Deficiency of ChPks and ChThr1 Inhibited DHN-Melanin Biosynthesis, Disrupted Cell Wall Integrity and Attenuated Pathogenicity in Colletotrichum higginsianum
Abstract
:1. Introduction
2. Results
2.1. Identification and Characterization of ChPks and ChThr1 in C. higginsianum
2.2. Expression of Genes ChPKS and ChTHR1 Is Up-Regulated during Melanization
2.3. ChPks and ChThr1 Are Involved in DHN-Melanin Biosynthesis
2.4. Melanin Is Essential for Cell Wall Integrity in C. higginsianum
2.5. Analysis of Conidiation, Appressorium Formation and Morphology, Turgor Pressure in ChpksΔ and Chthr1Δ Mutants
2.6. ChPks and ChThr1 Are Enssential for Pathogenicity in C. higginsianum
3. Discussion
4. Materials and Methods
4.1. Strains, Plant and Culture Conditions
4.2. Bioinformatics Analysis of ChPks and ChThr1
4.3. Expression Pattern Analysis of Genes ChPKS and ChTHR1
4.4. Deletion and Complementation of ChPKS and ChTHR1
4.5. Phenotypic Analysis
4.6. Transmission Electron and Confocal Microscopy
4.7. Pathogenicity Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Primer Name | Sequence (5′~3′) |
---|---|
ACTIN | CCCCAAGTCCAACAGAGAGA |
CATCAGGTAGTCGGTCAAGTCA | |
β-TUBULIN | GCCCTATTCTCGCTCGTCTTCC |
GGGCTCCAAATCGCAGTAAATG | |
PKS-qP | GTTCAAGTGGTTCTCATGGCTT |
CTGGCGATGGGGATGTAATTAG | |
THR1-qP | ACTTTGTCATCTCCAACTCGG |
ATGGAGGAGGTGAGGAGGAT | |
THNR-qP | GCACCATCGAGACATTTGT |
GGTACATGTCGGTCTTGATG | |
PKS-UP | ACGACGGCCAGTGCCAAGCTTCTGCTTTCAAACTCGTTTCACG |
GACCTGCAGGCATGCAAGCTCAAGAACAAGATTTCTGGATGTCG | |
PKS-DS | GACTCTAGAACTAGTGGATCCGGAACTTCATAGCGAAGCATATGA |
AGCTCGGTACCCGGGGGATCCGGAGTATTCGGGGAAGGAGAAT | |
THR1-UP | ACGACGGCCAGTGCCAAGCTTTTAAGCCAAAAAACGACTTGATAGA |
GACCTGCAGGCATGCAAGCTTAGTTGCTATGAAAAGACTGTGGCG | |
THR1-DS | CCGGGTACCGAGCTCGAATTCGTGCGATGATGCCACAAACTC |
TATGGAGAAACTCGAGAATTCGACACCGGGAGAGGGGAGG | |
PKSc-F1 | GAAACTCGAGCTCGAGAATTCATCGGTACCCTATCTGGCTGC |
CCGGGTACCGAGCTCGAATTCGCTCGCTGGTTACCTCGTTCG | |
PKSc-F2 | GACTCTAGATCTAGAGTCGACCTGCTTTCAAACTCGTTTCACG |
CTTGCATGCCTGCAGGTCGACTTACTCGCAAACCGCTTCACG | |
PKSc-F3 | GACCTGCAGGCATGCAAGCTTGGAACTTCATAGCGAAGCATATGA |
ACGACGGCCAGTGCCAAGCTTGTAACGACAAAAAAAGCCACAGG | |
THR1c-F1 | GAAACTCGAGCTCGAGAATTCCTCTAGCCGAAGGTATGAACCG |
CCGGGTACCGAGCTCGAATTCCCGCCGCCACGGAGGGGC | |
THR1c-F2 | ACTCTAGATCTAGAGTCGACCGGTGGCGGCGGCATGGTCGCG |
TCACACCAGATCCGCCTGTGCCGCCACCGGTGACC | |
THR1c-F3 | CACAGGCGGATCTGGTGTGAGCA |
GGCATCATCGCACTTACTTGTACAGCTCGTCCATGCC | |
THR1c-F4 | CAAGTAAGTGCGATGATGCCACAAACTC |
CTTGCATGCCTGCAGGTCGACCCGACATCCGGGGAACAC | |
PKS | GCACCGGTACCCAGG |
GGTAGATGGTGTCGCTGG | |
THR1 | ATGGCGCCCTCAGCGACTGAGAA |
CCTAGATATAAGCTATGTTAGGTATCCC | |
HPH1 | GGGCGTCGGTTTCCACTAT |
GATATGTCCTGCGGGTAAATAGC | |
NeoR | GATAGAAGGCGATGCGCT |
ACCCGGTCATACCTTCTTAAG | |
PKS-probe | CACAGGACATTACATGACGCA |
CACCCTTGCTTATAACTTCGCA | |
THR1-probe | ACATAGCTTATATCTAGGACAGGCG |
TGTTACTAAACCTAGCTTGGGAAGG |
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Duan, L.; Wang, L.; Chen, W.; He, Z.; Zhou, E.; Zhu, Y. Deficiency of ChPks and ChThr1 Inhibited DHN-Melanin Biosynthesis, Disrupted Cell Wall Integrity and Attenuated Pathogenicity in Colletotrichum higginsianum. Int. J. Mol. Sci. 2023, 24, 15890. https://doi.org/10.3390/ijms242115890
Duan L, Wang L, Chen W, He Z, Zhou E, Zhu Y. Deficiency of ChPks and ChThr1 Inhibited DHN-Melanin Biosynthesis, Disrupted Cell Wall Integrity and Attenuated Pathogenicity in Colletotrichum higginsianum. International Journal of Molecular Sciences. 2023; 24(21):15890. https://doi.org/10.3390/ijms242115890
Chicago/Turabian StyleDuan, Lingtao, Li Wang, Weilun Chen, Zhenrui He, Erxun Zhou, and Yiming Zhu. 2023. "Deficiency of ChPks and ChThr1 Inhibited DHN-Melanin Biosynthesis, Disrupted Cell Wall Integrity and Attenuated Pathogenicity in Colletotrichum higginsianum" International Journal of Molecular Sciences 24, no. 21: 15890. https://doi.org/10.3390/ijms242115890