The use of microRNAs as reference genes for quantitative polymerase chain reaction in soybean

Kulcheski, Franceli Rodrigues; Marcelino-Guimarães, Francismar Corrêa; Nepomuceno, Alexandre Lima; Abdelnoor, R. V.; Margis, Rogério

doi:10.1016/j.ab.2010.07.020

Cited by 129 publications

(118 citation statements)

References 32 publications

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“…Despite the different results from the geNorm and NormFinder rankings, the results of the present study indicated that the peach reference genes were stably expressed and suitable for normalizing RT-qPCR expression analysis, which is in agreement with the results observed by Kulcheski et al (2010). However, it is important to consider that not all of the genes tested were constitutively expressed under different conditions, and the universality of these results needs to be demonstrated in other plants and under other experimental conditions.…”

Section: Expression Stability Of All Candidate Reference Genes Under supporting

confidence: 80%

Identification of reference genes for RT-qPCR analysis in peach genotypes with contrasting chilling requirements

et al. 2017

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ABSTRACT. Selecting and validating reference genes are the first steps in studying gene expression by reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR). The present study aimed to evaluate the stability of five reference genes for the purpose of normalization when studying gene expression in various cultivars of Prunus persica with different chilling requirements. Flower bud tissues of nine peach genotypes from Embrapa's peach breeding program with different chilling requirements were used, and five candidate reference genes based on the RT-qPCR that were useful for studying the relative quantitative gene expression and stability were evaluated using geNorm, NormFinder, and bestKeeper software packages. The results indicated that among the genes tested, the most stable genes to be used as reference genes are Act and UBQ10. This study is the first survey of the stability of reference genes in peaches under chilling stress and provides guidelines for more accurate RT-qPCR results.

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Section: Expression Stability Of All Candidate Reference Genes Under supporting

confidence: 80%

Identification of reference genes for RT-qPCR analysis in peach genotypes with contrasting chilling requirements

et al. 2017

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“…Soybean miR1520d was used as a reference miRNA gene to normalize the samples as outlined previously (Kulcheski et al, 2010). The expression pattern of all soybean miR172 family members was assessed in leaves, roots, and nodules 28 DAI with B. japonicum.…”

Section: Stem-loop Qrt-pcrmentioning

confidence: 99%

“…Stem-loop-specific reverse transcription for miRNAs was performed as described previously (Chen et al, 2005;Kulcheski et al, 2010). Soybean miR1520d was used as a reference miRNA gene to normalize the samples as outlined previously (Kulcheski et al, 2010).…”

Section: Stem-loop Qrt-pcrmentioning

confidence: 99%

Soybean miR172c Targets the Repressive AP2 Transcription Factor NNC1 to Activate ENOD40 Expression and Regulate Nodule Initiation

et al. 2014

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MicroRNAs are noncoding RNAs that act as master regulators to modulate various biological processes by posttranscriptionally repressing their target genes. Repression of their target mRNA(s) can modulate signaling cascades and subsequent cellular events. Recently, a role for miR172 in soybean (Glycine max) nodulation has been described; however, the molecular mechanism through which miR172 acts to regulate nodulation has yet to be explored. Here, we demonstrate that soybean miR172c modulates both rhizobium infection and nodule organogenesis. miR172c was induced in soybean roots inoculated with either compatible Bradyrhizobium japonicum or lipooligosaccharide Nod factor and was highly upregulated during nodule development. Reduced activity and overexpression of miR172c caused dramatic changes in nodule initiation and nodule number. We show that soybean miR172c regulates nodule formation by repressing its target gene, Nodule Number Control1, which encodes a protein that directly targets the promoter of the early nodulin gene, ENOD40. Interestingly, transcriptional levels of miR172c were regulated by both Nod Factor Receptor1α/5α-mediated activation and by autoregulation of nodulation-mediated inhibition. Thus, we established a direct link between miR172c and the Nod factor signaling pathway in addition to adding a new layer to the precise nodulation regulation mechanism of soybean.

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“…gma-miR1520d was used as a reference miRNA gene for normalization, as described by Kulcheski et al (2010). All primers used in stem-loop qRT-PCR are listed Supplemental Table S3.…”

Section: Stem-loop Qrt-pcrmentioning

confidence: 99%

MicroRNA167-Directed Regulation of the Auxin Response Factors GmARF8a and GmARF8b Is Required for Soybean Nodulation and Lateral Root Development

et al. 2015

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The use of microRNAs as reference genes for quantitative polymerase chain reaction in soybean

Cited by 129 publications

References 32 publications

Identification of reference genes for RT-qPCR analysis in peach genotypes with contrasting chilling requirements

Identification of reference genes for RT-qPCR analysis in peach genotypes with contrasting chilling requirements

Soybean miR172c Targets the Repressive AP2 Transcription Factor NNC1 to Activate ENOD40 Expression and Regulate Nodule Initiation

MicroRNA167-Directed Regulation of the Auxin Response Factors GmARF8a and GmARF8b Is Required for Soybean Nodulation and Lateral Root Development

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