Indian Society of Genetics & Plant Breeding


Validation of reference genes in pansy for accurate transcript normalization during its flowering stages

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Quantitative real-time PCR (qRT-PCR) is a perfect method
for rapid and accurate quantification of gene expression in
different organs or at different development stages in
plants. Suitable reference genes are important in this
method. In order to obtain more accurate genes expression
data in pansy (Viola × wittrockiana Gams.) flower, the
expression stability of four housekeeping genes, b-actin
(ACT), glyceraldehyde-3-phosphate dehydrogenase
(GAPDH), tubulin (TUB) and 18S ribosomal proteins (18S)
at seven different floral development stages in pansy were
studied in this experiment. The results showed that TUB
and 18S genes were the top two stable genes. Additionally,
the expression pattern of VwbHLH1 gene was studied with
the two most stable reference genesTUB and 18S, and the
worst gene GAPDH respectively. The gene expression data
were very different when various reference genes were
applied. The present study proved that selection of
reference genes was definitely important to obtain precise
experimental data in qRT-PCR even for the same organ but
at different development stages. This study would provide
guidelines to obtain more accurate gene expression results
for future molecular mechanism study in pansy.

Keywords: Quantitative real-time PCR, reference gene, pansy, normalization, gene expression


Year: 2016
Volume: 76
Issue: 2
Article DOI: 10.5958/0975-6906.2016.00034.1
Print ISSN: 0019-5200
Online ISSN: 0975-6906


Jing Li info_circle
Sheng Gong info_circle
Ke Zhang info_circle
Haiyan Sun info_circle
Guopeng Zhu info_circle
Xiqiang Song info_circle
Jian Wang info_circle

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