the expression of two or more genes. On the other hand it is well known that sunflower is a recalcitrant specie, highly difficult to be genetically transformed. In this context, the final aim of this project was to obtain sunflower plants expressing at least two antifungal genes, avoiding sequential transformation. The antifungal genes used encode for two enzymes that degrade the fungal wall (glucanase and chitinase), an osmotin and a ribosome inhibitor protein. Two types of transformation vectors were used: a more traditional system with a double cassette and a novel system producing a unique polyprotein with antifungal proteins released in equimolecular quantities. The polyprotein vector system generated hyperhydric shoots with necrotic areas and abnormal growth at the end of the tissue culture procedure, making impossible the use of this interesting vector in sunflower. Transformation assays carried out with the pHGC39 vector (including glucanase and chitinase genes) vector produced 0.83% efficiency, corresponding to 13 rooted shoots in kanamycin (Km) from a total of 1568 agroinfected shoots. T0 rooted shoots resulted positive by PCR analysis and were transferred to greenhouse to obtain their offspring. In addition, we corroborate the transformation protocol using Km as selective marker, previously described (Radonic et al. 2006) with a reporter gene, but in this opportunity with antifungal genes.
author	Radonic,Laura Mabel Zimmermann,Julián Marcos Zavallo,Diego López,Nilda López Bilbao,Marisa
author_facet	Radonic,Laura Mabel Zimmermann,Julián Marcos Zavallo,Diego López,Nilda López Bilbao,Marisa
author_sort	Radonic,Laura Mabel
title	Introduction of antifungal genes in sunflower via Agrobacterium
title_short	Introduction of antifungal genes in sunflower via Agrobacterium
title_full	Introduction of antifungal genes in sunflower via Agrobacterium
title_fullStr	Introduction of antifungal genes in sunflower via Agrobacterium
title_full_unstemmed	Introduction of antifungal genes in sunflower via Agrobacterium
title_sort	introduction of antifungal genes in sunflower via agrobacterium
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2008
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000500008
work_keys_str_mv	AT radoniclauramabel introductionofantifungalgenesinsunflowerviaagrobacterium AT zimmermannjulianmarcos introductionofantifungalgenesinsunflowerviaagrobacterium AT zavallodiego introductionofantifungalgenesinsunflowerviaagrobacterium AT lopeznilda introductionofantifungalgenesinsunflowerviaagrobacterium AT lopezbilbaomarisa introductionofantifungalgenesinsunflowerviaagrobacterium
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Introduction of antifungal genes in sunflower via Agrobacterium

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