OPTIMASI MEDIA PROPAGASI KALUS TOMAT (Solanum lycopersicum L. 'Permata') DAN KARAKTERISASI KANDUNGAN METABOLITNYA
Arkan Setiaji, Dr. Endang Semiarti, M.S., M.Sc.
2020 | Skripsi | S1 BIOLOGIEkstrak kalus dan suspensi sel tanaman banyak digunakan dalam produk perawatan kulit, sehingga produksi sel dan metabolit bahan tersebut menjadi penting untuk dipelajari. Penelitian ini bertujuan untuk optimasi produksi kalus menggunakan zat pengatur tumbuh (ZPT), analisis karakter perkembangan kalus secara anatomi, dan analisis metabolit yang terkandung di dalam kalus tomat (S. lycoperscium 'Permata'). Metode: biji ditanam pada medium 1/4 Murashige and Skoog (MS) selama 10-15 hari. Eksplan hipokotil ditanam pada medium MS + 2 mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D) selama 15 hari untuk induksi kalus. Kalus ditransfer ke medium MS dan B5 dengan 8 variasi ZPT kombinasi 6-Benzylaminopurine (BAP) + 1-Naphthaleneacetic acid (NAA), dan 2,4-D tunggal. Berat basah dan berat kering kalus ditimbang setiap 5 hari selama 60 hari untuk mengetahui efisiensi produksi dan kinetika pertumbuhan kalus. Pembuatan preparat anatomi pada kalus dilakukan dengan metode parafin dengan berbagai umur (0-24 hari). Kalus dan buah tomat dimaserasi dengan pelarut akuabides. Kandungan metabolit pada ekstrak dianalisis menggunakan teknik Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS). Hasil penelitian menunjukkan medium MS dengan penambahan 2 mg/L NAA + 0,2 mg/L BAP merupakan medium terbaik untuk perbanyakan kalus tomat dengan nilai tertinggi pada laju pertumbuhan mutlak (LPM) berat basah (73,77% per hari), berat kering (3,84% per hari), dan waktu inisiasi kalus (5,6 hari). Pertumbuhan kalus tomat pada medium optimal bertipe kompak dengan nodul akar. Secara anatomi kalus awalnya berkembang dari sel-sel perisikel. Ekstrak air kalus tomat mengandung 16 asam amino, terutama DL-Tryptophan, L-Norleucine, dan DL-Arginine. Keragaman jenis peptida dan asam lemak pada ekstrak air kalus tomat lebih tinggi dibandingkan dengan ekstrak air buah tomat. Berbagai metabolit pada ekstrak air kalus tomat berpotensi untuk diformulasikan dalam produk perawatan kulit.
Extract of plant callus and cell suspension culture has been used as materials in skincare products, including the production of cells and metabolites therefore it is important to be studied. Aims of this research are optimization of plant growth regulators (PGRs) in callus induction medium, to analyze callus development through anatomical character, and to analyze the metabolites content in the tomato (S. lycoperscium 'Permata') calli. Methods: seeds were grown on 1/4 MS medium for 10-15 days. Hypocotyl was excised and cultured on Murashige and Skoog (MS) medium + 2 mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D) for 15 days as the explants for callus induction. Callus was transferred to MS and B5 media with 8 variations of PGRs including the combination of 6-Benzylaminopurine (BAP) + 1-Naphthaleneacetic acid (NAA), and 2,4-D alone. Both fresh and dry weight of calli were measured every 5 days over 60 days to established the growth kinetics and production efficiency of callus. Histological characters of calli were examined anatomically through paraffin-embedded method at various ages (0-24 days). Extracts of tomato fruit and callus were prepared by using a process of maceration in the aquabidest solution. Metabolite content in the extract was analyzed using Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS) technique. Results: MS medium supplemented with 2.0 mg/L NAA + 0.2 mg/L BAP was the best PGRs for tomato callus induction, based on the highest number of absolute growth rate on fresh weight (73.77% per day), dry weight (3.84% per day), and callus initiation time (5.6 days). Type of tomato callus texture in the optimal medium is compact with root nodules. Based on anatomical studies, the callus initially develops from pericycle cells. Aqueous extract of tomato callus contains 16 amino acids, mainly DL-Tryptophan, L-Norleucine, and DL-Arginine. The diversity of peptides and fatty acids on aqueous extract of tomato callus was higher than the aqueous extract of tomato fruit. Various metabolites on aqueous extract of tomato callus have the potential to be formulated in skincare products.
Kata Kunci : tomat, kalus, media in vitro, ZPT, metabolit
