Identification and Validation of Reference Genes for qRT-PCR Studies of Gene Expression in Dioscorea opposita.
Quantitative real-time polymerase chain reaction (qRT-PCR) is one of the most common methods for gene expression studies. Data normalization based on reference genes is essential for obtaining reliable results for qRT-PCR assays. This study evaluated potential reference genes of Chinese yam (Dioscorea opposita Thunb.), which is an important tuber crop and medicinal plant in East Asia. The expression of ten candidate reference genes across 20 samples from different organs and development stages was assessed. We identified the most stable genes for qRT-PCR studies using combined samples from different organs. Our results also suggest that different suitable reference genes or combinations of reference genes for normalization should be applied according to different organs and developmental stages. To validate the suitability of the reference genes, we evaluated the relative expression of PE2.1 and PE53, which are two genes that may be associated with microtuber formation. Our results provide the foundation for reference gene(s) selection in D. opposita and will contribute toward more accurate gene analysis studies of the genus Dioscorea.
College of Life Sciences, Henan Normal University, Xinxiang 453007, China; Engineering Technology Research Center of Nursing and Utilization of Genuine Chinese Crude Drugs, University of Henan Province, Xinxiang 453007, China; Henan Province Engineering Laboratory of Green Medicinal Plant Biotechnology, Xinxiang 453007, China.
Xiting
X
Zhao
College of Life Sciences, Henan Normal University, Xinxiang 453007, China; Engineering Technology Research Center of Nursing and Utilization of Genuine Chinese Crude Drugs, University of Henan Province, Xinxiang 453007, China; Henan Province Engineering Laboratory of Green Medicinal Plant Biotechnology, Xinxiang 453007, China.
Xiaoli
X
Zhang
College of Life Sciences, Henan Normal University, Xinxiang 453007, China.
Xiaobo
X
Guo
College of Life Sciences, Henan Normal University, Xinxiang 453007, China.
Shujie
S
Li
College of Life Sciences, Henan Normal University, Xinxiang 453007, China.
Linlin
L
Han
College of Life Sciences, Henan Normal University, Xinxiang 453007, China.
Zhihui
Z
Song
College of Life Sciences, Henan Normal University, Xinxiang 453007, China.
Yunying
Y
Wang
College of Life Sciences, Henan Normal University, Xinxiang 453007, China; Engineering Technology Research Center of Nursing and Utilization of Genuine Chinese Crude Drugs, University of Henan Province, Xinxiang 453007, China; Henan Province Engineering Laboratory of Green Medicinal Plant Biotechnology, Xinxiang 453007, China.
Junhua
J
Li
College of Life Sciences, Henan Normal University, Xinxiang 453007, China; Engineering Technology Research Center of Nursing and Utilization of Genuine Chinese Crude Drugs, University of Henan Province, Xinxiang 453007, China; Henan Province Engineering Laboratory of Green Medicinal Plant Biotechnology, Xinxiang 453007, China.
Mingjun
M
Li
D027742:Dioscorea / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*