TY  - JOUR
AU  - Khamis, Galal
AU  - Chi, Jun
AU  - Zhao, Qingshun
PY  - 2024
DA  - 2024/06/18
TI  - Establishment of an <i>In Vitro</i> Propagation, Transformation, and Gene Editing System in Tomato (<i>Solanum lycopersicum</i>)
JO  - OBM Genetics
SP  - 244
VL  - 08
IS  - 02
AB  - Tomato (Solanum lycopersicum), a family of Solanaceae, is an annual crop grown in fields and greenhouses. Tomato is the fourth-largest agricultural crop in the world, with a market value of more than $50 billion. Numerous attempts were conducted on the tomato for further improvement via in vitro regeneration and transformation. This study aims to establish in vitro multiplication, transformation, and gene editing systems in tomatoes by using cotyledon explants. Cotyledon explants were placed on the MS medium supported with 6-benzyl adenine (BA) and kinetin (Kin) at four different concentrations for shoot induction. The shoot induction from cotyledon explants significantly increased, and the regeneration rate was 45% after using BA at 8.8 µM compared to Kin. For root induction, indole-3-butyric acid (IBA) at 4.8 µM concentration revealed the highest number of explants producing root (55%). Three Agrobacterium tumefaciens strains (EHA105, GV3101, and LBA4404) were used to establish and develop a tomato transformation method. Each Agrobacterium strain harbors the plasmid pCAMBIA2301, including the nptII marker. Agrobacterium strain LBA4404 showed the lowest number of dead explants, besides increasing the number of explant-produced shoots (30%) and the number of shoots per transformed explants compared to GV3101 and EHA105 strains. Our results revealed that the success of tomato transformation depends on selecting a suitable A. tumefaciens strain. Finally, the tomato gene editing system was successfully established by transferring the pCAMBIA1300-pCas9-sgRNA-U3 binary vector into the cotyledon using the selective Agrobacterium strain LBA4404, and the transformation effectivity was confirmed by PCR analysis.
SN  - 2577-5790
UR  - https://doi.org/10.21926/obm.genet.2402244
DO  - 10.21926/obm.genet.2402244
ID  - Khamis2024
ER  -