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Recovering triploid citrus hybrids from 2x × 2x sexual crosses with the aid of embryo rescue and flow cytometry in Turkey

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Abstract

Background

Turkey is one of the major exporters of mandarins in the Mediterranean region. Seedlessness in citrus, which is one of the most desired fruit quality traits, especially in fresh mandarin export markets, can be obtained via triploidy as in many fruit species. Triploid plants can be recovered by 2x × 2x hybridizations in citrus, as well as 2x × 4x and 4x × 2x crosses. Hence, we aimed to develop local triploid hybrids by using the embryo rescue technique in five crosses using eight different citrus varieties in the present study.

Methods and results

Embryos isolated from abortive seeds derived by 135 days after pollinations were cultured on modified Murashige and Tucker (MT) medium by adding different levels of GA3 to achieve a high germination rate. A population of 574 plants was developed as a result of embryo rescue. After screening the ploidy levels of this 574-plant population with the aid of flow cytometry, 4 triploids from ‘Encore’ × ‘Murcott’, 8 triploids from ‘Fortune’ × ‘Willow leaf’, 1 triploid from ‘Kiyomi’ × ‘Murcott’, and 1 triploid from ‘Ortanique’ × ‘Murcott’ hybridization were recovered. Triploid hybrid plants and related parents were analyzed with SSR markers heterozygotic for parental mandarin varieties. In addition, we evaluated stomatal characteristics of diploid and triploid hybrids obtained from different crosses. Stomatal traits of diploid and triploid hybrids in all crosses significantly differed except the stomata index.

Conclusions

Genotyping of triploid plants confirmed using Simple Sequence Repeat (SSR) molecular markers and five SSRs were able to identify three alleles of triploid hybrids. Selected triploid mandarin hybrids have been grafted on several rootstocks for field trials and are in the process of yield and quality performances.

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Acknowledgements

The authors are grateful that the manuscript has been read and critically corrected by Dr. Ben FABER who works at the Division of Agriculture and Natural Resources, University of California Cooperative Extension. The authors also would like to thank the ‘Atlas Tarim’ and ‘Incesulu Tarim’ companies for supplying parental plant materials used in hybridizations.

Funding

This research was funded by the Scientific Research Projects Coordination Unit of Cukurova University (Grant Number FBA-2018-11126).

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Authors and Affiliations

  1. Department of Horticulture, Faculty of Agriculture, Cukurova University, 01330, Adana, Turkey

    Berken Cimen, Turgut Yesiloglu & Yildiz Aka Kacar

  2. Biotechnology Research and Application Center, Cukurova University, 01330, Adana, Turkey

    Dicle Donmez & Yildiz Aka Kacar

  3. Department of Horticulture, Faculty of Agriculture, Ataturk University, 25240, Erzurum, Turkey

    Sezai Ercisli

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  1. Berken Cimen
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Contributions

BC and TY performed hybridization studies and in vitro embryo culture, DD and YAK performed in SSR screening and result interpretation. BC and SE helped in data analysis and preparation of this manuscript.

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Correspondence to Berken Cimen.

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Cimen, B., Yesiloglu, T., Donmez, D. et al. Recovering triploid citrus hybrids from 2x × 2x sexual crosses with the aid of embryo rescue and flow cytometry in Turkey. Mol Biol Rep 49, 5625–5634 (2022). https://doi.org/10.1007/s11033-022-07555-2

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