Micropropagation and Genetic Stability of Neolamarckia cadamba (Kelampayan)

Mok, Pei Kieng (2019) Micropropagation and Genetic Stability of Neolamarckia cadamba (Kelampayan). Masters thesis, Universiti Malaysia Sarawak (UNIMAS).

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Abstract

Rapid clonal propagation of Neolamarckia cadamba (Roxb.) Bosser is significant for large-scale production of planting materials to support the development of planted forests and reduce reliance on natural forests in Malaysia. The genetic stability of regenerated plantlets is crucial to ascertain the quality and uniformity of planting materials for planted forests development. An efficient protocol was established for micropropagation of N. cadamba through direct shoot organogenesis using nodal explants. An average of 5.4 shoots per explant was obtained in B5 medium fortified with 1.0 mg/L BAP. The axillary shoot proliferation was achieved with the number of shoots ranging from 3.5 to 5.4 after subcultured on B5 medium augmented with 0.8 mg/L BAP. Half-strength B5 medium enriched with 0.1 mg/L PBZ was found suitable for rooting with 100% of root induction. The rooted plantlets were successfully acclimatised and transplanted to the greenhouse with more than 95% survival rate. The genetic stability of the regenerants from first, second, third and sixth subcultures of four stock plants was verified by using five random amplified polymorphism DNA (RAPD) and two inter-simple sequence repeats (ISSR) primers along with single nucleotide polymorphism (SNP) analysis by using a gene-specific marker. By using the RAPD and ISSR markers, a total of 9,334 bands and 2,760 bands were amplified respectively, from a total number of 164 tested plants. All of these bands were monomorphic across all in vitro regenerated plantlets. The consensus sequences of Susy gene from plantlets were aligned and compared, however no SNP was detected. These results ascertained the efficiency and reliability of this micropropagation protocol for commercial propagation and germplasm conservation of selected N. cadamba, as well as to facilitate the development of planted forests.

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