Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

The effect of an elevated cytokinin level using the ipt gene and N 6-benzyladenine on single node and intact potato plant tuberization in vitro


Two models of potato (Solanum tuberosum L.) tuberization in vitro (intact plants and single nodes) were used to study the role of cytokinins in this process. We applied hormone in two different ways. The exogenous addition of 10 mg · L-1 N 6-benzyladenine (BA) into the tuberization medium resulted in advanced tuber formation in intact plants, and microtubers appeared 10–20 days earlier than in the experiments in which no cytokinin was supplied. Transformation with the Agrobacterium tumefaciens ipt gene provided potato clones with endogenously elevated cytokinin levels (3–20 times higher zeatin riboside content in different clones). The onset of tuberization in intact ipt-transformed plants with low transgene expression was advanced in comparison with control material, and exogenously applied BA further promoted the tuberization process. On the contrary, tuberization was strongly inhibited in ipt-transformed nodes, and an external increase of the cytokinin level caused complete inhibition of expiant growth. In untransformed (control) nodes cytokinin application resulted in primary and secondary tuber formation, which depended on the BA concentration in cultivation media.

This is a preview of subscription content, log in to check access.





polymerase chain reaction


high performance liquid chromatography


enzyme-linked immunosorbent assay


α-naphthylacetic acid


  1. Ainley WM, McNeil KJ, Hill JW, Lingle WL, Simpson RB, Brenner ML, Nagao RT, Key JL (1993) Regulatable endogenous production of cytokinins up to “toxic” levels in transgenic plants and plant tissues. Plant Mol Biol 22:13–23

  2. Amasino RM, Powell ALT, Gordon MP (1984) Changes in T-DNA methylation and expression are associated with phenotypic variation and plant regeneration in a crown gall tumor line. Mol Gen Genet 197:437–446

  3. Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: Version II. Plant Mol Biol Rep 1:19–21

  4. Ewing EE (1978) Critical photoperiod for tuberization: A screening technique with potato cuttings. Am Potato J 55:43–53

  5. Ewing EE (1990) Induction of tuberization in potato. In: Vayda ME, Park WD (eds) The molecular and cellular biology of the potato. CAB International, Redwood Press, Melksham, pp 25–41

  6. Hamill JD (1993) Alterations in auxin and cytokinin metabolism of higher plants due to expression of specific genes from pathogenic bacteria: A review. Aust J Plant Physiol 20: 405–423

  7. Hare PD, Van Staden J (1994) Cytokinin oxidase: Biochemical features and physiological significance. Physiol Plant 91: 128–136

  8. Harvey BMR, Crothers SH, Evans NE, Selby C (1991) The use of growth retardants to improve microtuber formation by potato (Solanum tuberosum). Plant Cell Tiss Org Cult 27: 59–64

  9. Horsch RB, Fry JE, Hoffmann N, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231

  10. Kado CI (1991) Molecular mechanisms of crown gall tumorogenesis. Crit Rev Plant Sci 10:1–32

  11. Levy D, Seabrook JEA, Coleman S (1993) Enhancement of tuberization of axillary shoot buds of potato (Solanum tuberosum L.) cultivars cultured in vitro. J Exp Bot 44:381–386

  12. Medford JI, Horgan R, El-Sawi Z, Klee HJ (1989) Alterations of endogenous cytokinins in transgenic plants using a chimeric isopentenyl transferase gene. Plant Cell 1:403–413

  13. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

  14. Neuteboom STC, Hulleman E, Schilperoort RA, Hoge HC (1993) In planta analysis of the Agrobacterium tumefaciens T-cyt gene promoter: Identification of an upstream region essential for promoter activity in leaf, stem and root cells of transgenic tobacco. Plant Mol Biol 22: 923–929

  15. Ooms G, Lenton JR (1985) T-DNA genes to study plant development: Precocious tuberisation and enhanced cytokinins in A. tumefaciens transformed potato. Plant Mol Biol 5: 205–212

  16. Pavingerová D, Hrouda M (1991) Meiotic transmission of T-DNA genes in Arabidopsis thaliana plants and their expression after 5-azacytidine treatment. Biol Plant 33: 461–467

  17. Pelacho AM, Mingo-Castel AM (1991) Effects of photoperiod on kinetin-induced tuberization of isolated potato stolons cultured in vitro. Am Potato J 68:533–541

  18. Skoog F, Miller CO (1957) Chemical regulation of growth and organ formation in plant tissues cultured in vitro. Soc Exp Bot Symp 11:118–131

  19. Slimmon T, Machado VS, Coffin R (1989) The effect of light on in vitro microtuberization of potato cultivars. Am Potato J 66:843–848

  20. Smart CM, Scofield SR, Bevan MW, Dyer TA (1991) Delayed leaf senescence in tobacco plants transformed with tmr, a gene for cytokinin production in Agrobacterium. Plant Cell 3:647–656

  21. Stallknecht GF, Farnsworth S (1982) General characteristics of coumarin-induced tuberization of axillary shoots of Solanum tuberosum L. cultured in vitro. Am Potato J 59:17–31

  22. Strnad M, Vaněk T, Binarová P, Kamínek M, Hanuš J (1990) Enzyme immunoassays for cytokinins and their use for immunodetection of cytokinins in alfalfa cell culture. In: Kutáček M, Elliott MC, Macháčková I (eds) Molecular aspects of hormonal regulation of plant development. SPB Academic Publishing bv, The Hague, pp 41–54

  23. Van Den Berg JH, Struik PC, Ewing EE (1990) One-leaf cuttings as a model to study second growth in potato (Solanum tuberosum) plant. Ann Bot 66:273–280

  24. Van Staden J, Upfold SJ, Altman A (1994) Metabolism of [14C] trans-zeatin and [14C]benzyladenine by detached yellow, green and variegated leaves of Schefflera. Physiol Plant 90:73–78

  25. Vlasák J, Ondřej M (1992) Construction and use of Agrobacterium tumefaciens binary vectors with A. tumefaciens T-DNA genes. Folia Microbiol 37:227–230

  26. Vreugdenhil D, Helder H (1992) Hormonal and metabolic control of tuber formation. In: Karssen CM, Van Loon LC, Vreugdenhil D (eds) Progress in plant growth regulation. Proceedings of the 14th international conference on plant growth substances, Amsterdam, July 21–26, 1991, Kluwer Academic Publishers, Dordrecht, pp 393–400

  27. Vreugdenhil D, Struik PC (1989) An integrated view of the hormonal regulation of tuber formation in potato (Solanum tuberosum). Physiol Plant 75:525–531

Download references

Author information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Gális, I., Macas, J., Vlasák, J. et al. The effect of an elevated cytokinin level using the ipt gene and N 6-benzyladenine on single node and intact potato plant tuberization in vitro . J Plant Growth Regul 14, 143–150 (1995).

Download citation


  • Tuber Formation
  • Potato Plant
  • Zeatin
  • Intact Plant
  • Zeatin Riboside