Dorota Olszewska

Department of Agricultural Biotechnology, University of Science and Technology, Bernardynska 6, 85-029 Bydgoszcz, Poland

Paweł Nowaczyk

Department of Agricultural Biotechnology, University of Science and Technology, Bernardynska 6, 85-029 Bydgoszcz, Poland


The results of the investigation proved the gametophytic origin of diploids derived from two-embryonic seeds of pepper C. annuum L. hybrid. During the germination of seeds harvested from the red fruited hybrid (C. annuum L. ATZ × C. annuum L.`Sono`) F1, additional embryos have been found. Four of diploid twins F2 generation were different in their phenotype within the pair and/or from the F1 mother plant. Plants of three pairs: 1A-1B, 2A-2B, 4A-4B were significantly different with regard to the average fruit weight, length and seeds number. Yellow colour of ripe fruit was characteristic for 2A, 3A and 3B plants. In RAPD molecular analysis, twenty-three primers were used and six of them enabled polymorphic products to be obtained in reactions. The results of the analysis confirmed phenotypic differentiation of the twins and their parental forms. The phenotypic and molecular analyses proved that spontaneous diploids from a two-embryonic seeds are ready for the production of genetically stable, sexual progeny.


polyembryony, twin plants, spontaneous diploidisation, phenotypic marker, RAPD analysis

Barcaccia, G., Albertini, E. (2013). Apomixis in plant reproduction: a novel perspective on an old dilemma. Plant Reprod., 26, 159–179, DOI: 10.1007/s00497-013-0222-y.

Campos, F., Morgan, D.,T., Jr. (1958). Haploid pepper from a sperm. Androgenetic haploid of Capsicum frutescens L. J. Heredity, 49, 135–137, DOI: 10.1093/oxfordjournals.jhered.a106786.

Chauhan, H., Khurana, P. (2011). Use of doubled haploid technology for development of stable drought-tolerant bread wheat (Triticum aestivum L.) transgenics. Plant Biotech. J., 9, 408–417, DOI: 10.1111/j.1467-7652.2010.00561.x.

Dolcet-Sanjuan R., Claveria E., Huera, A. (1997). Androgenesis in Capsicum annuum L.: effect of carbohydrate and carbon dioxide enrichment. J. Amer. Soc. Hort. Sci., 122, 468–475, DOI: 10.21273/JASHS.122.4.468.

Dumas de Vaulx, R., Chambonnet, D., Pochard, E. (1981). Culture in vitro d’anthéres de piment (Capsicum annuum L.) amélioration des taux d’obtention de plantes chez différents génotypes par des traitements á +35EC. Agronomie, 1, 859-864. Dunwell, J.M. (2010). Haploids in flowering plants: origins and exploitation. Plant Biotech. J., 8, 377–424.

Forster, B.P., Thomas, W.T.B. (2005). Doubled haploids in genetics and plant breeding. Plant Breed. Rev., 25, 57–88, DOI: 10.1002/9780470650301.ch3.

Forster, B.P., Heberle-Bors, E., Kasha, K.J., Touraev, A. (2007). The resurgence of haploids in higher plants. Trends Plant Sci., 12, 368–375, DOI: 10.1016/j.tplants.2007.06.007.

Galbraith, D.W., Harkins, K.R., Maddox, J.M., Ayres, N.M., Sharma, D.P., Firoozabady, E. (1983). Rapid flow cytometry analysis of the cell cycle in intact plant tissues. Science, 220, 1049–1051, DOI: 10.1126/science.220.4601.1049.

Germana, M.A. (2011). Anther culture for haploid and doubled haploid production. Plant Cell Tiss. Organ Cult., 104, 283–300, DOI: 10.1007/s11240-010-9852-z.

Ilbi, H. (2003). RAPD markers assisted varietal identification and genetic purity test in pepper Capsicum annuum L. Scientia Hortic., 97, 211-221, DOI: 10.1016/S0304-4238(02)00158-9.

Irikova, T., Grozeva, S., Rodeva, V. (2011). Anther culture in pepper (Capsicum annuum L.) in vitro. Acta Physiol. Plant., 33, 1559–1570, DOI: 10.3906/BIY-1506-79.

Jedrzejczyk, I., Nowaczyk, P. (2009). In vivo polyembryony induction in species of Capsicum. Acta Biol. Cracoviensia, 51(1), 55–60.

Kim, M., Kim, J., Yoon, M., Choi, D.I., Lee, K.M. (2004). Origin of multicellular pollen and pollen embryos in cultured anthersof pepper (Capsicum annuum L.). Plant Cell Tissue Organ. Cult., 77, 63–72, DOI: 10.1023/B:TICU.0000016506.02796.6a.

Kothari, S.L., Joshi A., Kachhwaha, S., Ochoa-Alejo, N. (2010). Chilli peppers-areview on tissue culture and transgenis. Biotechnol. Adv., 28, 35–48, DOI: 10.1016/j.biotechadv.2009.08.005.

Kristiansen, K., Andersen, S.B. (1993). Effects of donor plant, temperature, photoperiod and age on anther culture response of Capsicum annuum L. Euphytica, 67, 105–109, DOI:

Lantos, C., Gemes, A.J., Somogyi, G., Otvos, K., Vagi, P., Mithaly, R., Kristof, Z., Somogyi, N., Pauk, J. (2009). Improvement of isolated microspore culture of pepper (Capsicum annuum L.) via co-culture with ovary tissues of pepper or wheat. Plant Cell Tissue Organ Cult., 97, 285–293, DOI : 10.1007/s11240-009-9527-9.

Mityko, J., Fari, M. (1997). Problems and results of doubled haploid plant production in pepper (Capsicum annuum L.) via anther and microspore culture. Acta Hortic., 447, 281–287, DOI: 10.17660/ActaHortic.1997.447.57.

Naumova, T. (1992). Apomixis in angiosperms: nucellar and integumentary embryony. CRC Press, Boca Raton, 1–144, DOI: 105586/asbp.1981.034.

Nei, M., Li., W.H. (1979). Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc. Natl. Acad. Sci. USA, 76, 5269–5273, DOI: 10.1073/pnas.76.10.5269.

Niemirowicz-Szczytt, K. (1997). Excessive homozygosity in doubled haploids – advantages and disadvantages for plant breeding and fundamental research. Acta Physiol. Plant., 19, 155–167, DOI: 10.1007/s11738-997-0032-7.

Nikolova, V., Niemirowicz-Szczytt, K. (1996). Dihaploidization of cucumber (Cucumis sativus L.) haploids by colchicine treatment. Acta Soc. Bot. Pol., 65, 311–317, DOI: 10.5586/asbp.1996.048.

Nowaczyk, L., Nowaczyk, P. (2006). Genetic analysis of tomato (Lycopersicon esculentum Mill.) twin forms. Acta Biol. Cracoviensia, 48(1), 53–58.

Nowaczyk, L., Nowaczyk, P., Olszewska, D. (2015). Genetic analysis of anther culture-derived diploids of Capsicum spp. J. Hortic. Sci. Biotech., 90(6), 747–752, DOI: 10.1080/14620316.2015.11668741.

Olszewska, D., Jedrzejczyk, I., Nowaczyk, P., Sendel, S., Gaczkowska, B. (2015). In vitro colchicine treatment of anther-derived pepper haploids. Bulgarian J. Agri. Sci., 21(4), 806–810.

Rao, R.K.G., Harini, I., Kumar, A.O. (1987). Colchicine induced chromosome mosaicism in chili pepper (Capsicum annuum L.). Proc. Indian Acad. Sci., 97, 55–61,

Rodeva, V., Irikova, T., Todorova, V. (2004). Anther culture of pepper (Capsicum annuum L.): comparative study on effect of the genotype. Biotechnol. Equip., 3, 34–38, DOI: 10.1080/13102818.2004.10817117.



Dorota Olszewska 
Department of Agricultural Biotechnology, University of Science and Technology, Bernardynska 6, 85-029 Bydgoszcz, Poland
Paweł Nowaczyk 
Department of Agricultural Biotechnology, University of Science and Technology, Bernardynska 6, 85-029 Bydgoszcz, Poland



Articles are made available under the conditions CC BY 4.0 (until 2020 under the conditions CC BY-NC-ND 4.0).
Submission of the paper implies that it has not been published previously, that it is not under consideration for publication elsewhere.

The author signs a statement of the originality of the work, the contribution of individuals, and source of funding.


Most read articles by the same author(s)