Garuda - Garba Rujukan Digital

Sulistiani, Erina

Unknown Affiliation

Author-ID : 1123057

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Immunology & microbiology Veterinary

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CALLUS INDUCTION OF COTTONI SEAWEED (Kappaphycus alvarezii (Doty) Doty) COLLECTED FROM NATUNA ISLANDS, RIAU ISLANDS PROVINCE Sulistiani, Erina
BIOTROPIA Vol. 19 No. 2 (2012): BIOTROPIA Vol. 19 No. 2 December 2012
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11598/btb.2012.19.2.254

The objective of this study was to obtain the optimal medium for callus induction from thallus explants off Kappaphycus alvarezii (Doty) and to regenerate filamentous callus from induced callus. Before cultured cottonii seaweeds collected from the Natuna Islands (Riau Islands Province) were acclimatized in greenhouse and in semi-sterile culture in the laboratory. Sterilized explants were cultured on PES and Conwy media solidified with 0.8% Bacto Agar. In each of these media two combinations of plant growth regulators i.e. BA+IAA and BA+NAA were added. The concentrations of BA used were 0, 0.5, 1 mg/l, the concentrations of IAA were 0, 2.5, 5 mg/l, whereas the concentration of NAA were 0, 0.5, 1 mg/l. The result indicated that the optimal medium for callus induction was PES solidified medium supplemented with BA 1 mg/l. Types of callus formed were (a) white compact callus, (b) white filamentous callus, (c) greenish/brownish callus. Regeneration of callus into clumps offilament had been done by subculturing the callus into PES solidified medium supplemented with BA 1 mg/l + IAA 2.5 mg/l

AGROBACTERIUM-MEDIATED GENETIC TRANSFORMATION OF SEAWEED Kappaphycus alvarezii USING Gα GENE AND CALLUS CULTURES Sulistiani, Erina; Suharsono, Suharsono; Supena, Ence Darmo Jaya; Miftahudin, Miftahudin
BIOTROPIA Vol. 26 No. 1 (2019): BIOTROPIA Vol. 26 No. 1 April 2019
Publisher : SEAMEO BIOTROP

Cottonii seaweed (Kappaphycus alvarezii Doty) is one of the most important commercial sources of carrageenan, which is widely used in the pharmaceutical and food industries. A major problem in the cultivation of this seaweed is ice-ice disease, which is caused by extreme changes in environmental conditions such as temperature and seawater salinity. Gene transformation to produce transgenic Kappaphycus lines that are tolerant to environmental stress is a potential solution to this problem. The Gα gene, which encodes the heterotrimeric G-protein alpha subunit, plays an important role in tolerance to biotic and abiotic environmental stress. This study aimed to: (a) introduce the Gα gene into the callus cells of K. alvarezii and regenerate transformed callus cells into transgenic plantlets; and (b) determine the appropriate concentration of acetosyringone and Agrobacterium tumefaciens strain for successful 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 under the control of the CaMV-35S promoter. The calli and A. tumefaciens were co-cultivated in different concentrations of acetosyringone (20, 40, and 60 mg/L). The regeneration of transformed callus cells into transgenic plantlets was successfully achieved using the somatic embryogenesis technique. The results showed that the highest percentage of putative transgenic micropropagule formation occurred at acetosyringone concentrations of 20–40 mg/L. Polymerase chain reaction (PCR) analysis of twenty regenerated plantlets indicated that the Gα gene was successfully introduced into the genomic DNA of all samples. The highest transformation efficiency was obtained from the 20–40 mg/L acetosyringone co-cultivation treatment (22–28%). The transformation efficiency produced by Agrobacterium tumefaciens EHA105 (23%) was not significantly different from that produced using strain LBA4404 (15%).

OVEREXPRESSION OF Gα GENE INCREASES GROWTH AND HYPOSALINE TOLERANCE IN Kappaphycus alvarezii TRANSGENIC PLANTLETS Sulistiani, Erina
BIOTROPIA Vol. 28 No. 3 (2021): BIOTROPIA Vol. 28 No. 3 December 2021
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11598/btb.2021.28.3.1047

G proteins are membrane proteins that play roles in signal transduction in living organisms. They consist of α, β and γ subunits. The G protein α subunit (Gα) plays a role in plant resistance toward biotic and abiotic environmental stresses. Transgenic plantlets of Kappaphycus alvarezii carrying the Gα gene (derived from soybean) have been successfully obtained through Agrobacterium tumefaciens-mediated transformation. The present study aimed to: 1. compare the growth of non-transgenic and transgenic plantlets of K. alvarezii in vitro using Provasoli enriched seawater (PES) medium with normal salinity and hyposalinity and 2. analyze the expression level of the Gα gene in transgenic plantlets using quantitative Polymerase Chain Reaction (qPCR). The results showed that all transgenic plantlets (six clones) had significantly higher daily growth rate (DGR, % / d) than that of non-transgenic under the condition of normal salinity (30 ppt) and hyposalinity (15 and 20 ppt) for 5 weeks of observation. At 15 ppt, transgenic plantlets were more tolerant than non-transgenic ones, as most thalli of transgenic plantlets remained brown in color, whereas most thalli of non-transgenic plantlets were bleached. The results of the qPCR analysis showed that the expression of the Gα gene in transgenic plantlets increased by 6.43 - 8.03 times compared with that of non-transgenic plantlets. The result of Pearson correlation analysis showed that relative expression of Gα gene had a strong correlation, both with DGRs in normal salinity and hyposalinity of transgenic plantlets (correlation coefficient > 0.7). The correlation was linearly positive, where increased expression of the Gα gene was strongly associated with an increase in DGRs.

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