Androgenic responses of winter wheat (Triticum aestivum L.) combinations in in vitro anther culture

The androgenic parameters were investigated by in vitro anther culture (AC) on nine F25 breeding combinations of winter wheat (Triticum aestivium L.). Each combination produced embryo-like structures (ELS), green plantlets, albino plantlets, transplanted plantlets and acclimatized plantlets, with respect to the number of anthers in AC. The number of AC-derived ELS was between 11.73 and 52.76 ELS/100 anthers with the mean of 26.22 ELS/100 anthers, out of which the number of regenerated green plantlets varied from 3.20 to 26.40 green plantlets/100 anthers and the mean was 9.76 green plantlets/100 anthers, while the number of transplanted plantlets ranged from 2.16 to 21.77 transplanted plantlets/100 anthers. Furthermore, the number of albinos/100 anthers was mitigated and varied between 0.72 and 6.20 albinos/100 anthers. We also studied the number of green and albino plantlets per 100 ELS. The rate of green plantlets per 100 ELS ranged between 14.81% and 64.01%, with the overall mean 33.59%, while the rate of albinos per 100 ELS ranged from 4.82% to 20.70% with the overall mean 11.93%. In our experiment, the rate of acclimatized plantlets (70.15-91.57%) depended mostly on the combination. This study asserted the importance of AC method in wheat for in vitro production of green plants. Although albinism was found in each combination, it did not hinder the production of green plantlets. The satisfying results were achieved in green plantlets production compared to the previously published data, but further improvement will be needed continuously, experiment by experiment. The generated acclimatized plantlets will be used in the wheat breeding program as doubled haploid (DH) lines

Generation of doubled haploid lines from winter wheat (Triticum aestivum L.) breeding material using in vitro anther culture

We investigated the anther culture (AC) efficiency of thirteen F4 combinations of winter wheat (Triticum aestivum L.). The genotype dependency was assessed during the induction of the androgenic entities, i.e. embryo-like structures (ELS), regenerated-, green-, albino-, and transplanted plantlets. The number of green plantlets per 100 anthers (GP/100A) varied from 0.36 to 24.74 GP/100A with a mean of 8.31 GP/100A. Albino plantlets (AP) occurred in each combination, ranging from 0.20 to 22.80 AP/100A with an average value of 5.59 AP/100A. Between 25–87.76 doubled haploid (DH) plants per 100 acclimatised plantlets (DH/100ADP), depending on the combination, with a mean of 59.74% were recovered. We have found the highest DH production in the combinations Béres/Midas, Kalász/Tacitus, Béres/Pamier, and Premio/5009. This improves remarkably the choice of basic genetic material in subsequent crossing programmes. These observations emphasise the usability and efficiency of in vitro AC in producing a large number of DH lines for breeding and the applied researches of winter wheat. Although albinism was found in each combination, it was mitigated by the in vitro AC application

Efficient in vitro anther culture for androgenic plant production in F3-6 winter wheat (Triticum aestivum L.) bulk combinations

284-293The efficiency of in vitro anther culture (AC) method on six common wheat (Triticum aestivum L.) F3-6 bulk combinations was investigated for the AC-induced androgenic traits, such as embryo-like structures (ELSs), regenerated plantlets, green plantlets, albinos, transplanted plantlets, and acclimatized plantlets. Moreover, genotype dependency was tracked among the tested combinations in the AC method. The results showed that all androgenic traits were produced in each tested combination and non-significant differences were found among combinations for all androgenic traits except albinos. The number of green plantlets ranged between 8.00 and 19.00 green plantlets/100 anthers based on the combination, with an overall mean of 14.37 green plantlets/100 anthers. The total number of acclimatized plantlets was 496 plantlets out of the 604 transplanted plantlets, with a rate of 82.12%. This investigation confirms the efficiency and utility of in vitro AC method in winter wheat to produce doubled haploid (DH) lines based on androgenesis for the breeding program and genetic studies. AC- induced DH lines produced subsequently from acclimatized plantlet individuals in the nursery will be inserted in breeding programs for wheat drought tolerance evaluation and in crossing programs for more improvement in green plants production in the nursery. Albinism and genotype dependency, limiting-factors for wheat AC-induced DH production, were mitigated by the application of some modifications of the wheat AC protocol. The modifications included a 2-week cold pre-treatment of donor tiller at 4C in constant dim light and the use of the W14 mf induction medium instead of P4 mf