scholarly journals Evaluation and Selection of Suitable qRT-PCR Reference Genes for Light Responses in Tea Plant (Camellia Sinensis)

2020 ◽  
Author(s):  
Jiaxin Wang ◽  
Qianhui Tang ◽  
Kang Sun ◽  
Liang Zeng ◽  
Zhijun Wu
Keyword(s):  
Gene Expression ◽  
Light Intensity ◽  
Camellia Sinensis ◽  
Reference Genes ◽  
Light Quality ◽  
Family Members ◽  
Tea Plant ◽  
Regulation Mechanism ◽  
Qrt Pcr ◽  
Selection Of

Abstract Background: Tea plant (Camellia sinensis) is an important woody economic crop used for processing leaf-type beverages. Tea has been proved to be beneficial to human health because it is rich in tea polyphenols and other active ingredients. Numerous studies have shown that light is a necessary environmental condition to control the growth and metabolism of C. sinensis. Gene expression experiments are always performed to explore the transcriptional regulation mechanism of plants widely based on the technique of quantitative real time polymerase chain reaction (qRT-PCR). The screening and application of reference genes are necessary for the normalization of gene expression under specific conditions. However, the reference genes for systematic analysis of light-induced transcription mechanisms are still not available in C. sinensis.Results: In this research, we identified actin family genes that are always used as reference genes with high frequency and without distinction for various expression experiments in C. sinensis. Six pairs of distinctive primers (corresponding to CsACT1, CsACT2, CsACT(3-4), CsACT(5-6), CsACT(7-8), and CsACT(9-10) genes) were designed to evaluate their expression stability in response to light quality (LQ), light intensity (LI), and photoperiod (PD). Simultaneously, six other family members (CsUBC1, CsCLATHRIN1, CsGAPDH, CsTBP, CsTIP41, and CseIF-4α) of C. sinensis commonly used as reference genes were also investigated. The stability rankings of gene expression were calculated by the statistical algorithms of geNorm, BestKeeper, NormFinder, and RefFinder softwares. Conclusions: CsACT(5-6), CsTIP41, and CsACT(3-4) were the most stable genes for light quality (LQ), light intensity (LI), and photoperiod (PD) treatments, respectively. This study provides a basis for the selection of reference genes for future research on the transcription mechanism of light response in C. sinensis. Moreover, the analysis of actin family members of C. sinensis will help to understand the individual transcription mechanism of housekeeping family.

Evaluation and selection of suitable qRT-PCR reference genes for light responses in tea plant (Camellia sinensis)

2021 ◽  
Vol 289 ◽  
pp. 110488
Author(s):  
Jiaxin Wang ◽  
Linlin Liu ◽  
Qianhui Tang ◽  
Kang Sun ◽  
Liang Zeng ◽  
...  
Keyword(s):  
Camellia Sinensis ◽  
Reference Genes ◽  
Tea Plant ◽  
Light Responses ◽  
Qrt Pcr ◽  
Selection Of

scholarly journals Selection of Reference Genes for qRT-PCR Analysis of Gene Expression in Stipa grandis during Environmental Stresses

PLoS ONE ◽  
2017 ◽  
Vol 12 (1) ◽  
pp. e0169465 ◽  
Author(s):  
Dongli Wan ◽  
Yongqing Wan ◽  
Qi Yang ◽  
Bo Zou ◽  
Weibo Ren ◽  
...  
Keyword(s):  
Gene Expression ◽  
Reference Genes ◽  
Environmental Stresses ◽  
Pcr Analysis ◽  
Qrt Pcr ◽  
Selection Of

scholarly journals Selection and Validation of Appropriate Reference Genes for Real-Time Quantitative PCR Analysis in Needles of Larix olgensis under Abiotic Stresses

Forests ◽  
2020 ◽  
Vol 11 (2) ◽  
pp. 193
Author(s):  
Dandan Li ◽  
Sen Yu ◽  
Minzhen Zeng ◽  
Xiao Liu ◽  
Jia Yang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Heat Stress ◽  
Real Time ◽  
Reference Genes ◽  
Pcr Analysis ◽  
Stable Gene ◽  
Larix Olgensis ◽  
Qrt Pcr ◽  
Study Gene Expression ◽  
Selection Of

Larix olgensis Henry is an important afforestation species in northeastern China because of its fast juvenile growth, high-quality timber, and significant economic and ecological values. The selection of appropriate reference genes is necessary for the normalization of gene expression determination during quantitative real-time polymerase chain reaction (qRT-PCR) experiments. In this study, qRT-PCR was used to study gene expression. Three software packages geNorm, NormFinder, BestKeeper were used, and a comprehensive ranking of candidate reference genes was produced based on their output to evaluate the expression stability of 16 candidate reference genes from L. olgensis under drought, salt, cold, and heat stress. PP2A-1 and GAPDH ranked as the most stable reference genes under drought and cold stress, PP2A-1 and UBQ10 were most stable under salt stress, and TIP41 and ACT2 were most stable under heat stress. The least stable gene was ADP, which ranked the last under all treatments. Expression profile analysis of the antioxidant gene CAT using the two most stable and the single least stable reference genes under each stress further verified that the selected reference genes were suitable for gene expression normalization. This study provides an important foundation for the selection of suitable reference genes for the normalization and quantification of L. olgensis gene expression under abiotic stress conditions.

scholarly journals Validation of reference genes for gene expression studies in post-harvest leaves of tea plant (Camellia sinensis)

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6385 ◽  
Author(s):  
Zi-wei Zhou ◽  
Hui-li Deng ◽  
Qing-yang Wu ◽  
Bin-bin Liu ◽  
Chuan Yue ◽  
...  
Keyword(s):  
Gene Expression ◽  
Reference Genes ◽  
Economic Value ◽  
Tea Plant ◽  
Alcoholic Beverages ◽  
Post Harvest ◽  
Tea Plants ◽  
Suitable Reference ◽  
Turn Over ◽  
Selection Of

Tea is one of three major non-alcoholic beverages that are popular all around the world. The economic value of tea product largely depends on the post-harvest physiology of tea leaves. The utilization of quantitative reverse transcription polymerase chain reaction is a widely accepted and precise approach to determine the target gene expression of tea plants, and the reliability of results hinges on the selection of suitable reference genes. A few reliable reference genes have been documented using various treatments and different tissues of tea plants, but none has been done on post-harvest leaves during the tea manufacturing process. The present study selected and analyzed 15 candidate reference genes: Cs18SrRNA, CsGADPH, CsACT, CsEF-1α, CsUbi, CsTUA, Cs26SrRNA, CsRuBP, CsCYP, CselF-4α, CsMON1, CsPCS1, CsSAND, CsPPA2, CsTBP. This study made an assessment on the expression stability under two kinds of post-harvest treatment, turn over and withering, using three algorithms—GeNorm, Normfinder, and Bestkeeper. The results indicated that the three commonly used reference genes, CsTUA, Cs18SrRNA, CsRuBP, together with Cs26SrRNA, were the most unstable genes in both the turn over and withering treatments. CsACT, CsEF-1α, CsPPA2, and CsTBP were the top four reference genes in the turn over treatment, while CsTBP, CsPCS1, CsPPA2, CselF-4α, and CsACT were the five best reference genes in the withering group. The expression level of lipoxygenase genes, which were involved in a number of diverse aspects of plant physiology, including wounding, was evaluated to validate the findings. To conclude, we found a basis for the selection of reference genes for accurate transcription normalization in post-harvest leaves of tea plants.

scholarly journals Validation of reference genes for gene expression studies in post-harvest leaves of tea plant (Camellia sinensis)

2018 ◽  
Author(s):  
Ziwei Zhou ◽  
Huili Deng ◽  
Qingyang Wu ◽  
Binbin Liu ◽  
Chuan Yue ◽  
...  
Keyword(s):  
Gene Expression ◽  
Reference Genes ◽  
Economic Value ◽  
Tea Plant ◽  
Alcoholic Beverages ◽  
Post Harvest ◽  
Tea Plants ◽  
Suitable Reference ◽  
Turn Over ◽  
Selection Of

Tea is one of three major non-alcoholic beverages that are popular all around the world. The economic value of the final tea product largely depends on the post-harvest physiology of tea leaves. The utilization of quantitative reverse transcription polymerase chain reaction (RT-qPCR) is a widely accepted and precise approach to determine the target gene expression of tea plants, and the reliability of results hinges on the selection of suitable reference genes. A few reliable reference genes have been documented using various treatments and different tissues of tea plants, but none have been done on post-harvest leaves during the tea manufacturing process. The present study selected and analyzed 15 candidate reference genes: Cs18SrRNA, CsGADPH, CsACT, CsEF-1α, CsUbi, CsTUA, Cs26SrRNA, CsRuBP, CsCYP, CselF-4α, CsMON1, CsPCS1, CsSAND, CsPPA2, CsTBP. This study made an assessment on the expression stability under two kinds of post-harvest treatment, turn over and withering, using three algorithms—geNorm, Normfinder and Bestkeeper. The results indicated that the three commonly used reference genes, CsTUA, Cs18SrRNA, CsRuBP, together with Cs26SrRNA, were the most unstable genes in both the turn over and witheringtreatments. CsACT, CsEF-1α, CsPPA2, and CsTBP were the top four reference genes in the turn over treatment, while CsTBP, CsPCS1, CsPPA2, CselF-4α, and CsACT were the four best reference genes in the witheringgroup. The expression level of lipoxygenase (LOX) genes, which were involved in a number of diverse aspects of plant physiology, including wounding, was evaluated to validate the findings. To conclude, we found a basis for the selection of reference genes for accurate transcription normalization in post-harvest leaves of tea plants.

scholarly journals Selection of Reliable Reference Genes for Real-time qRT-PCR Analysis of Zi Geese (<italic>Anser anser domestica</italic>) Gene Expression

2013 ◽  
Vol 26 (3) ◽  
pp. 423-432 ◽  
Author(s):  
Hong Ji ◽  
Jianfa Wang ◽  
Juxiong Liu ◽  
Jingru Guo ◽  
Zhongwei Wang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Reference Genes ◽  
Pcr Analysis ◽  
Anser Anser ◽  
Qrt Pcr ◽  
Selection Of
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