Validation of reference genes for real-time quantitative PCR normalization in soybean developmental and germinating seeds.
Plant Cell Rep
; 31(10): 1789-98, 2012 Oct.
Article
em En
| MEDLINE
| ID: mdl-22588479
ABSTRACT
Most of traditional reference genes chosen for real-time quantitative PCR normalization were assumed to be ubiquitously and constitutively expressed in vegetative tissues. However, seeds show distinct transcriptomes compared with the vegetative tissues. Therefore, there is a need for re-validation of reference genes in samples of seed development and germination, especially for soybean seeds. In this study, we aimed at identifying reference genes suitable for the quantification of gene expression level in soybean seeds. In order to identify the best reference genes for soybean seeds, 18 putative reference genes were tested with various methods in different seed samples. We combined the outputs of both geNorm and NormFinder to assess the expression stability of these genes. The reference genes identified as optimums for seed development were TUA5 and UKN2, whereas for seed germination they were novel reference genes Glyma05g37470 and Glyma08g28550. Furthermore, for total seed samples it was necessary to combine four genes of Glyma05g37470, Glyma08g28550, Glyma18g04130 and UKN2 [corrected] for normalization. Key message We identified several reference genes that stably expressed in soybean seed developmental and germinating processes.
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Sementes
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Glycine max
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Genes de Plantas
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Germinação
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Regulação da Expressão Gênica de Plantas
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Reação em Cadeia da Polimerase em Tempo Real
Tipo de estudo:
Prognostic_studies
Idioma:
En
Revista:
Plant Cell Rep
Assunto da revista:
BOTANICA
Ano de publicação:
2012
Tipo de documento:
Article