|Table of Contents|

Selection of reference genes for RT-qPCR in red callus of Vitis davidii (Rom. Caill.) Fo?x under different light qualit ies*(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

Issue:
2019 06
Page:
1-10
Research Field:
Articles
Publishing date:

Info

Title:
Selection of reference genes for RT-qPCR in red callus of Vitis davidii (Rom. Caill.) Fo?x under different light qualit ies*
Author(s):
PAN Hong1 2 3 LAI Chengchun2 3** ZHANG Jing1 2 3 HUANG Xiangui2 3 LIN Yuling & LAI Zhongxiong1**
1 Institute of Horticultural Biotechnology, Fujian Agriculture and Forestry University, Fuzhou 350002, China
2 Institute of Agricultural Engineering and Technology, Fujian Academy of Agricultural Sciences , Fuzhou 350003, China
3 Fujian Key Laboratory of Agricultural Product (Food) Processing, Fuzhou 350003, China
Keywords:
Vitis davidii (Rom. Caill.) Fo?x red callus Light quality reference genes Real-time quantitative PCR
CLC:
-
PACS:
DOI:
10.19675/j.cnki.1006-687x.2019.01038
DocumentCode:

Abstract:
It is an important influence on the accumulation of anthocyanins and cell growth of red callus of Vitis davidii (Rom. Caill.) Fo?x (DLR) by different light qualities. The expression of functional genes of DLR cell cultures under different light quality can be analyzed quickly and accurately using RT-qPCR. In order to improve experimental data accuracy, it is crucial to screen suitable reference genes for RT-qPCR in this specific experiment. The DLR callus were cultured under nine light qualities, including dark, white light, red light, yellow light, blue light, green light, purple light, warm white light, warm yellow light. Then these cell cultures were taken as the materials. Ten reference genes, which were commonly used in grape, were selected as candidate reference genes. And expression stabilities of ten ref erence genes were analyzed by RT-qPCR under nine light qualities.The expression stability ranking of ten candidate reference genes was evaluated by using three soft wares, such as GeNorm, Normfinder and BestKeeper. These soft wares have different algorithms and lead to variation of reference genes rankings . . The top five reference genes of geNorm evaluation were α-Tubulin = AP-2> 60SRP > EF1-α > UBQ. The top five reference genes of Normfinder calculation were 60SRP > AP-2 > α-Tubulin > EF1-α > GAPDH. The top five reference genes of BestKeeper assessment were UBQ > α-Tubulin > 60SRP > EF1-α > SAND. Considering that the stable rankings of ten reference genes by three soft wares, α-Tubulin and 60SRP were the most stable reference genes for normalizing the RT-qPCR data of genes expression of the DLR cell cultures under different light qualities, and NAD5 was the lest stable reference gene. The reliability of α-Tubulin and 60SRP were validated by using the expression of target gene UFGT. The results showed that the expression profiles of UFGT were similar and had high stability using α-Tubulin or 60SRP to normalize , and α-Tubulin + 60SRP combination can achieve more accurate and reliable results . The present study has provided an important reference for analyzing the expression of critical genes in red callus of Vitis davidii under different light qualities.

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Last Update: 2019-05-09