Original Research Production of Haploids and Doubled Haploids from Unfertilized Ovule Culture of Various Wild Species of Gentians (Gentiana spp.)

The production of haploids and doubled haploids (DHs) on unfertilized ovule culture was examined in 19 wild species of gentians ( Gentiana ssp.) classified into eight sections. Of the 19 species including 35 strains, embryo-like structures (ELSs) were obtained in 15 species, and regenerated plants were produced in 11 species. ELS production has varied greatly among the15 species, i.e., 0.5%-79.2% frequency of responding flower buds and 0.01-1.99 ELSs per flower bud. Of the ELS-producing species, almost all were classified into the sects. Pneumonanthe or Cruciata . Species in sect. Pneumonanthe showed higher responses than those in sect. Cruciata . In examining the effect of flower bud stage on ELS production, more than twice as many ELSs were observed at the anther-dehiscent stage than that at the anther-indehiscent stage. Ploidy level was determined in 117 randomly selected regenerated plantlets, which suggests that most were haploid (32.5%) and diploid (46.2%). When 12 diploid plants were examined using simple sequence repeat (SSR) markers, 8 (66.7%) were DH. This study revealed that unfertilized ovule culture can be applied not only on cultivated gentian species but also on a number of wild species. The production of haploids and DHs in wild gentians provides novel prospects for ornamental and/or medicinal gentian breeding.

Development of breeding lines by zigotic ovule culture in Nigella sativa L.

This study was carried out in order to develop an embryo culture technique and to establish breeding lines with this technique. Ovule culture was preferred in the study because the seeds were too small and embryo isolation was difficult. Embryo cultures were done by using populations obtained from producers in Samsun, Denizli, Isparta, Mersin in Turkey and ?ameli black cumin variety. Hybridizations were done according to the semi-diallel hybridization method to gain zygotic embryos. LS2.5 and MS (Sigma Aldrich Catalog number: M5519) mediums were used for ovule culture and MSD4 medium was used growing the plants obtained from the ovules. As a result of the research, a total of 2904 ovules were cultured in LS2.5 medium. 148 of them showed callus development; callus formation rate was 5.10%. The highest callus formation rate in the investigated combinations was obtained from ?ameli x Denizli combination at 7.26%. Plant regeneration could not be obtained from these calluses. A total of 3526 ovules were cultured in MS medium. Sixty plantlets were obtained from these ovules and plant formation rate was determined as 1.70%. 41 plants from these matured and were harvested.