Regulation of in vitro morphogenesis in maize inbreds of the Lancaster group

  • K. V. Derkach Institute of Grain Crops of National Academy of Agrarian Science of Ukraine
  • O. E. Abraimova Institute of Grain Crops of National Academy of Agrarian Science of Ukraine
  • T. M. Satarova Institute of Grain Crops of National Academy of Agrarian Science of Ukraine
Keywords: Zea mays, 6-benzilaminopurine, indolylbutyric acid, cefotaxim, callusogenesis, regeneration


The aim of this work is the comparative evaluation of the influence of physiologically active substances (PAS) on the ratio of morphogenesis types in vitro and plant regeneration in callus tissues of maize inbreds of the Lancaster heterotic group. Lancaster inbreds of Ukrainian selection DК267, DК6080 and DК298 and lines PLS61, A188 and Chi31 of foreign selection which represented eponymous heterotic groups were selected for into the research. For callusogenesis induction immature embryos of 1.0–1.5 mm in length were planted on a modified N6 medium with 30 g/l sucrose. For regeneration the 30-day callus tissue was transplanted to a modified MS medium containing 20 g/l sucrose and various PAS: 6-benzylaminopurine (BAP) (0.1 mg/l) or indolylbutyric acid (IBA) (1.0 mg/l), or cefotaxime (CT) (150 mg/l). Morphogenesis and plant regeneration were obtained in callus tissue by organogenesis through the formation of leaf-like structures or shoots only without roots (hemmogenesis) and by embryogenesis through the development of embryos with the cooperative laying of the apexes of the shoot and the root. The predominant type of in vitro morphogenesis in both groups of genotypes was organogenesis. A tendency to increasing the embryogenesis level among Lancaster inbreds in comparison with the other investigated groups was observed. A certain tendency to increase the level of embryogenesis on a medium for regeneration with IBA, compared to other PAS, was marked. So, for the Lancaster group the embryogenesis level on the medium with IBA was 33.3%, while for the other investigated genotypes it was about 10.3%. Regeneration frequency was 45.8 plantlets/100 calli for Lancaster inbred DК298, 42.2 plantlets/100 calli for DК267 and 5.6 plantlets/100 calli for DК6080. Regeneration frequency of inbred PLS61 was 204.0 plantlets/100 calli, 100.0 plantlets/100 calli for Chi31 and 11.1 plantlets/100 calli for А188. Observed reductions in capacityfor plant regeneration of Lancaster inbreds may have been due to their pedigree. These inbreds belong to the commercial heterotic group that was not specially selected for increased regenerative ability, unlike the model inbreds of other heterotic groups. Overall, the level of regeneration frequency in the Lancaster group was 44.8 plantlets/100 calli on the medium under BAP, 41.4 plantlets/100 calli under IBA, and 20.7 plantlets/100 calli with CT. In general, the level of regeneration frequency for non-Lancaster heterotic groups reached 89.7 plantlets/100 calli on the medium with BAP, 96.7 plantlets/100 calli under IBA, and 93.1 plantlets/100 calli under CT. To enhance embryogenesis and frequency of regeneration for maize inbreds of the Lancaster group 30 g/l sucrose in the callusogenesis medium and physiologically active substance indolylbutyric acid (1.0 mg/l) in the medium for regeneration can be recommended.


Akoyi, J., Mgutu, A.J., Machuka, J., van Lijsebettens, M., Taracha, C., Anami, S.E., 2013. Dicamba growth regulator promotes genotype independent somatic embryogenesis from immature zygotic embryos of tropical maize inbred lines. J. Life Sci. 7(7), 677–689.

Ali, F., Ahsan, M., Saeed, N.A., Ahmed, M., Ali, Q., Kanwal, N., Tehseen, M.M., Ijaz, U., Bibi, I., Niazi, N.K., 2014. Establishment and optimization of callus-to-plant regeneration system using mature and immature embryos of maize (Zea mays). Int. J. Agric. Biol. 16, 111–117.

Assem, S.K., 2015. Maize, tropical (Zea mays L.). Methods Mol. Biol. 1223, 119–134. >>

Batygina, T.B., Kruglova, N.N., Gorbunova, V.Y.,Titova, G.Y., Seldimirova, O.A., 2010. Ot mikroskopa k raznoobraziju [From microspore to variety]. Nauka, Moskow (in Russian).

Bedada, L.T., Seth, M., Runo, S.M., Tefera, W., Jesse, M., 2012. Regenerability of elite tropical maize (Zea mays L.) inbred lines using immature zygotic embryo explants. Afr. J. Biotechnol. 11(8), 598–605. >>

Bennetzen, J.L., Hake, E.S.A., 2009. Handbook of maize. Genetics and Genomics. Springer Science, New York. >>

Chu, C.C., Wang, J.J., Sun, J.S., 1975. Establishment of an efficient medium for anther culture of rice through comparative experiments of the nitrogen sources. Sci. Sinica 18(5), 659–668.

Danilova, S.A., Dolgikh, Y.I., 2004. The stimulatory effect of the antibiotic cefotaxime on plant regeneration in maize tissue culture. Russ. J. Plant Physl. 51(4), 559–562. >>

Derkach, K.V., Abraimova, O.E., Satarova, T.M., 2011. Kalusogennyj potencial linij kukurudzy grupy Lankaster v umovah in vitro [Callusogenic potential of maize lines of Lancaster group in vitro]. Vìsn. Dnìpropetr. Unìv. Ser. Bìol. Ekol. 19(1), 16–21 (in Ukrainian).

González, G.A., Pacheco, M.G., Oneto, C.D., Etchart, V.J., Kandus, M.V., Salerno, J.C., Eyherabide, G., Presello, D., Lewi, D.M., 2012. Somatic embryogenesis and plant regeneration capacity in Argentinean maize (Zea mays L.) inbred lines. Electron J. biotechn. 15(1), 1–7. >>

Green, C.E., Phillips, H.L., 1975. Plant regeneration from tissue cultures of maize. Crop Sci. 15(5), 417–421. >>

Guruprasad, M., Sridevi, T.V., Udaya Sri, A.P.P., Kumar, M.S., 2015. Plant regeneration through callus initiation from mature and immature embryos of maize (Zea mays L.). Journal of Multidisciplinary Advanced Research Trends 2(1), 195–202. >>

Guruprasad, M., Sridevi, Т., Vijayakumar, G., Kumar, M.S., 2016. Plant regeneration through callus initiation from mature and immature embryos of maize (Zea mays L.). Indian J. Agr. Res. 50(2), 135–138. >>

Jia, X.X., Zhang, J.W., Wang, H.N., Kong, W.P., 2008. Efficient maize (Zea mays L.) regeneration derived from mature embryos in vitro. Maydica 53, 239–248.

Liu, B., Su, S., Wu, Y., Li, Y., Shan, X., Li, S., Liu, H., Dong, H., Ding, M., Han, J., Yuan, Y., 2015. Histological and transcript analyses of intact somatic embryos in an elite maize (Zea mays L.) inbred line Y423. Plant Physiol. Biochem. 92, 81–91. >>

Malini, N., Anandakumar, C.R., Hari Ramakrishnan, S., 2015. Regeneration of Indian maize genotypes (Zea mays L.) from immature embryo culture through callus induction. Journal of Applied and Natural Science 7(1), 131–137.

Muoma, J., Ombori, O., Machuka, J., 2011. Improvement in inheritance of somatic embryogenesis and plantlet regeneration in tropical maize lines from friable callus. Maize Genet. Coop. News Lett. 85, 1–2.

Murashige, T., Skoog, F., 1962. A revised media for rapid growth and bioassay with tobacco tissue culture. Physiol. Plant. 15, 473–497.

Oduor, R.O., Njagi, E.N.M., Machuka, J.S., 2006. In vitro regeneration of dryland Kenyan maize genotypes through somatic embryogenesis. Int. J. Bot. 2(2), 146–151. >>

Ombori, O., Gitonga, N.M., Machuka, J., 2008. Somatic embryogenesis and plant regeneration from immature embryos of tropical maize (Zea mays L.) inbred lines. Biotechnology 7, 224–232. >>

Qamar, Z., Aaliyah, K., Nasir, I.A., Farooq, A.M., Tabassum, B., Ali, Q., Ali, A., Awan, M.F., Tariq, M., Hasnain, T., 2015. An overview of genetic transformation of glyphosate resistant gene in Zea mays. Nature and Science 13(3), 80–90.

Satarova, T.M., Cherchel, V.Y., Cherenkov, A.V., 2013. Kukuruza: Biotehnologicheskie i selekcionnye aspekty gaploidii [Maize: Biotechnological and breeding aspects of haploidy]. New ideology, Dniepropetrovsk (in Russian).

Seth, M.S., Bedada, L.T., Mneney, E.E., Oduor, R.O., Machuka, J.S., 2012. In vitro regeneration of selected commercial Tanzanian open pollinated maize varieties. Afr. J. Biotechnol. 11(22), 6043–6049. >>

Shohael, A.M., Akanda, M.A.L., Parvez, S., Mahfuja, S., 2003. Somatic embryogenesis and plant regeneration from immature embryoderived callus of inbred mays (Zea mays L.). Biotechnology 2(2), 154–161. >>

Welham, S.J., Gezan, S.A., Clark, S.J., Mead, A., 2015. Statistical methods in biology: Design and analysis of experiments and regression. CRC Press, Boca Raton.