Agrobacterium-MEDIATED GENETIC TRANSFORMATION OF SEAWEED Kappaphycus alvarezii USING Gα GENE AND CALLUS CULTURES
DOI:
https://doi.org/10.11598/btb.2019.26.1.989Keywords:
Agrobacterium tumefaciens, Gα gene, Kappaphycus alvarezii, transgenic plantletsAbstract
Cottonii seaweed (Kappaphycus alvarezii Doty) is one of the most important commercial sources of carrageenans which are widely used in the pharmaceuticals and food industries. A problem in the cultivation of this seaweed is the ice-ice disease, which is caused by extreme changes in environmental conditions such as temperature and seawater salinity. Gene transformation to produce Cottonii seaweed transgenics that are tolerant to environmental stress is a potential solution to this problem. Gα gene encodes for the heterotrimeric G protein α subunit is a gene that plays a role in tolerance to biotic and abiotic environmental stresses. This study aimed to: (a) introduce the Gα gene into the callus cells of K. alvarezii and regenerate transformed callus cells to transgenic plantlets; (b) determine the appropriate concentration of acetosyringone and Agrobacterium tumefaciens strain for gene transfer into the callus of K. alvarezii. The callus cells of K. alvarezii were transformed using Agrobacterium tumefaciens strains LBA4404 and EHA105 carrying the expression vector pGWB502-Gα with a CaMV-35S promoter. The calli and A. tumefaciens were co-cultivated in several concentrations of acetosyringone (20, 40, 60 mg/L). The regeneration of transformed callus cells into transgenic plantlets was successfully performed using the somatic embryogenesis technique. The results showed that the highest percentage of putative transgenic micropropagule formation occurred at the 20-40 mg/L concentration of acetosyringone. Polymerase chain reaction (PCR) analysis on the twenty transgenic plantlets indicated that the Gα gene was successfully introduced into the genomic DNA of all of them. The highest transformation efficiency was in the co-cultivation treatment of 20-40 mg/L acetosyringone (22-28%). The transformation efficiency produced by Agrobacterium tumefaciens EHA105 (23%) was not significantly different from that produced by the LBA4404 (15%).Downloads
Download data is not yet available.
Downloads
Published
2018-11-16
How to Cite
Sulistiani, E., Suharsono, S., Supena, E. D. J., & Miftahudin, M. (2018). Agrobacterium-MEDIATED GENETIC TRANSFORMATION OF SEAWEED Kappaphycus alvarezii USING Gα GENE AND CALLUS CULTURES. BIOTROPIA - The Southeast Asian Journal of Tropical Biology, 26(1), 41–51. https://doi.org/10.11598/btb.2019.26.1.989
Issue
Section
Research Paper
License
Authors who publish with this journal agree with the following terms:
- Authors retain copyright and grant the journal right of first publication, with the work 1 year after publication simultaneously licensed under a Creative Commons attribution-noncommerical-noderivates 4.0 International License that allows others to share, copy and redistribute the work in any medium or format, but only where the use is for non-commercial purposes and an acknowledgement of the work's authorship and initial publication in this journal is mentioned.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).