DOI: https://doi.org/10.21498/2518-1017.15.3.2019.181084

Elaboration of an effective method of callusogenesis induction from mature germs of Triticum spelta L. and T. aestivum L.

А. В. Кирієнко, М. Ф. Парій, М. В. Кучук, Ю. В. Симоненко, Н. Л. Щербак

Abstract


Purpose. Introduction to in vitro culture and obtaining of callus from mature embryos of 3 spelt samples and comparing the effectiveness of their callusogenesis with 2 soft wheat samples.

Methods. Five samples of hexaploid wheat (three of spelt and two of soft wheat) were taken for experiments. Surface sterilization of grains was carried out in 96% ethanol and 5% sodium hypochlorite solution. Mature embryos were used as explants. Three types of MS culture media with different component compositions were used for callusogenesis. Explants were cultivated in the dark for 21 days.

Results. The optimal conditions for the induction of tissue culture of Triticum spelta L. and T. aestivum L. from mature embryos were selected. Received calli from different samples, which were grown on three types of culture media MS, did not differ morphologically from each other. A genetic predisposition to callus formation was observed for specimens of ‘Evropa’ spelt variety and soft wheat ‘Bunchuk’ and ‘Elehiia Myronivska’ wheat samples regardless of the composition of the MS medium, while callus formation was slow on the explants of ‘Zoria Ukrainy’ cultivar.

Conclusions. A tissue culture of 3 spelt samples and 2 soft wheat samples was obtained using mature embryos as explants. It was found that a nutrient medium containing 3% sucrose and supplemented with 2 mg/L 2,4-D, 10 ml/L silver nitrate was the most effective for callus formation from mature germ explants of soft wheat and spelt. The efficiency of the calluso­genesis on the 21st day of cultivation, depending on the sample, varied in the range of 80.2–100.0%. The studied samples differed among themselves in their ability to form calli on nutrient media with different component composition. The efficiency of spelt callusogenesis was first studied in Ukraine.


Keywords


spelt; soft wheat; in vitro culture; callus; explants

References


Kumar, A., Priya, Sharma, S., & Yadav, M. K. (2019). Plant tissue culture technology to Improve crop species – a comprehensive approach. Acta Sci. Agric., 3(2), 7–80.

Sticklen, B. M., & Oraby, F. H. (2005). Invited review: shoot apical meristem: a sustainable explant for genetic transformation of cereal crops. In Vitro Cell. Dev. Biol. Plant., 41(3), 187–200. doi: 10.1079/IVP2004616

Ahloowalia, B. S. (1982). Plant regeneration from callus culture in wheat. Crop Sci., 22(2), 405–410. doi: 10.2135/cropsci1982.0011183X002200020047x

Nasircilar, G. A., Turgut, K., & Fiskin, K. (2006). Callus induction and plant regeneration from mature embryos of different wheat genotypes. Pak. J. Bot., 38(3), 637–635.

McHughen, A. (1983). Rapid regeneration of wheat in vitro. Ann. Bot., 51(6), 851–853. doi: 10.1093/oxfordjournals.aob.a086535

Al-Kaaby, H. K., Abdul-Qadir, L. H., & Kareem, M. A. (2015). Effect of silver nitrate on callus induction, somatic embryos formation and plantlets regeneration in two local wheat (Triticum aestivum L.) cultivars. Thi-Qar Univ. J. Agric. Res., 4(2), 51–60.

Mahmood, I., Razzaq, A., Qureshi, A. A., Qayyum, A., & Qadeer Beig, M. M. (2018). Comparative morpho-physiological response of in vitro selected somaclones of wheat (Triticum aestivum L.) and explant donor parent to drought stress. J. Natn. Sci. Foundation Sri Lanka., 46(3), 293–302 doi: 10.4038/jnsfsr.v46i3.8481

Kopertekh, L. G., & Stribnaya, L. A. (2003). Plant regeneration from wheat leaf explants. Russ. J. Plant Physiol., 50(3), 365–368. doi: 10.1023/A:1023826304989

Konieczny, R., Czaplicki, A. Z., Golczyk, H., & Przywara, L. (2003). Two pathways of plant regeneration in wheat anther culture. Plant Cell Tissue Organ Cult., 73(2), 177–187. doi: 10.1023/A:1022877807237

Symillides, Y., Henry, Y., & De Buyser, J. (1995). Analysis of Chinese Spring regenerants obtained from short- and long-term wheat somatic embryogenesis. Euphytica, 82(3), 263–268. doi: 10.1007/BF00029569

Bommineni, V. R., & Jauhar, P. P. (1996). Regeneration of plantlets through isolated scutellum culture of durum wheat. J. Plant Sci., 116(2), 197–203. doi: 10.1016/0168-9452(96)84541-9

Przetakiewicz, A., & Nadolska-Orczyk, A. (2003). The effect of auxin on plant regeneration of wheat, barley and Triticale. Plant Cell Tissue Organ Cult., 73(3), 245–256. doi: 10.1023/A:1023030511800

Ihsan, S. M., Jabeen, M., & Ilahi, I. (2003). In vitro callus induction, its proliferation and regeneration in seed explants of wheat (Triticum aestivum L.) Var.lu-26s. Pak. J. Bot., 35(2), 209–217.

Haliloglu, K. (2002). Wheat immature embryo culture for embryogenic callus induction. J. Biol. Sci., 2(8), 520–521. doi: 10.3923/jbs.2002.520.521

Baday, S. J. S. (2018). In vitro study of the callus induction of two varieties of wheat seeds by plant growth regulators. Agric. J., 13(3), 67–71. doi: 10.3923/aj.2018.67.71

Fernandez, S., Michaux-Ferriere, N., & Coumans, M. (1999). The embryogenic response of immature embryo cultures of durum wheat (Triticum durum Desf.): histology and improvement by AgNO3. J. Plant Growth Regul., 28(3), 147–155. doi: 10.1023/A:1006142504577

Murashige, T., & Skoog, F. (1962). A revised medium for ra­pid growth and bio assays with tobacco tissue cultures. Phys. Plant., 15(3), 473–497. doi: 10.1111/j.1399-3054.1962.tb08052.x

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

Gouranga, U., Moutushi, S., & Amitava, R. (2015). In vitro callus induction and plant regeneration of rice (Oryza sativa L.) var. ‘Sita’, ‘Rupali’ and ‘Swarna Masuri’. Asian J. Plant Sci. Res., 5(5), 24–27.

Abd El-Fatah, B. E. S. (2018). Genetic studies of response to mature embryo culture and relationship with agro-morpholo­gical traits and molecular markers in wheat. Plant Breed. Biotech., 6(3), 267–284. doi: 10.9787/PBB.2018.6.3.267

Parmar, S. S., Sainger, M., Chaudhary, D., & Jaiwal, P. K. (2012). Plant regeneration from mature embryo of commercial Indian bread wheat (Triticum aestivum L.) cultivars. Physiol. Mol. Biol. Plants., 18(2), 177–183. doi: 10.1007/s12298-012-0101-2

Wu, L. M., Wei, Y. M., & Zheng, Y. L. (2006). Effects of silver nitrate on the tissue culture of immature wheat embryos. Russ. J. Plant Physiol., 53(4), 530–534. doi: 10.1134/S1021443706040157


GOST Style Citations


Kumar A., Priya, Sharma S., Yadav M. K. Plant tissue culture technology to Improve crop species – a comprehensive approach. Acta Sci. Agric. 2019. Vol. 3, Iss. 2. P. 76–80.

Sticklen B. M., Oraby F. H. Invited review: shoot apical meristem: a sustainable explant for genetic transformation of cereal crops. In Vitro Cell. Dev. Biol. Plant. 2005. Vol. 41, Iss. 3. P. 187–200. doi: 10.1079/IVP2004616

Ahloowalia B. S. Plant regeneration from callus culture in wheat. Crop Sci. 1982. Vol. 22, Iss. 2. Р. 405–410. doi: 10.2135/cropsci1982.0011183X002200020047x

Nasircilar G. A., Turgut K., Fiskin K. Callus induction and plant regeneration from mature embryos of different wheat genotypes. Pak. J. Bot. 2006. Vol. 38, Iss. 3. P. 637–635.

McHughen A. Rapid regeneration of wheat in vitro. Ann. Bot. 1983. Vol. 51, Iss. 6. P. 851–853. doi: 10.1093/oxfordjournals.aob.a086535

Al-Kaaby H. K., Abdul-Qadir L. H., Kareem M. A. Effect of silver nitrate on callus induction, somatic embryos formation and plantlets regeneration in two local wheat (Triticum aestivum L.) cultivars. Thi-Qar Univ. J. Agric. Res. 2015. Vol. 4, Iss. 2. P. 51–60.

Mahmood I., Razzaq A., Qureshi A. A. et al. Comparative morpho-physiological response of in vitro selected somaclones of wheat (Triticum aestivum L.) and explant donor parent to drought stress. J. Natn. Sci. Foundation Sri Lanka. 2018. Vol. 46, Iss. 3. P. 293–302 doi: 10.4038/jnsfsr.v46i3.8481

Kopertekh L. G., Stribnaya L. A. Plant regeneration from wheat leaf explants. Russ. J. Plant Physiol. 2003. Vol. 50, Iss. 3. P. 365–368. doi: 10.1023/A:1023826304989

Konieczny R., Czaplicki A. Z., Golczyk H., Przywara L. Two pathways of plant regeneration in wheat anther culture. Plant Cell Tissue Organ Cult. 2003. Vol. 73, Iss. 2. P. 177–187. doi: 10.1023/A:1022877807237

Symillides Y., Henry Y., De Buyser J. Analysis of Chinese Spring regenerants obtained from short- and long-term wheat somatic embryogenesis. Euphytica. 1995. Vol. 82, Iss. 3. P. 263–268. doi: 10.1007/BF00029569

Bommineni V. R., Jauhar P. P. Regeneration of plantlets through isolated scutellum culture of durum wheat. Plant Sci. 1996. Vol. 116, Iss. 2. P. 197–203. doi: 10.1016/0168-9452(96)84541-9

Przetakiewicz A., Nadolska-Orczyk A. The effect of auxin on plant regeneration of wheat, barley and Triticale. Plant Cell Tissue Organ Cult. 2003. Vol. 73, Iss. 3. P. 245–256. doi: 10.1023/A:1023030511800

Ihsan S. M., Jabeen M., Ilahi I. In vitro callus induction, its proliferation and regeneration in seed explants of wheat (Triticum aestivum L.) Var.lu-26s. Pak. J. Bot. 2003. Vol. 35, Iss. 2. P. 209–217.

Haliloglu K. Wheat immature embryo culture for embryogenic callus induction. J. Biol. Sci. 2002. Vol. 2, Iss. 8. P. 520–521. doi: 10.3923/jbs.2002.520.521 

Baday S. J. S. In vitro study of the callus induction of two varieties of wheat seeds by plant growth regulators. Agric. J. 2018. Vol. 13, Iss. 3. P. 67–71. doi: 10.3923/aj.2018.67.71

Fernandez S., Michaux-Ferriere N., Coumans M. The embryogenic response of immature embryo cultures of durum wheat (Triticum durum Desf.): histology and improvement by AgNO3. Plant Growth Regul. 1999. Vol. 28, Iss. 3. P. 147–155. doi: 10.1023/A:1006142504577

Murashige T., Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Phys. Plant. 1962. Vol. 15, Iss. 3. P. 473–497. doi: 10.1111/j.1399-3054.1962.tb08052.x

Gamborg O. L., Miller R. A., Ojima K. Nutrient requirements of suspension cultures of soybean root cells. J. Exp. Cell Res. 1968. Vol. 50, Iss. 1. P. 151–158. doi: 10.1016/0014-4827(68)90403-5

Gouranga U., Moutushi S., Amitava R. In vitro callus induction and plant regeneration of rice (Oryza sativa L.) var. ‘Sita’, ‘Rupali’ and ‘Swarna Masuri’. Asian J. Plant Sci. Res. 2015. Vol. 5, Iss. 5. P. 24–27.

Abd El-Fatah B. E. S. Genetic studies of response to mature embryo culture and relationship with agro-morphological traits and molecular markers in wheat. Plant Breed. Biotech. 2018. Vol. 6, Iss. 3. P. 267–284. doi: 10.9787/PBB.2018.6.3.267

Parmar S. S., Sainger M., Chaudhary D., Jaiwal P. K. Plant regene­ration from mature embryo of commercial Indian bread wheat (Triticum aestivum L.) cultivars. Physiol. Mol. Biol. Plants. 2012. Vol. 18, Iss. 2. P. 177–183. doi: 10.1007/s12298-012-0101-2

Wu L. M., Wei Y. M., Zheng Y. L. Effects of silver nitrate on the tissue culture of immature wheat embryos. Russ. J. Plant Physiol. 2006. Vol. 53, Iss. 4. P. 530–534. doi: 10.1134/S1021443706040157







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DOI: 10.21498/2518-1017

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