Calluses from Tomato CV. Tymoti.… Every treatment was conducted in a series of 4 were observed and photographed after 5 weeks of bottles and each bottle contained 3 explants. Each incubation. treatment was divided into two culture conditions, i.e. dark condition (24h dark) and light condition (16h light Photograph collection and characterization of callus period per day). Incubator was set at 23 ± 2ºC and 80% Calluses were photographed with Canon EOS 60D humidity. The light intensity was regulated at 1.000 lux 18-55 mm. The photographs were then characterized with 16/8 h (light/dark) cycle. The development of callus based on the morphological features, i.e. compact, friable, embryonic, and organogenic (shooty and rooty). RESULTS Table 1. Effect of Various Plant Growth Regulator on Callus Development Callus Formation Treatments Callus Type* Callus Color Callus Score** (%) (ppm) Dark Light Dark Light Dark Light Dark Light 0.5 BAP + 0.1 IAA 100 83.3 f,o c,o brownish green 4 2 1.0 BAP + 0.1 IAA 100 100 f,o f,o brownish green 4 2 1.5 BAP + 0.1 IAA 100 100 f,o c,o brownish green 4 1 2.0 BAP + 0.1 IAA 100 100 f,o c,o brownish green 4 2 2.5 BAP + 0.1 IAA 100 66.7 f,o c,o brownish green 4 1 0.5 TDZ + 0.1 IAA 100 83.3 f,e c,o brownish green 4 1 1.0 TDZ + 0.1 IAA 100 100 f,o c,o brownish green 4 1 1.5 TDZ + 0.1 IAA 100 100 f c,o brownish green 4 4 2.0 TDZ + 0.1 IAA 100 66.7 f,e f,c brownish green 3 3 2.5 TDZ + 0.1 IAA 100 100 c,o c,o brownish green 3 3 * Callus Type : f, friable; c, compact; o, organogenic; e, embryogenic ** Callus Score: 0, no callus formed; 1, quarter of explant formed callus; 2, half of explant formed callus; 3, entire explant formed callus; 4, callus size is twice of the initial explant A B Figure 1. Friable callus. A, light condition (MS + 1.0 ppm BAP + 0.1 ppm IAA); B, dark condition (MS + 2.5 ppm BAP + 0.1 ppm IAA) A B Figure 2. Compact callus. A, light condition (MS + 1.0 ppm TDZ + 0.1 ppm IAA); B, dark condition (MS + 2.5 ppm TDZ + 0.1 ppm IAA) Prosiding Seminar Nasional Biologi 2016_ ISBN: 978‐602‐0951‐11‐9 294 Calluses from Tomato CV. Tymoti.… A B Figure 3. Rooty callus from leaf explants incubated in dark condition. A, on MS + 0.5 ppm BAP + 0.1 ppm IAA; B, on on MS + 0.5 ppm TDZ + 0.1 ppm IAA Figure 4. Shooty callus. A, light condition (MS + 2.5 ppm TDZ + 0.1 ppm IAA); B, dark condition (MS + 1.5 ppm BAP + 0.1 ppm IAA) Figure 5. Embryonic callus from leaf explant cultured in dark condition (MS + 0.5 ppm TDZ + 0.1 ppm IAA) Prosiding Seminar Nasional Biologi 2016_ ISBN: 978‐602‐0951‐11‐9 295 Calluses from Tomato CV. Tymoti.… DISCUSSION All treatments incubated in dark condition The characterization of callus morphology was successfully resulted maximum callus formation (100%). hardly accomplished only by naked eye. Stereo However, cultures in light condition produced lower microscope was needed to make sure that a callus feature frequency (66.7-83.3%) due to contamination. If the was actually an organ primordia or an embryo primordia. contamination could be avoided, there was a probability Furthermore, high resolution of image was also helpful to to achieve 100% callus frequency as well because the characterize the callus types. In this case, callus types explants already showed response but no callus formed could obviously observed only after the photographs yet. So it was likely that there was no correlation between were taken. lower callus frequency and treatment in light condition. Friable callus mostly produced in dark condition, CONCLUSION whereas compact callus were more frequent on cultures Callus formation from leaf discs of tomato cv. incubated in light condition. Friable and compact callus Tymoti by using combination of TDZ/IAA and BAP/IAA are calluses which do not exhibit any obvious organ is better in dark condition than in light. (Ikeuchi et al, 2013). They are differ physically, that friable callus is soft, fragile and watery, while compact ACKNOWLEDGEMENTS callus is hard (Akaneme and Eneobong, 2008). In this We thank to Lembaga Penelitian dan research, friable and compact callus is shown on Figure 1 Pengambidan kepada Masyarakat (LPPM), University of and 2. In dark condition, callus color was brownish, when Surabaya (Ubaya) for funding this research. it was green in light condition. This fact indicated that no chlorophyll produced on cultures incubated in dark REFERENCES condition. 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