Genetic Stability of Micropropagated White Kelampayan (Neolamarckia cadamba) plantlets using ISSR Markers

Mok, Pei Kieng (2016) Genetic Stability of Micropropagated White Kelampayan (Neolamarckia cadamba) plantlets using ISSR Markers. [Final Year Project Report / IMRAD] (Unpublished)

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Abstract

Micropropagation of Neolamarckia cadamba (Roxb.) Bosser is important for large scale commercial production while maintaining its elite genetic information and for forest plantation in Sarawak: in order to achieve the goal of of 1.0 million hectares of forest plantations with fast-growing tree species in 2020. Occurrence of soma clonal variations is one of the major problems in micropropagation due to repeated subcultures of plantlets. Five stock plants of N. cadamba were successfully regenerated from tissue culture through direct organogenesis by using shoot tips and nodal explants. By using 1.0 mglL of BAP alone in the Gamborg B5 medium, the highest shoot multiplication rate of 5.0 of shoots per explants was observed. Elongation of shoots and root formation in the presence of plant growth regulator was not essential in this study. The mUltiplied shoots were subsequently subcultured two to three times. In this study, genetic stability of the plantlets after several cycles of subcultures was successfully assessed through Inter-Simple Sequence Repeats (ISSR) marker. ISSR profiles with scorable and clear banding pattern were generated. A total of 13 loci and total of 1378 bands that ranged from 600 bp to 3500 bp were detected from a total samples of 106 by using the primer. All the bands from the plantlets of two stock plants were monomorphic whereas some polymorphisms from the other three stock plants had been detected with PPB ranging from 7.69 % to 38.46 % at each subculture and Shannon index ranging from 0.0661 to 0.1314 respectively. The number of subculture passages has showed effect on the genetic stability of micropropagated plantlets. Therefore, more studies will be required to find out and overcome the problems in order to produce plantlets that are able to maintain elite genetic traits that suit commercial requirements.

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