scholarly journals Assessment of suitable reference genes for qRT-PCR analysis in Adelphocoris suturalis

2018 ◽  
Vol 17 (12) ◽  
pp. 2745-2757 ◽  
Author(s):  
Jing LUO ◽  
Chao MA ◽  
Zhe LI ◽  
Bang-qin ZHU ◽  
Jiang ZHANG ◽  
...  
Keyword(s):  
Reference Genes ◽  
Pcr Analysis ◽  
Qrt Pcr ◽  
Suitable Reference

scholarly journals Comparison of Reliable Reference Genes Following Different Hormone Treatments by Various Algorithms for qRT-PCR Analysis of Metasequoia

2018 ◽  
Vol 20 (1) ◽  
pp. 34 ◽  
Author(s):  
Jing-Jing Wang ◽  
Shuo Han ◽  
Weilun Yin ◽  
Xinli Xia ◽  
Chao Liu
Keyword(s):  
Gene Expression ◽  
Reference Genes ◽  
Elongation Factor ◽  
Pcr Analysis ◽  
Qrt Pcr ◽  
Gene Normalization ◽  
Accurate Normalization ◽  
Suitable Reference ◽  
Different Tissues ◽  
Gene Expression Levels

Quantitative reverse transcription polymerase chain reaction (qRT-PCR) is the most sensitive technique for evaluating gene expression levels. Choosing appropriate reference genes for normalizing target gene expression is important for verifying expression changes. Metasequoia is a high-quality and economically important wood species. However, few systematic studies have examined reference genes in Metasequoia. Here, the expression stability of 14 candidate reference genes in different tissues and following different hormone treatments were analyzed using six algorithms. Candidate reference genes were used to normalize the expression pattern of FLOWERING LOCUS T and pyrabactin resistance-like 8. Analysis using the GrayNorm algorithm showed that ACT2 (Actin 2), HIS (histone superfamily protein H3) and TATA (TATA binding protein) were stably expressed in different tissues. ACT2, EF1α (elongation factor-1 alpha) and HIS were optimal for leaves treated with the flowering induction hormone solution, while Cpn60β (60-kDa chaperonin β-subunit), GAPDH (glyceraldehyde-3-phosphate dehydrogenase) and HIS were the best reference genes for treated buds. EF1α, HIS and TATA were useful reference genes for accurate normalization in abscisic acid-response signaling. Our results emphasize the importance of validating reference genes for qRT-PCR analysis in Metasequoia. To avoid errors, suitable reference genes should be used for different tissues and hormone treatments to increase normalization accuracy. Our study provides a foundation for reference gene normalization when analyzing gene expression in Metasequoia.

scholarly journals Evaluation of potential reference genes for quantitative RT-PCR analysis in spotted sea bass (Lateolabrax maculatus) under normal and salinity stress conditions

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e5631 ◽  
Author(s):  
Haolong Wang ◽  
Haishen Wen ◽  
Yun Li ◽  
Kaiqiang Zhang ◽  
Yang Liu
Keyword(s):  
Salinity Stress ◽  
Reference Genes ◽  
18S Rrna ◽  
Sea Bass ◽  
Stress Conditions ◽  
Pcr Analysis ◽  
Stable Gene ◽  
Qrt Pcr ◽  
Lateolabrax Maculatus ◽  
Suitable Reference

The aim of this study was to select the most suitable reference genes for quantitative real-time polymerase chain reaction (qRT-PCR) of spotted sea bass (Lateolabrax maculatus), an important commercial marine fish in Pacific Asia, under normal physiological and salinity stress conditions. A total of 9 candidate reference genes (HPRT, GAPDH, EF1A, TUBA, RPL7, RNAPol II, B2M, ACTB and 18S rRNA) were analyzed by qRT-PCR in 10 tissues (intestine, muscle, stomach, brain, heart, liver, gill, kidney, pectoral fins and spleen) of L. maculatus. Four algorithms, geNorm, NormFinder, BestKeeper, and comparative ΔCt method, were used to evaluate the expression stability of the candidate reference genes. The results showed the 18S rRNA was most stable in different tissues under normal conditions. During salinity stress, RPL7 was the most stable gene according to overall ranking and the best combination of reference genes was RPL7 and RNAPol II. In contrast, GAPDH was the least stable gene which was not suitable as reference genes. The study showed that different algorithms might generate inconsistent results. Therefore, the combination of several reference genes should be selected to accurately calibrate system errors. The present study was the first to select reference genes of L. maculatus by qRT-PCR and provides a useful basis for selecting the appropriate reference gene in L. maculatus. The present study also has important implications for gene expression and functional genomics research in this species or other teleost species.

scholarly journals Selection and validation of suitable reference genes for qRT-PCR analysis in pear leaf tissues under distinct training systems

PLoS ONE ◽  
2018 ◽  
Vol 13 (8) ◽  
pp. e0202472 ◽  
Author(s):  
Zheng Liu ◽  
Kexin Cheng ◽  
Zhongqi Qin ◽  
Tao Wu ◽  
Xianming Li ◽  
...  
Keyword(s):  
Reference Genes ◽  
Pcr Analysis ◽  
Training Systems ◽  
Qrt Pcr ◽  
Leaf Tissues ◽  
Suitable Reference

Selection and validation of reference genes for quantitative real-time PCR analysis of Nitraria tangutorum

2020 ◽  
Author(s):  
Bo Wang ◽  
Lirong WANG ◽  
Huirong Duan ◽  
Peifang Chong ◽  
Shiping Su ◽  
...  
Keyword(s):  
Real Time ◽  
Reference Genes ◽  
Pcr Analysis ◽  
Expression Stability ◽  
Experimental Conditions ◽  
Qrt Pcr ◽  
Nitraria Tangutorum ◽  
Desert Areas ◽  
Polymerase Chain ◽  
Suitable Reference

Abstract Background: Suitable reference genes can be used to calibrate the error in quantitative real‑time polymerase chain reaction (qRT-PCR) experiments and make the results more credible. However, reference genes suitable for different species and different experimental conditions do not exist. Nitraria tangutorum Bobr. is a typical plant in desert areas and desert plains, which is drought-resistant, saline-alkali resistant, barren-resistant, and has extremely strong adaptability. Due to insufficient understanding of the importance of this germplasm in the past, it is still unclear which genes can be used as reference genes to calibrate qRT-PCR data of N. tangutorum .Results: In this study, we analyzed the expression levels of 10 candidate reference genes (ACT, GAPDH, TUA, TUB, CYP, UBC, His, PP2A, HSP, and EF1-α) in three tissues (root, stem and leaf) and under five abiotic stresses (salt, drought, heat, cold, and ABA) of N. tangutorum seedlings by qRT-PCR. Three analysis software programs (geNorm, NormFinder, and BestKeeper) were used to evaluate expression stability of ten genes. Comprehensive analysis showed that EF1-α and His had the best expression stability, whereas HSP was the least suitable as a reference gene. The expression profile of NtCER7, a gene related to the regulation of the waxy synthesis of N. tangutorum, verified the accuracy of the experimental results.Conclusion: Based on this study, we recommend EF1-α and His as suitable reference genes for N. tangutorum. This study provides the first data on stable reference genes in N. tangutorum, which will be beneficial to study of the gene expression of N. tangutorum and other Nitraria species in the future.

scholarly journals Identification of Suitable Reference Genes for Gene Expression Normalization in qRT-PCR Analysis in Watermelon

PLoS ONE ◽  
2014 ◽  
Vol 9 (2) ◽  
pp. e90612 ◽  
Author(s):  
Qiusheng Kong ◽  
Jingxian Yuan ◽  
Lingyun Gao ◽  
Shuang Zhao ◽  
Wei Jiang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Reference Genes ◽  
Pcr Analysis ◽  
Qrt Pcr ◽  
Suitable Reference

scholarly journals Identification And Evaluation of Reference Genes For Cynanchum Auriculatum Under Various Stress Conditions

2022 ◽  
Author(s):  
Zhi-Peng Zhu ◽  
Jian-Xiang Yu ◽  
Ke-Xin Wu ◽  
Qin-Yi Xu ◽  
Yi-Jun Kang ◽  
...  
Keyword(s):  
Reference Genes ◽  
Herbal Medicines ◽  
Stress Conditions ◽  
Pcr Analysis ◽  
Expression Stability ◽  
Qrt Pcr ◽  
Chinese Herbal ◽  
Cynanchum Auriculatum ◽  
Screening Study ◽  
Suitable Reference

Abstract Baishouwu (Cynanchum auriculatum) is a kind of critical Chinese herbal medicine. However, compared with the studies of other Chinese herbal medicines, the screening study on the reference genes of C. auriculatum is still the blank. Deterioration of the natural environment severely affects the growth and development of C. auriculatum. This study screened and identified suitable reference genes of C. auriculatum under various stress conditions. Based on qRT-PCR, geNorm, NormFinder, BestKeeper, and RefFinder were used for the expression stability evaluation of 12 potential reference genes from C. auriculatum. The ranking table showed that optimal reference genes included EF2 and SAMDC (heat stress), CYP and TUB-β (cold stress), TUB-α and GAPDH (drought stress), SAMDC and TUB-α (waterlogging stress), along with EF2 and ACT7 (salt stress). These results also demonstrated that under different abiotic stresses, suitable reference genes of plants should be selected for qRT-PCR analysis.

Selection of suitable reference genes for qRT-PCR analysis of Begonia semperflorens under stress conditions

2019 ◽  
Vol 46 (6) ◽  
pp. 6027-6037 ◽  
Author(s):  
Yanman Li ◽  
Ying Qu ◽  
Yang Wang ◽  
Xue Bai ◽  
Geng Tian ◽  
...  
Keyword(s):  
Reference Genes ◽  
Stress Conditions ◽  
Pcr Analysis ◽  
Qrt Pcr ◽  
Suitable Reference ◽  
Selection Of ◽  
Begonia Semperflorens

scholarly journals Evaluation of suitable reference genes for normalization of quantitative real-time PCR analysis in rice plants under Xanthomonas oryzae pv. oryzae--infection and melatonin supplementation

2020 ◽  
Vol 2 (1) ◽  
Author(s):  
Xian Chen ◽  
Pedro Laborda ◽  
Yan Dong ◽  
Fengquan Liu
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Reference Genes ◽  
Housekeeping Genes ◽  
Suitable Reference Gene ◽  
Pcr Analysis ◽  
Quantitative Real Time Pcr ◽  
Rice Plants ◽  
Qrt Pcr ◽  
Suitable Reference

Abstract Exogenous melatonin (MT) was found to be an interesting tool for enhancing the resistance of rice to Xanthomonasoryzaepv. oryzae (Xoo)-caused bacterial blight (BB). However, the accurate comparison of the expression levels across samples was a challenging task. In this work, the stability of 10 common used housekeeping genes under Xoo-infection and MT supplementation in rice was analyzed using quantitative real-time PCR (qRT-PCR), and algorithms geNorm, NormFinder and BestKeeper. Our results indicated that most reference genes remained stable in Xoo-infected rice plants, while a number of reference genes were affected by MT supplementation. Among all studied genes, the transcript levels of 18S(18S ribosomal RNA) and UBC (Ubiquitin-conjugating enzyme E2) remained unaltered by Xoo infection, while UBC and UBQ5(Ubiquitin 5) were the most stable genes when examining simultaneous Xoo-infection and MT supplementation, demonstrating that UBC is a suitable reference gene for qRT-PCR data normalization in rice under Xoo-infection and MT supplementation.

Sign in / Sign up

Export Citation Format

Share Document