Selection and Verification of Candidate Reference Genes for Mature MicroRNA Expression by Quantitative RT-PCR in the Tea Plant (Camellia sinensis)

Song, Hui; Xiao, Zhigang; Shi, Cong; Wang, Shuangshuang; Wu, Ailin; Wei, Chaoling

doi:10.3390/genes7060025

Cited by 31 publications

(17 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A similar study was conducted to compare the 5.8S rRNA gene in the carmine spider mite, Tetranychus cinnabarinus (Boisduval) (Acarina: Tetranychidae), with other commonly used HkGs and validated its stability of expression under specific experimental conditions [53]. Another study rectified the stability evaluated from miRNAs and non-coding small RNAs [54] and thus showed uniformity among all cell types and experimental systems. Furthermore, the linearized plasmids of wsp and 5.8S rRNA and genomic DNA of B. longissima were amplified within the acceptable range of efficiency [55].…”

Section: Discussionmentioning

confidence: 98%

Infection Density Dynamics and Phylogeny of Wolbachia Associated with Coconut Hispine Beetle, Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), by Multilocus Sequence Type (MLST) Genotyping

Ali

Muhammad

Hou

2018

Journal of Microbiology and Biotechnology

View full text Add to dashboard Cite

The intracellular bacterium is widespread in arthropods. Recently, possibilities of novel-mediated hosts, their distribution, and natural rate have been anticipated, and the coconut leaf beetle (Gestro) (Coleoptera: Chrysomelidae), which has garnered attention as a serious pest of palms, was subjected to this interrogation. By adopting surface protein () and multilocus sequence type (MLST) genotypic systems, we determined the infection density within host developmental stages, body parts, and tissues, and the results revealed that all the tested samples of were infected with the same strain (wLog), suggesting complete vertical transmission. The MLST profile elucidated two new alleles (-234 and coxA-266) that define a new sequence type (ST-483), which indicates the particular genotypic association of and. The quantitative real-time PCR analysis revealed a higher infection density in the eggs and adult stage, followed by the abdomen and reproductive tissues, respectively. However, no significant differences were observed in the infection density between sexes. Moreover, the and concatenated MLST alignment analysis of this study with other known Wolbachia-mediated arthropods revealed similar clustering with distinct monophyletic supergroup B. This is the first comprehensive report on the prevalence, infection dynamics, and phylogeny of the endosymbiont in , which demonstrated that is ubiquitous across all developmental stages and distributed in the entire body of . Understanding the infection dynamics would provide useful insight to build a framework for future investigations, understand its impacts on host physiology, and exploit it as a potential biocontrol agent.

show abstract

Section: Discussionmentioning

confidence: 98%

Infection Density Dynamics and Phylogeny of Wolbachia Associated with Coconut Hispine Beetle, Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), by Multilocus Sequence Type (MLST) Genotyping

Ali

Muhammad

Hou

2018

Journal of Microbiology and Biotechnology

View full text Add to dashboard Cite

show abstract

“…For example, some house-keeping genes, such as ACT 7 , UBQ , GAPDH and TUB , were widely used for gene expression analysis of mRNAs in diverse plants, and several noncoding RNAs, such as U6 snRNA and 5.8S rRNA , were typically chosen for normalizing miRNA quantification data. As miRNA research continues to expand, the potential use of miRNAs as reference genes has attracted increasing attention, and some small RNAs were obtained from plant species, such as grapevine [7], wheat [38], peach [39], soybean [40], and tea [41] demonstrating that miRNAs are more stable than the currently used reference genes under specific conditions. However, there are no reports on universal reference genes for the quantitative expression of both mRNA and miRNA, which may be necessary to calibrate both miRNA and its target gene(s) in a given sample.…”

Section: Discussionmentioning

confidence: 99%

Selection and validation of reference genes for quantitative expression analysis of miRNAs and mRNAs in Poplar

et al. 2019

View full text Add to dashboard Cite

Background: Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) is a rapid and sensitive approach to identify miRNA and protein-coding gene expression in plants. However, because of the specially designated reverse transcription and shorter PCR products, very few reference genes have been identified for the quantitative analysis of miRNA expression in plants, and different internal reference genes are needed to normalize the expression of miRNAs and mRNA genes respectively. Therefore, it is particularly important to select the suitable common reference genes for normalization of quantitative PCR of miRNA and mRNA. Results: In this study, a modified reverse transcription PCR protocol was adopted for selecting and validating universal internal reference genes of mRNAs and miRNAs. Eight commonly used reference genes, four stably expressed novel genes in Populus tremula, three small noncoding RNAs and three conserved miRNAs were selected as candidate genes, and the stability of their expression was examined across a set of 38 tissue samples from four developmental stages of poplar clone 84K (Populus alba × Populus glandulosa). The expression stability of these candidate genes was evaluated systematically by four algorithms: geNorm, NormFinder, Bestkeeper and DeltaCt. The results showed that Eukaryotic initiation factor 4A III (EIF4A) and U6-2 were suitable for samples of the callus stage; U6-1 and U6-2 were best for the seedling stage; Protein phosphatase 2A-2 (PP2A-2) and U6-1 were best for the plant stage; and Protein phosphatase 2A-2 (PP2A-2) and Oligouridylate binding protein 1B (UBP) were the best reference genes in the adventitious root (AR) regeneration stage. Conclusions: The purpose of this study was to identify the most appropriate reference genes for qRT-PCR of miRNAs and mRNAs in different tissues at several developmental stages in poplar. U6-1, EIF4A and PP2A-2 were the three most appropriate reference genes for qRT-PCR normalization of miRNAs and mRNAs during the plant regeneration process, and PP2A-2 and UBP represent the best reference genes in the AR regeneration stage of poplar. This work will benefit future studies of expression and function analysis of miRNAs and their target genes in poplar.

show abstract

“…Thus, quantification of these miRNAs may be very important from the viewpoint of understanding their prevalence and regulatory role in different systems . Previously, several studies have conducted in‐depth analyses of miRNAs and their roles in diverse plant growth and developmental processes of the tea plant …”

Section: Discussionmentioning

confidence: 99%

Absolute quantification of microRNAs in green tea (Camellia sinensis) by stem‐loop quantitative real‐time PCR

et al. 2016

Self Cite

View full text Add to dashboard Cite

show abstract

Selection and Verification of Candidate Reference Genes for Mature MicroRNA Expression by Quantitative RT-PCR in the Tea Plant (Camellia sinensis)

Cited by 31 publications

References 47 publications

Infection Density Dynamics and Phylogeny of Wolbachia Associated with Coconut Hispine Beetle, Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), by Multilocus Sequence Type (MLST) Genotyping

Infection Density Dynamics and Phylogeny of Wolbachia Associated with Coconut Hispine Beetle, Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), by Multilocus Sequence Type (MLST) Genotyping

Selection and validation of reference genes for quantitative expression analysis of miRNAs and mRNAs in Poplar

Absolute quantification of microRNAs in green tea (Camellia sinensis) by stem‐loop quantitative real‐time PCR

Contact Info

Product

Resources

About