Abstract
Production of doubled haploid (DH) cereals is becoming increasingly important in crop breeding programs, but the methods currently applied still remain inefficient. In this study, we present the procedure for obtaining haploid and DH oat plants by pollination with maize. Thirty-three oat genotypes were used in the experiments. Oat plants (14,543 florets) were pollinated with maize pollen 2 days after emasculation and treated with auxin analogues: 2,4-dichlorophenoxyacetic acid (2,4-D) or 3,6-dichloro-2-methoxybenzoic acid (dicamba), at a concentration of 100 mg dm−3. These auxins had no significant influence on the number of haploid embryos developed, but they significantly affected their germination ability, and thus haploid and DH plant production. After application of 2,4-D, 5.06 % of haploid embryos developed per emasculated florets, 1.37 % of haploid plants and 0.54 % of DH lines, whereas after dicamba treatment, 4.3 % of haploid embryos, 0.64 % of haploid plants and 0.25 % of DH lines. Haploid embryos were obtained from all genotypes tested, however, their frequency differed between individual genotypes. The highest number of embryos per emasculated florets (9.0 %) was obtained from the DC09040 genotype after dicamba treatment, and from STH123 × Skorpion (8.9 %) after 2,4-D treatment. The genotype did not significantly affect the development of haploid plants, nevertheless the highest number of DH lines was obtained from the Arab × Typhon genotype. There were 52 DH lines acquired from 28 genotypes, which produced a total of 5227 seeds. The number of seeds varied between the DH lines from 2 to 595. Seeds of all the DH lines produced fertile next generation. DH lines are currently included in breeding programs.
Highlights
Haploids are plants that contain the gametic number of chromosomes (n)
Two-way ANOVA for production traits of oat doubled haploids indicated no significant differences in the efficiency of haploid embryos
The differences were highly significant for haploid plants and doubled haploid (DH) lines per florets when the type of auxin was the source of variance (Table 1)
Summary
Haploids are plants that contain the gametic number of chromosomes (n). In nature, the spontaneous emergence of haploids in higher plants is very rare (a frequency of0.001–0.01 %), currently there are several techniques available enabling their production. Haploids are produced in vivo and in vitro by androgenesis (culture of anthers or microspores), gynogenesis (culture of unfertilized ovules), interspecific or intergeneric crosses, followed by chromosome elimination, or pollination with irradiated pollen (Ferrie et al.2014). The time required to obtain them using in vitro techniques is very short when compared to traditional breeding schemes, based on multiple self-pollination generations, which take 6–10 years. Haploid plants are very important in various research disciplines, such as plant biotechnology, molecular genetics or traditional plant breeding. They have many applications in basic plant research, such as cytogenetics, crop evolution, mutagenesis induction, genetic transformation and in breeding programs
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