Agrobacterium-mediated transient transformation of Flaveria bidentis leaves: a novel method to examine the evolution of C4 photosynthesis
Abstract The genus Flaveria has been studied extensively as a model for the evolution of C4 photosynthesis. Thus far, molecular analyses in this genus have been limited due to a dearth of genomic information and the lack of a rapid and efficient transformation protocol. Since their development, Agro...
Saved in:
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2024-12-01
|
| Series: | Plant Methods |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13007-024-01306-z |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850086640875208704 |
|---|---|
| author | Christopher J. Baros Jeremy Beerkens Martha Ludwig |
| author_facet | Christopher J. Baros Jeremy Beerkens Martha Ludwig |
| author_sort | Christopher J. Baros |
| collection | DOAJ |
| description | Abstract The genus Flaveria has been studied extensively as a model for the evolution of C4 photosynthesis. Thus far, molecular analyses in this genus have been limited due to a dearth of genomic information and the lack of a rapid and efficient transformation protocol. Since their development, Agrobacterium-mediated transient transformation protocols have been instrumental in understanding many biological processes in a range of plant species. However, this technique has not been applied to the genus Flaveria. Here, an efficient protocol for the Agrobacterium-mediated transient transformation of the leaves of the C4 species Flaveria bidentis is presented. This technique has the distinct advantages of rapid turnaround, the ability to co-transform with multiple constructs, and the capacity to assay coding and non-coding regions of Flaveria genomes in a homologous context. To illustrate the utility of this protocol, the quantitative transcriptional regulation of phosphoenolpyruvate carboxylase, the primary carboxylase of C4 plants, was investigated. A 24 bp region in the ppcA1 proximal promoter was found to elicit high levels of reporter gene expression. The Agrobacterium-mediated transient transformation of F. bidentis leaves will accelerate the understanding of the biology and evolution of C4 photosynthesis in the genus Flaveria as well as in other C4 lineages. |
| format | Article |
| id | doaj-art-b042e97864b6473190ce8fc179e65e88 |
| institution | DOAJ |
| issn | 1746-4811 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Plant Methods |
| spelling | doaj-art-b042e97864b6473190ce8fc179e65e882025-08-20T02:43:25ZengBMCPlant Methods1746-48112024-12-0120111410.1186/s13007-024-01306-zAgrobacterium-mediated transient transformation of Flaveria bidentis leaves: a novel method to examine the evolution of C4 photosynthesisChristopher J. Baros0Jeremy Beerkens1Martha Ludwig2School of Molecular Sciences, University of Western AustraliaSchool of Molecular Sciences, University of Western AustraliaSchool of Molecular Sciences, University of Western AustraliaAbstract The genus Flaveria has been studied extensively as a model for the evolution of C4 photosynthesis. Thus far, molecular analyses in this genus have been limited due to a dearth of genomic information and the lack of a rapid and efficient transformation protocol. Since their development, Agrobacterium-mediated transient transformation protocols have been instrumental in understanding many biological processes in a range of plant species. However, this technique has not been applied to the genus Flaveria. Here, an efficient protocol for the Agrobacterium-mediated transient transformation of the leaves of the C4 species Flaveria bidentis is presented. This technique has the distinct advantages of rapid turnaround, the ability to co-transform with multiple constructs, and the capacity to assay coding and non-coding regions of Flaveria genomes in a homologous context. To illustrate the utility of this protocol, the quantitative transcriptional regulation of phosphoenolpyruvate carboxylase, the primary carboxylase of C4 plants, was investigated. A 24 bp region in the ppcA1 proximal promoter was found to elicit high levels of reporter gene expression. The Agrobacterium-mediated transient transformation of F. bidentis leaves will accelerate the understanding of the biology and evolution of C4 photosynthesis in the genus Flaveria as well as in other C4 lineages.https://doi.org/10.1186/s13007-024-01306-zC4 photosynthesisEvolution of C4 photosynthesisFlaveriaPhosphoenolpyruvate carboxylaseTransient transformation |
| spellingShingle | Christopher J. Baros Jeremy Beerkens Martha Ludwig Agrobacterium-mediated transient transformation of Flaveria bidentis leaves: a novel method to examine the evolution of C4 photosynthesis Plant Methods C4 photosynthesis Evolution of C4 photosynthesis Flaveria Phosphoenolpyruvate carboxylase Transient transformation |
| title | Agrobacterium-mediated transient transformation of Flaveria bidentis leaves: a novel method to examine the evolution of C4 photosynthesis |
| title_full | Agrobacterium-mediated transient transformation of Flaveria bidentis leaves: a novel method to examine the evolution of C4 photosynthesis |
| title_fullStr | Agrobacterium-mediated transient transformation of Flaveria bidentis leaves: a novel method to examine the evolution of C4 photosynthesis |
| title_full_unstemmed | Agrobacterium-mediated transient transformation of Flaveria bidentis leaves: a novel method to examine the evolution of C4 photosynthesis |
| title_short | Agrobacterium-mediated transient transformation of Flaveria bidentis leaves: a novel method to examine the evolution of C4 photosynthesis |
| title_sort | agrobacterium mediated transient transformation of flaveria bidentis leaves a novel method to examine the evolution of c4 photosynthesis |
| topic | C4 photosynthesis Evolution of C4 photosynthesis Flaveria Phosphoenolpyruvate carboxylase Transient transformation |
| url | https://doi.org/10.1186/s13007-024-01306-z |
| work_keys_str_mv | AT christopherjbaros agrobacteriummediatedtransienttransformationofflaveriabidentisleavesanovelmethodtoexaminetheevolutionofc4photosynthesis AT jeremybeerkens agrobacteriummediatedtransienttransformationofflaveriabidentisleavesanovelmethodtoexaminetheevolutionofc4photosynthesis AT marthaludwig agrobacteriummediatedtransienttransformationofflaveriabidentisleavesanovelmethodtoexaminetheevolutionofc4photosynthesis |