α-l-Arabinofuranosidase from strawberry fruit: Cloning of three cDNAs, characterization of their expression and analysis of enzymatic activity in cultivars with contrasting firmnes...

Autores
Rosli, Hernan Guillermo; Civello, Pedro Marcos; Martinez, Gustavo Adolfo
Año de publicación
2009
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Softening of fleshy fruits during ripening is associated to catabolism of cell wall components. In strawberry, pectin degradation, as well as loss of neutral sugars (mainly arabinose), increases during ripening, and probably contributes to fruit softening. In this work, we report the activity of a-L-arabinofuranosidase (a-L-arafase) and the expression of related genes in strawberry. Activity of a-L-arafase was measured during ripening of cultivars with contrasting firmness. An important increment in the specific activity of a-L-arafase was detected during ripening in both cultivars. However, in the softest one (Toyonoka) the specific activities were higher than in the firmest (Camarosa). A combination of semiquantitative reverse transcriptase-PCR (RT-PCR) with degenerate primers and a screening of a cDNA library allowed the isolation and cloning of three cDNAs encoding putative a-L-arafases (FaAra1, FaAra2 and FaAra3). The deduced proteins revealed that FaAras belong to the glycoside hydrolase family 51 and not to glycoside hydrolase family 3. Expression studies, carried out by means of Northern-blot and semiquantitative RT-PCR, revealed that FaAras were predominantly expressed in fruit tissue and detected over the entire ripening process. Due to similarity of FaAras sequences, Northern-blot analysis probably grouped the expression of the three genes. The expression was high at small green stage, decreased at white stage and increased thereafter. The increment of the expression from white to 50% red stage was more evident in the softest cultivar (Toyonoka). Semi quantitative RT-PCR analysis allowed determining the expression of individual FaAras. The expression of the three genes was detected in all developmental and ripening stages. However, differences in expression levels could be detected between cultivars. In the softest cultivar, the expression of the three FaAras was higher at 50% and 75% red stages, and in the case of FaAra3 a higher expression was found also at 100% red stage. Overall, specific activity of a-L-arafase was higher in the softest cultivar; such activity reflects the expression of at least three putative FaAra genes.
Fil: Rosli, Hernan Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina
Fil: Civello, Pedro Marcos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina
Fil: Martinez, Gustavo Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina
Materia
FRAGARIA X ANANASSA
RIPENING
SOFTENING
CELL WALL
PECTIN
HEMICELLULOSES
ARABINOFURANOSIDASE
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/256768
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/256768
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling α-l-Arabinofuranosidase from strawberry fruit: Cloning of three cDNAs, characterization of their expression and analysis of enzymatic activity in cultivars with contrasting firmnessRosli, Hernan GuillermoCivello, Pedro MarcosMartinez, Gustavo AdolfoFRAGARIA X ANANASSARIPENINGSOFTENINGCELL WALLPECTINHEMICELLULOSESARABINOFURANOSIDASEhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Softening of fleshy fruits during ripening is associated to catabolism of cell wall components. In strawberry, pectin degradation, as well as loss of neutral sugars (mainly arabinose), increases during ripening, and probably contributes to fruit softening. In this work, we report the activity of a-L-arabinofuranosidase (a-L-arafase) and the expression of related genes in strawberry. Activity of a-L-arafase was measured during ripening of cultivars with contrasting firmness. An important increment in the specific activity of a-L-arafase was detected during ripening in both cultivars. However, in the softest one (Toyonoka) the specific activities were higher than in the firmest (Camarosa). A combination of semiquantitative reverse transcriptase-PCR (RT-PCR) with degenerate primers and a screening of a cDNA library allowed the isolation and cloning of three cDNAs encoding putative a-L-arafases (FaAra1, FaAra2 and FaAra3). The deduced proteins revealed that FaAras belong to the glycoside hydrolase family 51 and not to glycoside hydrolase family 3. Expression studies, carried out by means of Northern-blot and semiquantitative RT-PCR, revealed that FaAras were predominantly expressed in fruit tissue and detected over the entire ripening process. Due to similarity of FaAras sequences, Northern-blot analysis probably grouped the expression of the three genes. The expression was high at small green stage, decreased at white stage and increased thereafter. The increment of the expression from white to 50% red stage was more evident in the softest cultivar (Toyonoka). Semi quantitative RT-PCR analysis allowed determining the expression of individual FaAras. The expression of the three genes was detected in all developmental and ripening stages. However, differences in expression levels could be detected between cultivars. In the softest cultivar, the expression of the three FaAras was higher at 50% and 75% red stages, and in the case of FaAra3 a higher expression was found also at 100% red stage. Overall, specific activity of a-L-arafase was higher in the softest cultivar; such activity reflects the expression of at least three putative FaAra genes.Fil: Rosli, Hernan Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); ArgentinaFil: Civello, Pedro Marcos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; ArgentinaFil: Martinez, Gustavo Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); ArgentinaElsevier France-Editions Scientifiques Medicales Elsevier2009-12-24info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/256768Rosli, Hernan Guillermo; Civello, Pedro Marcos; Martinez, Gustavo Adolfo; α-l-Arabinofuranosidase from strawberry fruit: Cloning of three cDNAs, characterization of their expression and analysis of enzymatic activity in cultivars with contrasting firmness; Elsevier France-Editions Scientifiques Medicales Elsevier; Plant Physiology and Biochemistry; 47; 4; 24-12-2009; 272-2810981-94281873-2690CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0981942808002441info:eu-repo/semantics/altIdentifier/doi/10.1016/j.plaphy.2008.12.009info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:57:57Zoai:ri.conicet.gov.ar:11336/256768instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-15 14:57:57.812CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv α-l-Arabinofuranosidase from strawberry fruit: Cloning of three cDNAs, characterization of their expression and analysis of enzymatic activity in cultivars with contrasting firmness
title α-l-Arabinofuranosidase from strawberry fruit: Cloning of three cDNAs, characterization of their expression and analysis of enzymatic activity in cultivars with contrasting firmness
spellingShingle α-l-Arabinofuranosidase from strawberry fruit: Cloning of three cDNAs, characterization of their expression and analysis of enzymatic activity in cultivars with contrasting firmness
Rosli, Hernan Guillermo
FRAGARIA X ANANASSA
RIPENING
SOFTENING
CELL WALL
PECTIN
HEMICELLULOSES
ARABINOFURANOSIDASE
title_short α-l-Arabinofuranosidase from strawberry fruit: Cloning of three cDNAs, characterization of their expression and analysis of enzymatic activity in cultivars with contrasting firmness
title_full α-l-Arabinofuranosidase from strawberry fruit: Cloning of three cDNAs, characterization of their expression and analysis of enzymatic activity in cultivars with contrasting firmness
title_fullStr α-l-Arabinofuranosidase from strawberry fruit: Cloning of three cDNAs, characterization of their expression and analysis of enzymatic activity in cultivars with contrasting firmness
title_full_unstemmed α-l-Arabinofuranosidase from strawberry fruit: Cloning of three cDNAs, characterization of their expression and analysis of enzymatic activity in cultivars with contrasting firmness
title_sort α-l-Arabinofuranosidase from strawberry fruit: Cloning of three cDNAs, characterization of their expression and analysis of enzymatic activity in cultivars with contrasting firmness
dc.creator.none.fl_str_mv Rosli, Hernan Guillermo
Civello, Pedro Marcos
Martinez, Gustavo Adolfo
author Rosli, Hernan Guillermo
author_facet Rosli, Hernan Guillermo
Civello, Pedro Marcos
Martinez, Gustavo Adolfo
author_role author
author2 Civello, Pedro Marcos
Martinez, Gustavo Adolfo
author2_role author
author
dc.subject.none.fl_str_mv FRAGARIA X ANANASSA
RIPENING
SOFTENING
CELL WALL
PECTIN
HEMICELLULOSES
ARABINOFURANOSIDASE
topic FRAGARIA X ANANASSA
RIPENING
SOFTENING
CELL WALL
PECTIN
HEMICELLULOSES
ARABINOFURANOSIDASE
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Softening of fleshy fruits during ripening is associated to catabolism of cell wall components. In strawberry, pectin degradation, as well as loss of neutral sugars (mainly arabinose), increases during ripening, and probably contributes to fruit softening. In this work, we report the activity of a-L-arabinofuranosidase (a-L-arafase) and the expression of related genes in strawberry. Activity of a-L-arafase was measured during ripening of cultivars with contrasting firmness. An important increment in the specific activity of a-L-arafase was detected during ripening in both cultivars. However, in the softest one (Toyonoka) the specific activities were higher than in the firmest (Camarosa). A combination of semiquantitative reverse transcriptase-PCR (RT-PCR) with degenerate primers and a screening of a cDNA library allowed the isolation and cloning of three cDNAs encoding putative a-L-arafases (FaAra1, FaAra2 and FaAra3). The deduced proteins revealed that FaAras belong to the glycoside hydrolase family 51 and not to glycoside hydrolase family 3. Expression studies, carried out by means of Northern-blot and semiquantitative RT-PCR, revealed that FaAras were predominantly expressed in fruit tissue and detected over the entire ripening process. Due to similarity of FaAras sequences, Northern-blot analysis probably grouped the expression of the three genes. The expression was high at small green stage, decreased at white stage and increased thereafter. The increment of the expression from white to 50% red stage was more evident in the softest cultivar (Toyonoka). Semi quantitative RT-PCR analysis allowed determining the expression of individual FaAras. The expression of the three genes was detected in all developmental and ripening stages. However, differences in expression levels could be detected between cultivars. In the softest cultivar, the expression of the three FaAras was higher at 50% and 75% red stages, and in the case of FaAra3 a higher expression was found also at 100% red stage. Overall, specific activity of a-L-arafase was higher in the softest cultivar; such activity reflects the expression of at least three putative FaAra genes.
Fil: Rosli, Hernan Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina
Fil: Civello, Pedro Marcos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina
Fil: Martinez, Gustavo Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina
description Softening of fleshy fruits during ripening is associated to catabolism of cell wall components. In strawberry, pectin degradation, as well as loss of neutral sugars (mainly arabinose), increases during ripening, and probably contributes to fruit softening. In this work, we report the activity of a-L-arabinofuranosidase (a-L-arafase) and the expression of related genes in strawberry. Activity of a-L-arafase was measured during ripening of cultivars with contrasting firmness. An important increment in the specific activity of a-L-arafase was detected during ripening in both cultivars. However, in the softest one (Toyonoka) the specific activities were higher than in the firmest (Camarosa). A combination of semiquantitative reverse transcriptase-PCR (RT-PCR) with degenerate primers and a screening of a cDNA library allowed the isolation and cloning of three cDNAs encoding putative a-L-arafases (FaAra1, FaAra2 and FaAra3). The deduced proteins revealed that FaAras belong to the glycoside hydrolase family 51 and not to glycoside hydrolase family 3. Expression studies, carried out by means of Northern-blot and semiquantitative RT-PCR, revealed that FaAras were predominantly expressed in fruit tissue and detected over the entire ripening process. Due to similarity of FaAras sequences, Northern-blot analysis probably grouped the expression of the three genes. The expression was high at small green stage, decreased at white stage and increased thereafter. The increment of the expression from white to 50% red stage was more evident in the softest cultivar (Toyonoka). Semi quantitative RT-PCR analysis allowed determining the expression of individual FaAras. The expression of the three genes was detected in all developmental and ripening stages. However, differences in expression levels could be detected between cultivars. In the softest cultivar, the expression of the three FaAras was higher at 50% and 75% red stages, and in the case of FaAra3 a higher expression was found also at 100% red stage. Overall, specific activity of a-L-arafase was higher in the softest cultivar; such activity reflects the expression of at least three putative FaAra genes.
publishDate 2009
dc.date.none.fl_str_mv 2009-12-24
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/256768
Rosli, Hernan Guillermo; Civello, Pedro Marcos; Martinez, Gustavo Adolfo; α-l-Arabinofuranosidase from strawberry fruit: Cloning of three cDNAs, characterization of their expression and analysis of enzymatic activity in cultivars with contrasting firmness; Elsevier France-Editions Scientifiques Medicales Elsevier; Plant Physiology and Biochemistry; 47; 4; 24-12-2009; 272-281
0981-9428
1873-2690
CONICET Digital
CONICET
url http://hdl.handle.net/11336/256768
identifier_str_mv Rosli, Hernan Guillermo; Civello, Pedro Marcos; Martinez, Gustavo Adolfo; α-l-Arabinofuranosidase from strawberry fruit: Cloning of three cDNAs, characterization of their expression and analysis of enzymatic activity in cultivars with contrasting firmness; Elsevier France-Editions Scientifiques Medicales Elsevier; Plant Physiology and Biochemistry; 47; 4; 24-12-2009; 272-281
0981-9428
1873-2690
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0981942808002441
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.plaphy.2008.12.009
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier France-Editions Scientifiques Medicales Elsevier
publisher.none.fl_str_mv Elsevier France-Editions Scientifiques Medicales Elsevier
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
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instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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