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Characterization of Ogura CMS fertility-restored interspecific hybrids and backcross progenies from crosses between broccoli and rapeseed

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Abstract

Interspecific F1 hybrids were obtained from crosses between two Ogura cytoplasmic male sterility (Ogura CMS) broccoli lines (B14 and 137) and a rapeseed restorer line to introgress the Ogura CMS’s fertility-restoration gene Rfo from rapeseed into broccoli. Ten Rfo-positive interspecific, triploid plant progenies, phenotypically between rapeseed and broccoli, were obtained but still contained multiple rapeseed genes, with reduced fertility and seed setting potential under natural pollination. For fertility and seed setting improvement, successive backcrosses with broccoli were carried out to yield BC2 progenies. Screening revealed six plants with Rfo among 25 BC2 progenies, which were investigated for agronomic traits, ploidy determination, pollen viability and SSR (simple sequence repeat) markers to evaluate for ideal individuals closer to broccoli. Pollen viability tests were carried out at different stages for various hybrid parents. The results were as follows: 137-F1 (68.54%) > B14-F1 (65.32%) > 137-BC1 (48.15%) > B14-BC1 (32.41%). Although there were significant differences among the BC1 and BC2 individuals, the genetic background of BC2 was closer to the parent broccoli compared with that of BC1 plants. The existence of the Rfo gene in broccoli not only facilitates broccoli germplasm innovation, combination of interspecific hybridization and backcrossing, but may also shed light on trait segregation in different generations.

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Acknowledgements

The experiment was performed at Institute of Horticulture, Shanghai Academy of Agricultural Sciences, National Center for Vegetable Improvement (Shanghai Branch), Shanghai Key Laboratory of Protected Horticultural Technology. We thank Liyong Yang from Institute of Crop Breeding and Cultivation, Shanghai Academy of Agricultural Sciences for providing the rapeseed restorer material.

Funding

This research was funded by the National Key R&D Program of China (Grant No. 2017YFD0101805), Shanghai Agriculture Applied Technology Development Program, China (Grant No. G2016060105), The Youth Talent Development Plan of Shanghai Municipal Agricultural System, China (Grant No. 20180116).

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    Authors and Affiliations

    1. Institute of Horticulture, Shanghai Academy of Agricultural Sciences, National Center for Vegetable Improvement (Shanghai Branch), Shanghai Key Laboratory of Protected Horticultural Technology, Shanghai, 201403, China

      Chun-qing Liu, Guang-qing Li, Xue-qin Yao, Lei Huang & Zhu-jie Xie

    2. School of Ecological Technology and Engineering, Shanghai Institute of Technology, Shanghai, 201418, China

      Xin-yan Wu

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    1. Chun-qing Liu
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    Contributions

    Conceptualization, Z.X.; methodology, C.L.; validation, C.L., G.L. and X.Y.; formal analysis, C.L., L.H. and X.W.; resources, X.Y. and C.L.; data curation, G.L.; writing—original draft preparation, C.L.; writing—review and editing, C.L.; visualization, G.L.; supervision, Z.X.; project administration, Z.X.; funding acquisition, Z.X. All authors have read and agreed to the published version of the manuscript.”

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    Figure S1

    PCR amplification with the primers BnRfo-AS2F/BnRfo-AS2R (a,c) and BnRfo-AS2F/BnRFO-NEW-R (b,d). Lanes 1-22: interspecific hybrids F1 from crosses of B14 as female parents, lanes 25-32: interspecific hybrids F1 from crosses of 137 as female parents, lanes 23 and 33: parent rapeseed, lanes 24 and 34: parent broccoli. Lanes 1-11: BC1 plants from crosses of B14 as female parents, lanes 25-32: BC1 plants from crosses of 137 as female parents, lanes 12 and 33: parent rapeseed, lanes 13 and 34: parent broccoli, M: marker (PNG 338 kb)

    Figure S2

    PCR amplification with the primers BnRfo-AS2F/BnRfo-AS2R (a,c) and BnRfo-AS2F/BnRFO-NEW-R (b,d) (PNG 162 kb)

    Figure S3

    PCR amplification with the primers BnRfo-AS2F/BnRfo-AS2R (a,c) and BnRfo-AS2F/BnRFO-NEW-R (b,d)). Lanes 1-9: BC2 plants from crosses of B14 as female parents, lanes 12-27: BC2 plants from crosses of 137 as female parents, lanes 10 and 28: parent rapeseed, lanes 11 and 29: parent broccoli, M: marker (PNG 231 kb)

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    Liu, Cq., Li, Gq., Yao, Xq. et al. Characterization of Ogura CMS fertility-restored interspecific hybrids and backcross progenies from crosses between broccoli and rapeseed. Euphytica 216, 194 (2020). https://doi.org/10.1007/s10681-020-02721-8

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