期刊
PLANT BIOTECHNOLOGY
卷 35, 期 3, 页码 225-235出版社
JAPANESE SOC PLANT CELL & MOLECULAR BIOLOGY
DOI: 10.5511/plantbiotechnology.18.0525a
关键词
fruit development; internal control gene; Micro-Tom; qRT-PCR; reference gene
资金
- University of Tsukuba, Gene Research Center, through the National Bio-Resource Project (NBRP) of the Japan Agency for Research and Development (AMED), Japan [TOMJP00001]
- Cooperative Research Grant of the Plant Transgenic Design Initiative (PTradD) by Gene Research Center, Tsukuba-Plant Innovation Research Center, University of Tsukuba [16A1-15, 17A-14, 18A-16]
- Kato Memorial Bioscience Foundation
- Building of Consortia for the Development of Human Research in Science and Technology, MEXT, Japan
Quantitative real-time PCR (qRT-PCR) is widely used to analyze the expression profiles of the genes of interest. In order to obtain accurate quantification data, normalization by using reliable internal control genes is essential. In this study, we evaluated the stability and applicability of eight internal control gene candidates for analyzing gene expression during fruit development in dwarf tomato cultivar Micro-Tom. We collected seventeen different samples from flowers and fruits at different developmental stages, and estimated the expression stability of the candidate genes by two statistical algorithms, geNorm and NormFinder. The combined ranking order and qRT-PCR analyses for expression profiles of SlYABBY2a, SlYABBY1a, FRUITFULL1 and APETALA2c suggested that EXPRESSED was the most stable and reliable internal control gene among the candidates. Our analysis also suggested that RPL8 was also suitable if the sample group is limited to fruits at different maturation stages. In addition to EXPRESSED, GAPDH was also applicable for relative quantitation to monitor gene expression profiles through fruit development from pistil to pericarp.
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