Selection and validation of reference genes for quantitative gene expression analyses in persimmon (Diospyros kaki Thunb.) using real-time quantitative PCR.
Diospyros kaki Thunb.
NormFinder
RT-qPCR
geNorm
reference genes
Journal
Biologia futura
ISSN: 2676-8607
Titre abrégé: Biol Futur
Pays: Switzerland
ID NLM: 101738236
Informations de publication
Date de publication:
Dec 2019
Dec 2019
Historique:
received:
07
11
2018
accepted:
20
05
2019
entrez:
23
9
2021
pubmed:
1
12
2019
medline:
1
12
2019
Statut:
ppublish
Résumé
Persimmon (Diospyros kaki) is an economically important fruit tree species with complex flowering characteristics. To obtain accurate expression pattern analysis results, it is vital to select a reliable gene for the normalization of real-time quantitative polymerase chain reaction data. The aim of this study was to identify the optimal internal control gene among six candidate genes for gene expression analysis in different persimmon organs and developmental stages. This analysis was conducted using geNorm and NormFinder software to show differences in the stability of the six reference genes among tissues and floral developmental stages of the same plant. Although genes that exhibited moderate expression in NormFinder revealed slightly different expression stabilities than those obtained by geNorm, both sets of results showed that GAPDH was the best reference gene in different organs and floral buds at different developmental stages, whereas 18SrRNA was the least stable gene. Based on the overall ranking, GAPDH is the most suitable reference gene and is highly recommended for gene expression studies in different organs and different developmental stages of persimmon. This study provides useful reference data for future gene expression studies and will contribute to improving the accuracy of gene expression results in persimmon.
Sections du résumé
BACKGROUND AND AIMS
OBJECTIVE
Persimmon (Diospyros kaki) is an economically important fruit tree species with complex flowering characteristics. To obtain accurate expression pattern analysis results, it is vital to select a reliable gene for the normalization of real-time quantitative polymerase chain reaction data. The aim of this study was to identify the optimal internal control gene among six candidate genes for gene expression analysis in different persimmon organs and developmental stages.
MATERIALS AND METHODS
METHODS
This analysis was conducted using geNorm and NormFinder software to show differences in the stability of the six reference genes among tissues and floral developmental stages of the same plant.
RESULTS
RESULTS
Although genes that exhibited moderate expression in NormFinder revealed slightly different expression stabilities than those obtained by geNorm, both sets of results showed that GAPDH was the best reference gene in different organs and floral buds at different developmental stages, whereas 18SrRNA was the least stable gene.
CONCLUSIONS
CONCLUSIONS
Based on the overall ranking, GAPDH is the most suitable reference gene and is highly recommended for gene expression studies in different organs and different developmental stages of persimmon. This study provides useful reference data for future gene expression studies and will contribute to improving the accuracy of gene expression results in persimmon.
Identifiants
pubmed: 34554547
doi: 10.1556/019.70.2019.24
pii: 10.1556/019.70.2019.24
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
261-267Informations de copyright
© 2019. Akadémiai Kiadó Zrt.
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