Regeneration and micropropagation of panax vietnamensis ha et grushv. using thin cell layer technology

sis NAA 0.2 0 e 0 e 0 d 0 d Necrosis NAA 0.5 0 e 0 e 31.3 c 2.0 c White short root NAA 1.0 86.6 a 0 e 75.5 b 6.4 b Black callus. Some white lateral root NAA 2.0 76.6 b 46.6 c 89.9 a 15.5 a Yellowish callus. A lot of white lateral root * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. 3.1.1.3. Effect of single PGRs on morphogenesis of lTCL_C petiole explant in light and dark conditions The indicators recorded after 10 weeks of culture are shown in tables 3.4, 3.5 and 3.4, 3.5. 7 Table 3.4. Effect of single PGRs on morphogenesis of lTCL_C petiole explant at photoperiod of 16 hours/day. PGRs (mg/l) Embryogenesis (%) Callogenesis (%) Root formation (%) Root number Remarks Control 0 0 e 0 e 0 f 0 c Necrosis TDZ 0.0 1 0 e 0 e 0 f 0 c Necrosis TDZ 0.0 5 0 e 0 e 0 f 0 c Necrosis TDZ 0.1 0 e 0 e 0 f 0 c Necrosis TDZ 0.2 0 e 0 e 0 f 0 c Necrosis TDZ 0.5 0 e 0 e 0 f 0 c Necrosis TDZ 1.0 0 e 0 e 0 f 0 c Necrosis BA 0.1 0 e 0 e 0 f 0 c Necrosis BA 0.2 0 e 0 e 0 f 0 c Necrosis BA 0.5 0 e 0 e 0 f 0 c Necrosis BA 1.0 0 e 0 e 0 f 0 c Necrosis BA 2.0 0 e 0 e 0 f 0 c Necrosis 2,4-D 0.1 0 e 0 e 0 f 0 c Necrosis 2,4-D 0.2 23 d 24.4 d 25.6 d 0.8 c Few embryos. Black compact callus 2,4-D 0.5 59.6 c 89.9 b 79.9 a 4.7 ab Cluster of global, heart shaped and cotyledon embryos Yellow compact callus. Green elongated root 2,4-D 1.0 86.3 a 97.7 a 85.5 a 6.2 a Many global, heart, torpedo shaped and cotyledon embryos Brown callus. Yellowish roots 2,4-D 2.0 69.6 b 98.8 a 71 b 4.0 b Heart and global shaped embryos only at leaf edges Brown compact callus Few transparent white root NAA 0.1 0 e 0 e 0 f 0 c Necrosis NAA 0.2 0 e 0 e 0 f 0 c Necrosis NAA 0.5 0 e 0 e 0 f 0 c Necrosis NAA 1.0 58.6 c 84.4 c 10.6 e 3.1 b Large amount of callus. Purple embryo cluster NAA 2.0 84.0 a 96.6 a 37.7 c 4.0 b milky white heart and global shaped embryos. Little brown calli * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. 8 Table 3.5. Effect of single PGRs on morphogenesis of lTCL_C petiole explant in dark. PGRs (mg/l) Embryogenesis (%) Callogenesis (%) Root formation (%) Root number Remarks Contr ol 0 0 c 0 e 0 g 0 f Necrosis TDZ 0.01 0 c 0 e 0 g 0 f Necrosis TDZ 0.05 0 c 0 e 0 g 0 f Necrosis TDZ 0.1 0 c 0 e 0 g 0 f Necrosis TDZ 0.2 0 c 0 e 0 g 0 f Necrosis TDZ 0.5 0 c 0 e 0 g 0 f Necrosis TDZ 1.0 0 c 0 e 0 g 0 f Necrosis BA 0.1 0 c 0 e 0 g 0 f Necrosis BA 0.2 0 c 0 e 0 g 0 f Necrosis BA 0.5 0 c 0 e 0 g 0 f Necrosis BA 1.0 0 c 0 e 0 g 0 f Necrosis BA 2.0 0 c 0 e 0 g 0 f Necrosis 2,4-D 0.1 0 c 46.6 d 11.0 f 0.34 ef Embryogenic calli. Few yellow roots 2,4-D 0.2 0 c 67.7 c 35.5 de 0.91 d Large amount of yellow and white calli. White root 2,4-D 0.5 49.6 b 84.4 b 41.1 cd 0.51 e Few global embryos. Brown callus. Few white roots 2,4-D 1.0 69.6 a 94.4 a 46.6 c 1.87 c White soft calli. Few embryos. Transparent white root 2,4-D 2.0 0 c 0 e 0 g 0 f Necrosis NAA 0.1 0 c 0 e 0 g 0 f Necrosis NAA 0.2 0 c 0 e 0 g 0 f Necrosis NAA 0.5 0 c 0 e 31.1 e 0.94 d Few white roots NAA 1.0 0 c 45.5 d 61.1 b 6.09 b Little yellow callus. Milky white short roots. NAA 2.0 0 c 81 b 94.4a 19.2 a Little yellow callus Many milky white short roots. * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. 9 3.1.1.4. Effect of the combination of auxin and cytokinin on the morphogenesis of the rhizome explant tTCL_R in light and dark conditions. After 10 weeks, we observe and record the indicators, the results are shown in tables 3.6, 3.7 and Figures 3.6 and 3.7. Table 3.6. Effect of the combination of auxin and cytokinin on the morphogenesis of the rhizome explant tTCL_R at photoperiod of 16 hours/day. PGRs (mg/l) Embryogenesis (%) Callogenesis (%) Root formation (%) Root number Remarks Contr ol 0 0 g 0 f 0 c 0 c Necrosis TDZ 0.01 0 g 0 f 0 c 0 c Necrosis TDZ 0.05 0 g 0 f 0 c 0 c Necrosis TDZ 0.1 0 g 0 f 0 c 0 c Necrosis TDZ 0.2 0 g 0 f 0 c 0 c Necrosis TDZ 0.5 0 g 0 f 0 c 0 c Necrosis TDZ 1.0 0 g 0 f 0 c 0 c Necrosis BA 0.1 0 g 0 f 0 c 0 c Necrosis BA 0.2 0 g 0 f 0 c 0 c Necrosis BA 0.5 0 g 0 f 0 c 0 c Necrosis BA 1.0 0 g 0 f 0 c 0 c Necrosis BA 2.0 0 g 0 f 0 c 0 c Necrosis 2,4-D 0.1 0 g 75.5 c 0 c 0 c Little white callus 2,4-D 0.2 31.0 f 87.7 b 0 c 0 c Large amount white callus. Few global shaped embryos. 2,4-D 0.5 57.3 d 89.9 b 0 c 0 c Large amount of brown calli. Cluster of global, and heart shaped embryos 2,4-D 1.0 79.6 b 97.7 a 0 c 0 c Large amount of dark brown calli. Global, heart shaped and cotyledon embryos. 2,4-D 2.0 91.0 a 88.8 b 0 c 0 c Large amount of dark brown calli. Many global, heart shaped and cotyledon embryos. NAA 0.1 0 g 0 f 0 c 0 c Necrosis NAA 0.2 71.0 c 0 f 0 c 0 c Embryo shaped global, heart, cotyledon, torpedo 10 NAA 0.5 70.6 c 0 f 0 c 0 c Global, heart, torpedo shaped and cotyledon embryos. NAA 1.0 48.6 e 44.4 e 74.4 b 5.09 b Global and heart shaped embryos. Few white roots NAA 2.0 47.3 e 67.7 d 83.3 a 9.24 a Global shaped and cotyledon embryos. Yellowish callus. Short white roots. * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. Table 3.7. Effect of the combination of auxin and cytokinin on the morphogenesis of the rhizome explant tTCL_R in dark. PGRs (mg/l) Embryogenesis (%) Callogenesis (%) Root formation (%) Root number Remarks Control 0 0 d 0 f 0 d 0 c Necrosis TDZ 0.01 0 d 0 f 0 d 0 c Necrosis TDZ 0.05 0 d 0 f 0 d 0 c Necrosis TDZ 0.1 0 d 0 f 0 d 0 c Necrosis TDZ 0.2 0 d 0 f 0 d 0 c Necrosis TDZ 0.5 0 d 0 f 0 d 0 c Necrosis TDZ 1.0 0 d 0 f 0 d 0 c Necrosis BA 0.1 0 d 0 f 0 d 0 c Necrosis BA 0.2 0 d 0 f 0 d 0 c Necrosis BA 0.5 0 d 0 f 0 d 0 c Necrosis BA 1.0 0 d 0 f 0 d 0 c Necrosis BA 2.0 0 d 0 f 0 d 0 c Necrosis 2,4-D 0.1 56.3 b 41.1 c 0 d 0 c Global shaped embryos. Yellowish callus. 2,4-D 0.2 84.0 a 61 b 24.4 c 0.93 b Global, heart shaped, and cotyledon embryos. Yellow callus 2,4-D 0.5 46.3 c 91 a 27.7 c 0.69 bc Few embryo Large amount of yellow calli 2,4-D 1.0 0 d 95.5 a 44.4 b 0.58 bc Brown compact callus Few white roots 2,4-D 2.0 0 d 31 d 0 d 0 c Yellow callus NAA 0.1 0 d 0 f 0 d 0 c Explants turned yellow NAA 0.2 0 d 0 f 28.2 c 1.28 b Short white roots 11 NAA 0.5 0 d 24.4 e 48.8 b 0.59 bc Few roots NAA 1.0 0 d 0 f 52.2 b 1.37 b Few yellow short roots NAA 2.0 0 d 0 f 98.8 a 21.7 a Many white roots and lateral roots * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. 3.1.1.5. Effect of the combination of auxin and cytokinin on the morphogenesis of the leaf explants tTCL_L in light and dark conditions The results obtained after 10 weeks of culture are shown in tables 3.8, 3.9, 3.10, 3.11, 3.12, 3.13; Figures 3.8, 3.9, 3.10. Table 3.8. Effect of the combination of 2,4-D and BA on the morphogenesis of the leaf explants tTCL_L at photoperiod of 16 hours/day. PGRs (mg/l) Morphogenesis Remarks 2,4-D BA Callogenesis (%) 1.0 0.1 100 a White green compact callus 1.0 0.2 100 a Many greenis white callus 1.0 0.5 100 a Little yellow and milky white soft callus 1.0 1.0 93.3 b Little yellowis green compact callus 1.0 2.0 90 c Light yellow soft callus 0.1 1.0 47.7 f Little yellowish brown callus 0.2 1.0 60.0 e Little green and brown compact callus 0.5 1.0 80.0 d Little brown compact callus 2.0 1.0 100 a Large amount of yellow and milky white soft callus * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. Table 3.9. Effect of the combination of 2,4-D and BA on the morphogenesis of the leaf explants tTCL_L in dark PGRs (mg/l) Morphogenesis Remarks 2,4-D BA Callogenesis (%) Root formation (%) 1.0 0.1 100 a 27.3 a Little yellow and milky white soft callus 1.0 0.2 100 a 25.5 b Large amount of milky white soft callus 1.0 0.5 100 a 0 c Large amount of transparent and milky white soft callus 1.0 1.0 100 a 0 c White and milky white soft callus 1.0 2.0 93.3 b 0 c Brownish yellow soft callus 0.1 1.0 73.3 d 0 c Brownish yellow compact callus 12 0.2 1.0 80.0 c 0 c Little brownish yellow and transparent compact callus 0.5 1.0 93.3 b 0 c White and milky white soft callus 2.0 1.0 93.3 b 0 c White and brownish red soft callus * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. Table 3.10. Effect of the combination of 2,4-D and TDZ on the morphogenesis of the leaf explants tTCL_L at photoperiod of 16 hours/day. PGRs (mg/l) Morphogenesis Remarks 2,4-D TDZ Callogenesis (%) 1.0 0.01 100 a White and yellow soft callus 1.0 0.05 100 a Greenish white and brownish yellow compact callus 1.0 0.1 100 a Large amount of greenish white and redish yellow compact callus 1.0 0.2 100 a Little greenish white and yellow soft callus 1.0 0.5 100 a Little greenish white and redish yellow compact callus 1.0 1.0 100 a White and yellow soft callus 0.1 0.2 80 b Little greenish white and redish yellow compact callus 0.2 0.2 80 b Little green compact callus 0.5 0.2 100 a White and brown soft callus 2.0 0.2 73.3 c Brownish yellow soft callus * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. Table 3.11. Effect of the combination of 2,4-D and TDZ on the explants tTCL_L in dark. PGRs (mg/l) Morphogenesis Morphogenesis of the leaf Remarks 2,4-D TDZ Callogenesis (%) 1.0 0.01 100 a Brownish yellow soft callus 1.0 0.05 100 a Little yellow and milky white soft callus 1.0 0.1 100 a Large amount of milky white soft callus 1.0 0.2 100 a Little white and brownish yellow soft callus 1.0 0.5 100 a Transparent white and milky white soft callus 1.0 1.0 86.6 c Little milky white and brownish yellow soft callus 0.1 0.2 80.0 d Little transparent white and brown soft callus 0.2 0.2 93.3 b White and brown soft callus 0.5 0.2 93.3 b Little milky white and brownish yellow soft callus 2.0 0.2 0 e Necrosis * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. 13 Table 3.12. Effect of the combination of NAA and BA on the morphogenesis of the leaf explants tTCL_L at photoperiod of 16 hours/day. PGRs (mg/l) Morphogenesi s Remarks NAA BA Callogenesis (%) 1.0 0.1 13.3 c Little green compact callus 1.0 0.2 0 d Necrosis 1.0 0.5 0 d Necrosis 1.0 1.0 33.3 b Little brown compact callus 1.0 2.0 0 d Survival but no development 0.1 1.0 0 d Necrosis 0.2 1.0 0 d Necrosis 0.5 1.0 0 d Necrosis 2.0 1.0 60 a Little green compact callus * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. Table 3.13. Effect of the combination of NAA and BA on the morphogenesis of the leaf explants tTCL_L in dark. PGRs (mg/l) Morphogenesis Remarks NAA BA Callogenesis (%) 1.0 0.1 40 e Little transparent white and brownish yellow callus 1.0 0.2 33.3 f Little brown compact callus 1.0 0.5 53.3d Little brown compact callus 1.0 1.0 93.3 b White and brownish yellow soft callus 1.0 2.0 13.3 g Little brownish yellow compact callus 0.1 1.0 0 h Survival but no development 0.2 1.0 0 h Necrosis 0.5 1.0 90.0 c Little greenish white and brownish yellow soft callus 2.0 1.0 100 a White and brown soft callus * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. 3.1.1.6. Effect of the combination of auxin and cytokinin on the morphogenesis of the tTCL_C petiole explant in light and dark conditions. After 10 weeks of culture, the results were observed, recorded and compared the effect of each pair of 2,4-D in combination with BA, 2,4-D in combination with TDZ and BA in combination with NAA. present in tables 3.14, 3.15, 3.16, 3.17, 3.18, 3.19 and Figures 3.11, 3.12, 3.13. 14 Table 3.14. Effect of the combination of 2,4-D and BA on the morphogenesis of the tTCL_C petiole explant at photoperiod of 16 hours/day PGRs (mg/l) Morphogenesis Remarks 2,4-D BA Callogenesis (%) 1.0 0.1 98.8 a Little green and brownish yellow soft callus 1.0 0.2 97.7 a Yellowish white soft callus 1.0 0.5 98.8 a Yellow soft callus 1.0 1.0 97.7 a Milky white, green, and brown compact callus 1.0 2.0 74.4 c Little light yellow compact callus 0.1 1.0 96.6 a Little green compact callus 0.2 1.0 98.8 a Green and white compact callus 0.5 1.0 98.8 a Large amount of green and purple compact callus 2.0 1.0 86.6 b Little yellow soft callus * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. Table 3.15. Effect of the combination of 2,4-D and BA on the morphogenesis of the tTCL_C petiole explant in dark. PGRs (mg/l) Morphogenesis Remarks 2,4-D BA Callogenesis (%) 1.0 0.1 97.7 a Light yellow soft callus 1.0 0.2 98.8 a Large amount of yellow and milky white soft callus 1.0 0.5 97.7 a Milky white and light yellow soft callus 1.0 1.0 97.7 a Little light brown and light yellow soft callus 1.0 2.0 98.8 a Low light brown and light yellow soft callus 0.2 1.0 96.6 a Yellow brown soft callus 0.5 1.0 97.7 a Milky white and light yellow soft callus 2.0 1.0 96.6 a Little yellow soft callus * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. Table 3.16. Effect of the combination of 2,4-D and TDZ on the morphogenesis of the tTCL_C petiole explant at photoperiod of 16 hours/day PGRs (mg/l) Morphogenesis Remarks 2,4-D TDZ Callogenesis (%) 1.0 0.01 100 a Green and white soft callus 1.0 0.05 100 a Dark yellow soft callus 1.0 0.1 100 a Large amount of yellow and milky white soft callus 1.0 0.2 100 a Little yellow and milky white soft callus 1.0 0.5 100 a Little green, brown, and white soft callus 15 1.0 1.0 100 a Little yellow and milky white soft callus 0.1 0.2 100 a Small green compact callus 0.2 0.2 86.6 b Little green, and brownish yellow compact callus 0.5 0.2 100 a Little yellow and milky white soft callus 2.0 0.2 0 c Necrosis * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. Table 3.17. Effect of the combination of 2,4-D and TDZ on the morphogenesis of the tTCL_C petiole explant in dark. PGRs (mg/l) Morphogenesis Remarks 2,4-D TDZ Callogenesis (%) 1.0 0.01 100 a Transparent white and brownish yellow callus 1.0 0.05 100 a Brownish yellow soft callus 1.0 0.1 100 a Large amount of yellow and milky white soft callus 1.0 0.2 100 a Transparent yellow soft callus 1.0 0.5 100 a Brownish redish and yellow soft callus 1.0 1.0 100 a Brownish red soft callus 0.1 0.2 93.3 c Little brown compact callus 0.2 0.2 93.3 c Little yellow soft callus 0.5 0.2 79.9 c White and yellow soft callus 2.0 0.2 100 a Little milky white soft callus * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. Table 3.18. Effect of the combination of NAA and BA on the morphogenesis of the tTCL_C petiole explant at photoperiod of 16 hours/day. PGRs (mg/l) Morphogenesis Remarks NAA BA Callogenesis (%) 1.0 0.1 100 a Necrosis 1.0 0.2 100 a Necrosis 1.0 0.5 100 a Large amount of yellow and white compact callus 1.0 1.0 100 a Yellow and white compact callus 1.0 2.0 100 a Brownish yellow compact callus 0.1 1.0 0 c Necrosis 0.2 1.0 0 c Necrosis 0.5 1.0 0 c Necrosis 2.0 1.0 86.6 b Little brown compact callus * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. 16 Table 3.19. Effect of the combination of NAA and BA on the morphogenesis of the tTCL_C petiole explant in dark. PGRs (mg/l) Morphogenesis Remarks NAA BA Callogenesis (%) 1.0 0.1 100 a Very little brownish yellow soft callus 1.0 0.2 100 a Little yellow and milky white soft callus 1.0 0.5 100 a Large amount dark yellow soft callus 1.0 1.0 100 a Large amount dark yellow soft callus 1.0 2.0 100 a Large amount dark yellow soft callus 0.1 1.0 0 b Necrosis 0.2 1.0 100 a Large amount dark yellow soft callus 0.5 1.0 100 a Large amount dark yellow soft callus 2.0 1.0 100 a Large amount dark yellow soft callus * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. 3.1.1.7. Effect of the combination of auxin and cytokinin on the morphogenesis of the lTCL_C petiole explants in light and dark conditions. The results recorded after 10 weeks of culture shown in tables 3.20 to 3.25 and Figures 3.14, 3.15, 3.16. Table 3.20. Effect of the combination of 2,4-D and BA on the morphogenesis of the lTCL_C petiole explants at photoperiod of 16 hours/day PGRs (mg/l) Morphogenesis Remarks 2,4- D BA Callog enesis (%) Embryogene sis (%) 1.0 0.1 100 a 24.4 c Little brownish red, green and yellow compact callus 1.0 0.2 93.3 b 48.8 a Red and milky white soft callus 1.0 0.5 86.6 c 31.0 b Little white, green and red callus 1.0 1.0 66.7 d 0 d Red, brownish yellow, and light green callus 1.0 2.0 13.3 e 0 d Little light yellow, and red callus 0.1 1.0 86.6 c 0 d Very tittle callus 0.2 1.0 100 a 0 d Little red brown compact callus 0.5 1.0 93.3 b 0 d Green, and yellow callus on cut surface 2.0 1.0 100 a 0 d Little yellow and milky white soft callus on explant surface and two cut surfaces * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. 17 Table 3.21. Effect of the combination of 2.4-D and BA on the morphogenesis of the lTCL_C petiole explants in dark PGRs (mg/l) Morphogenesis Remarks 2,4- D BA Callogen esis (%) Shoot formation (%) 1.0 0.1 100 a 0 b Brownish yellow, and milky white soft callus 1.0 0.2 100 a 21.0a Large amount of milky white soft callus 1.0 0.5 100 a 0 b Milky white soft callus 1.0 1.0 100 a 0 b Little yellow and milky white soft callus 1.0 2.0 100 a 0 b Light yellow soft callus 0.1 1.0 46.7 d 0 b Little brownish red compact callus 0.2 1.0 80.0 c 0 b Little yellow and brown compact callus on surface and two cut explants 0.5 1.0 93.3 b 0 b Milk white soft callus on explant surface and two cut surfaces 2.0 1.0 100 a 0 b Little yellow and brown soft callus * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. Table 3.22. Effect of the combination of 2.4-D and TDZ on the morphogenesis of the lTCL_C petiole explants at photoperiod of 16 hours/day PGRs (mg/l) Morphogenesis Remarks 2,4-D TDZ Callogenesis (%) 1.0 0.01 86.6 c Little milky white, and transparent white soft callus on one cut surface 1.0 0.05 93.3 b Little milky white soft callus on explant surface and two cut surfaces 1.0 0.1 93.3 b Greenish white compact callus 1.0 0.2 100 a Large amount of milky white soft callus on explant surface and two cut surfaces 1.0 0.5 100 a Greenish white compact callus on one cut surface 1.0 1.0 93.3 b Brownish red and milky white soft callus on two cut surfaces 0.1 0.2 73.3 d Little green compact callus on one cut surface 0.2 0.2 73.3 d Little milky white soft callus on one cut surface 0.5 0.2 93.3 b Large amount of milky white soft callus on one cut surface 2.0 0.2 0 e Necrosis * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. 18 Table 3.23. Effect of the combination of 2.4-D and TDZ on the morphogenesis of the lTCL_C petiole explants in dark PGRs (mg/l) Morphogenesis Remarks 2,4-D TDZ Callogenesis (%) 1.0 0.01 93.3 b Milky white, and brown soft callus on one cut surface 1.0 0.05 100 a Little yellow and milky white soft callus on one cut surface 1.0 0.1 100 a Large amount of transparent and milky white soft callus on two cut surfaces 1.0 0.2 100 a Milky white and brownish yellow soft callus 1.0 0.5 93.3 b Large amount of yellow and milky white soft callus on one cut surface 1.0 1.0 86.6 c Little milky white and yellow soft callus on two cut surfaces 0.1 0.2 0 e Survival but no development 0.2 0.2 80.0 d White and brownish yellow soft callus on explant surface and two cut surfaces 0.5 0.2 100 a White and brownish yellow soft callus on explant surface and two cut surfaces 2.0 0.2 86.6 c Little brownish yellow soft callus on one cut surfaces * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. Table 3.24. Effect of the combination of NAA and BA on the morphogenesis of the lTCL_C petiole explants at photoperiod of 16 hours/day PGRs (mg/l) Morphogenesis Remarks NAA BA Callogenesis (%) 1.0 0.1 60.0 c Little brownish green compact callus on two cut surfaces 1.0 0.2 53.3 d Little brownish green compact callus on one cut surface 1.0 0.5 13.3 f Little brown compact callus 1.0 1.0 46.6 e Little brown compact callus on two cut surfaces 1.0 2.0 80.0 b Little green and brown callus on explant surface and two cut surfaces 0.1 1.0 0 g Necrosis 0.2 1.0 0 g Necrosis 0.5 1.0 0 g Necrosis 2.0 1.0 93.3 a Little white and brown callus on explant surface and one cut surfaces * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. 19 Table 3.25. Effect of the combination of NAA and BA on the morphogenesis of the lTCL_C petiole explants in dark PGRs (mg/l) Morphogenesis Remarks NAA BA Callog enesis (%) Adventitious root Root formation (%) Number of roots 1.0 0.1 86.6 c 80.0 b 1.1 b Large amount of brownish yellow soft callus on explant surface and one cut surface. Long white root 1.0 0.2 100 a 93.3 a 3.9 a Little yellow and brown soft callus on explant surface and two cut surfaces Long white root 1.0 0.5 93.3 b 7.7 c 0 c Little yellow and white soft callus Long white root 1.0 1.0 86.6 c 0 d 0 c Little yellow soft callus on one cut surface 1.0 2.0 86.6 c 0 d 0 c Brownish yellow soft callus on one cut surface 0.1 1.0 0 d 0 d 0 c Necrosis 0.2 1.0 0 d 0 d 0 c Necrosis 0.5 1.0 0 d 0 d 0 c Survival but no development 2.0 1.0 100 a 0 d 0 c Little milky white soft callus on explant surface and two cut surfaces Long white root * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. 3.1.1.8. Effect of the combination of auxin and cytokinin on the morphogenesis of the tTCL_R rhizome explants in light and dark conditions. After 10 culture weeks, results are shown in Tables 3.26 to 3.31 and Figures 3.17, 3.18, 3.19. Table 3.26. Effect of the combination of 2.4-D and BA on the morphogenesis of the tTCL_R rhizome explants at photoperiod of 16 hours/day PGRs (mg/l) Morphogenesis Remarks 2,4-D BA Callogenesis (%) 1.0 0.1 100 a Large amount of white and brownish yellow compact callus 1.0 0.2 100 a Large amount of white and brownish yellow compact callus 1.0 0.5 100 a Large amount of milky white soft callus 20 1.0 1.0 100 a Large amount of milky white soft callus 1.0 2.0 100 a Little light yellow and greenish white soft callus 0.1 1.0 34.4 e Little brown compact callus 0.2 1.0 66.6 c Little brown compact callus 0.5 1.0 50 d Little brownish yellow soft callus 2.0 1.0 72.2 b Little milky white and brownish yellow compact callus * Different letters within a column indicate significant differences at P<0.05 by Duncan’s multiple range tests. Table 3.27. Effect of the combination of 2.4-D and BA on the morphogenesis of the tTCL_R rhizome explants in dark. PGRs (mg/l) Morphogenesi s Remarks 2,4-D BA Callogenesis (%) 1.0 0.1 100 a Large amount of white and yellow compact callus 1.0 0.2 100 a Large amount of milky white and yellow compact callus 1.0 0.5 100 a Large a