Shedding light on AT1G29480 of Arabidopsis thaliana—An enigmatic locus restricted to Brassicacean genomes
Abstract A key feature of C4 Kranz anatomy is the presence of an enlarged, photosynthetically highly active bundle sheath whose cells contain large numbers of chloroplasts. With the aim to identify novel candidate regulators of C4 bundle sheath development, we performed an activation tagging screen...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wiley
2022-10-01
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| Series: | Plant Direct |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/pld3.455 |
| _version_ | 1797989742753611776 |
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| author | Kumari Billakurthi Stefanie Schulze Eva Lena Marie Schulz Tammy L. Sage Tina B. Schreier Julian M. Hibberd Martha Ludwig Peter Westhoff |
| author_facet | Kumari Billakurthi Stefanie Schulze Eva Lena Marie Schulz Tammy L. Sage Tina B. Schreier Julian M. Hibberd Martha Ludwig Peter Westhoff |
| author_sort | Kumari Billakurthi |
| collection | DOAJ |
| description | Abstract A key feature of C4 Kranz anatomy is the presence of an enlarged, photosynthetically highly active bundle sheath whose cells contain large numbers of chloroplasts. With the aim to identify novel candidate regulators of C4 bundle sheath development, we performed an activation tagging screen with Arabidopsis thaliana. The reporter gene used encoded a chloroplast‐targeted GFP protein preferentially expressed in the bundle sheath, and the promoter of the C4 phosphoenolpyruvate carboxylase gene from Flaveria trinervia served as activation tag because of its activity in all chlorenchymatous tissues of A. thaliana. Primary mutants were selected based on their GFP signal intensity, and one stable mutant named kb‐1 with a significant increase in GFP fluorescence intensity was obtained. Despite the increased GFP signal, kb‐1 showed no alterations to bundle sheath anatomy. The causal locus, AT1G29480, is specific to the Brassicaceae with its second exon being conserved. Overexpression and reconstitution studies confirmed that AT1G29480, and specifically its second exon, were sufficient for the enhanced GFP phenotype, which was not dependent on translation of the locus or its parts into protein. We conclude, therefore, that the AT1G29480 locus enhances the GFP reporter gene activity via an RNA‐based mechanism. |
| first_indexed | 2024-04-11T08:25:18Z |
| format | Article |
| id | doaj.art-131462a5ce0c4889a4e854e118e2614f |
| institution | Directory Open Access Journal |
| issn | 2475-4455 |
| language | English |
| last_indexed | 2024-04-11T08:25:18Z |
| publishDate | 2022-10-01 |
| publisher | Wiley |
| record_format | Article |
| series | Plant Direct |
| spelling | doaj.art-131462a5ce0c4889a4e854e118e2614f2022-12-22T04:34:47ZengWileyPlant Direct2475-44552022-10-01610n/an/a10.1002/pld3.455Shedding light on AT1G29480 of Arabidopsis thaliana—An enigmatic locus restricted to Brassicacean genomesKumari Billakurthi0Stefanie Schulze1Eva Lena Marie Schulz2Tammy L. Sage3Tina B. Schreier4Julian M. Hibberd5Martha Ludwig6Peter Westhoff7Institute of Plant Molecular and Developmental Biology Universitätsstrasse 1, Heinrich‐Heine‐University Duesseldorf GermanyInstitute of Plant Molecular and Developmental Biology Universitätsstrasse 1, Heinrich‐Heine‐University Duesseldorf GermanyInstitute of Plant Molecular and Developmental Biology Universitätsstrasse 1, Heinrich‐Heine‐University Duesseldorf GermanyDepartment of Ecology and Evolutionary Biology The University of Toronto Toronto Ontario CanadaDepartment of Plant Sciences, Downing Street University of Cambridge Cambridge UKDepartment of Plant Sciences, Downing Street University of Cambridge Cambridge UKSchool of Molecular Sciences University of Western Australia Perth Western Australia AustraliaInstitute of Plant Molecular and Developmental Biology Universitätsstrasse 1, Heinrich‐Heine‐University Duesseldorf GermanyAbstract A key feature of C4 Kranz anatomy is the presence of an enlarged, photosynthetically highly active bundle sheath whose cells contain large numbers of chloroplasts. With the aim to identify novel candidate regulators of C4 bundle sheath development, we performed an activation tagging screen with Arabidopsis thaliana. The reporter gene used encoded a chloroplast‐targeted GFP protein preferentially expressed in the bundle sheath, and the promoter of the C4 phosphoenolpyruvate carboxylase gene from Flaveria trinervia served as activation tag because of its activity in all chlorenchymatous tissues of A. thaliana. Primary mutants were selected based on their GFP signal intensity, and one stable mutant named kb‐1 with a significant increase in GFP fluorescence intensity was obtained. Despite the increased GFP signal, kb‐1 showed no alterations to bundle sheath anatomy. The causal locus, AT1G29480, is specific to the Brassicaceae with its second exon being conserved. Overexpression and reconstitution studies confirmed that AT1G29480, and specifically its second exon, were sufficient for the enhanced GFP phenotype, which was not dependent on translation of the locus or its parts into protein. We conclude, therefore, that the AT1G29480 locus enhances the GFP reporter gene activity via an RNA‐based mechanism.https://doi.org/10.1002/pld3.455activation taggingArabidopsis thalianaAT1G29480GFP reporter linenon‐coding RNA |
| spellingShingle | Kumari Billakurthi Stefanie Schulze Eva Lena Marie Schulz Tammy L. Sage Tina B. Schreier Julian M. Hibberd Martha Ludwig Peter Westhoff Shedding light on AT1G29480 of Arabidopsis thaliana—An enigmatic locus restricted to Brassicacean genomes Plant Direct activation tagging Arabidopsis thaliana AT1G29480 GFP reporter line non‐coding RNA |
| title | Shedding light on AT1G29480 of Arabidopsis thaliana—An enigmatic locus restricted to Brassicacean genomes |
| title_full | Shedding light on AT1G29480 of Arabidopsis thaliana—An enigmatic locus restricted to Brassicacean genomes |
| title_fullStr | Shedding light on AT1G29480 of Arabidopsis thaliana—An enigmatic locus restricted to Brassicacean genomes |
| title_full_unstemmed | Shedding light on AT1G29480 of Arabidopsis thaliana—An enigmatic locus restricted to Brassicacean genomes |
| title_short | Shedding light on AT1G29480 of Arabidopsis thaliana—An enigmatic locus restricted to Brassicacean genomes |
| title_sort | shedding light on at1g29480 of arabidopsis thaliana an enigmatic locus restricted to brassicacean genomes |
| topic | activation tagging Arabidopsis thaliana AT1G29480 GFP reporter line non‐coding RNA |
| url | https://doi.org/10.1002/pld3.455 |
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