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Efficient resynthesis of oilseed rape (Brassica napus L.) from crosses of winter types B. rapa × B. oleracea via simple ovule culture and early hybrid verification

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Abstract

Resynthesis of Brassica napus is an important tool for the broadening of genetic diversity as well as crop cultivar improvement in oilseed rape. To the best of our knowledge, this is the first study focusing on the crossability, regeneration, and early verification of hybrids between diploid winter turnip rapes (Brassica rapa ssp. oleifera f. biennis), as the female component, and diploid winter curly kales (Brassica oleracea convar. acephalla var. sabellica). Pollination efficiency, mean embryo regeneration per bud and siliqua, as well as their relationships were evaluated in ovule cultures of twenty-four different one-sided crosses between six accessions of B. rapa and four accessions of B. oleracea. Successful germination of the embryos was achieved in 23 combinations. The mean number of embryos per bud reached 0.34. The most productive cross, Svalöfs Duro × Kapral, yielded 2.7 embryos per bud in the best replication. A significant and positive correlation (r = 0.26*) was detected between the pollination efficiency (i.e., setting of siliquae) and the number of embryos per siliqua. In the majority of crosses, procedures implemented exhibited better results than in similar published experiments on spring Brassica genotypes, and provides a simplified and less laborious method. The hybrid nature of all regenerants was verified via flow cytometry. The use of bulked samples for analysis was reliable due to highly significant differences in relative DNA content between all hybrid combinations and their respective parental components. Chromosome counting and morphological assessment further corroborated the results of the flow cytometric analysis.

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Abbreviations

BA:

N6-Benzyladenine

IAA:

Indole-3-acetic acid

FCM:

Flow cytometry

HPR:

Hybrid production rate

HSR:

Hybrid siliqua ratio

RS:

Resynthesized

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Acknowledgments

This work was supported by the Ministry of Agriculture of the Czech Republic, Projects No 0002700604, RO0414; and by the Czech University of Life Sciences in Prague, Project No IGA FTA 20145022. We thank Petra Bartosová, Václava Stresková, and MSc. Vlasta Jarolímová for their assistance with plant material processing; René Miller and Peter Lemkin for critical reviewing of the manuscript and linguistic revision.

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Authors and Affiliations

  1. Division of Crop Genetics and Breeding, Crop Research Institute, Drnovská 507, 161 06, Ruzyne, Prague 6, Czech Republic

    Alois Hilgert-Delgado, Miroslav Klíma, Milan Oldrich Urban & Miroslava Vyvadilová

  2. Faculty of Tropical AgriSciences, Czech University of Life Sciences Prague, Kamýcká 129, 165 21, Suchdol, Prague 6, Czech Republic

    Alois Hilgert-Delgado, Iva Viehmannová & Eloy Fernández-Cusimamani

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  1. Alois Hilgert-Delgado
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  2. Miroslav Klíma
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Correspondence to Alois Hilgert-Delgado.

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Hilgert-Delgado, A., Klíma, M., Viehmannová, I. et al. Efficient resynthesis of oilseed rape (Brassica napus L.) from crosses of winter types B. rapa × B. oleracea via simple ovule culture and early hybrid verification. Plant Cell Tiss Organ Cult 120, 191–201 (2015). https://doi.org/10.1007/s11240-014-0593-2

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