Sohi Maryam Ghaderi, Gürcan Kahraman, Teber Saffet, Akbulut Mikail, Tunç Yazgan, Yaman Mehmet, Khadivi Ali, Nikbakht-Dehkordi Azam, Karcı Harun, Özgören Burak, Roumi Vahid
Genome and Stem Cell Research Center, Erciyes University, Kayseri, 38280, Türkiye.
Department of Biology, Erciyes University, Kayseri, 38280, Türkiye.
Sci Rep. 2025 Apr 10;15(1):12362. doi: 10.1038/s41598-025-97546-7.
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.
李痘病毒（PPV）是全球核果类果树最具毁灭性的病毒病害。传统的PPV防治措施效率低下，加之其在全球广泛传播以及核果类果树的经济重要性，凸显了PPV抗性计划的必要性和重要性。在本研究中，利用源自PPV-T分离株KyEsAp301的UTR/P1、HCPro、HCPro/P3和CP区域的四个反向重复构建体，通过农杆菌介导法转化本氏烟草（Nicotiana benthamiana Domin）。通过接种PPV-D、-M和-T株系，评估构建体在转基因植物中诱导病毒抗性的效果。通过小RNA高通量测序比较野生型和转基因植物中发夹结构在产生siRNA和miRNA方面的潜力。尽管在先前的实验中已使用PPV的四个基因组区域进行转基因抗性研究，但本研究首次使用小RNA高通量测序来证明PPV构建体的效果，并确定siRNA和miRNA的表达谱。结果表明，发夹构建体产生siRNA的潜力及其在靶区域的积累存在显著差异。在野生型和转基因植物中，几种已知和新的miRNA的表达谱在PPV感染后均发生了显著变化，表明这些miRNA可能参与了植物与病毒的相互作用。基于在含有UTR/P1和CP发夹构建体的转基因植物中siRNA的丰度以及PPV病毒积累的缺乏，我们得出结论，UTR/P1和CP可能是诱导对PPV抗性的最佳病毒区域。
