scholarly journals Identification and Validation of Reference Genes for qRT-PCR Studies of Gene Expression inDioscorea opposita

2016 ◽  
Vol 2016 ◽  
pp. 1-13 ◽  
Author(s):  
Xiting Zhao ◽  
Xiaoli Zhang ◽  
Xiaobo Guo ◽  
Shujie Li ◽  
Linlin Han ◽  
...  
Keyword(s):  
Gene Expression ◽  
Reference Genes ◽  
Developmental Stages ◽  
Chain Reaction ◽  
Qrt Pcr ◽  
Expression Studies ◽  
Tuber Crop ◽  
Polymerase Chain ◽  
Gene Expression Studies ◽  
Suitable Reference

Quantitative real-time polymerase chain reaction (qRT-PCR) is one of the most common methods for gene expression studies. Data normalization based on reference genes is essential for obtaining reliable results for qRT-PCR assays. This study evaluated potential reference genes of Chinese yam (Dioscorea oppositaThunb.), which is an important tuber crop and medicinal plant in East Asia. The expression of ten candidate reference genes across 20 samples from different organs and development stages was assessed. We identified the most stable genes for qRT-PCR studies using combined samples from different organs. Our results also suggest that different suitable reference genes or combinations of reference genes for normalization should be applied according to different organs and developmental stages. To validate the suitability of the reference genes, we evaluated the relative expression ofPE2.1andPE53, which are two genes that may be associated with microtuber formation. Our results provide the foundation for reference gene(s) selection inD. oppositaand will contribute toward more accurate gene analysis studies of the genusDioscorea.

scholarly journals Identification of Suitable Reference Genes for gene Expression Studies by qRT-PCR in the Blister BeetleMylabris cichorii

2014 ◽  
Vol 14 (94) ◽  
pp. 1-10 ◽  
Author(s):  
Yu Wang ◽  
Zhong-Kang Wang ◽  
Yi Huang ◽  
Yu-Feng Liao ◽  
You-Ping Yin
Keyword(s):  
Gene Expression ◽  
Reference Genes ◽  
Qrt Pcr ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
Suitable Reference

Determination of Suitable Candidate Genes as Endogenous Control for Gene Expression Analysis in Local Capsicum Annuum MC11

2021 ◽  
Vol 12 (3) ◽  
pp. 1011-1017
Author(s):  
Marina Mokhtar Et.al
Keyword(s):  
Gene Expression ◽  
Candidate Genes ◽  
Capsicum Annuum ◽  
Reference Genes ◽  
Data Normalization ◽  
Plant Parts ◽  
Qrt Pcr ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
Suitable Reference

Quantitative real-time polymerase chain reaction (qRT-PCR) is one of the most common methods for gene expression studies. Data normalization based on reference genes is essential for qRT-PCR assays. This study identifies suitable reference genes for local chilli, Capsicum annuum var MC11 under incident of Cucumber mosaic virus infection. Six candidate genes actin, tub, EF1α, GAPDH, TEF1α and 18SrRNA and three validated Capsicum reference genes UBI-3 ref, β-tub ref and gapdhref were tested against five chilli plant parts stem, shoot, leave, flower and root.  The PCR/qRT-PCR results demonstrate only five candidate references genes actin, EF1α, GAPDH, 18SrRNA, and TEF1α that show specific single band of amplicon, without primer dimers and at the targeted sizes. Through qRT-PCR, GAPDH gives single peak in dissociation curve in all plant parts used further fulfilling the characteristic of reference genes.Previous work on validation of reference genes in pepper shows that only UBI-3 suits to C. annuum var MC11 infected CMV, thus we suggest that GAPDH has a potential to be a validated reference gene for C. annuum var MC11 and can be used together UBI-3 for the purpose of data normalization. 

Screening of reference genes using real-time quantitative PCR for gene expression studies inNeoseiulus barkeriHughes (Acari: Phytoseiidae)

2018 ◽  
Vol 109 (4) ◽  
pp. 443-452 ◽  
Author(s):  
C. Wang ◽  
J. Yang ◽  
Q. Pan ◽  
S. Yu ◽  
R. Luo ◽  
...  
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Reference Genes ◽  
Developmental Stages ◽  
Quantitative Polymerase Chain Reaction ◽  
Real Time Quantitative Pcr ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
The Stability ◽  
Suitable Reference

AbstractA stable reference gene is a key prerequisite for accurate assessment of gene expression. At present, the real-time reverse transcriptase quantitative polymerase chain reaction has been widely used in the analysis of gene expression in a variety of organisms.Neoseiulus barkeriHughes (Acari: Phytoseiidae) is a major predator of mites on many important economically crops. Until now, however, there are no reports evaluating the stability of reference genes in this species. In view of this, we used GeNorm, NormFinder, BestKeeper, and RefFinder software tools to evaluate the expression stability of 11 candidate reference genes in developmental stages and under various abiotic stresses. According to our results, β-ACTandHsp40were the top two stable reference genes in developmental stages. TheHsp60andHsp90were the most stable reference genes in various acaricides stress. For alterations in temperature,Hsp40and α-TUBwere the most suitable reference genes. About UV stress,EF1α and α-TUBwere the best choice, and for the different prey stress, β-ACTand α-TUBwere best suited. In normal conditions, the β-ACT and α-TUB were the two of the highest stable reference genes to respond to all kinds of stresses. The current study provided a valuable foundation for the further analysis of gene expression inN. barkeri.

scholarly journals Selection of suitable reference genes for gene expression studies in myxosporean (Myxozoa, Cnidaria) parasites

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Anush Kosakyan ◽  
Gema Alama-Bermejo ◽  
Pavla Bartošová-Sojková ◽  
Ana Born-Torrijos ◽  
Radek Šíma ◽  
...  
Keyword(s):  
Gene Expression ◽  
Rna Polymerase Ii ◽  
Ribosomal Rna ◽  
Reference Genes ◽  
Developmental Stages ◽  
Elongation Factor ◽  
Eukaryotic Translation ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
Suitable Reference

Abstract Myxozoans (Cnidaria: Myxozoa) are an extremely diversified group of endoparasites some of which are causative agents of serious diseases in fish. New methods involving gene expression studies have emerged over the last years to better understand and control myxozoan diseases. Quantitative RT-PCR is the most extensively used approach for gene expression studies. However, the accuracy of the results depends on the normalization of the data to reference genes. We studied the expression of eight commonly used reference genes, adenosylhomocysteinase (AHC1), beta actin (ACTB), eukaryotic translation elongation factor 2 (EF2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine-guanine phosphoribosyltransferase 1 (HPRT1), DNA-directed RNA polymerase II (RPB2), 18S ribosomal RNA (18S), 28S ribosomal RNA (28S) across different developmental stages of three myxozoan species, Sphaerospora molnari, Myxobolus cerebralis and Ceratonova shasta, representing the three major myxozoan linages from the largest class Myxosporea. The stable reference genes were identified using four algorithms: geNorm, NormFinder, Bestkeeper and ΔCq method. Additionally, we analyzed transcriptomic data from S. molnari proliferative and spore-forming stages to compare the relative amount of expressed transcripts with the most stable reference genes suggested by RT-qPCR. Our results revealed that GAPDH and EF2 are the most uniformly expressed genes across the different developmental stages of the studied myxozoan species.

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