ekin journal Ekin Journal of Crop Breeding and Genetics 2149-1275 2459-069X Bitki Islahçıları Alt Birliği Agronomy Agronomi Effects of Different Nutrient Media on Embryo Induction and Plant Regeneration in Cucumber (Cucumis sativus L.) Ovary Cultures Doğan Selcen Hektaş Seed Şimşek Cansu Hektaş Seed Murat Selçuk Murat X Hektaş Seed Pinar Hasan Erciyes University, Faculty of Agriculture, Department of Horticulture Mutlu Nedim Akdeniz University, Faculty of Agriculture, Department of Agricultural Biotechnology Ellialtioğlu Ş. Şebnem Doqutech Academy Llc. Co. 07 31 2025 11 2 60 67 04 27 2025 06 25 2025 Copyright © 2015, Ekin Journal of Crop Breeding and Genetics 2015 Ekin Journal of Crop Breeding and Genetics

Haploids and doubled haploids play a crucial role in the process of plant breeding. Ovary culture has proven to be aneffective method for generating double haploid (DH) plants in cucumbers. The objective of this study was to optimizeculture media conditions for gynogenesis induction in vitro to broaden morphogenetic possibilities and accelerate thecreation of homozygous lines. The effects of seven different induction culture media on embryo and plant developmentin the ovary culture of Beith Alpha-type and Silor-type cucumber (Cucumis sativus L.) were investigated. Ovary cultureexperiments were conducted during the fall and spring seasons with 26 Beith Alpha-type and 11 Silor-type cucumbergenotypes at the F2 and F3 generations. The results revealed that the most successful outcomes were obtained fromgenotypes coded HTB20, HTB24 and HTB25 of the Beith Alpha-type and HTS4 and HTS5 of the Silor-type. The findingsalso demonstrated that explants cultured in the autumn season showed higher regeneration efficiency compared to thosecultured in spring. Overall when compared to Silor types, Beith Alpha types exhibited a more favorable response in termsof plant regeneration and embryo development, depending on genotype.

 Gynogenesis medium compositions haploid Beith Alpha Silor-Type Anonymous, (2024). Bitkisel üretim istatistikleri. https:// biruni.tuik.gov.tr/medas/?locale=tr Access Date: 16.06.2025 (in Turkish) Baktemur G, Keleş D, Kara E, Yıldız S & Taşkın H, (2022). Effects of genotype and nutrient medium on obtaining haploid plants through ovary culture in cucumber. Molecular Biology Reports, 49:5451- 5458. https://doi.org/10.1007/s11033-022-07238-y Beharav A & Cohen Y, (1995). Effect of kinetin and GA3 on in vitro ovule embryo culture of Cucumis melo L. Plant Growth Regulation, 16(3):267-269. https://doi.org/10.1007/BF00024785 Chambonnet D & Vaulx RD, (1985). Obtention of embryos and plants from in vitro culture of unfertilized ovules of Cucurbita pepo. Cucurbit Genetics Cooperative, 8:66 Çağlar G & Şensoy S, (2021). Hıyar ıslahı. In K. Abak, A. Balkaya, Ş. Ş. Ellialtıoğlu & E. Düzyaman (Eds.), Sebze Islahı. Cilt II: Cucurbitaceae (Kabakgiller) (ss. 87-201). BİSAB Yayınları, Gece Kitaplığı Yayınevi (in Turkish) Çetinkaya E, (2015). Farklı besi ortamı kombinasyonlarının bazı hıyar (Cucumis sativus L.) genotiplerinde gynogenesis yolu ile embriyo ve haploid bitki oluşumu üzerine etkisi [Yüksek lisans tezi, Akdeniz Üniversitesi]. Antalya (in Turkish) Diao WP, Jia YY, Song H, Zhang XQ, Lou QF & Chen JF, (2009). Efficient embryo induction in cucumber ovary culture and homozygous identification of the regenerants using SSR markers. Scientia Horticulturae, 119(3):246-251. https://doi. org/10.1016/j.scienta.2008.08.016 Domblides E, Shmykova N, Khimich G, Korotseva I, Kan L, Domblides A, Pivovarov V & Soldatenko A, (2020). Production of doubled haploid plants of Cucurbitaceae family crops through unpollinated ovule culture in vitro. In VI International Symposium on Cucurbits (Vol. 1294, pp. 19-28) https://doi.org/10.17660/ActaHortic.2020.1294.4 Dong YQ, Zhao WX, Li XH, Liu XC, Gao NN, Huang JH, Wang WY, Xu XL & Tang ZH, (2016). Androgenesis, gynogenesis, and parthenogenesis haploids in cucurbit species. Plant Cell Reports, 35, 1991-2019. https://doi.org/10.1007/s00299- 016-2018-7 Dumas de Vaulx R & Chambonnet D, (1986). Obtention of embryos and plants from in vitro culture of unfertilized ovules of Cucurbita pepo. In W. Gruyter (Ed.), Genetic Manipulation in Plant Breeding (pp. 295-297). De Gruyter. FAOSTAT, (2023). http://www.fao.org/faostat/en/#data/ QC (Access Date: 16.06.2025) Ficcadenti N, Sestili S, Annibali S, Marco MD & Schiavi M, (1999). In vitro gynogenesis to induce haploid plants in melon (Cucumis melo L.). Journal of Genetics and Breeding, 53(3):255-257. Gémes Juhász A, Balogh P, Ferenczy A & Kristo´f Z, (2002). Effect of optimal stage of female gametophyte and heat treatment on in vitro gynogenesis induction in cucumber (Cucumis sativus L.). Plant Cell Reports, 21(2):105-111. https://doi.org/10.1007/s00299-002-0482-8 Gémes Juhász A & Jakše M, (2005). Haploids in the improvement of miscellaneous crop species (Cucurbitaceae, Liliaceae, Asparagaceae, Chenopodiaceae, Araceae and Umbelliferae). In C. E. Don Palmer, W. A. Keller, & K. J. Kasha (Eds.), Biotechnology in Agriculture and Forestry: Haploids in Crop Improvement II (pp. 259-278). Springer Gémesné Juhász A, Venczel G & Balogh P, (1997). Haploid plant induction in zucchini (Cucurbita pepo L. convar. giromontiina Duch) and in cucumber (Cucumis sativus L.) lines through in vitro gynogenesis. Acta Horticulturae, 447, 623- 625 Günay A, (2005). Sebze Yetiştiriciliği (Cilt II). Meta Basımevi (in Turkish) Kwack SN & Fujieda K, (1988). Somatic embryogenesis in cultured unfertilized ovules of Cucurbita moschata. Journal of the Japanese Society for Horticultural Science, 57(1):34-42. https://doi. org/10.2503/jjshs.57.34 Li JW, Si SW, Cheng JY, Li JX & Liu JQ, (2013). Thidiazuron and silver nitrate enhanced gynogenesis of unfertilized ovule cultures of Cucumis sativus. Biologia Plantarum, 57(1):164- 168. https://doi.org/10.1007/s10535-012-0269-x Metwally EI, Moustafa S A, EI-Sawy BI & Shalaby TA, (1998a). Haploid plantlets derived by anther culture of Cucurbita pepo. Plant Cell, Tissue and Organ Culture, 52(3):171-176. https://doi. org/10.1023/A:1005908326663 Metwally EI, Moustafa SA, El-Sawy BI, Haroun SA & Shalaby TA, (1998b). Production of haploid plants from in vitro culture of unpollinated ovules of Cucurbita pepo L. Plant Cell, Tissue and Organ Culture, 52(3):117-121. https://doi. org/10.1023/A:1005948809825 Min ZY, Li H, Zou T, Tong L, Cheng J & Sun XW, (2016). Studies of in vitro culture and plant regeneration of unfertilized ovary of pumpkin. Chinese Bulletin of Botany, 51(1):74-80. https:// doi.org/10.11983/CBB15004 Moqbeli E, Peyvast GH, Hamidoghli Y & Olfati JA, (2013). In vitro cucumber haploid line generation in several new cultivars. Asia-Pacific Journal of Molecular Biology and Biotechnology, 21(1): 18- 25 Murashige T & Skoog F, (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15(3):473-497 Nyirahabimana F & Solmaz İ, (2021). Haploid plant production technology in cucumber (Cucumis sativus L.). In M. Tutuncu (Ed.), Proceedings Book. 10th Int. Mol. Biol. & Biotech. Congress (pp. 28-37). Online Nyirahabimana F & Solmaz İ, (2024). Haploid induction through ovary culture in cucumber. In Vitro Cellular & Developmental Biology-Plant, 60(1):122-130. https://doi.org/10.1007/s11627- 023-10406-9 Özsan T, Gozen V & Onus A, (2017). Cucumber gynogenesis: Effects of 8 different media on embryo and plant formation. International Journal of Agriculture Innovations and Research, 6(2):419- 422 Paris HS, Daunay MC & Janick J, (2012). Occidental diffusion of cucumber (Cucumis sativus) 500- 1300 CE: Two routes to Europe. Annals of Botany, 109(1):117-126. https://doi.org/10.1093/aob/ mcr281 Pınar H, Toprak S, Hancı F & Şimsek D, (2021). Effects of genotypic variation and some new protocols on gynogenesis efficiency of cucumber. Fresenius Environmental Bulletin, 30(5):4763-4770 Plapung P, Khamsukdee S, Potapohn N & Smitamana P, (2014). Screening for cucumber mosaic resistant lines from the ovule culture derived double haploid cucumbers. American Journal of Agricultural and Biological Sciences, 9(3):261-269. https://doi. org/10.3844/ajabssp.2014.261.269 Przyborowski J & Nlemirowicz‐Szgzytt K, (1994). Main factors affecting cucumber (Cucumis sativus L.) haploid embryo development and haploid plant characteristics. Plant Breeding, 112(1):70- 75. https://doi.org/10.1111/j.1439-0523.1994. tb01278.x Rakha MT, Metwally EI, Moustafa SA, Etman AA & Dewir YH, (2012). Evaluation of regenerated strains from six ‘Cucurbita’ interspecific hybrids obtained through anther and ovule ‘in vitro’ cultures. Australian Journal of Crop Science, 6(1):23-30 Renner SS, Schaefer H & Kocyan A, (2007). Phylogenetics of Cucumis (Cucurbitaceae): Cucumber (C. sativus) belongs in an Asian/ Australian clade far from melon (C. melo). BMC Evolutionary Biology, 7(1):58. https://doi. org/10.1186/1471-2148-7-58 Renner SS & Schaefer H, (2008). Phylogenetics of Cucumis (Cucurbitaceae) as understood in 2008. In M. Pitrat (Ed.), Cucurbitaceae 2008, Proceedings of the IXth EUCARPIA Meeting on Genetics and Breeding of Cucurbitaceae (pp. 53-58). Avignon, France Shalaby TA, (2007). Factors affecting haploid induction through in vitro gynogenesis in summer squash (Cucurbita pepo L.). Scientia Horticulturae, 115(1):1-6. https://doi.org/10.1016/j. scienta.2007.07.008 Sun SR, Zhai QH, Hu JB, Chen JF & Zhang P, (2009). Effects of several physiological factors on embryo formation in unpollinated ovary culture of pumpkin (Cucurbita moschata Duch. Ex Poiret). Plant Physiology Communications, 45(10):977- 980 Suprunova T & Shmykova N, (2008). In vitro induction of haploid plants in unpollinated ovules, anther and microspore culture of Cucumis sativus. In M. Pitrat (Ed.), Proceedings of the IXth EUCARPIA Meeting on Genetics and Breeding of Cucurbitaceae (pp. 371-374). Avignon, France Staub JE, Robbins MD & Wehner TC, (2008). Cucumber. In J. Prohens & F. Nuez (Eds.), Vegetables I: Asteraceae, Brassicaceae, Chenopodicaceae, and Cucurbitaceae, Handbook of Plant Breeding (Vol. 1, pp. 241-282). Springer Tantasawat PA, Sorntip A, Poolsawat O, Chaowiset W & Pornbungkerd P, (2015). Evaluation of factors affecting embryo-like structure and callus formation in unpollinated ovary culture of cucumber (Cucumis sativus). International Journal of Agriculture and Biology, 17(3):613-618. https:// doi.org/10.17957/IJAB.17.3.14.257 Xie M, Qin LY, Pan JS, He HL, Wu AZ & Cai R, (2005). Flower morphogenesis and microspore development versus anther culture of cucumber. Acta Botanica Boreali-Occidentalia Sinica, 25(6):1096-1100