Studying the Function of Phytoplasma Effector Proteins Using a Chemical-Inducible Expression System in Transgenic Plants
Phytoplasmas are bacterial pathogens that live mainly in the phloem of their plant hosts. They dramatically manipulate plant development by secreting effector proteins that target developmental proteins of their hosts. Traditionally, the effects of individual effector proteins have been studied by e...
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MDPI AG
2021-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/24/13582 |
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| author | Keziah M. Omenge Florian Rümpler Subha Suvetha Kathalingam Alexandra C. U. Furch Günter Theißen |
| author_facet | Keziah M. Omenge Florian Rümpler Subha Suvetha Kathalingam Alexandra C. U. Furch Günter Theißen |
| author_sort | Keziah M. Omenge |
| collection | DOAJ |
| description | Phytoplasmas are bacterial pathogens that live mainly in the phloem of their plant hosts. They dramatically manipulate plant development by secreting effector proteins that target developmental proteins of their hosts. Traditionally, the effects of individual effector proteins have been studied by ectopic overexpression using strong, ubiquitously active promoters in transgenic model plants. However, the impact of phytoplasma infection on the host plants depends on the intensity and timing of infection with respect to the developmental stage of the host. To facilitate investigations addressing the timing of effector protein activity, we have established chemical-inducible expression systems for the three most well-characterized phytoplasma effector proteins, SECRETED ASTER YELLOWS WITCHES’ BROOM PROTEIN 11 (SAP11), SAP54 and TENGU in transgenic <i>Arabidopsis thaliana</i>. We induced gene expression either continuously, or at germination stage, seedling stage, or flowering stage. mRNA expression was determined by quantitative reverse transcription PCR, protein accumulation by confocal laser scanning microscopy of GFP fusion proteins. Our data reveal tight regulation of effector gene expression and strong upregulation after induction. Phenotypic analyses showed differences in disease phenotypes depending on the timing of induction. Comparative phenotype analysis revealed so far unreported similarities in disease phenotypes, with all three effector proteins interfering with flower development and shoot branching, indicating a surprising functional redundancy of SAP54, SAP11 and TENGU. However, subtle but mechanistically important differences were also observed, especially affecting the branching pattern of the plants. |
| first_indexed | 2024-03-10T03:54:20Z |
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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T03:54:20Z |
| publishDate | 2021-12-01 |
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| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-1705e7d02ae14521bf2e8d64f8f2bd7c2023-11-23T08:48:30ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-12-0122241358210.3390/ijms222413582Studying the Function of Phytoplasma Effector Proteins Using a Chemical-Inducible Expression System in Transgenic PlantsKeziah M. Omenge0Florian Rümpler1Subha Suvetha Kathalingam2Alexandra C. U. Furch3Günter Theißen4Department of Genetics, Matthias Schleiden Institute for Genetics, Bioinformatics and Molecular Botany, Friedrich Schiller University Jena, Philosophenweg 12, 07743 Jena, GermanyDepartment of Genetics, Matthias Schleiden Institute for Genetics, Bioinformatics and Molecular Botany, Friedrich Schiller University Jena, Philosophenweg 12, 07743 Jena, GermanyDepartment of Genetics, Matthias Schleiden Institute for Genetics, Bioinformatics and Molecular Botany, Friedrich Schiller University Jena, Philosophenweg 12, 07743 Jena, GermanyDepartment of Plant Physiology, Matthias Schleiden Institute for Genetics, Bioinformatics and Molecular Botany, Friedrich Schiller University Jena, Dornburger Str. 159, 07743 Jena, GermanyDepartment of Genetics, Matthias Schleiden Institute for Genetics, Bioinformatics and Molecular Botany, Friedrich Schiller University Jena, Philosophenweg 12, 07743 Jena, GermanyPhytoplasmas are bacterial pathogens that live mainly in the phloem of their plant hosts. They dramatically manipulate plant development by secreting effector proteins that target developmental proteins of their hosts. Traditionally, the effects of individual effector proteins have been studied by ectopic overexpression using strong, ubiquitously active promoters in transgenic model plants. However, the impact of phytoplasma infection on the host plants depends on the intensity and timing of infection with respect to the developmental stage of the host. To facilitate investigations addressing the timing of effector protein activity, we have established chemical-inducible expression systems for the three most well-characterized phytoplasma effector proteins, SECRETED ASTER YELLOWS WITCHES’ BROOM PROTEIN 11 (SAP11), SAP54 and TENGU in transgenic <i>Arabidopsis thaliana</i>. We induced gene expression either continuously, or at germination stage, seedling stage, or flowering stage. mRNA expression was determined by quantitative reverse transcription PCR, protein accumulation by confocal laser scanning microscopy of GFP fusion proteins. Our data reveal tight regulation of effector gene expression and strong upregulation after induction. Phenotypic analyses showed differences in disease phenotypes depending on the timing of induction. Comparative phenotype analysis revealed so far unreported similarities in disease phenotypes, with all three effector proteins interfering with flower development and shoot branching, indicating a surprising functional redundancy of SAP54, SAP11 and TENGU. However, subtle but mechanistically important differences were also observed, especially affecting the branching pattern of the plants.https://www.mdpi.com/1422-0067/22/24/13582<i>Arabidopsis thaliana</i>effector proteininducible gene expressionMADS-box genephytoplasmatranscription factor |
| spellingShingle | Keziah M. Omenge Florian Rümpler Subha Suvetha Kathalingam Alexandra C. U. Furch Günter Theißen Studying the Function of Phytoplasma Effector Proteins Using a Chemical-Inducible Expression System in Transgenic Plants International Journal of Molecular Sciences <i>Arabidopsis thaliana</i> effector protein inducible gene expression MADS-box gene phytoplasma transcription factor |
| title | Studying the Function of Phytoplasma Effector Proteins Using a Chemical-Inducible Expression System in Transgenic Plants |
| title_full | Studying the Function of Phytoplasma Effector Proteins Using a Chemical-Inducible Expression System in Transgenic Plants |
| title_fullStr | Studying the Function of Phytoplasma Effector Proteins Using a Chemical-Inducible Expression System in Transgenic Plants |
| title_full_unstemmed | Studying the Function of Phytoplasma Effector Proteins Using a Chemical-Inducible Expression System in Transgenic Plants |
| title_short | Studying the Function of Phytoplasma Effector Proteins Using a Chemical-Inducible Expression System in Transgenic Plants |
| title_sort | studying the function of phytoplasma effector proteins using a chemical inducible expression system in transgenic plants |
| topic | <i>Arabidopsis thaliana</i> effector protein inducible gene expression MADS-box gene phytoplasma transcription factor |
| url | https://www.mdpi.com/1422-0067/22/24/13582 |
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