MoDHX35, a DEAH-Box Protein, Is Required for Appressoria Formation and Full Virulence of the Rice Blast Fungus, Magnaporthe oryzae
Abstract
:1. Introduction
2. Results
2.1. Isolation and Analysis of the Gene Locus Inserted by T-DNA
2.2. MoDHX35 Is Up-Regulated during Appressoria Formation
2.3. Gene Replacement of MoDHX35 and Mutant Recovery
2.4. Loss of MoDHX35 Increases the Sensitivity of the Mutant to Doxorubicin
2.5. MoDHX35 Contributes to M. oryzae Appressorium Formation
2.6. MoDHX35 Is Required for M. oryzae Pathogenicity
2.7. Deletion of MoDHX35 Does Not Affect the Sexual Development of the Fungus
2.8. MoDHX35 Mutants Are Not Temperature Sensitive
2.9. Deletion of MoDHX35 Does Not Alter the Nutritional Utilization, Osmic STRESS or Resistance to Chemicals of the Fungus
3. Discussion
4. Material and Methods
4.1. Fungal Strains and Growth Conditions
4.2. MoDHX35 Isolation and Sequence Analysis
4.3. Vector Construction, Gene Deletion and Mutant Complementation
4.4. Vegetable Growth and Conidiation on Culture Media
4.5. Assay for Conidial Germination and Appressorial Formation
4.6. Pathogenicity Tests
4.7. Nucleic Acid Manipulations and Quantitative RT-PCR
4.8. Sexual Reproduction
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Strains | Description |
---|---|
Guy11 | Wild type |
MoDHX35-6 | Null mutant |
MoDHX35-7 | Null mutant |
MoDHX35-8 | Null mutant |
MoDHX35-9 | Null mutant |
MoDHX35-5 | Random-inserted-transformant |
MoDHX35-9-10 | Complementary transformant |
MoDHX35-9-16 | Complementary transformant |
Primer Name | Primer Sequence |
---|---|
MoDHX35-CDS-F1 | CGGAATTCTTTTACCTGGTAAAAGTVGGGCGC |
MoDHX35-CDS-R1 | TCCCCCGGGATGGCCGACTTTGATCTGGGTGCA |
MoDHX35-Com-F1 | GCTCTAGAAGCTCAAGCAGACAGACCTAGTTG |
MoDHX35-Com-R1 | TCCCCCGGGGACGATAATGTCAATCTCCGAGGA |
MoDHX35-Up-F1 | GATGGRGCGCCGGATGTGATGAC |
MoDHX35-Up-R1 | GTATTGATTATTTGGGAGGCTTCT |
MoDHX35-Down-F1 | CGGGATCCAAGTACCCTGCGGAAACATC |
MoDHX35-Down-R1 | AAGTTTCTCTCCTCCAAAAAGCTT |
MoDHX35-Genecheck-F1 | CAGTGACAGTTGTCGTTGGA |
MoDHX35-Genecheck-R1 | GTGCTGGATTCTAGTTCTAG |
HPH-Check-F1 | TGGAGGTCAACACATCAATGCTATT |
HPH-Check-R1 | CTACTCTATTCCTTTGCCCTCGGAC |
Sequence-Up | ACTCGCCGATAGTGGAAACC |
MoDHX35-Upcheck-F1 | CGGTCTGTATTTGGCATACG |
Sequence-Down | CCAGTTGCCTAAATGAACCA |
MoDHX35-Downcheck-R1 | TCCTCGTTGTGCAGAGTCTT |
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Ying, S.; Zhang, Z.; Zhang, Y.; Hao, Z.; Chai, R.; Qiu, H.; Wang, Y.; Zhu, X.; Wang, J.; Sun, G.; et al. MoDHX35, a DEAH-Box Protein, Is Required for Appressoria Formation and Full Virulence of the Rice Blast Fungus, Magnaporthe oryzae. Int. J. Mol. Sci. 2022, 23, 9015. https://doi.org/10.3390/ijms23169015
Ying S, Zhang Z, Zhang Y, Hao Z, Chai R, Qiu H, Wang Y, Zhu X, Wang J, Sun G, et al. MoDHX35, a DEAH-Box Protein, Is Required for Appressoria Formation and Full Virulence of the Rice Blast Fungus, Magnaporthe oryzae. International Journal of Molecular Sciences. 2022; 23(16):9015. https://doi.org/10.3390/ijms23169015
Chicago/Turabian StyleYing, Shumin, Zhen Zhang, Yanan Zhang, Zhongna Hao, Rongyao Chai, Haiping Qiu, Yanli Wang, Xueming Zhu, Jiaoyu Wang, Guochang Sun, and et al. 2022. "MoDHX35, a DEAH-Box Protein, Is Required for Appressoria Formation and Full Virulence of the Rice Blast Fungus, Magnaporthe oryzae" International Journal of Molecular Sciences 23, no. 16: 9015. https://doi.org/10.3390/ijms23169015