Explant, Medium, and Plant Growth Regulator (PGR) Affect Induction and Proliferation of Callus in Abies koreana

Abstract

Korean fir (Abies koreana E.H. Wilson) is a unique Pinaceae tree species endemic in Korea. In recent years, it is believed that climate change has caused many of them to die. Therefore, it has become extremely important to protect and preserve this tree species. In this study, the possibility of callus induction using different explants, media, and plant growth regulators (PGRs) was studied. After the dormancy period in May 2020, needles and stem segments that grew from the leaf buds as the explants were collected from one-year-old shoots. The explants were disinfected and subsequently transferred to culture media supplemented with different combinations of auxins and cytokinins. These explants were cultured in the dark in a culture room with a 16 h photoperiod, day/night temperature of 24/18 & DEG;C, and 80% relative humidity. After 8 weeks, significant differences were observed in the callus induction and proliferation, as affected by the explant type, basic medium, and PGR. The stem segments were more suitable as the explants for callus induction than needles were. Furthermore, fluffy calli suitable for differentiating the regeneration buds were observed on the calli induced from stem segments. The Murashige and Skoog (MS) medium was the most effective of the three media used in this study, namely MS, Douglas fir cotyledon revised (DCR), and Quoirin and Lepoivre (LP) media, with the highest callus induction ratio of stem segments being 100.0%. The highest fresh callus weight was also observed on the MS medium (819.3 mg). Moreover, the PGR combinations of alpha-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), and 6-benzylaminopurine (6-BA) consistently exerted a positive influence on callus induction throughout this study. In addition, the advantages of these two kinds of PGR were reflected in callus proliferation. The callus proliferation ratio reached 1147.6% as compared to the initial fresh weight, with a high concentration of 2,4-D (3.0 mg & BULL;L-1). In conclusion, the MS medium was optimal for callus induction on the stem segment explants, and 2,4-D promoted callus induction as well as an increased proliferation ratio of callus in A. koreana.</p>