Selection of appropriate reference genes for RT-qPCR analysis under abiotic stress and hormone treatment in celery
Celery is one of the most important vegetable crop and its yield and quality is influenced by many environmental factors. Researches on gene expression not only help to unravel the molecular regulatory mechanism but also identify the key genes in the biological response. RT-qPCR is a commonly used t...
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PeerJ Inc.
2019-10-01
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Online Access: | https://peerj.com/articles/7925.pdf |
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record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Kai Feng Jie-xia Liu Guo-Ming Xing Sheng Sun Sen Li Ao-Qi Duan Feng Wang Meng-Yao Li Zhi-Sheng Xu Ai-Sheng Xiong |
spellingShingle |
Kai Feng Jie-xia Liu Guo-Ming Xing Sheng Sun Sen Li Ao-Qi Duan Feng Wang Meng-Yao Li Zhi-Sheng Xu Ai-Sheng Xiong Selection of appropriate reference genes for RT-qPCR analysis under abiotic stress and hormone treatment in celery PeerJ Abiotic stress Celery Expression stability Hormone stimuli Reference gene RT-qPCR |
author_facet |
Kai Feng Jie-xia Liu Guo-Ming Xing Sheng Sun Sen Li Ao-Qi Duan Feng Wang Meng-Yao Li Zhi-Sheng Xu Ai-Sheng Xiong |
author_sort |
Kai Feng |
title |
Selection of appropriate reference genes for RT-qPCR analysis under abiotic stress and hormone treatment in celery |
title_short |
Selection of appropriate reference genes for RT-qPCR analysis under abiotic stress and hormone treatment in celery |
title_full |
Selection of appropriate reference genes for RT-qPCR analysis under abiotic stress and hormone treatment in celery |
title_fullStr |
Selection of appropriate reference genes for RT-qPCR analysis under abiotic stress and hormone treatment in celery |
title_full_unstemmed |
Selection of appropriate reference genes for RT-qPCR analysis under abiotic stress and hormone treatment in celery |
title_sort |
selection of appropriate reference genes for rt-qpcr analysis under abiotic stress and hormone treatment in celery |
publisher |
PeerJ Inc. |
series |
PeerJ |
issn |
2167-8359 |
publishDate |
2019-10-01 |
description |
Celery is one of the most important vegetable crop and its yield and quality is influenced by many environmental factors. Researches on gene expression not only help to unravel the molecular regulatory mechanism but also identify the key genes in the biological response. RT-qPCR is a commonly used technology to quantify the gene expression. Selecting an appropriate reference gene is an effective approach to improve the accuracy of RT-qPCR assay. To our knowledge, the evaluation of reference genes under different treatments in celery has not been reported yet. In this study, the expression stabilities of eight candidate reference genes (ACTIN, eIF-4α, GAPDH, TBP, TUB-A, UBC, TUB-B, and EF-1α) under abiotic stresses (heat, cold, drought, and salt) and hormone treatments (SA, MeJA, GA, and ABA) were detected. The expression stabilities of candidate genes were compared and ranked by geNorm, NormFinder, BestKeeper, ΔCt, and RefFinder programs. The results calculated by different programs were not completely consistent. Considering the comprehensive analysis results, ACTIN was the most stable reference gene and TUB-B showed the worst expression stabilities under the selected abiotic stress and hormone treatments in celery. The reliability of reference genes was further confirmed by the normalization of CAT1 gene under drought stress. This study presented evidences and basis to select the appropriate reference genes under different treatments in celery. |
topic |
Abiotic stress Celery Expression stability Hormone stimuli Reference gene RT-qPCR |
url |
https://peerj.com/articles/7925.pdf |
work_keys_str_mv |
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doaj-bb15812ce76c49a69953a255f6fca2d92020-11-25T02:15:22ZengPeerJ Inc.PeerJ2167-83592019-10-017e792510.7717/peerj.7925Selection of appropriate reference genes for RT-qPCR analysis under abiotic stress and hormone treatment in celeryKai Feng0Jie-xia Liu1Guo-Ming Xing2Sheng Sun3Sen Li4Ao-Qi Duan5Feng Wang6Meng-Yao Li7Zhi-Sheng Xu8Ai-Sheng Xiong9State Key Laboratory of Crop Genetics and Germplasm Enhancement, Ministry of Agriculture and Rural Affairs Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, College of Horticulture, Nanjing Agricultural University, Nanjing, ChinaState Key Laboratory of Crop Genetics and Germplasm Enhancement, Ministry of Agriculture and Rural Affairs Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, College of Horticulture, Nanjing Agricultural University, Nanjing, ChinaCollaborative Innovation Center for Improving Quality and Increase Profits of Protected Vegetables in Shanxi, Taigu, ChinaCollaborative Innovation Center for Improving Quality and Increase Profits of Protected Vegetables in Shanxi, Taigu, ChinaCollaborative Innovation Center for Improving Quality and Increase Profits of Protected Vegetables in Shanxi, Taigu, ChinaState Key Laboratory of Crop Genetics and Germplasm Enhancement, Ministry of Agriculture and Rural Affairs Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, College of Horticulture, Nanjing Agricultural University, Nanjing, ChinaState Key Laboratory of Crop Genetics and Germplasm Enhancement, Ministry of Agriculture and Rural Affairs Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, College of Horticulture, Nanjing Agricultural University, Nanjing, ChinaState Key Laboratory of Crop Genetics and Germplasm Enhancement, Ministry of Agriculture and Rural Affairs Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, College of Horticulture, Nanjing Agricultural University, Nanjing, ChinaState Key Laboratory of Crop Genetics and Germplasm Enhancement, Ministry of Agriculture and Rural Affairs Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, College of Horticulture, Nanjing Agricultural University, Nanjing, ChinaState Key Laboratory of Crop Genetics and Germplasm Enhancement, Ministry of Agriculture and Rural Affairs Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, College of Horticulture, Nanjing Agricultural University, Nanjing, ChinaCelery is one of the most important vegetable crop and its yield and quality is influenced by many environmental factors. Researches on gene expression not only help to unravel the molecular regulatory mechanism but also identify the key genes in the biological response. RT-qPCR is a commonly used technology to quantify the gene expression. Selecting an appropriate reference gene is an effective approach to improve the accuracy of RT-qPCR assay. To our knowledge, the evaluation of reference genes under different treatments in celery has not been reported yet. In this study, the expression stabilities of eight candidate reference genes (ACTIN, eIF-4α, GAPDH, TBP, TUB-A, UBC, TUB-B, and EF-1α) under abiotic stresses (heat, cold, drought, and salt) and hormone treatments (SA, MeJA, GA, and ABA) were detected. The expression stabilities of candidate genes were compared and ranked by geNorm, NormFinder, BestKeeper, ΔCt, and RefFinder programs. The results calculated by different programs were not completely consistent. Considering the comprehensive analysis results, ACTIN was the most stable reference gene and TUB-B showed the worst expression stabilities under the selected abiotic stress and hormone treatments in celery. The reliability of reference genes was further confirmed by the normalization of CAT1 gene under drought stress. This study presented evidences and basis to select the appropriate reference genes under different treatments in celery.https://peerj.com/articles/7925.pdfAbiotic stressCeleryExpression stabilityHormone stimuliReference geneRT-qPCR |