The Retromer Subunit CfVps29 Is Involved in the Growth, Development, and Pathogenicity of Colletotrichum fructicola
Abstract
:1. Introduction
2. Materials and Methods
2.1. Strains and Culture Conditions
2.2. Phylogenetic Analysis and Domain Prediction
2.3. Targeted Gene Deletion and Complementation
2.4. Growth, Conidiation, and Appressoria Formation
2.5. Pathogenicity Assays
2.6. Cell Wall Integrity Assays
2.7. Real-Time Quantitative Reverse Transcription PCR Analysis
2.8. Statistical Analysis
3. Results
3.1. Identification and Phylogenetic Analysis of CfVps29
3.2. Generation of VPS29 Gene Deletion and Complemented Strains
3.3. CfVPS29 Is Required for Development and Pathogenicity
3.4. The CfVPS29 Gene Is Involved in the Maintenance of Cell Wall Integrity
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Primer | Sequence (5′–3′) | Purpose |
---|---|---|
CfVPS29-1F | GGACAGAAGATTACACTGAG | amplify CfVPS29 5′ flank sequence |
CfVPS29-2R | TTGACCTCCACTAGCTCCAGCCAAGCCGGTCGTTAGGGGTGTGTATA | amplify CfVPS29 5′ flank sequence |
CfVPS29-3F | CAAAGGAATAGAGTAGATGCCGACCGGCCGTCATCATGCAAGACGA | amplify CfVPS29 3′ flank sequence |
CfVPS29-4R | ACTGTCACGATCAAGCGCGA | amplify CfVPS29 3′ flank sequence |
CfVPS29-5F | CATGTGTGCCTATGGCGTCA | validation of CfVPS29 gene deletion |
H855R | GCTGATCTGACCAGTTGC | validation of CfVPS29 gene deletion |
CfVPS29-7F | GCGCTCTTGATATCCCCCAA | validation of CfVPS29 gene deletion |
CfVPS29-8R | CACGGGCTTCGTGTAAGTCA | validation of CfVPS29 gene deletion |
CfVPS29-9F | ACTCACTATAGGGCGAATTGGGTACTCAAATTGGTTGATAACACGGACCTGTAGTG | amplify complemented sequence |
CfVPS29-10R | CACCACCCCGGTGAACAGCTCCTCGCCCTTGCTCACTGATGTTGCAGACGGCTCCA | amplify complemented sequence |
Hyg F | GGCTTGGCTGGAGCTAGTGGAGGTCAA | amplify HPH sequence |
Hyg R | CGGTCGGCATCTACTCTATTCCTTTG | amplify HPH sequence |
GFP-R | GACACGCTGAACTTGTGGCCGTT | validation of complemented sequence |
CHS2-F | TCCGCCCCTCTGATTCCTAA | RT-qPCR |
CHS2-R | ACATGAAGGAAGCCGCGTAA | RT-qPCR |
CHS4-F | GAACATCGAGATGGCGCAAC | RT-qPCR |
CHS4-R | CTCGCCGGACTCAGGTATTC | RT-qPCR |
CHS5-F | CCCACAAGATGACGGACCTC | RT-qPCR |
CHS5-R | GCGTCGAGGTAGAACTTGGT | RT-qPCR |
CHS6-F | CAGTCTTGCCGCCTACATCA | RT-qPCR |
CHS6-R | GTCGGCGTAGGAGTAAGCTC | RT-qPCR |
CHS7-F | GCAAATTCACCGCTGTTGGT | RT-qPCR |
CHS7-R | CAGCATACACGGAGAAGCCA | RT-qPCR |
ACTIN-F | CCCCATCTACGAGGGTTTCG | RT-qPCR |
ACTIN-R | CGTCAGGAAGCTCGTAGGAC | RT-qPCR |
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Li, S.; Li, X.; Li, H. The Retromer Subunit CfVps29 Is Involved in the Growth, Development, and Pathogenicity of Colletotrichum fructicola. J. Fungi 2022, 8, 835. https://doi.org/10.3390/jof8080835
Li S, Li X, Li H. The Retromer Subunit CfVps29 Is Involved in the Growth, Development, and Pathogenicity of Colletotrichum fructicola. Journal of Fungi. 2022; 8(8):835. https://doi.org/10.3390/jof8080835
Chicago/Turabian StyleLi, Sizheng, Xiya Li, and He Li. 2022. "The Retromer Subunit CfVps29 Is Involved in the Growth, Development, and Pathogenicity of Colletotrichum fructicola" Journal of Fungi 8, no. 8: 835. https://doi.org/10.3390/jof8080835