Version 1
: Received: 23 September 2024 / Approved: 23 September 2024 / Online: 24 September 2024 (09:24:17 CEST)
How to cite:
Molina, D.; Acuña, R. Optimizing the Genetic Transformation of Coffea arabica Using Agrobacterium tumefaciens. Preprints2024, 2024091827. https://doi.org/10.20944/preprints202409.1827.v1
Molina, D.; Acuña, R. Optimizing the Genetic Transformation of Coffea arabica Using Agrobacterium tumefaciens. Preprints 2024, 2024091827. https://doi.org/10.20944/preprints202409.1827.v1
Molina, D.; Acuña, R. Optimizing the Genetic Transformation of Coffea arabica Using Agrobacterium tumefaciens. Preprints2024, 2024091827. https://doi.org/10.20944/preprints202409.1827.v1
APA Style
Molina, D., & Acuña, R. (2024). Optimizing the Genetic Transformation of <em>Coffea arabica</em> Using <em>Agrobacterium tumefaciens</em>. Preprints. https://doi.org/10.20944/preprints202409.1827.v1
Chicago/Turabian Style
Molina, D. and Ricardo Acuña. 2024 "Optimizing the Genetic Transformation of <em>Coffea arabica</em> Using <em>Agrobacterium tumefaciens</em>" Preprints. https://doi.org/10.20944/preprints202409.1827.v1
Abstract
The genetic transformation of Coffea arabica L. is an alternative strategy for obtaining plants with agronomic traits of interest that is less time-consuming than conventional breeding methods. Given the importance of coffee cultivation in Colombia, this study evaluated the main factors interfering with the genetic transformation of C. arabica using Agrobacterium tumefaciens. An effi-cient and reproducible method was accordingly established that involved the propagation of “early” embryogenic tissue in a liquid proliferation medium, supplemented with 3 mg l−1 BAP for eight months, which was subsequently sonicated for 300 sec in a suspension of LBA4405 OD600 of 0.5 harboring pC1301 and incubated in this same suspension for 1 hr. This plasmid contained the uidA gene under control of the 35S promoter. The Agrobacterium suspen-sion was subsequently removed from the embryogenic tissue using a micropipette, after which the tissue was deposited on filter paper to remove the remaining Agrobacterium suspension. The embryogenic tissue was co-cultured for four days in a solid differentiation medium supple-mented with 100 µM acetosyringone on filter paper. Subsequently, the tissue was post-cultured for four days in liquid differentiation medium under orbital shaking at 100 rpm with 300 mg l−1 Claforan (Hoechst), followed by selection with 50 mg l-1 hygromycin at 26 °C in the dark, with subcultures at 20-day intervals until somatic embryos were formed for subsequent culturing in germination medium. Molecular analysis confirmed the presence of the uidA gene in coffee seedlings transformed with strains LBA4405 and EHA105 and vectors pC1301 and pC2301. This method successfully enables the stable integration of genes of interest in the coffee plant ge-nome
Biology and Life Sciences, Biology and Biotechnology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
