Establishment of a Serology- and Molecular-Combined Detection System for Youcai Mosaic Virus and Its Application in Various Host Plants
Abstract
:1. Introduction
2. Materials and Methods
2.1. Construction of Recombinant CPYoMV-YZ Protein Expression Vector
2.2. Prokaryotic Expression and Purification of Recombinant CPYoMV-YZ Protein
2.3. Production of Polyclonal Antibody against His-CPYoMV-YZ Protein
2.4. Western Blotting, Immuno-Dot Blotting, and ELISA Detection of YoMV
2.5. RT-PCR for YoMV
2.6. S-RT-LAMP Assay for YoMV Using Prepared PAb-CPYoMV-YZ
2.7. Multiple Sequence Alignment and Phylogenetic Tree Construction of the CPYoMV Gene.
3. Results
3.1. Production of Recombinant CPYoMV-YZ Protein
3.2. Antiserum Production Using Recombinant His-CPYoMV-YZ Protein
3.3. Evaluation of Purified PAb-CPYoMV-YZ by Western Blotting and Immuno-Dot Blotting
3.4. Screening of YoMV-Positive and Negative Radish Plants by Molecular and Serological Methods
3.5. Establishment of ELISA and Immuno-Dot Blotting Assays for YoMV in Radish
3.6. Establishment of an S-RT-LAMP Assay Based on the CPYoMV-YZ Gene
3.7. Purified PAb-CPYoMV-YZ for YoMV Detection in a Wide Range of Plants
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Feng, C.; Hua, Y.; Liu, D.; Chen, H.; Wu, M.; Hua, J.; Zhang, K. Establishment of a Serology- and Molecular-Combined Detection System for Youcai Mosaic Virus and Its Application in Various Host Plants. Agronomy 2024, 14, 1900. https://doi.org/10.3390/agronomy14091900
Feng C, Hua Y, Liu D, Chen H, Wu M, Hua J, Zhang K. Establishment of a Serology- and Molecular-Combined Detection System for Youcai Mosaic Virus and Its Application in Various Host Plants. Agronomy. 2024; 14(9):1900. https://doi.org/10.3390/agronomy14091900
Chicago/Turabian StyleFeng, Chenwei, Yanhong Hua, Duxuan Liu, Haoyu Chen, Mingjie Wu, Jing Hua, and Kun Zhang. 2024. "Establishment of a Serology- and Molecular-Combined Detection System for Youcai Mosaic Virus and Its Application in Various Host Plants" Agronomy 14, no. 9: 1900. https://doi.org/10.3390/agronomy14091900