Ankita Rajendra Parab1, Chew Bee Lynn1, Sreeramanan Subramaniam2,3,4,5. 1. School of Biological Sciences, Universiti Sains Malaysia (USM), 11800, Georgetown, Penang, Malaysia. 2. School of Biological Sciences, Universiti Sains Malaysia (USM), 11800, Georgetown, Penang, Malaysia. sreeramanan@gmail.com. 3. Chemical Centre Biology (CCB), Universiti Sains Malaysia (USM), 11900, Bayan Lepas, Penang, Malaysia. sreeramanan@gmail.com. 4. School of Chemical Engineering Technology, Universiti Malaysia Perlis (UNIMAP), 02600, Arau, Perlis, Malaysia. sreeramanan@gmail.com. 5. National Poison Centre, Universiti Sains Malaysia (USM), 11800, Georgetown, Penang, Malaysia. sreeramanan@gmail.com.
Abstract
BACKGROUND: Clonal propagation is one of the attributes of plant tissue culture. Therefore, analysis of genetic stability among the in vitro cultured plants is a crucial step. It helps to signify the clonal propagation of the micropropagated plants. Regenerated Ficus carica var. Black Jack plantlets were established using woody plant medium supplemented with 20 μM 6-Benzylaminopurine and 8 μM Indole-3-acetic acid under different light treatments such as normal fluorescent white light (60 μmol m-2 s-1), and four different LED spectra, white (400-700 nm), blue (440 nm), red (660 nm) and blue + red (440 nm + 660 nm). Genetic stability analysis was performed on the in vitro and ex vitro plants of Ficus carica var. Black Jack. METHODS AND RESULTS: Ten primers of each, ISSR and DAMD molecular markers, were used to assess the genetic stability of the eight samples of Ficus carica var. Black Jack. ISSR markers showed 97.87% of monomorphism whereas DAMD markers showed 100% monomorphism. Polymorphism of 2.13% was observed for the UBC840 ISSR-DNA primer which was negated under the genetic similarity index analysis for the eight samples. The findings of this study revealed that ISSR and DAMD markers are efficient in determining the polymorphism and monomorphism percentage among the in vitro and ex vitro samples of Ficus carica var. Black Jack. CONCLUSION: Monomorphism of 100% obtained using DAMD markers and more than 95% of monomorphism obtained using ISSR markers indicate that the regenerated plants are significantly genetically stable. These molecular markers can be used to test the genetic stability of in vitro regenerated plants. It is recommended that genetic stability analysis should be performed for long-term maintenance of such micropropagated plants.
BACKGROUND: Clonal propagation is one of the attributes of plant tissue culture. Therefore, analysis of genetic stability among the in vitro cultured plants is a crucial step. It helps to signify the clonal propagation of the micropropagated plants. Regenerated Ficus carica var. Black Jack plantlets were established using woody plant medium supplemented with 20 μM 6-Benzylaminopurine and 8 μM Indole-3-acetic acid under different light treatments such as normal fluorescent white light (60 μmol m-2 s-1), and four different LED spectra, white (400-700 nm), blue (440 nm), red (660 nm) and blue + red (440 nm + 660 nm). Genetic stability analysis was performed on the in vitro and ex vitro plants of Ficus carica var. Black Jack. METHODS AND RESULTS: Ten primers of each, ISSR and DAMD molecular markers, were used to assess the genetic stability of the eight samples of Ficus carica var. Black Jack. ISSR markers showed 97.87% of monomorphism whereas DAMD markers showed 100% monomorphism. Polymorphism of 2.13% was observed for the UBC840 ISSR-DNA primer which was negated under the genetic similarity index analysis for the eight samples. The findings of this study revealed that ISSR and DAMD markers are efficient in determining the polymorphism and monomorphism percentage among the in vitro and ex vitro samples of Ficus carica var. Black Jack. CONCLUSION: Monomorphism of 100% obtained using DAMD markers and more than 95% of monomorphism obtained using ISSR markers indicate that the regenerated plants are significantly genetically stable. These molecular markers can be used to test the genetic stability of in vitro regenerated plants. It is recommended that genetic stability analysis should be performed for long-term maintenance of such micropropagated plants.