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      Chitosan Coating Enriched With Ruta graveolens L. Essential Oil Reduces Postharvest Anthracnose of Papaya ( Carica papaya L.) and Modulates Defense-Related Gene Expression

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          Abstract

          Anthracnose of papaya ( Carica papaya L.) caused by the fungus Colletotrichum spp. is one of the most economically important postharvest diseases. Coating with chitosan (CS) and Ruta graveolens essential oil (REO) might represent a novel eco-friendly method to prevent postharvest anthracnose infection. These compounds show both antimicrobial and eliciting activities, although the molecular mechanisms in papaya have not been investigated to date. In this study, the effectiveness of CS and REO alone and combined (CS-REO) on postharvest anthracnose of papaya fruit during storage were investigated, along with the expression of selected genes involved in plant defense mechanisms. Anthracnose incidence was reduced with CS, REO, and CS-REO emulsions after 9 days storage at 25°C, by 8, 21, and 37%, respectively, with disease severity reduced by 22, 29, and 44%, respectively. Thus, McKinney’s decay index was reduced by 22, 30, and 44%, respectively. A protocol based on reverse transcription quantitative real-time PCR (RT-qPCR) was validated for 17 papaya target genes linked to signaling pathways that regulate plant defense, pathogenesis-related protein, cell wall-degrading enzymes, oxidative stress, abiotic stress, and the phenylpropanoid pathway. CS induced gene upregulation mainly at 6 h posttreatment (hpt) and 48 hpt, while REO induced the highest upregulation at 0.5 hpt, which then decreased over time. Furthermore, CS-REO treatment delayed gene upregulation by REO alone, from 0.5 to 6 hpt, and kept that longer over time. This study suggests that CS stabilizes the volatile and/or hydrophobic substances of highly reactive essential oils. The additive effects of CS and REO were able to reduce postharvest decay and affect gene expression in papaya fruit.

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          Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

          The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data. Copyright 2001 Elsevier Science (USA).
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            The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments.

            Currently, a lack of consensus exists on how best to perform and interpret quantitative real-time PCR (qPCR) experiments. The problem is exacerbated by a lack of sufficient experimental detail in many publications, which impedes a reader's ability to evaluate critically the quality of the results presented or to repeat the experiments. The Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines target the reliability of results to help ensure the integrity of the scientific literature, promote consistency between laboratories, and increase experimental transparency. MIQE is a set of guidelines that describe the minimum information necessary for evaluating qPCR experiments. Included is a checklist to accompany the initial submission of a manuscript to the publisher. By providing all relevant experimental conditions and assay characteristics, reviewers can assess the validity of the protocols used. Full disclosure of all reagents, sequences, and analysis methods is necessary to enable other investigators to reproduce results. MIQE details should be published either in abbreviated form or as an online supplement. Following these guidelines will encourage better experimental practice, allowing more reliable and unequivocal interpretation of qPCR results.
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              The plant immune system.

              Many plant-associated microbes are pathogens that impair plant growth and reproduction. Plants respond to infection using a two-branched innate immune system. The first branch recognizes and responds to molecules common to many classes of microbes, including non-pathogens. The second responds to pathogen virulence factors, either directly or through their effects on host targets. These plant immune systems, and the pathogen molecules to which they respond, provide extraordinary insights into molecular recognition, cell biology and evolution across biological kingdoms. A detailed understanding of plant immune function will underpin crop improvement for food, fibre and biofuels production.
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                Author and article information

                Contributors
                Journal
                Front Plant Sci
                Front Plant Sci
                Front. Plant Sci.
                Frontiers in Plant Science
                Frontiers Media S.A.
                1664-462X
                11 November 2021
                2021
                : 12
                : 765806
                Affiliations
                [1] 1Department of Agricultural, Food and Environmental Sciences, Marche Polytechnic University , Ancona, Italy
                [2] 2Faculty of Bioscience and Technology for Food, Agriculture and Environment, University of Teramo , Teramo, Italy
                [3] 3Facultad de Ingeniería, Programa de Ingeniería Agroindustrial, Universidad del Atlántico , Puerto Colombia, Colombia
                Author notes

                Edited by: María Serrano, Miguel Hernández University of Elche, Spain

                Reviewed by: Pradeep Kumar, North Eastern Regional Institute of Science and Technology, India; Ghulam Khaliq, Lasbela University of Agriculture, Water and Marine Sciences, Pakistan; Simona Marianna Sanzani, International Centre for Advanced Mediterranean Agronomic Studies, Italy; Periyar Selvam Sellamuthu, SRM Institute of Science and Technology, India

                *Correspondence: Gianfranco Romanazzi, g.romanazzi@ 123456univpm.it

                These authors have contributed equally to this work

                This article was submitted to Crop and Product Physiology, a section of the journal Frontiers in Plant Science

                Article
                10.3389/fpls.2021.765806
                8632526
                c88a7b2a-4e80-4a68-aae5-828ad5d40efc
                Copyright © 2021 Landi, Peralta-Ruiz, Chaves-López and Romanazzi.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 01 September 2021
                : 05 October 2021
                Page count
                Figures: 4, Tables: 3, Equations: 4, References: 93, Pages: 14, Words: 12353
                Categories
                Plant Science
                Original Research

                Plant science & Botany
                chitosan,essential oils,gene expression,induced resistance,rt-qpcr
                Plant science & Botany
                chitosan, essential oils, gene expression, induced resistance, rt-qpcr

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