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In vitro selection and characterization of buffelgrass somaclones with different responses to water stress

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Buffelgrass is a forage grass that reproduces mainly by apomixis. In species with this reproduction mode, in vitro selection allows the incorporation of alternatives in a breeding program. The aims were to define a protocol for in vitro selection, provide a molecular and morphological characterization of the progenies of regenerated plants, and evaluate them under water stress conditions. In the embryogenic callus induction medium (IM), the highest values of the variables fresh weight of embryogenic calli, proportion of embryogenic calli and number of regenerated seedlings (NRS) were obtained in the 25 mM mannitol treatment. The remaining concentrations of the osmotic agent (50, 75, 100 and 150 mM) had a negative effect on these variables. In the regeneration medium (RM), NRS was reduced at all mannitol concentrations. When embryogenic calli were induced and seedlings were regenerated maintaining mannitol concentrations in IM and RM, the highest NRS values were recorded at 25 mM mannitol. In vitro regenerated seedlings transplanted to an experimental plot exhibited different morphological characteristics from those of the anther donor plant. ISSR primers detected 22% of polymorphic bands and divergence between 0.20 and 0.37 in in vitro regenerated plants. Finally, water stress assays confirmed that S1 progenies exhibited a differential behavior from that of the parent material. Under 100 mM of mannitol used as selection pressure in IM or in both IM and RM, S1 progenies of two regenerated materials had higher height, fresh weight and dry weight at the end of water stress assay.

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Induction medium


Regeneration medium


Fresh weight of embryogenic calli


Proportion of embryogenic calli


Number of regenerated seedlings


Induction stage


Regeneration stage

R1 :

Regenerated seedlings

S1 :

Progenies of in vitro regenerated plants


Inter simple sequence repeat


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This work was supported financially by The Instituto Nacional de Tecnología Agropecuaria (INTA), Argentina, Project PNPA-1126072 and CONICET PIP 112 201101 0031.

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Correspondence to Edgardo Carloni.

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Carloni, E., Tommasino, E., López Colomba, E. et al. In vitro selection and characterization of buffelgrass somaclones with different responses to water stress. Plant Cell Tiss Organ Cult 130, 265–277 (2017). https://doi.org/10.1007/s11240-017-1220-9

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  • Anther culture
  • Apomixis
  • Polyploids
  • Somatic embryogenesis
  • Abiotic stress