Production of Microspore-Derived Plants by Anther Culture of Cyclamen coum

Year 2024, , 720 - 728, 15.11.2024

Mahdi Al-khafaji , Mehtap Yıldız , Metin Koçak , Mehmet Tütüncü

https://doi.org/10.47115/bsagriculture.1554595

Abstract

Cyclamen is one of the most important ornamental crops sold worldwide as a potted flower for winter production. Cyclamen species take up a wide swathe of habitats across Türkiye. Ten wild Cyclamen species grow naturally in Türkiye and some of them are endemic. This study aimed to produce haploid plants of C. coum using anther culture. The microspore developmental stage was evaluated by staining anther with acetocarmine (%2), and then the stage was correlated with bud size. It was determined that the buds between 7.64 and 8.23 mm had the appropriate bud size for the late uninuclear stage. Anthers were cultured in B5 medium containing different levels of 1-Naphthaleneacetic acid (NAA) (0.1, 1, 2 mgL-1), 2,4-Dichlorophenoxyacetic acid (2,4-D) (0.1, 1, 2 mgL-1), and kinetin (0, 1 mgL-1), 90 gL-1 sucrose and 3 gL-1 gelrite for haploid embryo production. Anthers were kept at 4°C for 4 days after culture. The explants were incubated at 24°C in a completely dark condition until the embryo was formed, then embryos were transferred to hormone-free media in 16:8 hours (light (75 µmolm-2s-1): dark) photoperiod. The experiment was carried out for two years. In the first year, 12 different media were examined in view of regeneration and the experiments were continued with selected 7 media in the second year. The highest callus regeneration rates were %5.71 and 14.5% and the highest embryo induction rates varied between 8.57% and 4.0% in the first and second year respectively. Embryo/callus formation was observed in 7 of a total of 12 different media tested for haploid plant production, and the best media were kinetin (1 mgL-1) + NAA (1, 2 mgL-1) and kinetin (1 mgL-1) +2,4-D (2 mgL-1). Our findings indicated that cold pre-treated anther explants collected at appropriate flower bud size resulted in embryo production. Additionally, B5 medium supplemented with NAA and kinetin ensured successful embryo regeneration from anther explants in wild C. coum.

Keywords

Androgenesis, Anther culture, Cyclamen coum, Haploid plants

Supporting Institution

Van Yuzuncu Yil University- Scientific Research Projects Coordinating Office

Project Number

FYL-2020-8797

Thanks

The authors are grateful to Van Yuzuncu Yil University- Scientific Research Projects Coordinating Office for financial support

References

• Amaury MAF. 1997. Production of doubled-haploid plants from Lilium longiflorum Thunb. anther culture. Plant Sci, 123: 179-187.
• Amssa M, Buyser JD, Henry Y. 1980. Origin of the diploid plants obtained by in vitro anther culture of bread wheat (Triticum aestivum L.): effect of cold pretreatment and of in vitro culture on doubling, 290: 1095-1097.
• Atalay İ. 1994. Vegetation geography of Turkey. Ege Üniversitesi Basımevi, İzmir, Türkiye, pp: 58.
• Başar S. 2018. Obtaining haploid plant via anther and ovule culture techniques in cyclamen. PhD Thesis, Çukurova University, Institue of Natural and Applied Sciences, Adana, Türkiye, pp: 204.
• Canhoto JM, Ludovina M, Guimaraes S, Cruz GS. 1990. In vitro induction of haploid, diploid and triploid plantlets by anther culture of Iochroma warscewiczii Regel. PCTOC, 21: 171-177.
• Çürük P. 2013. Morphological and molecular characterization of cyclamen species growing naturally in Adana and its surroundings. PhD Thesis, Çukurova University, Institue of Natural and Applied Sciences, Adana, Türkiye, pp: 216.
• Custódio L, Carneiro MF, Romano A. 2005. Microsporogenesis and anther culture in carob tree (Ceratonia siliqua L.). Sci Hortic, 104(1): 65-77.
• de Vaulx RD, Chambonnet D. 1982. Culture in vitro d'anthères d'aubergine (Solanum melongena L.): stimulation de la production de plantes au moyen de traitements à+ 35° C associés à de faibles teneurs en substances de croissance. Agronomie, 2(10): 983-988.
• Dole JM, Wilkins HF. 1999. Floriculture principles and species. Prentice Hall Inc., New York, US. ‏ Ekim T, Koyuncu M, Güner A, Erik S, Yıldız B, Vural M. 1991. Taxonomic and ecologic researches on Econnomically important Geofits in Turkey. Publication Publicity, 669(65): 111.
• Ellialtıoğlu Ş, Sarı N, Abak K. 2000. Haploid bitki üretimi. Bitki Biyoteknol, 1: 137-189.
• Fehr WR. 1993. Principles of cultivar devеlоpment: Theory and technique, 1.
• Ferrie AMR, Caswell KL. 2011. Isolated microspore culture techniques and recent progress for haploid and doubled haploid plant production. PCTOC, 104: 301-309.
• Finnie SJ, Powll W, Dyer AF. 1989. The effect of carbohydrate composition and concentration on anther culture response in barley (Hordeum vulgare L.). Plant Breed, 103(2): 110-118.
• Folling L. 2002. Transformation of wheat (Triticum aesticum L.) microspore-derived callus and microspores by particle bombardment. Plant Cell Rep, 20: 1098-1105.
• Forster BP, Heberle-Bors E, Kasha KJ, Touraev A. 2007. The resurgence of haploids in higher plants. Trends Plant Sci, 12(8): 368-375.
• Gamborg OL. 1968. Nutrient requirements of suspension cultures of soybean root cultures. Exp Cell Res, 50: 148-158.
• Germana MA. 2011. Anther culture for haploid and doubled haploid production. PCTOC, 104: 283-300.
• Goyer A. 2010. Thiamine in plants: aspects of its metabolism and functions. Phytochem, 71(14-15): 1615-1624.
• Grey-Wilson C. 1988. The genus Cyclamen (A Kew Magazine Monograph). The Royal Botanic Gardens, Kew in association with Christopher Helm and Timber Press. Chromosome association and fertility in interspecific hybrid of Cyclamen, pp: 371.
• Heberle-Bors E. 1985. In vitro haploid formation from pollen: a critical review. Theor Appl Genet, 71: 361-374.
• Heyne EG. 1987. Wheat and wheat improvement. American Society of Agronomy, Madison, US, pp: 322.
• Irikova T, Grozeva S, Rodeva V. 2011. Anther culture in pepper (Capsicum annuum L.) in vitro. Acta Physiol Plant, 33: 1559-1570.
• Ishizaka H, Uematsu J. 1993. Production of plants from pollen in Cyclamen persicum Mill. through anther culture. Jpn J Breed, 43(2): 207-218.
• Ishizaka H. 1998. Production of microspore-derived plants by anther culture of an interspecific F1 hybrid between Cyclamen persicum and C. purpurascens. PTOC, 54(1): 21-28.
• Jain SM, Sopory SK, Veilleux R. 1996. In vitro haploid production in higher plants. Springer Sci Busin Media, 2(24): 442.
• Jalali N, Naderi R, Shahi-Gharahlar A, da Silva JAT. 2012. Tissue culture of Cyclamen spp. Sci Hortic, 137: 11-19.
• Kasha KJ, Maluszynski M. 2003. Production of doubled haploids in crop plants. An introduction. In: Maluszynski M, Kasha KJ, Forster BP, Szarejko (editors), Doubled Haploid Production in Crop Plants. Springer, Dordrecht, the Nederland, pp: 1-4.
• Keleş D, Pınar H, Ata A, Taşkın H, Yıldız S, Büyükalaca S. 2015. Effect of pepper types on obtaining spontaneous doubled haploid plants via anther culture. Hort Sci, 50(11): 1671-1676.
• Kocak M, Izgu T, Sevindik B, Tutuncu M, Curuk P, Simsek O, Mendi YY. 2014. Somatic embryogenesis of Turkish Cyclamen persicum Mill. Sci Hortic, 172: 26-33.
• Nitsch JP. 1969. Experimental andro-genesis in Nicotiana. Phytomorphology, 19: 389-404.
• Olsen FL. 1992. Isolation and cultivation of embryogenic microspores from barley (Hordeum vulgare L.). Hereditas, 115(3): 255-266.
• Pelletier G, Ilami M. 1972. Les facteurs de l’androgenese in vitro chez Nicotiana tabacum. Zeitschrift für Pflanzenphysiologie, 68(2): 97-114.
• Powell W. 1990. Environmental and genetical aspects of pollen embryogenesis. Haploids Crop Improv, I: 45-65.
• Pulido A, Bakos F, Castillo A, Vallés MP, Barnabas B, Olmedilla A. 2005. Cytological and ultrastructural changes induced in anther and isolated-microspore cultures in barley: Fe deposits in isolated-microspore cultures. J Struct Biol, 149(2): 170-181.‏
• Reed SM. 2005. Haploid culture. In: Trigiano RN, Gray DJ, (editors), Plant Development and Biotechnology. CRC Press LLC, NW, US, pp: 225-234.
• Reinert J, Bajaj YPS. 1977. Anther culture: haploid production and its significance. In: Reinert J, Bajaj YPS (editors). Applied and fundamental aspects of plant cell, tissue, and organ culture. Springer-Verlag, Berlin, Germany, pp: 251-267.
• Schwartz-Tzachor R, Eisikowitch D, Dafni A. 2008. Flower characteristics and breeding system of two phenological ecotypes of Cyclamen persicum Mill. (Myrsinaceae) in Israel. Pl Syst Evol, 274: 127-134.
• Seguí‐Simarro JM, Nuez F. 2008. How microspores transform into haploid embryos: changes associated with embryogenesis induction and microspore‐derived embryogenesis. Physiol Plant, 134(1): 1-12.
• Seyring M, Ewald A, Mueller A, Haensch KT. 2009. Screening for propagation suitability in vitro of different Cyclamen species. Electron J Biotechnol, 12(4): 10-11.
• Snape JW. 1989. Doubled haploid breeding: theoretical basis and practical applications. Rev Advan Plant Biotechnol, 2: 19-31.
• Takamura T, Miyajima I. 1997. Micropropagation of Cyclamen persicum Mill. In: Bajaj YPS (editor). High-tech and micropropagation VI. biotechnology in agriculture and forestry, 40th ed. Springer, Berlin, Germany, pp: 96-112.
• Takamura T, Sakamoto K, Horikawa M. 2011. Effect of carbon source on in vitro plant regeneration in anther culture of Cyclamen (Cyclamen persicum Mill.). In XXVIII International Horticultural Congress on science and horticulture for people (IHC2010): International Symposium on Micro, December 31, Lisbon, Portugal, pp: 129-134.
• Testillano PS, Coronado MJ, Seguı JM, Domenech J, González-Melendi P, Raška I, Risueño, MC. 2000. Defined nuclear changes accompany the reprogramming of the microspore to embryogenesis. J Struct Biol, 129(2-3): 223-232.
• Trejo-Tapia G, Maldonado Amaya U, Salcedo Morales G, De Jesús Sánchez A, Martínez Bonfil B, Rodríguez-Monroy M, Jiménez-Aparicio A. 2002. The effects of cold-pretreatment, auxins and carbon source on anther culture of rice. PTOC, 71: 41-46.
• URL1: http://www.cyclamen.org/indexCS.html (accessed date: July 15, 2024).
• Wehr WH. 1976. Plant production from cultured anthers of Hordeum vulgare L., Zeitschrift fuer Pflanzenzuechtung. 77: 198- 204.
• Yesson C, Culham A. 2006. A phyloclimatic study of Cyclamen. BMC Evol Biol, 6:1-23.
• Yilmaz R, Butt SJ, Korkut A. 2003. Plant diversity and effects of environmental problems in Turkey. J Environ Protect Ecol, 4: 924-930.
• Zhang XQ, Yang JS, Zheng ZX. 2009. Study on induction of embryoid through in vitro anther of Cyclamen persicum. JAAS, 1: 57-58.