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dc.contributor.author
Fernández, María Belén
dc.contributor.author
Lukaszewicz, Germán
dc.contributor.author
Lamattina, Lorenzo
dc.contributor.author
Cassia, Raul Oscar
dc.date.available
2021-12-17T10:36:33Z
dc.date.issued
2021-03-22
dc.identifier.citation
Fernández, María Belén; Lukaszewicz, Germán; Lamattina, Lorenzo; Cassia, Raul Oscar; Selection and optimization of reference genes for RT-qPCR normalization: a case study in Solanum lycopersicum exposed to UV-B; Elsevier France-Editions Scientifiques Medicales Elsevier; Plant Physiology and Biochemistry; 160; 22-3-2021; 269-280
dc.identifier.issn
0981-9428
dc.identifier.uri
http://hdl.handle.net/11336/148920
dc.description.abstract
Quantitative RT- PCR is one of the most common methods to study gene expression in response to stress. Therefore, it is crucial to have suitable reference genes (RGs) for result normalization. Although several reports describe UV-B-modulated gene expression in Solanum lycopersicum, there are no suitable RGs identified until now. The aim of this work was to evaluate the suitability of seven traditional genes: actin (ACT), tubulin (TUB), ubiquitin (UBI), glyceraldehyde- 3 phosphate dehydrogenase (GAPDH), elongation factor 1α (EF1α), phosphatase 2A catalytic subunit (PP2A) and GAGA binding transcriptional activator (GAGA); and two non-traditional genes: thioredoxin h1 (TRX h1) and UV-B RESISTANCE LOCUS 8 (UVR8), as candidate RGs for their potential use as reliable internal controls in leaves, stems and roots of tomato seedlings exposed to acute and chronic UV-B. The stability of these genes expression was evaluated using five statistical algorithms: geNorm, NormFinder, BestKeeper, Delta Ct and ANOVA. Considering the comprehensive stability ranking, we recommend ACT+TUB as the best pair of RGs for leaves, PP2A+GAPDH+TRX h1 for stems and TUB+UVR8 for roots. The reliability of the selected RGs for each tissue was verified amplifying tomato chalcone synthase 1 (CHS1) and cyclobutane pyrimidine dimer (CPD) photolyase (PHR1-LIKE). Under UV-B treatment, CHS1 was upregulated in leaves, stems and roots whereas PHR1-LIKE was only upregulated in leaves and stems. This interpretation differs when the most and least stable RGs are chosen. This is the first report regarding suitable RGs selection for accurate normalization of gene expression in tomato seedlings exposed to UV-B irradiation.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Elsevier France-Editions Scientifiques Medicales Elsevier
dc.rights
info:eu-repo/semantics/restrictedAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
CHS1
dc.subject
PHR1-LIKE
dc.subject
QUANTITATIVE RT-PCR
dc.subject
REFERENCE GENE
dc.subject
SOLANUM LYCOPERSICUM
dc.subject
UV-B
dc.subject
UVR8
dc.subject.classification
Bioquímica y Biología Molecular
dc.subject.classification
Ciencias Biológicas
dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
dc.title
Selection and optimization of reference genes for RT-qPCR normalization: a case study in Solanum lycopersicum exposed to UV-B
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2021-12-03T19:48:12Z
dc.journal.volume
160
dc.journal.pagination
269-280
dc.journal.pais
Francia
dc.description.fil
Fil: Fernández, María Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
dc.description.fil
Fil: Lukaszewicz, Germán. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
dc.description.fil
Fil: Lamattina, Lorenzo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
dc.description.fil
Fil: Cassia, Raul Oscar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
dc.journal.title
Plant Physiology and Biochemistry
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://linkinghub.elsevier.com/retrieve/pii/S0981942821000371
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1016/j.plaphy.2021.01.026
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