TY - EJOU
AU - Li, Gang
AU - Zhou, Yao
AU - Zhao, Yaqi
AU - Liu, Yaxue
AU - Ke, Yuwei
AU - Jin, Xiaoqing
AU - Ma, Haoli
TI - Internal Reference Gene Selection for Quantitative Real-Time RT-PCR Normalization in Potato Tissues
T2 - Phyton-International Journal of Experimental Botany
PY - 2020
VL - 89
IS - 2
SN - 1851-5657
AB - Quantitative real-time PCR (qRT-PCR) is widely used for investigating
gene expression patterns and has many advantages, including its high sensitivity,
fidelity, and specificity. Selecting a satisfactory internal reference gene is crucial
for obtaining precise gene expression results in qRT-PCR analyses. In this study,
the transcriptomic data of 2 potato varieties were screened for housekeeping genes
with stable expression patterns. A total of 77 putative genes were selected, which
were highly and stably expressed. Then, qRT-PCR analyses were performed to
examine the expression levels of these 77 candidate reference genes in various
potato tissues, including leaves, flowers, stolons, and tubers. Gene expression
was represented by analyzing the Ct values at given threshold. Through geNorm
and NormFinder program analyses, 10 candidate genes with the most stable
expression patterns were obtained, including RPL19, RPS15, RPS9, EF1α, TrxP1,
RPS8, NTF, CAM, AACM, and RPS28. Moreover, through the comprehensive
analyses of 4 statistical algorithms (i.e., geNorm, NormFinder, BestKeeper, and
RefFinder), results indicated that the most appropriate internal reference genes
were RPL19 and EF1α. The obtained stable reference genes will contribute to
future qRT-PCR analyses on potato tissue-related gene expression.
KW - qRT-PCR; potato tissues; reference genes; geNorm; NormFinder; BestKeeper; RefFinder
DO - 10.32604/phyton.2020.08874