Shoot organogenesis in three Miscanthus species and evaluation for genetic uniformity using AFLP analysis

A simple, efficient protocol for direct in vitro shoot organogenesis and regeneration was established for three species of Miscanthus including two clones of Miscanthus x giganteus, one clone of M. sinensis and one clone of M. sacchariflorus. Shoots were induced from the axillary nodes of both M. x giganteus and M. sacchariflorus and from apical meristems of both M. sinensis and M. sacchariflorus. A tillering method was used to accelerate shoot proliferation. Shoots were rooted in a wet perlite substrate in pots in the greenhouse. Subsequently, rooted plants were transferred to the field. The genetic uniformity of regenerated plants was evaluated using amplified fragment length polymorphism analysis and compared to that of rhizome-propagated plants. A total of 33,443 fragments were generated, representing 869 markers. There were 21 fragments (0.06 % of the fragments) or 19 markers (2.19 % of the markers) that were polymorphic, and almost all of these were singletons. The three species showed similar polymorphisms. Genetic variability was also found in the rhizome-propagated plants, sometimes at a higher rate than in the in vitro culture, indicating that the genetic uniformity was not altered by the protocol. This protocol may help breeders produce new clones of Miscanthus in the future.