Identification and testing of reference genes for gene expression analysis in pollen of Pyrus bretschneideri

Chen, Jianqing; Li, Xinyue; Wang, Danqi; Li, Leiting; Zhou, Hongsheng; Liu, Zhe; Wu, Jun; Wang, Peng; Jiang, Xiaoxue; Fabrice, Musana R.; Zhang, Shaoling; Wu, Juyou

doi:10.1016/j.scienta.2015.04.010

Cited by 36 publications

(18 citation statements)

References 50 publications

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“…All analyses were achieved by three independently biological replicates. A pear TUBULIN gene was used as a constitutive control (Chen et al ). All primer sequences are listed in Table S1 (Supporting information).…”

Section: Methodsmentioning

confidence: 99%

Transcriptome analysis unravels an ethylene response factor involved in regulating fruit ripening in pear

Hao

Wang

Cheng

et al. 2018

Physiologia Plantarum

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Ethylene response factor (ERF) has been widely studied in regulating fruit ripening in tomato, apple, banana and kiwifruit, but little is known in pear. In this study 1-methylcyclopropene (1-MCP) treatment, an inhibitor of ethylene perception, was conducted at approximately 30 days before harvest to delay fruit ripening in a climacteric white pear cultivar Yali. Transcriptome libraries were constructed and sequenced in pre-ripening, ripening, and 1-MCP treated fruits. Data analysis showed that 73 candidate genes related to fruit ripening were induced by 1-MCP, among which two were positively related, namely 1-aminocyclopropane-1-carboxyla oxidase and an ERF gene (designated as ACO54 and ERF24). Transient transformations in pear fruit revealed that over-expression of ACO54 enhance transcription level of ERF24 and most ripening-related genes. Meanwhile, over-expression of ERF24 raises expression level of ACO54 and partially ripening-related genes. Moreover, dual-luciferase and yeast-one-hybrid assays unravel an interaction between ERF24 and the ACO54 promoter. Therefore, the ERF24 could directly regulate ACO54 expression by binding to its promoter. These results suggested that the first identified ERF24 is involved in regulating fruit ripening in Chinese white pear.

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Section: Methodsmentioning

confidence: 99%

Transcriptome analysis unravels an ethylene response factor involved in regulating fruit ripening in pear

Hao

Wang

Cheng

et al. 2018

Physiologia Plantarum

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“…1 In addition, to determine the expression stability, qRT-PCR data were analyzed using geNorm (Vandesompele et al 2002), NormFinder (Andersen et al 2004), and BestKeeper (Pfaffl et al 2004). The details were identical to the previous report (Chen et al 2015).…”

Section: Plant Materials and Quantitative Real-time Polymerase Chain mentioning

confidence: 99%

“…Reference genes for gene expression studies should be expressed constantly and stably throughout various conditions, such as developmental stages (Thellin et al 1999;Schmittgen and Zakrajsek 2000). Presently, reference genes selected for fruit development and ripening were varied in different genus, such as 18S RNA in tomato (Martel et al 2011;Lee et al 2012;Nguyen et al 2014), TEF2, RP, and UBQ in peach (Tong et al 2009;Zhou et al 2015), ACTIN in strawberry, apple, and orange (Yu et al 2012;Ireland et al 2013;Jia et al 2013), and PbTUB in pear (Wu et al 2012;Chen et al 2015). In this study, we found that four genes including PbPDI.F1, PbPDI.E, and PbPDI.D2 had more stable expression than PbTUB in pear fruits (Fig.…”

Section: Phylogeny and Duplication Events Of Pdi Genes In Fruit Treesmentioning

confidence: 99%

“…In fruit trees, for example, the genes encoding 18S ribosomal RNA (18S RNA), β-actin (ACT), cyclophilin (CYP), translation elongation factor (TEF), glyceraldehydes-3-phosphate dehydrogenase (GAPDH), phospholipase A (PLA), RNA polymerase (RPII), ribosomal protein L (RPL), tubblin alpha (TUA), tubblin beta (TUB), ubiquitin (UBQ), vacuolar H+-ATPase (VHA), and phosphoprotein phosphatase (PP) have been tested whether they could be used as reference genes for gene expression analysis (Tong et al 2009;Chen et al 2015). In pear, as a result, TUB is the optimal gene for gene expression analysis, while GAPDH, PP, CYP, and UBQ were suitable for pollen tube growth underlying different treatments, respectively (Chen et al 2015). In peach fruit, TEF, UBQ, and RPII were the three most stable genes in expression level underlying various treatments (Tong et al 2009).…”

Section: Introductionmentioning

confidence: 99%

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Phylogenetic and expression analysis of protein disulfide isomerase unravels good reference genes for gene expression studies in pear and peach fruits

Ke¹,

Cheng²,

Liu³

et al. 2018

Can. J. Plant Sci.

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Protein disulfide isomerase (PDI) is an important enzyme for protein folding in endoplasmic reticulum. The PDI gene family has been systematically studied in Arabidopsis, barley, rice, and bread wheat; however, little is known about this gene family and their roles during fruit development and ripening in fruit trees. In this study, 63 PDI genes were isolated from 8 fruit trees. Phylogenetic trees showed that these genes were clustered into six different groups, designated as A to F. In the groups, the PDI genes had significant differences in gene structure and conserved regions. The chromosome location of each PDI gene was determined in complete genome-assembly fruit trees and the synteny of chromosome segments was detected among peach, pear, and strawberry. Expression profiles of PDI genes in peach, pear, and strawberry showed that nearly all genes in group D and E were more highly expressed in developmental and ripening fruit tissues than those in other groups, while all genes in group A and B presented the lowest levels of expression in fruits of each stage. Moreover, qRT-PCR analyses revealed that these expressed genes were stable expressed in pear and peach fruits, as well as the reported reference genes. Eventually, PbPDI.F1 presented the highest expression stability in pear fruit while PpPDI.F displayed stronger stability than other genes in peach fruit. Thus, these two genes, which were clustered in group F, are good reference genes for gene expression studies during fruit development and ripening.

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“…The raw data were analyzed with iQ5 software (version 2.0, Bio-Rad), and the relative expression levels were calculated with the 2 −△△CT method (Livak and Schmittgen 2001). Tubulin (AY338250) was used as the internal control (Chen et al, 2015). qPCR data are technical replicates (n=3) with error bars, representing means±standard error (SE).…”

Section: Qrt-pcr Analysismentioning

confidence: 99%

Calcium treatments promote the aroma volatiles emission of pear (Pyrus ussuriensis ‘Nanguoli’) fruit during post-harvest ripening process

Wei

Qin

Zhang

et al. 2017

Scientia Horticulturae

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Aroma is an important factor affecting pear fruit quality. This study was undertaken to assess whether pre-harvest calcium sprays (15 d before harvest, applications at 4%, w/v, 120 d after full bloom) could improve aroma of Pyrus ussuriensis 'Nanguoli' pear fruit at harvest and post-harvest, and analyse the mechanism of metabolic regulation. Most compounds contributing to overall flavor in ripe 'Nanguoli' fruit at harvest and post-harvest were enhanced in response to pre-harvest calcium applications. Ester, especially acetyl ester, content was enhanced by calcium treatment; however, contents of alcohols, aldehydes and hydrocarbons were little affected. These effects were consistent with higher pyruvate decarboxylase and alcohol dehydrogenase activities in treated fruit, suggesting that calcium treatment increased supply of alcohols and acyl CoAs for ester biosynthesis. We also found that activities of β-glucosidases (involved in the emission of bound volatile compounds) were improved and the contents of bound volatiles were reduced by calcium treatment. At the same time, relative contents of linoleic and linolenic acids were improved by calcium treatment, expression of genes encoding fatty acid desaturase involved in the biosynthesis of unsaturated fatty acid precursors (linoleic and linolenic acids) for volatile aroma were improved. Calcium

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Identification and testing of reference genes for gene expression analysis in pollen of Pyrus bretschneideri

Cited by 36 publications

References 50 publications

Transcriptome analysis unravels an ethylene response factor involved in regulating fruit ripening in pear

Transcriptome analysis unravels an ethylene response factor involved in regulating fruit ripening in pear

Phylogenetic and expression analysis of protein disulfide isomerase unravels good reference genes for gene expression studies in pear and peach fruits

Calcium treatments promote the aroma volatiles emission of pear (Pyrus ussuriensis ‘Nanguoli’) fruit during post-harvest ripening process

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