Produksi gerbera umumnya dilakukan melalui pemuliaan konvensional sehingga memerlukan waktu relatif lama dan tunas/anakan yang dihasilkan jumlahnya terbatas. Tujuan penelitian adalah mengoptimalkan metode perbanyakan massal gerbera secara in vitro. Penelitian dilaksanakan di Laboratorium Kultur Jaringan Kebun Percobaan Tanaman Hias Cipanas dari bulan Januari 2019 – Februari 2020. Bahan tanaman yang digunakan adalah gerbera varietas Chandramaya Agrihorti, Zsofia Agrihorti, Nirwasita Agrihorti, Awandini Agrihorti, dan Arkadewi Agrihorti. Eksplan berupa capitulum bunga. Penelitian menggunakan Rancangan Acak Lengkap (RAL), dengan 40 kombinasi perlakuan. Hasil penelitian menunjukkan bahwa formulasi media MS + BAP 0,5 mg/l + TDZ 0,3 mg/l + Sukrosa 20 g/l + Gelrite 2 g/l merupakan media yang paling baik untuk menginduksi pembentukan kalus dan tunas lima varietas gerbera (varietas Nirwasita, Chandramaya, Arkadewi, Zsopia, dan Awandini).KeywordsGerbera; Kultur in vitro; TDZ; Produksi benihAbstractGerbera is usually produced through conventional breeding, which is time consuming and produce limited number shoots and explants. The objectives of the research was to optimize mass propagation method of gerbera in vitro. The research was conducted in Indonesian Ornamental Crops Research Institute, January 2019 until February 2020. Gerbera’s capitulum of Chandramaya Agrihorti, Zsopia Agrihorti, Nirwasita Agrihorti, Awandini Agrihorti and Arkadewi Agrihorti cultivars were used as explants. Design of the research was a completely randomized design, with eight treatment combinations. MS + BAP 0.5 mg/l + TDZ 0.3 mg/l + Sucrose 20 g/l + Gelrite 2 g/l was best media for callus induction and shoot formation in five gerbera cultivars (Nirwasita, Chandramaya, Arkadewi, Zsopia and Awandini cultivars).