Overexpression of a 3-ketoacyl-CoA thiolase in Leptosphaeria maculans causes reduced pathogenicity on Brassica napus
- PMID: 16776292
- DOI: 10.1094/MPMI-19-0588
Overexpression of a 3-ketoacyl-CoA thiolase in Leptosphaeria maculans causes reduced pathogenicity on Brassica napus
Abstract
Agrobacterium tumefaciens-mediated random mutagenesis was used to generate insertional mutants of the fungus Leptosphaeria maculans. Of 91 transformants screened, only one (A3) produced lesions of reduced size on cotyledons of canola (Brassica napus). Genes flanking the T-DNA insertion had the best matches to an alcohol dehydrogenase class 4 (ADH4)-like gene (Adh4L) and a 3-ketoacyl-CoA thiolase gene (Thiol) and were expressed in mutant A3 in vitro and in planta at significantly higher levels than in the wild type. This is the first report of a T-DNA insertion in fungi causing increased gene expression. Transformants of the wild-type isolate expressing both Adh4L and Thiol under the control of a heterologous promoter had similar pathogenicity to mutant A3. Ectopic expression of only thiolase resulted in loss of pathogenicity, suggesting that thiolase overexpression was primarily responsible for the reduced pathogenicity of the A3 isolate. The thiolase gene encoded a functional protein, as shown by assays in which a nontoxic substrate (2, 4 dichlorophenoxybutyric acid) was converted to a toxic product. The use of a translational fusion with a reporter gene showed thiolase expressed in organelles that are most likely peroxisomes.
Similar articles
-
A key enzyme of the Leloir pathway is involved in pathogenicity of Leptosphaeria maculans toward oilseed rape.Mol Plant Microbe Interact. 2009 Jun;22(6):725-36. doi: 10.1094/MPMI-22-6-0725. Mol Plant Microbe Interact. 2009. PMID: 19445597
-
The Lmpma1 gene of Leptosphaeria maculans encodes a plasma membrane H+-ATPase isoform essential for pathogenicity towards oilseed rape.Fungal Genet Biol. 2008 Jul;45(7):1122-34. doi: 10.1016/j.fgb.2008.04.008. Fungal Genet Biol. 2008. PMID: 18538267
-
Sit4-Associated Protein is Required for Pathogenicity of Leptosphaeria maculans on Brassica napus.Curr Microbiol. 2017 Dec;74(12):1438-1446. doi: 10.1007/s00284-017-1338-3. Epub 2017 Aug 24. Curr Microbiol. 2017. PMID: 28840344
-
Leptosphaeria maculans, the causal agent of blackleg disease of Brassicas.Fungal Genet Biol. 2001 Jun;33(1):1-14. doi: 10.1006/fgbi.2001.1274. Fungal Genet Biol. 2001. PMID: 11407881 Review.
-
Genome analysis of the fungal plant pathogen, Leptosphaeria maculans using pulsed field gel electrophoresis.Electrophoresis. 1997 Aug;18(9):1544-7. doi: 10.1002/elps.1150180910. Electrophoresis. 1997. PMID: 9378119 Review.
Cited by
-
Peroxisome dynamics determines host-derived ROS accumulation and infectious growth of the rice blast fungus.mBio. 2023 Dec 19;14(6):e0238123. doi: 10.1128/mbio.02381-23. Epub 2023 Nov 15. mBio. 2023. PMID: 37966176 Free PMC article.
-
Large-scale transcriptomics to dissect 2 years of the life of a fungal phytopathogen interacting with its host plant.BMC Biol. 2021 Mar 23;19(1):55. doi: 10.1186/s12915-021-00989-3. BMC Biol. 2021. PMID: 33757516 Free PMC article.
-
The fungal gene cluster for biosynthesis of the antibacterial agent viriditoxin.Fungal Biol Biotechnol. 2019 Jul 1;6:2. doi: 10.1186/s40694-019-0072-y. eCollection 2019. Fungal Biol Biotechnol. 2019. PMID: 31304040 Free PMC article.
-
Analysis of Repeat Induced Point (RIP) Mutations in Leptosphaeria maculans Indicates Variability in the RIP Process Between Fungal Species.Genetics. 2019 Jan;211(1):89-104. doi: 10.1534/genetics.118.301712. Epub 2018 Nov 2. Genetics. 2019. PMID: 30389803 Free PMC article.
-
Spontaneous and CRISPR/Cas9-induced mutation of the osmosensor histidine kinase of the canola pathogen Leptosphaeria maculans.Fungal Biol Biotechnol. 2017 Dec 16;4:12. doi: 10.1186/s40694-017-0043-0. eCollection 2017. Fungal Biol Biotechnol. 2017. PMID: 29270298 Free PMC article.
Publication types
MeSH terms
Substances
Associated data
- Actions
LinkOut - more resources
Full Text Sources
Other Literature Sources