Optimization of Barley Mature Embryo Regeneration and Comparison with Immature Embryos of Local Cultivars

Optimization of Barley Mature Embryo Regeneration and Comparison with Immature Embryos of Local Cultivars Regeneration ability of plant cells or tissues in explant culture is one of the key factors affecting success of genetic transformation. In experiments, the effect of explant type (whole embryo, scutellum embryonic axis, meristematic/central zone of embryonic axis) and plant growth regulators (BAP or TDZ) on mature embryo regeneration was determined. Explant type significantly affected regeneration efficiency. While no regenerants were observed using mature scutella, whole embryos or embryonic axes produced the highest number of regenerants. Using embryonic axes with discarded apical and basal parts, regeneration efficiency dramatically decreased. No statistical differences in regeneration were observed between BAP and TDZ added to the regeneration medium in concentration 0.1 or 1 mg l-1. At last, regeneration ability of mature embryos of nine Slovak spring barley cultivars (Donaris, Ezer, Levan, Ludan, Nitran, Pribina Sladar, Orbit, Pax) and Golden Promise as a model cultivar was examined and compared with regeneration ability of immature embryos which have been usually used for genetic transformation of barley. Although the regeneration from mature embryos was very weak, the same cultivars Golden Promise, Pribina and Levan showed the best regeneration ability by using both, immature and mature embryos. On the other hand cultivars Ezer and Pax belonged to the weakest ones in both experiments.

[1]  D. Mihálik,et al.  REGENERATION EFFICIENCY OF SLOVAK SPRING BARLEY CULTIVARS AND WINTER WHEAT CULTIVARS , 2011 .

[2]  J. Snape,et al.  High-throughput Agrobacterium-mediated barley transformation , 2008, Plant Methods.

[3]  J. Schulze,et al.  Mature embryo axis-based high frequency somatic embryogenesis and plant regeneration from multiple cultivars of barley (Hordeum vulgare L.). , 2005, Journal of experimental botany.

[4]  J. Schulze,et al.  A highly efficient plant regeneration system through multiple shoot differentiation from commercial cultivars of barley (Hordeum vulgare L.) using meristematic shoot segments excised from germinated mature embryos , 2004, Plant Cell Reports.

[5]  G. Scoles,et al.  Production of multiple shoots from thidiazuron-treated mature embryos and leaf-base/apical meristems of barley (Hordeum vulgare) , 2003, Plant Cell, Tissue and Organ Culture.

[6]  L. Dahleen Improved plant regeneration from barley callus cultures by increased copper levels , 1995, Plant Cell, Tissue and Organ Culture.

[7]  R. Qu,et al.  Thidiazuron promotes in vitro regeneration of wheat and barley , 2000, In Vitro Cellular & Developmental Biology - Plant.

[8]  L. Dahleen Donor-plant environment effects on regeneration from barley embryo-derived callus , 1999 .

[9]  P. Lemaux,et al.  Transformation of recalcitrant barley cultivars through improvement of regenerability and decreased albinism , 1998 .

[10]  R. Kalla,et al.  Agrobacterium tumefaciens‐mediated barley transformation , 1997 .

[11]  P. Lemaux,et al.  Generation of Large Numbers of Independently Transformed Fertile Barley Plants , 1994, Plant physiology.

[12]  F. Skoog,et al.  A revised medium for the growth and bioassay with tobacco tissue culture , 1962 .

[13]  L. Klčová,et al.  Cultivar and environmental conditions affect the morphogenic ability of barley (Hordeum vulgare) scutellum-derived calli , 2022 .