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Evaluation of the androgenic competence of 66 wild Turkish Vaccaria hispanica (Mill.) Rauschert genotypes through microspore culture

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Abstract

One of the most remarkable natural plants is Vaccaria hispanica (Mill.) Rauschert, which has a high economic, medicinal and industrial potential due to its valuable compounds. However, it is mostly an underused plant worldwide. To implement doubled haploid technology in plant breeding programs and exploit its potential, first knowing the particulars of the species is necessary. This study was aimed to explore the androgenic potential of different wild Turkish V. hispanica genotypes collected from different Turkish regions, as a starting point to identify bottlenecks to solve in future studies. As to induction of microspore embryogenesis, nearly all of them responded, with efficiencies reaching 300 embryos/100 buds in some cases. However, we also found three main bottlenecks. First, the presence of foam-producing saponins in flowers prevented an efficient isolation of microspores. Second, embryos showed a reduced ability to germinate. Third, a dense network of hairy roots prevented the formation of a true, fully functional root system. Together, these drawbacks led to the production of very few DH plants. The presence of rhizogenic endophytes may be the cause of most of these problems, which opens the door for possible solutions.

Key message

Vaccaria hispanica is a highly responsive species to microspore embryogenesis. However, the two main bottlenecks, possibly caused by endophytes, were the formation of dense abnormal embryos and short hairy roots.

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Data availability

All data generated or analyzed during this study are included in this article and supplementary materials.

Abbreviations

DH:

Doubled haploid

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Acknowledgements

The authors would like to thank the TUBITAK (The Scientific and Technological Research Council of Turkey) (TOVAG 1001 Project No: 112O136) for financial support. Authors also grateful to Assoc. Prof. Ozer Calis (Akdeniz University, Turkey) for the help in the detection of fungi and bacteria, and Prof. Stefaan Werbrouck (Ghent University, Belgium) for opening his laboratory without any expectation, sharing his invaluable professional experiences, and helping in the seed germination studies.

Funding

The study was funded from TUBITAK - TOVAG 1001 - Project 112O136 financed by The Scientific and Technological Research Council of Turkey (TUBITAK).

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Authors and Affiliations

  1. Department of Agricultural Biotechnology, Faculty of Agriculture, Akdeniz University, Antalya, Turkey

    Esin Ari & Hilal Bedir

  2. Department of Biology Education Antalya, Faculty of Education, Akdeniz University, Antalya, Turkey

    Ismail Gokhan Deniz

  3. Department of Pharmaceutical Botany, Faculty of Pharmacy, Istanbul University, Istanbul, Turkey

    Ilker Genc

  4. Cell Biology Group - COMAV Institute, Universitat Politècnica de València, Valencia, Spain

    Jose M. Seguí-Simarro

Authors
  1. Esin Ari
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  2. Hilal Bedir
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  3. Ismail Gokhan Deniz
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  4. Ilker Genc
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  5. Jose M. Seguí-Simarro
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Contributions

EA conceived and designed the research, conducted experiments, analyzed the data and wrote the manuscript. IGD and IG collected wild plant materials. HB performed the experiments and collected the data. JMSS analyzed the data, reviewed and edited the manuscript. All authors read and approved the final manuscript

Corresponding author

Correspondence to Esin Ari.

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The authors declare no conflict of interest.

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The authors declare consent for publication.

Research involving human and/or animal participants

This research did not involve experiments with human or animal participants.

Additional information

Communicated by Alison M. R. Ferrie.

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Supplementary Information

Below is the link to the electronic supplementary material.

Suppl. Fig. S1.

Massive hairy root formation in Vaccaria hispanica. A, B: Hairy roots formed while keeping backup seedling trays for 3 weeks in the greenhouse to renew the planted seedlings if required. C: Hairy roots formed after 1 week seed germination on wet filter paper in culture plates. D: Hairy roots formed after 2 weeks seed germination in agar-solidified in vitro culture medium. E: Hairy roots formed during in vitro seed germination in 3-month culture media containing Gelrite (from left to right, the first three tubes), vermiculite, peat and perlite (the 4th, 5th and 6th tubes, respectively). F: Hairy roots formed during microspore-derived embryo regeneration in 6-month culture medium containing activated charcoal. Supplementary material 1 (JPG 1325.4 kb)

Suppl. Table S1.

Experiments of different medium modifications performed on different V. hispanica genotypes to increase the efficiency of isolated microspore culture and embryo germination. Supplementary material 2 (PDF 1033.3 kb)

Suppl. Table S2

. Experiments with different chemicals added to cultures and different light sources and their effects on seed germination, hairy root formation and shoot development in small-scale in vitro seed germination assays in V. hispanica genotype 35. Root and shoot growth densities were determined according to 0–4 visual rating scale [0: none (−), 1: very little (*), 2: little (**), 3: moderate (***), and 4: abundant (****)]. Supplementary material 3 (PDF 48.9 kb)

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Ari, E., Bedir, H., Deniz, I.G. et al. Evaluation of the androgenic competence of 66 wild Turkish Vaccaria hispanica (Mill.) Rauschert genotypes through microspore culture. Plant Cell Tiss Organ Cult 148, 209–214 (2022). https://doi.org/10.1007/s11240-021-02169-1

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  • DOI: https://doi.org/10.1007/s11240-021-02169-1

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