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

Abstract

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.

Direct and indirect in vitro regeneration of Miscanthus × giganteus cultivar Freedom: effects of explant type and medium on regeneration efficiency

“…The individuals were similar for all characteristics except plant height, which might have been attributed to environmental differences. This is consistent with the few Mxg regeneration studies that analyzed field-transplanted micropropagules, although their primary focus was on comparison to rhizome-propagated plants (Lewandowski 1998;Kim et al 2010;Rambaud et al 2013). We also conducted ISSR PCR analysis on field-transplanted regenerants, the first time that this procedure has been used to assess somaclonal variation among/between Mxg tissue culture and rhizomepropagated regenerants.…”

“…We also conducted ISSR PCR analysis on field-transplanted regenerants, the first time that this procedure has been used to assess somaclonal variation among/between Mxg tissue culture and rhizomepropagated regenerants. Rambaud et al (2013) conducted amplified fragment length polymorphism (AFLP) analysis on their Mxg cv. Floridulus regenerants and noted one polymorphic fragment in their regenerants, but that primer pair also yielded a polymorphic fragment in the rhizome-propagated plants.…”

“…However, the genotypes/cultivars utilized were not often identified; Mxg genotypes were provided by Głowacka et al (2010;cv. MG1) and Rambaud et al (2013;cv. Floridulus).…”

“…In addition to indirect regeneration, a few direct regeneration procedures have been reported via axillary shoot tillering from shoot apices (Lewandowski 1997) and nodal sections (Gubišová et al 2013;Rambaud et al 2013) on initial media containing either 4.4-22.2 μM BA alone or with 2.6 μM indole-3-acetic acid (IAA).…”

Bioenergy Crops and Natural Enemies: Host Plant-Mediated Effects of Miscanthus on the Aphid Parasitoid Lysiphlebus testaceipes

“…Sac, 2n=2x=38), and M. sinensis (var. Goliath, 2n=4x=76) [35], were obtained by in vitro multiplication as described by Rambaud et al [36].…”