Somatic embryogenesis from in vitro anther culture of apomictic buffel grass genotypes and analysis of regenerated plants using flow cytometry

E. CARLONI; A. RIBOTTA; E LÓPEZ COLOMBA; GRIFFA, S.; QUIROGA, M.; TOMASSINO, E.; GRUNBERG, K.

PLANT CELL TISSUE AND ORGAN CULTURE

SPRINGER

Lugar: Berlin; Año: 2014

0167-6857

Buffel grass is a fodder grass that reproducesmainly via apomixis. Using in vitro tissue culture in apomicticspecies provides alternatives that can be applied in abreeding program. The aims of this work were to establisha protocol for in vitro generation of apomictic buffel grassgenotypes using anthers as explants and to evaluate thegenetic stability of regenerated plants via flow cytometry(FCM). Three genotypes were able to induce embryogeniccalli in Murashige and Skoog (MS) medium supplementedwith 6 mg/l of 2,4-dichlorophenoxyacetic acid. Seedlingregeneration occurred in a MS medium supplemented with0.5 mg/l napthaleneacetic acid ? 1 mg/l 6-benzylaminopurin.Seedlings were derived from somatic embryos and themorphogenic process was induced using the somatic tissueof the stamens. Induction and regeneration efficiencydepended on the genotype and was affected by date of tillercollection, different pretreatments or the interaction ofthese variables. FCM analyses in in vitro regenerated plantsshowed genetic instability in their nuclear DNA content.Plants with lower nuclear DNA content may indicate DNAaneuploids (8.6 %), whereas plants that had twice the valueof nuclear DNA content (4.7 %) suggest in vitro polyploidization.This variation observed in apomictic genotypesprovides an opportunity to obtain new variants, which maythen be included as sources of genetic variability in buffelgrass breeding programs.