An advanced protocol for the establishment of plantlets originating from somatic embryos in Pinus massoniana
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ORIGINAL ARTICLE
An advanced protocol for the establishment of plantlets originating from somatic embryos in Pinus massoniana Ruiling Yao1 · Yin Wang2 Received: 25 September 2019 / Accepted: 8 August 2020 © King Abdulaziz City for Science and Technology 2020
Abstract This study describes critical factors affecting germination of somatic embryos and plantlet regeneration in Pinus massoniana. Somatic embryos from the same embryogenic line 27 of P. massoniana were used as test materials. The supplementation of activated charcoal (AC) in the medium was essential for the germination of mature somatic embryos, while the addition of excessive AC to the medium was prohibitive for somatic embryo germination. The highest germination rate was found on the medium containing 10 g/l AC, and the addition of 5 g/l AC to the medium was optimal to the growth of germinating somatic embryos. Thidiazuron (TDZ) was linearly related to the number of sprouting axillary buds. However, the growth of sprouting buds was retarded when > 4 µmol/l TDZ was added into culture medium. Exogenous plant growth regulators added to the medium significantly improved the root regeneration capacity of shoots. The highest root regeneration rate was observed under the treatment of 1.2 µmol/l ɑ-naphthaleneacetic acid (NAA) plus 2 µmol/l paclobutrazol (PBZ), reaching 96.3%. One year after the field transfer, the growth performance of plant height, caliper, and survival rate for rooted shoots was significantly better than that of plantlets directly developed via somatic embryogenesis. The presented results provide useful instruction for the establishment of plantlets originating from somatic embryos, and would be able to make a great contribution to the clonal forestry of P. massoniana. Keywords Clonal forestry · Indirect somatic embryogenesis · Masson pine · Paclobutrazol · Thidiazuron
Introduction Masson pine (Pinus massoniana), a native species of southern China, is one of the most important tree species in subtropical regions, and is cultivated for the production of timber and natural resin (Ding et al. 2006). To date, seedlings propagated by seeds are the main planting materials for the establishment of P. massoniana plantations, resulting in the low productivity of forests due to the great genetic variations among trees (Zhu et al. 2010). To enhance stand Ruiling Yao and Yin Wang contributed equally to this work. * Ruiling Yao [email protected] Yin Wang [email protected] 1
Tree Biotechnology Research Centre, Guangxi Forestry Research Institute, Nanning 530002, China
Pines Breeding Research Centre, Guangxi Forestry Research Institute, Nanning 530002, China
2
productivity, and promote the industrial development of P. massoniana, clonal forestry is urgent, with the aim of genetic gains with cloning. Concerning the clonal propagation of P. massoniana, the utility of cutting propagation has been indicated (Shen 2018). However, donor plant age is negatively related to the root regeneration capacities of P. massoniana cuttings (Ji et al. 1996), as
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