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Stevia ( Stevia rebaudiana Bert . ), a sweetener plant, has been mass propagated by tissue culture technique. Optimal conditions to increase vigor of stevia plantlets are needed to support the sustainability of in vitro plantlet stocks and increase plantlet survival rate during acclimatization. The aim of this research was to investigate the effect of different media, culture vessel sizes, and vessel closure types on the vigor of stevia plantlets. The plant material was derived from apical shoot cuttings of sterile stevia plantlets grown on WP medium without growth regulator. Several treatment s used in this study were solid or double layer media ; short or tall culture vessel ; and polypropile screw cap or plastic film closures.  Growth of plantlets was determined after 3 weeks of culture. Temperature and light intensity inside the vessels were also observed. The results showed that the best treatment to increase the vigor of stevia plantlets was a double-layer medium in a tall culture vessel (diameter 7 cm and height 11 cm) with either screw cap or plastic film. It was exhibited by significantly bigger stem diameter, more and bigger leaves, longer roots, and higher biomass fresh weight than those of other treatments. Higher temperature was observed on tall culture vessel, whereas all treatments did not significantly affect light intensity inside the vessels. [ Keywords: stevia, plantlet vigor, double-layer medium, culture vessel size, vessel closure ] Abstrak Stevia ( Stevia rebaudiana Bert.), tanaman pemanis, telah diperbanyak melalui teknologi kultur jaringan. Kondisi kultur optimal untuk meningkatkan vigor planlet stevia masih diperlukan untuk mendukung keberlanjutan tanaman stock in vitro dan untuk meningkatkan daya hidup planlet ketika diaklimatisasi. Penelitian yang dilakukan bertujuan untuk menentukan pengaruh penggunaan jenis media, ukuran botol kultur, dan jenis penutup botol yang berbeda terhadap vigor planlet stevia. Material tanaman yang digunakan didapat dari potongan tunas apikal plantlet stevia steril yang ditumbuhkan pada media WP tanpa zat pengatur tumbuh. Perlakuan jenis media terdiri atas media padat dan media dua-lapis ( double-layer ), ukuran botol pendek dan tinggi, serta jenis tutup ulir berbahan polipropilen dan lembaran plastik transparan. Pengamatan pertumbuhan planlet dilakukan setelah 3 minggu, juga dilakukan pengamatan terhadap suhu dan intensitas cahaya di dalam botol kultur. Hasil penelitian memperlihatkan bahwa perlakuan terbaik untuk meningkatkan vigor planlet stevia adalah dengan menggunakan media dua-lapis dalam botol kultur (diameter 7 cm, dan tinggi 11 cm), baik dengan tutup ulir maupun plastik. Hal ini ditunjukkan dari diameter batang lebih besar, daun lebih banyak dan besar, akar lebih panjang, serta bobot segar biomassa lebih tinggi dibandingkan dengan perlakuan lainnya. Suhu lebih tinggi terukur pada perlakuan botol tinggi, sedangkan semua perlakuan tidak mempengaruhi secara nyata intensitas cahaya di dalam botol kultur. [Kata kunci: stevia, vigor, media dua-lapis, ukuran botol kultur, tutup botol]

Modifikasi sistem kultur in vitro untuk meningkatkan vigor planlet stevia (Stevia rebaudiana Bert.) [Modification of in vitro culture system to increase the vigor of stevia (Stevia rebaudiana Bert.) plantlets]