HTS analysis of resistance induction against PPV by four hairpin constructs in Nicotiana benthamiana Domin

Abstract Plum pox virus (PPV) is the most devastating viral disease of the stone fruits worldwide. Inefficiency of the traditional control measures against PPV along with its globally widespread distribution and the economic importance of stone fruits, signify the necessity and importance of PPV res...

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Main Authors: Maryam Ghaderi Sohi, Kahraman Gürcan, Saffet Teber, Mikail Akbulut, Yazgan Tunç, Mehmet Yaman, Ali Khadivi, Azam Nikbakht-Dehkordi, Harun Karcı, Burak Özgören, Vahid Roumi
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Language:English
Published: Nature Portfolio 2025-04-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-97546-7
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author Maryam Ghaderi Sohi
Kahraman Gürcan
Saffet Teber
Mikail Akbulut
Yazgan Tunç
Mehmet Yaman
Ali Khadivi
Azam Nikbakht-Dehkordi
Harun Karcı
Burak Özgören
Vahid Roumi
author_facet Maryam Ghaderi Sohi
Kahraman Gürcan
Saffet Teber
Mikail Akbulut
Yazgan Tunç
Mehmet Yaman
Ali Khadivi
Azam Nikbakht-Dehkordi
Harun Karcı
Burak Özgören
Vahid Roumi
author_sort Maryam Ghaderi Sohi
collection DOAJ
description Abstract Plum pox virus (PPV) is the most devastating viral disease of the stone fruits worldwide. Inefficiency of the traditional control measures against PPV along with its globally widespread distribution and the economic importance of stone fruits, signify the necessity and importance of PPV resistance programs. In the present study, Agrobacterium-mediated transformation of Nicotiana benthamiana Domin was performed using four inverted repeat constructs derived from UTR/P1, HCPro, HCPro/P3, and CP regions of PPV-T isolate KyEsAp301. The efficacy of the constructs for inducing virus resistance in transgenic plants was evaluated by inoculation with PPV-D, -M, and -T strains. The potential of hairpin structures in the production of siRNAs and miRNAs in both wild-type and transgenic plants was compared by small RNA high-throughput sequencing. Although the four PPV genomic regions were used for transgenic resistance in previous experiments, small RNA high-throughput sequencing was first time used in this study to demonstrate the efficacy of the PPV constructs and to determine expression profiles of siRNAs and miRNAs. The results revealed that the potentials of hairpin constructs in producing siRNAs and their accumulation in target regions were significantly different. Expression profiles of several known and novel miRNAs were dramatically changed in response to PPV infection in both wild-type and transgenic plants, demonstrating plausible involvement of these miRNAs in plant-virus interactions. Based on the abundance of siRNAs and lack of PPV virus accumulation in transgenic plants harboring UTR/P1 and CP hairpin construct, we have concluded that UTR/P1 and CP are likely the best viral regions for induction of resistance against PPV.
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spelling doaj-art-e0a9f7d3d71c48cfbaba93391d618fa02025-08-20T03:06:57ZengNature PortfolioScientific Reports2045-23222025-04-0115111410.1038/s41598-025-97546-7HTS analysis of resistance induction against PPV by four hairpin constructs in Nicotiana benthamiana DominMaryam Ghaderi Sohi0Kahraman Gürcan1Saffet Teber2Mikail Akbulut3Yazgan Tunç4Mehmet Yaman5Ali Khadivi6Azam Nikbakht-Dehkordi7Harun Karcı8Burak Özgören9Vahid Roumi10Genome and Stem Cell Research Center, Erciyes UniversityGenome and Stem Cell Research Center, Erciyes UniversityGenome and Stem Cell Research Center, Erciyes UniversityDepartment of Biology, Erciyes UniversityRepublic of Türkiye, Ministry of Agriculture and Forestry, General Directorate of Agricultural Research and Policies, Hatay Olive Research Institute Directorate, Hassa StationDepartment of Horticulture, Faculty of Agriculture, Erciyes UniversityDepartment of Horticultural Sciences, Faculty of Agriculture and Natural Resources, Arak UniversityResearch and Technology Institute of Plant Production, Shahid Bahonar UniversityDepartment of Horticulture, Çukurova UniversityDepartment of Horticulture, Çukurova UniversityPlant Protection Department, Faculty of Agriculture, University of MaraghehAbstract Plum pox virus (PPV) is the most devastating viral disease of the stone fruits worldwide. Inefficiency of the traditional control measures against PPV along with its globally widespread distribution and the economic importance of stone fruits, signify the necessity and importance of PPV resistance programs. In the present study, Agrobacterium-mediated transformation of Nicotiana benthamiana Domin was performed using four inverted repeat constructs derived from UTR/P1, HCPro, HCPro/P3, and CP regions of PPV-T isolate KyEsAp301. The efficacy of the constructs for inducing virus resistance in transgenic plants was evaluated by inoculation with PPV-D, -M, and -T strains. The potential of hairpin structures in the production of siRNAs and miRNAs in both wild-type and transgenic plants was compared by small RNA high-throughput sequencing. Although the four PPV genomic regions were used for transgenic resistance in previous experiments, small RNA high-throughput sequencing was first time used in this study to demonstrate the efficacy of the PPV constructs and to determine expression profiles of siRNAs and miRNAs. The results revealed that the potentials of hairpin constructs in producing siRNAs and their accumulation in target regions were significantly different. Expression profiles of several known and novel miRNAs were dramatically changed in response to PPV infection in both wild-type and transgenic plants, demonstrating plausible involvement of these miRNAs in plant-virus interactions. Based on the abundance of siRNAs and lack of PPV virus accumulation in transgenic plants harboring UTR/P1 and CP hairpin construct, we have concluded that UTR/P1 and CP are likely the best viral regions for induction of resistance against PPV.https://doi.org/10.1038/s41598-025-97546-7PotyvirusVirus silencingRNAiSmall RNA sequencing
spellingShingle Maryam Ghaderi Sohi
Kahraman Gürcan
Saffet Teber
Mikail Akbulut
Yazgan Tunç
Mehmet Yaman
Ali Khadivi
Azam Nikbakht-Dehkordi
Harun Karcı
Burak Özgören
Vahid Roumi
HTS analysis of resistance induction against PPV by four hairpin constructs in Nicotiana benthamiana Domin
Scientific Reports
Potyvirus
Virus silencing
RNAi
Small RNA sequencing
title HTS analysis of resistance induction against PPV by four hairpin constructs in Nicotiana benthamiana Domin
title_full HTS analysis of resistance induction against PPV by four hairpin constructs in Nicotiana benthamiana Domin
title_fullStr HTS analysis of resistance induction against PPV by four hairpin constructs in Nicotiana benthamiana Domin
title_full_unstemmed HTS analysis of resistance induction against PPV by four hairpin constructs in Nicotiana benthamiana Domin
title_short HTS analysis of resistance induction against PPV by four hairpin constructs in Nicotiana benthamiana Domin
title_sort hts analysis of resistance induction against ppv by four hairpin constructs in nicotiana benthamiana domin
topic Potyvirus
Virus silencing
RNAi
Small RNA sequencing
url https://doi.org/10.1038/s41598-025-97546-7
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