Somaclonal variations were not induced by the cryopreservation: Levels of somaclonal variations of in vitro and thawed protocorms of Dendrobium Bobby Messina analysed by SCoT and TRAP DNA markers

• Published in: South African journal of botany 2015-09, Vol.100, p.148-157
• Main Authors: Antony, J.J.J., Shamshir, R.A., Poobathy, R., Chew, B.L., Subramaniam, S.
• Format: Article
• Language: English
• Subjects: Cryopreservation; Dendrobium Bobby Messina; SCoT; TRAP; Vitrification
• ISSN: 0254-6299; 1727-9321
• DOI: 10.1016/j.sajb.2015.04.012

The present study was conducted to develop targeted region amplification polymorphism (TRAP) and start codon targeted polymorphism (SCoT) DNA markers for the identification of somaclonal variation in cryopreserved Dendrobium Bobby Messina. With reference to previous orchid cryopreservation via PVS2 (plant vitrification solution) vitrification methods, regenerated explants were assessed in order to determine the genetic similarity in comparison to the mother plant. 3 different samples were selected involving stock culture PLBs (protocorm-like bodies), non-cryopreserved PLBs and thawed cryopreserved PLBs. During the study, eight pairs of (8) TRAP primers (TRAP CMS TRAP 8-4A, FLS TRAP 2-2A, TRAP 14-5A, TRAP 20-6B, TRAP 16-3B, ChIC TRAP 5-5B, TRAP3-1D and TRAP 9-5A) produced unambiguous and reproducible bands ranging from 100 to 2000bp. In cryopreserved PLBs, all TRAP primers displayed polymorphic bands. In the non-cryopreserved PLBs, primer TRAP 20-6B produced monomorphism and the remaining 7 showed both complete and partial polymorphism, respectively. SCoT markers indicated that four primers were able to generate reproducible and clear bands at the sizes of 500 to 3000bp. SCoT primers (S26, S32, S33 and S36) showed polymorphism for both cryopreserved and non-cryopreserved PLBs of Dendrobium Bobby Messina. The TRAP and SCoT DNA markers designed were found to be an efficient tool to evaluate potential rate of somaclonal variations of regenerated PLBs following cryopreservation. •Both TRAP and SCoT markers displayed somaclonal variation in regenerated cryopreserved PLBs of Dendrobium Bobby Messina.•These techniques offers an effective approach in detecting somaclonal variation following cryopreservation.•To our knowledge, this study is the first to be conducted for Dendrobium Bobby Messina utilizing TRAP and SCOT markers for the analysis of genetic variation in cryopreservation research in orchids.