Evaluation of reference genes for transcript normalization in Fragaria chiloensis fruit and vegetative tissues

Quantitative real-time PCR (RT-qPCR) is used extensively in gene expression studies. For adequate comparisons, the identification and use of reliable reference genes are crucial. Nevertheless, the availability of such genes in strawberry species is limited and has yet to be described for the Chilean...

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Bibliographic Details
Published in:Physiology and molecular biology of plants 2022-08, Vol.28 (8), p.1535-1544
Main Authors: Gaete-Eastman, Carlos, Mattus-Araya, Elena, Herrera, Raúl, Moya-León, María Alejandra
Format: Article
Language:English
Subjects:
Abscisic acid
Algorithms
Auxins
Biological and Medical Physics
Biomedical and Life Sciences
Biophysics
Cell Biology
Fragaria chiloensis
Fruits
Gene expression
Genes
Life Sciences
Plant Physiology
Plant Sciences
Research Article
Ripening
Stability
Strawberries
Tissues
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Summary:Quantitative real-time PCR (RT-qPCR) is used extensively in gene expression studies. For adequate comparisons, the identification and use of reliable reference genes are crucial. Nevertheless, the availability of such genes in strawberry species is limited and has yet to be described for the Chilean strawberry, Fragaria chiloensis . In this study, the expression dynamics of a set of 10 candidate reference genes were analyzed in various F. chiloensis vegetative tissues (root, runners, stem, leaf, and flower), and fruits at different ripening stages or subjected to different hormonal treatments (ABA, auxin). The expression stability of candidate genes was examined by a series of algorithms, such as geNorm, NormFinder, BestKeeper, and ΔCt, for comparisons and rankings. Finally, by using RefFinder, a comprehensive and comparative ranking of the four methods was achieved. The results highlight that the expression stability of candidate reference genes fluctuates depending on tissue type, fruit stage, and hormonal treatment. As reference genes, the use of FcCHP2 and FcACTIN1 is recommended for F. chiloensis vegetative tissues; FcDBP and FcCHP1 for fruit ripening stages; FcGAPDH and FcDBP for fruit subjected to ABA and NDGA treatments; FcCHP1 and FcCHP2 for fruit under AUXIN and TIBA treatments; and FcDBP and FcCHP2 when all fruit stages and hormonal treatments are compared. If just one reference gene is employed as a normalizer, FcDBP should be chosen as it is the most stable internal control in most conditions. Therefore, the present study delivers a set of reliable reference genes for RT-qPCR expression analysis in F. chiloensis tissues and fruits subjected to several hormonal treatments.
ISSN:0971-5894
0974-0430
DOI:10.1007/s12298-022-01227-y