Plants regeneration of Balfourodendron riedelianum (Engl.) Engl via adventitious organogénesis

DUARTE, EVELYN; SANSBERRO P; LUNA C

La Plata

Congreso; Fourth International Conference of the IUFRO Working Party 2.09.02. Development and application of vegetative propagation technologies in plantation forestry to cope with a changing climate and environment. La Plata (Buenos Aires) Argentina. September 19-; 2016

IUFRO

Balfourodendron riedelianum (White Guatambú) is a tree species of the family Rutaceae, which is distributed from 10° latitude South in Brazil to 30° latitude south of northern Argentina. It is a representative specie of seasonal semi-deciduous forest of the Atlantic Forest, appreciated for its wood used in luxury furniture, construction and carpentry in general, leading to the overexploitation of the resource, therefore it has been included in the Red Book of the International Union for Conservation of Nature in the category of "endangered species". The aim of the study was to develop an in vitro protocol method, to amplify the germplasm of B. riedelianum. It was evaluated the organogenic capacity of hypocotyl segments and cotyledons from germinated plants in vitro. The seeds were harvested in the Reserve of the Faculty of Forestry at the National University of Misiones (Guaraní - Misiones - Argentina) in September 2012. The seeds were germinated in vitro in tubes of 15 ml capacity containing 4 ml of basal medium composed by the salts and vitamins of Murashige and Skoog (MS), with a sucrose concentration of 30 g.L -1 , semisolid (6.5 g.L -1 agar), and incubated for one month to 27 ± 2 ° C and 14 h photoperiod (116 mol.m -2 .s -1 ). In order to induce organogenesis, hypocotyl and cotyledons sections from in vitro plants were cultured in MS medium supplemented with phytagel (3,5 g.L -1 ), and combinations auxin naphthalene acetic acid (NAA 0.05 µM), cytokinin 6-benzylaminopurine (BA 0.44 µM) and Thidiazuron (TDZ 0.4, 0.04, 0.004 µM). Callus with buds were transferred to temporary immersion bioreactors (TIB), containing 100 ml of MS added with gibberellic acid (GA 3 0.5; 1 µM), BA (0.8, 1.5 µM), to achieve elongation of shoots. Rooting stage was developed in MS medium with agar (6.5 g.L -1 ), indole-3-butyric acid (IBA 2.5, 5, 7 µM) and NAA (0.5, 1.4, 2.7 µM). The cultures were incubated in environmental conditions described above for two months. The best results were obtained when the cotyledon fragments were cultured in NAA 0.05 y TDZ 0.04 µM, where 40±10 of callus provided 8±6.15 adventitious buds. Accordingly 48.79±4.01% thereof provided shoot more than 5 mm in length in the medium with BA 0.8 µM, of which the 46.67±5.77% become seedlings. The results of this work are presented as a viable alternative for large-scale production of seedlings B. riedelianum.