Identification of potential internal control genes for real-time PCR analysis during stress response in Pyropia haitanensis

Xia Wang; Jianhua Feng; Aiyou Huang; Linwen He; Jianfeng Niu; Guangce Wang

doi:10.1007/s00343-017-6005-9

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Identification of potential internal control genes for real-time PCR analysis during stress response in Pyropia haitanensis

OriginalPaper | Updated：2023-05-16

- Identification of potential internal control genes for real-time PCR analysis during stress response in Pyropia haitanensis
- Identification of potential internal control genes for real-time PCR analysis during stress response in Pyropia haitanensis
- 海洋湖沼学报（英文） 2017年35卷第6期页码：1432-1441
- Affiliations：
  
  1. Key Laboratory of Experimental Marine Biology Institute of Oceanology Chinese Academy of Sciences
  2. Laboratory for Marine Biology and Biotechnology Qingdao National Laboratory for Marine Science and Technology
  3. University of Chinese Academy of Sciences
  4. Nantong Branch Institute of Oceanology Chinese Academy of Sciences
- Author bio：
- Funds：
  
  Supported by the National Natural Science Foundation of China (Nos. 41476140, 41306151, 41676157, 41506172), the Strategic Leading Science and Technology Projects of Chinese Academy of Sciences (No. XDA11020404), the China Postdoctoral Science Foundation (No. 2015M582153), and the Science and Technology Plan of Jiangsu Province (No. BE2016330)
- DOI：10.1007/s00343-017-6005-9
  中图分类号：
- 网络首发：2017-01-26，
  
  纸质出版：2017
- Accepted：
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Identification of potential internal control genes for real-time PCR analysis during stress response in Pyropia haitanensis[J]. 海洋湖沼学报（英文）, 2017,35(6):1432-1441.

Wang, X., Feng, J., Huang, A. et al. Identification of potential internal control genes for real-time PCR analysis during stress response in Pyropia haitanensis., Chin. J. Ocean. Limnol. 35, 1432–1441 (2017). https://doi.org/10.1007/s00343-017-6005-9
Identification of potential internal control genes for real-time PCR analysis during stress response in Pyropia haitanensis[J]. 海洋湖沼学报（英文）, 2017,35(6):1432-1441. DOI： 10.1007/s00343-017-6005-9.

Wang, X., Feng, J., Huang, A. et al. Identification of potential internal control genes for real-time PCR analysis during stress response in Pyropia haitanensis., Chin. J. Ocean. Limnol. 35, 1432–1441 (2017). https://doi.org/10.1007/s00343-017-6005-9 DOI：

摘要

Abstract

Pyropia haitanensis has prominent stress-resistance characteristics and is endemic to China. Studies into the stress responses in these algae could provide valuable information on the stress-response mechanisms in the intertidal Rhodophyta. Here

the effects of salinity and light intensity on the quantum yield of photosystem II in Py. haitanensis were investigated using pulse-amplitude-modulation fluorometry. Total RNA and genomic DNA of the samples under different stress conditions were isolated. By normalizing to the genomic DNA quantity

the RNA content in each sample was evaluated. The cDNA was synthesized and the expression levels of seven potential internal control genes were evaluated using qRT-PCR method. Then

we used geNorm

a common statistical algorithm

to analyze the qRT-PCR data of seven reference genes. Potential genes that may constantly be expressed under different conditions were selected

and these genes showed stable expression levels in samples under a salinity treatment

while tubulin

glyceraldehyde-3-phosphate dehydrogenase and actin showed stability in samples stressed by strong light. Based on the results of the pulse amplitude-modulation fluorometry

an absolute quantification was performed to obtain gene copy numbers in certain stress-treated samples. The stably expressed genes as determined by the absolute quantification in certain samples conformed to the results of the geNorm screening. Based on the results of the software analysis and absolute quantification

we proposed that elongation factor 3 and 18S ribosomal RNA could be used as internal control genes when the Py. haitanensis blades were subjected to salinity stress

and that α-tubulin and 18S ribosomal RNA could be used as the internal control genes when the stress was from strong light. In general

our findings provide a convenient reference for the selection of internal control genes when designing experiments related to stress responses in Py. haitanensis.

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references

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