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Identification of Optimal Reference Genes for Normalization of qPCR Analysis during Pepper Fruit Development

Overview of attention for article published in Frontiers in Plant Science, June 2017
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Title
Identification of Optimal Reference Genes for Normalization of qPCR Analysis during Pepper Fruit Development
Published in
Frontiers in Plant Science, June 2017
DOI 10.3389/fpls.2017.01128
Pubmed ID
Authors

Yuan Cheng, Xin Pang, Hongjian Wan, Golam J. Ahammed, Jiahong Yu, Zhuping Yao, Meiying Ruan, Qingjing Ye, Zhimiao Li, Rongqing Wang, Yuejian Yang, Guozhi Zhou

Abstract

Due to its high sensitivity and reproducibility, quantitative real-time PCR (qPCR) is practiced as a useful research tool for targeted gene expression analysis. For qPCR operations, the normalization with suitable reference genes (RGs) is a crucial step that eventually determines the reliability of the obtained results. Although pepper is considered an ideal model plant for the study of non-climacteric fruit development, at present no specific RG have been developed or validated for the qPCR analyses of pepper fruit. Therefore, this study aimed to identify stably expressed genes for their potential use as RGs in pepper fruit studies. Initially, a total of 35 putative RGs were selected by mining the pepper transcriptome data sets derived from the PGP (Pepper Genome Platform) and PGD (Pepper Genome Database). Their expression stabilities were further measured in a set of pepper (Capsicum annuum L. var. 007e) fruit samples, which represented four different fruit developmental stages (IM: Immature; MG: Mature green; B: Break; MR: Mature red) using the qPCR analysis. Then, based on the qPCR results, three different statistical algorithms, namely geNorm, Normfinder, and boxplot, were chosen to evaluate the expression stabilities of these putative RGs. It should be noted that nine genes were proven to be qualified as RGs during pepper fruit development, namely CaREV05 (CA00g79660); CaREV08 (CA06g02180); CaREV09 (CA06g05650); CaREV16 (Capana12g002666); CaREV21 (Capana10g001439); CaREV23 (Capana05g000680); CaREV26 (Capana01g002973); CaREV27 (Capana11g000123); CaREV31 (Capana04g002411); and CaREV33 (Capana08g001826). Further analysis based on geNorm suggested that the application of the two most stably expressed genes (CaREV05 and CaREV08) would provide optimal transcript normalization in the qPCR experiments. Therefore, a new and comprehensive strategy for the identification of optimal RGs was developed. This strategy allowed for the effective normalization of the qPCR analysis of the pepper fruit development at the whole pepper genome level. This study not only explored the optimal RGs specific for studying pepper fruit development, but also introduced a referable strategy of RG mining which could potentially be implicated in other plant species.

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Mendeley readers

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The data shown below were compiled from readership statistics for 23 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 23 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 5 22%
Student > Master 3 13%
Researcher 3 13%
Student > Bachelor 2 9%
Professor 1 4%
Other 2 9%
Unknown 7 30%
Readers by discipline Count As %
Agricultural and Biological Sciences 10 43%
Biochemistry, Genetics and Molecular Biology 5 22%
Unknown 8 35%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 2. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 01 October 2017.
All research outputs
#14,945,861
of 22,988,380 outputs
Outputs from Frontiers in Plant Science
#9,377
of 20,454 outputs
Outputs of similar age
#187,984
of 315,304 outputs
Outputs of similar age from Frontiers in Plant Science
#305
of 558 outputs
Altmetric has tracked 22,988,380 research outputs across all sources so far. This one is in the 32nd percentile – i.e., 32% of other outputs scored the same or lower than it.
So far Altmetric has tracked 20,454 research outputs from this source. They receive a mean Attention Score of 4.0. This one is in the 47th percentile – i.e., 47% of its peers scored the same or lower than it.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 315,304 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 37th percentile – i.e., 37% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 558 others from the same source and published within six weeks on either side of this one. This one is in the 39th percentile – i.e., 39% of its contemporaries scored the same or lower than it.