Identification of reference genes for quantitative real-time PCR in different developmental stages and under refrigeration conditions in soursop fruits (Annona muricata L.)

Berúmen-Varela, Guillermo; Palomino-Hermosillo, Yolotzin Apatzingán; Bautista-Rosales, Pedro Ulises; Peña-Sandoval, Gabriela R.; López-Guzmán, Graciela Guadalupe; Balois-Morales, Rosendo

doi:10.1016/j.scienta.2019.108893

Cited by 18 publications

(15 citation statements)

References 26 publications

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“…Bioinformatics analysis was carried out to identify PPO and SOD genes in soursop according to the method described by Berumen-Varela et al [ 48 ]. In this regard, a BLAST database was built from a soursop leaf transcriptome ( , accessed on 16 June 2021) and then a BLAST search was performed using PPO and SOD sequences from other species to identify homologous genes.…”

Section: Methodsmentioning

confidence: 99%

Differential Responses of Antioxidative System during the Interaction of Soursop Fruits (Annona muricata L.) and Nectria haematococca at Postharvest Storage

Rubio-Melgarejo

Balois-Morales

Ochoa-Jiménez

et al. 2021

Plants

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Soursop fruit (Annona muricata L.) production is diminished by the attack of pathogens such as Nectria haematococca. However, the fruit–pathogen interaction at the biochemical and molecular levels is still unknown. The objective of this study was to analyze the response of the soursop fruit to the presence of N. haematococca during postharvest storage. Soursop fruits were inoculated with the pathogen and total phenolic compounds, antioxidant capacity by Ferric reducing/antioxidant power (FRAP), 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonate) (ABTS•+), and 2,2′-diphenyl-1-picrylhydrazyl radical (DPPH•), as well as enzymatic activity and transcript levels of polyphenol oxidase (PPO) and superoxide dismutase (SOD), were evaluated at 1, 3, and 5 days of storage. The noninoculated fruits were the controls of the experiment. The highest total phenol content was recorded on day one in the inoculated fruits. FRAP, ABTS, and DPPH activity presented the highest values on day three in the control fruits. Inoculated fruits recorded the highest PPO activity on day five and a five-fold induction in the PPO transcript on day three. SOD activity showed a decrease during the days of storage and 10-fold induction of SOD transcript on day three in the inoculated fruits. Principal component analysis showed that total phenols were the variable that contributed the most to the observed variations. Furthermore, a positive correlation between total phenols and SOD activity, PPO expression, and SOD expression, as well as between DPPH and FRAP, was recorded. The results showed a differential response in antioxidant capacity, enzymatic activity, and gene expression during the interaction of soursop fruits–N. haematococca at postharvest storage.

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Section: Methodsmentioning

confidence: 99%

Differential Responses of Antioxidative System during the Interaction of Soursop Fruits (Annona muricata L.) and Nectria haematococca at Postharvest Storage

Rubio-Melgarejo

Balois-Morales

Ochoa-Jiménez

et al. 2021

Plants

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“…The quantitative real-time polymerase chain reaction (qRT-PCR) is a technique to quantify gene expression [ 88 , 89 ]. In the present study, the qRT-PCR method was used to quantify the miRNA expression level.…”

Section: Resultsmentioning

confidence: 99%

The Impact of Zinc Oxide Nanoparticles on Cytotoxicity, Genotoxicity, and miRNA Expression in Barley (Hordeum vulgare L.) Seedlings

Plaksenkova

Kokina

Petrova

et al. 2020

The Scientific World Journal

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Zinc oxide nanoparticles are one of the most commonly engineered nanomaterials and necessarily enter the environment because of the large quantities produced and their widespread application. Understanding the impacts of nanoparticles on plant growth and development is crucial for the assessment of probable environmental risks to food safety and human health, because plants are a fundamental living component of the ecosystem and the most important source in the human food chain. The objective of this study was to examine the impact of different concentrations of zinc oxide nanoparticles on barley Hordeum vulgare L. seed germination, seedling morphology, root cell viability, stress level, genotoxicity, and expression of miRNAs. The results demonstrate that zinc oxide nanoparticles enhance barley seed germination, shoot/root elongation, and H2O2 stress level and decrease root cell viability and genomic template stability and up- and downregulated miRNAs in barley seedlings.

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“…Related studies have been carried out in other fruit species, such as peach (Tong et al 2009), apple (Zhu et al 2019), strawberry (Liu et al 2020), pitaya (Chen et al 2019), soursop (Berumen‐Varela et al 2020) and olive (Hürkan et al 2018), which collectively confirm that reference genes should be selected in accordance with different cultivars, tissues or developmental stages. For the grapevine, however, related studies are relatively few, resulting in the non‐scientific selection of reference genes.…”

Section: Introductionmentioning

confidence: 80%

Identification of optimal and novel reference genes for quantitative real‐time polymerase chain reaction analysis in grapevine

Wei

Wang

Liu

et al. 2021

Australian Journal of Grape and Wine Research

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Background and Aims Quantitative real‐time polymerase chain reaction (qRT‐PCR) has the advantages of sensitivity, specificity and repeatability, and is commonly used to investigate the expression levels of genes. The selection of appropriate endogenous reference genes is the pre‐condition for acquiring the desired results of qRT‐PCR. This study aimed to identify and screen the most suitable reference genes for grapevines. Methods and Results Three novel genes from previous transcriptomic data were selected for their stable expression either at the developmental stages of the grape berry or under the various treatments. The expression stability of these three candidate genes and six commonly used reference genes were evaluated with geNorm, NormFinder and BestKeeper algorithms in several grapevine samples. The geNorm analysis indicated that two reference genes were sufficient to normalise expression. The optimal reference gene combinations were: VvEF1‐γ and PPR2 in fruit; RRM1 and EF1‐α in leaf; EF1‐α and Actin in tendril of several cultivars. The combination of EF1‐α and VvEF1‐α was the most appropriate reference gene for fruits at different developmental stages. When all the samples were considered, the combination of RRM1 and Ubiquitin was optimal. Conclusions Different reference genes should be employed when qRT‐PCR is performed in different grapevine samples. The gene RRM1 is a new appropriate reference gene for grapevine. Significance of the Study This study provided alternatives for the optimal reference genes in qRT‐PCR, and identified one novel and appropriate reference gene (RRM1) for grapevine.

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Identification of reference genes for quantitative real-time PCR in different developmental stages and under refrigeration conditions in soursop fruits (Annona muricata L.)

Cited by 18 publications

References 26 publications

Differential Responses of Antioxidative System during the Interaction of Soursop Fruits (Annona muricata L.) and Nectria haematococca at Postharvest Storage

Differential Responses of Antioxidative System during the Interaction of Soursop Fruits (Annona muricata L.) and Nectria haematococca at Postharvest Storage

The Impact of Zinc Oxide Nanoparticles on Cytotoxicity, Genotoxicity, and miRNA Expression in Barley (Hordeum vulgare L.) Seedlings

Identification of optimal and novel reference genes for quantitative real‐time polymerase chain reaction analysis in grapevine

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