ISHS Acta Horticulturae 494: II International Symposium on Chestnut
INDUCTION OF ADVENTITIOUS SHOOTS FROM INTERNODAL LONGITUDINAL SECTIONS OF CASTANEA SATIVA MILL. |
| Authors: | M. Mulin, M.S. Pais, R. da Costa Seabra |
| Keywords: | Adventitious shoots, Internodal longitudinal sections, Castanea sativa |
| Abstract:
The objective of this study is to achieve a reliable protocol of plant regeneration from Castanea sativa Mill. to be used in genetic transformation experiments. The explants used were internodal longitudinal sections (ils) or internodal segments (isg) of in vitro material established from seedlings of a single C. sativa var. Martainha tree and maintained in culture for 3 years. Adventitious shoots were induced from the explants on a modified medium of Murashige and Skoog containing BAP and IAA. Histological observations indicate that the adventitious shoots formed on the cut surface of ils were formed from the cambial tissue. Although a lower rate of shoots are regenerated from ils than from isg, the use of ils instead of isg presents an advantage of increasing the surface area for Agrobacterium contamination or for direct DNA transfer by particle bombardment. Severe endogenous bacterial contamination problems were encountered in the non-resistant Martainha variety. In order to reduce the rate of endogenous contamination, we sub-cultured the apical end of the in vitro shoots at monthly intervals. In addition, shoots obtained from nodal explants (nls) were free from endogenous bacterial contamination. When a second cycle of ils were isolated, from these shoots and cultured, the induced shoots were free from endogenous bacterial contamination. Nodal explants gave rise to shoots which rooted in vitro. These roots were functional once tranferred to soil. | |
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