ABSTRAK Penelitian ini bertujuan untuk mengetahui pengaruh komposisi 2,4-D dan BAP terhadap pembentukan kalus eksplan pucuk nilam (Pogostemon cablin Benth) secara in vitro dengan pemotongan secara vertikal. Komposisi 2,4-D dan BAP dilakukan pada media Murashige & Skoog (MS) dengan 16 perlakukan (P0-P15) dan lima kali pengulangan (U1-U5). Parameter yang diukur adalah persentase eksplan pembentuk kalus, waktu kemunculan kalus (HST), berat kalus dan morfologi kalus. Hasil rata-rata yang didapatkan bahwa pemotongan eksplan secara vertikal mampu menginduksi kalus sebesar 1,5%. Waktu muncul kalus paling cepat didapatkan pada perlakuan P6U5 (1 ppm 2,4-D : 0,5 ppm BAP) dan P8U1 (0 ppm 2,4-D : 1 ppm BAP) pada 12 HST. Kalus yang memiliki massa paling berat didapatkan pada perlakuan P12U1 (0 ppm 2,4-D : 1,5 ppm BAP) yaitu 0,05 gram. Hasil juga menunjukkan bahwa kalus yang dihasilkan memiliki tekstur kompak (non friable) dan remah (friable) dengan warna kalus putih kecoklatan, cokelat, hingga cokelat kehitaman Kata kunci : Pogostemon cablin Benth., Pemotongan eksplan secara vertikal, Kalus, 2,4-D, BAP ABSTRACT This study aims to determine the effect of 2,4-D and BAP composition for callus establishment from shoots of patchouly plant (Pogostemon cablin Benth) by in vitro with vertical cutting method. The composition of 2,4-D and BAP were performed on Murashige & Skoog (MS) media with 16 treatments (P0-P15) and five replications (U1-U5). The parameters that measured were percentage of callus forming explants, time appearance of callus (day after planting, DAP), weight of callus and the morphology of callus. The results indicated that the percentage showed that the vertical cutting method are able to induce callus formation by an average 1,5%. The fastest time appearance of callus (DAP) were obtained in P6U5 (1 ppm 2,4-D : 0.5 ppm BAP) and P8U1 (0 ppm 2,4-D : 1 ppm BAP) at 12 DAP. The heaviest callus were obtained in P12U1 (0 ppm 2,4-D : 1.5 ppm BAP) at 0.05 grams. The results also showed that the obtained callus had a non friable and friable texture with brownish-white, brown, to black-brown color. Keywords : Pogostemon cablin Benth., vertical cutting explant method, callus, 2,4-D, BAP