Skip to main content
SpringerLink
Log in

Comparison of six anther culture methods for the production of doubled haploids in eggplant (Solanum melongena L.)

  • Published:
Euphytica Aims and scope Submit manuscript

Abstract

Eggplant (Solanum melongena L.) is one of the most produced and high value vegetable crop in the world. Hybrid breeding toward high yielding variety of eggplants with many interesting traits are needed. To produce hybrid variety, production of pure lines as parents are needed. Pure lines in doubled haploid (DH) form can be produced through anther culture. The purpose of this study was to compare six anther culture methods to determine the most effective method to produce in vitro androgenic regenerated plants in eggplant. Anther culture methods used were 4 existing methods (DDV, TAS, ARI, and VUR), and two new method modified from ARI dan VUR, respectively. The anther donor plant genotypes used were Mustang F1 (long purple), Hitavi F1 (long green), and Provita F1 (striated round green) eggplant varieties. The result showed that androgenesis through the direct embryogenesis process in eggplant was still genotype dependent and the heat shock treatment at 35 °C is basically required, but the duration between 2 and 8 days were not significantly different to stimulate microspore embryogenesis. Modification 1 method was found as the most effective method as indicated by the fastest embryo initiation time, the highest percentage of anther producing embryo, the highest percentage of embryogenesis, the highest percentage of regeneration and the highest efficiency of anther culture. Ploidy level analysis by flow cytometry revealed the ploidy status of regenerants. Rate of spontaneous DH plant formation was 25.9%. The ploidy of regenerants has been validated by anatomy and plant morphology observations.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

References

  • Acquaah G (2012) Principles of plant genetics and breeding, 2nd edn. Wiley, New York

    Book  Google Scholar 

  • Ahmadi B, Ebrahimzadeh H (2020) In vitro androgenesis: spontaneous vs. artificial genome doubling and characterization of regenerants. Plant Cell Rep 39(3):299–316. https://doi.org/10.1007/s00299-020-02509-z

    Article  CAS  PubMed  Google Scholar 

  • Alpsoy H, Şeniz V (2007) Researches on the in vitro androgenesis and obtaining haploid plants in some eggplant genotype. Acta Horticulturae 729:137–141. https://doi.org/10.17660/ActaHortic.2007.729.21

    Article  CAS  Google Scholar 

  • Ari E, Yildirim T, Mutlu N, Büyükalaca S, Gökmen Ü, Akman E (2016) Comparison of different androgenesis protocols for doubled haploid plant production in ornamental pepper (Capsicum annuum L.). Turkish J Biol 40(4):944–954. https://doi.org/10.3906/biy-1509-36

    Article  CAS  Google Scholar 

  • Başay S, Ellialtioğlu ŞŞ (2013) Effect of genotypical factors on the effectiveness of anther culture in eggplant (Solanum melongena L.). Turkish J Biol 37(4):499–505. https://doi.org/10.3906/biy-1210-38

    Article  CAS  Google Scholar 

  • Bat H, Altindag FN, Yigit MA, Ellialtioglu ŞŞ, Comlekcioglu N (2021) Ploidy estimation in pepper and eggplant via stomata characteristics. Int J Agricul Forest Life Sci 5(2):139–146

    Google Scholar 

  • Bohanec B (2003) Ploidy determination using flow cytometry. In: Maluszynski M, Kasha K, Forster B et al (eds) Doubled haploid production in crop plants. Springer, Dordrecht, pp 397–403

    Chapter  Google Scholar 

  • Calabuig-Serna A, Porcel R, Corral-Martínez P, Seguí-Simarro JM (2020) Anther culture in eggplant (Solanum melongena L.). In: Bayer M (ed) Plant embryogenesis. Humana, New York, pp 283–293

    Chapter  Google Scholar 

  • Calabuig-Serna A, Camacho-Fernández C, Mir R, Porcel R, Carrera E, López-Díaz I, Seguí-Simarro JM (2022) Quantitative and qualitative study of endogenous and exogenous growth regulators in eggplant (Solanum melongena) microspore cultures. Plant Growth Regul 96(2):1–11. https://doi.org/10.1007/s10725-021-00780-y

    Article  CAS  Google Scholar 

  • Chu CC, Wang CC, Sun CS, Hsu C, Yin KC, Bi FY (1975) Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources. Sci Sin 18(5):659–668

    Google Scholar 

  • de Vaulx RD, Chambonnet D (1982) Culture in vitro d’anth`eres d’aubergine (Solanum melongena L.): stimulation de la production de plantes au moyen de traitements á+35 c associ´es `a de faibles teneurs en substances de croissance. Agronomie 2(10):983–988. https://doi.org/10.1051/agro:19821011

    Article  Google Scholar 

  • de Vaulx RD, Chambonnet D, Pochard E (1981) Culture in vitro d’anthères de piment (Capsicum annuum L.): amélioration des taux d’obtention de plantes chez différents génotypes par des traitements à+35°C. Agronomie 1(10):859–864. https://doi.org/10.1051/agro:19811006

    Article  Google Scholar 

  • Dehkehan EM, Moieni A, Movahedi Z (2017) Effects of zeatin riboside, mannitol and heat stress on eggplant (Solanum melongena L.) anther culture. Iran J Genet Plant Breed 6(1):16–26

    Google Scholar 

  • Domblides E, Ermolaev A, Belov S, Kan L, Skaptsov M, Domblides A (2022) Efficient methods for evaluation on ploidy level of Cucurbita pepo L. regenerant plants obtained in unpollinated ovule culture in vitro. Horticulturae 8(11), 1083. https://doi.org/10.3390/horticulturae8111083

  • Doležel J, Binarová P, Lcretti S (1989) Analysis of nuclear dna content in plant cells by flow cytometry. Biol Plant 31(2):113–120

    Article  Google Scholar 

  • Gamborg OL, Miller R, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50(1):151–158. https://doi.org/10.1016/0014-4827(68)90403-5

    Article  CAS  PubMed  Google Scholar 

  • George EF, Klerk GJD (2008) The components of plant tissue culture media I: Macro-and micro-nutrients. In: George EF, Hall MA, De Klerk GJ (eds) Plant propagation by tissue culture, 3rd edn. Springer, Dordrecht, pp 65–113

    Google Scholar 

  • Germana M (2011) Anther culture for haploid and doubled haploid production. Plant Cell Tissue Organ Cult 104(3):283–300

    Article  Google Scholar 

  • Hansson B (1978) Temperature shock a method of increasing the frequency of embryoid formation in anther culture of swedc rape (Brassica napus L.). Sver. Utsaedsfoeren Tidskr 88(3):141–148

    Google Scholar 

  • Hassan MF, Islam SS (2021) Effect of silver nitrate and growth regulators to enhance anther culture response in wheat (Triticum aestivum L.). Heliyon 7(5):1–6. https://doi.org/10.1016/j.heliyon.2021.e07075

    Article  CAS  Google Scholar 

  • Hewitt EJ, (1966) Sand and water culture methods used in the study of plant nutrition (2nd ed.). Technical communication no. 22. Commonwealth Agriculture Bureau, Farnham Royal, UK

  • Isouard G, Raquin C, Demarly Y (1979) Obtention de plantes haploides et diploides par culture in vitro d' antheres d' aubergine (Solanum melongena L.) [hystophysiologie]. Comptes Rendus Hebdomadaires des Seances de l’Academie des Sciences Serie D 288(12):987–989

  • Kakizaki Y (1931) Hybrid vigor in eggplants and its practical utilization. Genetics 16(2):1–25

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kartal C (2015) Microsporogenesis, microgametogenesis and in vitro pollen germination in the endangered species Fritillaria stribrnyi (liliaceae). Caryologia Int J Cytol Cytosyst Cytogen 68(1):36–43. https://doi.org/10.1080/00087114.2014.998947

    Article  Google Scholar 

  • Keller WA, Armstrong KG (1979) Stimulation of embryogenesis and haploid production in Brassica campestris anther cultures by elevated temperature treatments. Theor Appl Genet 55:65–67

    Article  CAS  PubMed  Google Scholar 

  • Kratky BA, Maehira GT, Cupples RJ, Bernabe CC (2005) Non-circulating hydroponic methods for growing tomatoes. Proc. Natl Agric Plast Congress 32:31–36. https://doi.org/10.13140/2.1.4948.6727

  • Kumar V, Parvatam G, Ravishankar G (2009) AgNO3: a potential regula tor of ethylene activity and plant growth modulator. Electron J Biotechnol 12(2):1–15. https://doi.org/10.2225/vol12-issue2-fulltext-1

    Article  CAS  Google Scholar 

  • Kumar A, Sharma V, Jain BT, Kaushik P (2020) Heterosis breeding in eggplant (Solanum melongena L.): Gains and provocations. Plants 9(3):403–418. https://doi.org/10.3390/plants9030403

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kumari P, Singh AK, Ahmed S, Sharma M (2017) Effect of heat pretreatment on anther culture in different brassica genotypes. Bios Int Q J Life Sci. 12(3): 1443–1447. www.thebioscan.com

  • Mir R, Calabuig-Serna A, Seguí-Simarro JM (2021) Doubled haploids in eggplant. Biology 10(7):1–16. https://doi.org/10.3390/biology10070685

    Article  CAS  Google Scholar 

  • Mishra R, Rao G (2016) In-vitro androgenesis in rice: advantages, constraints and future prospects. Rice Sci 23(2):57–68. https://doi.org/10.1016/j.rsci.2016.02.001

    Article  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15(3):473–497

    Article  CAS  Google Scholar 

  • Niazian M, Shariatpanahi M (2020) In vitro-based doubled haploid production: recent improvements. Euphytica 216(5):69–89. https://doi.org/10.1007/s10681-020-02609-7

    Article  CAS  Google Scholar 

  • Ochatt SJ, Seguí-Simarro JM (2021) Analysis of ploidy in haploids and doubled haploids. In: Segui-Simarro JM (ed) Doubled haploid technology. Methods in molecular biology, vol 2287. Humana, New York

    Google Scholar 

  • Olsen FL (1987) Induction of microspore embryogenesis in cultured anthers of Hordeum vulgare. The effects of ammonium nitrate, glutamine and asparagine as nitrogen sources. Carlsberg Res Commun 52(6):393–404

    Article  CAS  Google Scholar 

  • Raina S, Iyer R (1973) Differentiation of diploid plants from pollen callus in anther cultures of Solanum melongena L. Z Pflanzenzucht, 275–280

  • Rotino GL (2016) Anther culture in eggplant (Solanum melongena L.). In: Germana M, Lambardi M (eds) In vitro embryogenesis in higher plants Methods in molecular biology, vol 1359. Humana Press, New York, pp 453–466

    Google Scholar 

  • Salas P, Prohens J, Seguí-Simarro JM (2011) Evaluation of androgenic competence through anther culture in common eggplant and related species. Euphytica 182(2):261–274. https://doi.org/10.1007/s10681-011-0490-2

    Article  CAS  Google Scholar 

  • Salas P, Rivas-Sendra A, Prohens J, Seguí-Simarro JM (2012) Influence of the stage for anther excision and heterostyly in embryogenesis induction from eggplant anther cultures. Euphytica 184(2):235–250. https://doi.org/10.1007/s10681-011-0569-9

    Article  Google Scholar 

  • Seeja G, Sreekumar S (2021) Doubled haploids in genetic improvement: a review. Int J Recent Sci Res 11(01):36941–36949. https://doi.org/10.24327/ijrsr.2020.1101.5029

    Article  Google Scholar 

  • Seguí-Simarro JM (2015) Doubled haploidy in model and recalcitrant species. Front Plant Sci 6(2015):1175. https://doi.org/10.3389/fpls.2015.01175

    Article  PubMed  PubMed Central  Google Scholar 

  • Swamy MK, Sudipta KM, Balasubramanya S, Anuradha M (2010) Effect of different carbon sources on in vitro morphogenetic response of patchouli (Pogostemon cablin benth.). J Phytol 2(8):11–17

    Google Scholar 

  • Taşkin H, Büyükalaca S, Keles D, Ekbiç E (2011) Induction of microspore-derived embryos by anther culture in selected pepper genotypes. Afr J Biotechnol 10(75):17116–17121. https://doi.org/10.5897/AJB11.2023

    Article  CAS  Google Scholar 

  • Vural GE, Ari E (2020) Triple synergistic effect of maltose silver nitrate and activated charcoal on high embryo yield of eggplant (Solanum melongena L.) anther cultures. Scientia Horticulturae 272:109472. https://doi.org/10.1016/j.scienta.2020.109472

    Article  CAS  Google Scholar 

  • Vural GE, Ari E, Zengin S, Ellialtioglu SS (2019) Development of androgenesis studies on eggplant (Solanum melongena L.) in turkey from past to present. Sustain Crop Prod 2:1–26. https://doi.org/10.5772/intechopen.88299

    Article  Google Scholar 

  • Wang Y, Tong Y, Li Y, Zhang Y, Zhang J, Feng J, Feng H (2011) High frequency plant regeneration from microspore-derived embryos of ornamental kale (Brassica oleracea L. var. acephala). Sci Horticult 130(1):296–302. https://doi.org/10.1016/j.scienta.2011.06.029

    Article  CAS  Google Scholar 

Download references

Funding

The authors declare that no external funds, grants, or other support were received during the preparation of this manuscript.

Author information

Authors and Affiliations

  1. Study Program of Plant Breeding and Biotechnology, Department of Agronomy and Horticulture, Faculty of Agriculture, IPB University, Darmaga, Bogor, 16680, West Java, Indonesia

    Ade Mulyana

  2. Department of Agronomy and Horticulture, Faculty of Agriculture, IPB University, Darmaga, Bogor, 16680, West Java, Indonesia

    Bambang S. Purwoko & Awang Maharijaya

  3. Research Center of Food Crops, National Research and Innovation Agency, Cibinong Science Center, Jl. Raya Jakarta-Bogor Km 46, Cibinong, Bogor, 16911, West Java, Indonesia

    Iswari S. Dewi

  4. Center for Tropical Horticulture Studies, IPB University, Baranangsiang, Bogor, West Java, Indonesia

    Awang Maharijaya

Authors
  1. Ade Mulyana
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bambang S. Purwoko
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Iswari S. Dewi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Awang Maharijaya
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by Ade Mulyana, Bambang S. Purwoko, Iswari S. Dewi and Awang Maharijaya. The first draft of the manuscript was written by Ade Mulyana and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Bambang S. Purwoko.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interests.

Human or animal rights

This article does not contain any studies involving animals performed by any of the authors.

Informed consent

Informed consent was obtained from all participants included in the study.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Mulyana, A., Purwoko, B.S., Dewi, I.S. et al. Comparison of six anther culture methods for the production of doubled haploids in eggplant (Solanum melongena L.). Euphytica 219, 44 (2023). https://doi.org/10.1007/s10681-023-03171-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s10681-023-03171-8

Keywords

Search

Navigation