Complementation of CTB7 in the Maize Pathogen Cercospora zeina Overcomes the Lack of In Vitro Cercosporin Production
Autor: | J.J. Marion Meyer, Velushka Swart, John B. Ridenour, David Kenneth Berger, Nicholas Abraham Olivier, Burt H. Bluhm, B.G. Crampton |
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Rok vydání: | 2017 |
Předmět: |
0301 basic medicine
Transcription Genetic Physiology Agrobacterium 030106 microbiology Mutant Genes Fungal Zea mays Mass Spectrometry Microbiology Fungal Proteins 03 medical and health sciences Transformation Genetic Cercospora Ascomycota Gene Expression Regulation Fungal Gene cluster Computer Simulation Amino Acid Sequence DNA Fungal Gene Perylene Conserved Sequence biology Base Sequence Genetic Complementation Test Fungal genetics General Medicine biology.organism_classification Introns Biosynthetic Pathways Complementation Transformation (genetics) Alternative Splicing Multigene Family Oxidoreductases Agronomy and Crop Science Gene Deletion |
Zdroj: | Molecular plant-microbe interactions : MPMI. 30(9) |
ISSN: | 0894-0282 |
Popis: | Gray leaf spot (GLS), caused by the sibling species Cercospora zeina or Cercospora zeae-maydis, is cited as one of the most important diseases threatening global maize production. C. zeina fails to produce cercosporin in vitro and, in most cases, causes large coalescing lesions during maize infection, a symptom generally absent from cercosporin-deficient mutants in other Cercospora spp. Here, we describe the C. zeina cercosporin toxin biosynthetic (CTB) gene cluster. The oxidoreductase gene CTB7 contained several insertions and deletions as compared with the C. zeae-maydis ortholog. We set out to determine whether complementing the defective CTB7 gene with the full-length gene from C. zeae-maydis could confer in vitro cercosporin production. C. zeina transformants containing C. zeae-maydis CTB7 were generated by Agrobacterium tumefaciens–mediated transformation and were evaluated for in vitro cercosporin production. When grown on nitrogen-limited medium in the light—conditions conducive to cercosporin production in other Cercospora spp.—one transformant accumulated a red pigment that was confirmed to be cercosporin by the KOH assay, thin-layer chromatography, and ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry. Our results indicated that C. zeina has a defective CTB7, but all other necessary machinery required for synthesizing cercosporin-like molecules and, thus, C. zeina may produce a structural variant of cercosporin during maize infection. |
Databáze: | OpenAIRE |
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