Plant regeneration from the embryogenic calli of five major Miscanthus species, the non-food biomass crops

Abstract

With the growing shortage of oil, coal, and other traditional fossil fuels, scientists in various fields have been looking for new fuel sources to solve the energy crisis. The genus Miscanthus is an ideal biofuel crop due to its rapid vegetative growth and its potential for high biomass yields. Plant regeneration through somatic embryogenesis is a viable method to achieve large-scale production of plant biomass. Callus induction from immature inflorescences of five Miscanthus species was performed on two different media, and the relative rates of callus proliferation were calculated. The highest multiplication coefficient, 3.92, was obtained with M. sacchariflorus ssp. lutarioriparius when the concentration of 2,4-dichlorophenoxyacetic acid (2,4-D) in the medium was 4.0 mg/L, and this species also performed the best at the induction phase. The proliferation coefficient for M. floridulus increased to 3.19 when the concentration of 2,4-D was decreased from 4.0 to 2.0 mg/L. When the concentration of 2,4-D was 2.0 mg/L, the proliferation coefficient for M. sinensis was 2.47. In M. sacchariflorus, the proliferation coefficients were 2.89 and 2.49 for the lower and higher concentrations of 2,4-D, respectively. The multiplication coefficient of M. x giganteus was 2.60 on medium containing 4.0 mg/L 2,4-D. Three different regeneration media were tested to induce shoots in vitro. M. floridulus and M. sacchariflorus regenerated shoots at 100% frequency in three different regeneration media. The regeneration rate for M. sacchariflorus ssp. lutarioriparius reached 99.0% on medium containing 4.0 mg/L N ⁶-Benzylaminopurine (6-BA) and 0.1 mg/L α-Naphthaleneacetic acid (NAA). The best regeneration rate of M. sinensis was 35.2% using 2.0 mg/L 6-BA and 0.3 mg/L NAA, whereas the regeneration rate of M. x giganteus was 57.4% on medium supplemented with 3.0 mg/L 6-BA and 0.2 mg/L NAA. The in vitro-derived plantlets of all five species had 100% rooting rates on basal MS medium without supplementation. The survival rates of plantlets were above 90% for each species when subsequently grown outdoors.