Effects of light, sucrose concentration and repetitive subculture on callus growth and medically important production in Justicia gendarussa Burm.f.
Dwi Kusuma Wahyuni, Alamil Huda, Siti Inayatul Faizah, Hery Purnobasuki, Bambang Prajogo Eko Wardojo
Abstract
OBJECTIVE: Burm.f. calluses based on biomass, morphological characters and metabolic profiles. METHODS: Burm.f. were isolated and cultured in solid Murashige and Skoog (MS) medium with the addition of 1 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/L benzyl amino purine (BAP). Different sucrose concentrations, varying from 0 to 5%, were included in the medium. The cultures were incubated under light and dark conditions for 6 weeks. Repetitive subculture was carried out every 2 weeks for a total of four times, and 3% sucrose gave the best callus growth. Dry calluses were extracted with methanol, and their metabolic profiles were analyzed using gas chromatography/mass spectrometry (GC/MS). Compound identification was performed by comparing the mass spectra to references from the WILEY version 7n.1 library. RESULTS: Among the 12 conditions tested for the prolonged 6-week culture, the 2-5 % sucrose treatments under light and 3-5 % sucrose treatments under dark exhibited the highest dry weight. For repetitive subculture, the highest wet and dry weights were identically detected under both light and dark conditions after the second repetitive subculture. A total of 19 metabolites was identified by GC/MS, with major compounds being taraxasterol, monoplex D, 2-methoxy-4-vinylphenol, and palmitic acid. CONCLUSION: Burm.f. calluses. A concentration of 3% sucrose demonstrated the best growth of callus and could be further applied for mass production.