Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 136, Issue 1, pp 133–140 | Cite as

Telomere length in Agave tequilana Weber plants during the in vitro to ex vitro transition

  • A. Rescalvo-Morales
  • K. M. Monja-Mio
  • M. L. Robert
  • L. F. Sánchez-TeyerEmail author
Original Article


Acclimatization ex vitro is a key stage of the micropropagation process, in which the vitro plants leave the sterile, high humidity environment in which they originated and form new leaves and roots, during which they suffer different types of stress. Changes in the telomere length (shortening and lengthening) have been associated with age, the development of tissue, loss of cell replication and the ability of regeneration in different plant species. However, the genetic and biological factors that are involved in the process of shortening of telomeres across the ageing of plant species are still unknown. In this study, we used terminal restriction fragments (TRF) to examine the changes of telomere length during the in vitro to ex vitro transition in vitro plants of Agave tequilana, and their relationships with age in plants grown in commercial plantations. The results showed that in vitro grown plants present the longest telomeres and that a shortening occurs during the first 6 months of ex vitro acclimatization, (compared to the plantlets that were kept in vitro). A lengthening of the telomeres was observed in the acclimatized 1-year-old plants and that this was maintained in 2 and 3-year-old plants. We also observed TRF variations in the different tissues (leaves, stems and roots) of acclimatized plants. In field plants, we did not observe any important changes in the length of the telomeres. We suggest that agaves have a mechanism that maintains telomere length at the non-critical stages during development.


Agave tequilana Telomere length Age Acclimatization Ex vitro 



We thank the technicians Adriana Quiroz Moreno and Gaston Herrera Herrera for technical support. This work was funded by CONACyT through a research Grant (CB-2012-180757-Z) and a Ph.D.-fellowship (326680) for the first author.


  1. Aronen T, Ryynänen L (2012) Variation in telomeric repeats of Scots pine (Pinus sylvestris L.). Tree Genet Genomes 8:267–275. CrossRefGoogle Scholar
  2. Aronen T, Ryynänen L (2014) Silver birch telomeres shorten in tissue culture. Tree Genet Genomes 10:67–74. CrossRefGoogle Scholar
  3. Barnes RP, Fouquerel E, Opresko PL (2018) The impact of oxidative DNA damage and stress on telomere homeostasis. Mech Ageing Dev. Google Scholar
  4. Broun P, Ganal MW, Tanksley SD (1992) Telomeric arrays display high levels of heritable polymorphism among closely related plant varieties. Proc Natl Acad Sci USA 89:1354–1357. CrossRefGoogle Scholar
  5. Chakrabarty D, Datta SK (2008) Micropropagation of gerbera: lipid peroxidation and antioxidant enzyme activities during acclimatization process. Acta Physiol Plant 30:325–331. CrossRefGoogle Scholar
  6. de Lange T (2010) Telomere biology and DNA repair: enemies with benefits. FEBS Lett 584:3673–3674. CrossRefGoogle Scholar
  7. Doyle J, Doyle J (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15Google Scholar
  8. Epel ES, Blackburn EH, Lin J et al (2004) Accelerated telomere shortening in response to life stress. Proc Natl Acad Sci USA 101:17312–17315. CrossRefGoogle Scholar
  9. Faisal M, Anis M (2009) Changes in photosynthetic activity, pigment composition, electrolyte leakage, lipid peroxidation, and antioxidant enzymes during ex vitro establishment of micropropagated Rauvolfia tetraphylla plantlets. Plant Cell Tissue Organ Cult 99:125–132. CrossRefGoogle Scholar
  10. Fajkus J, Fulnečková J, Hulánová M et al (1998) Plant cells express telomerase activity upon transfer to callus culture, without extensively changing telomere lengths. Mol Gen Genet 260:470–474. CrossRefGoogle Scholar
  11. Flanary BE, Kletetschka G (2005) Analysis of telomere length and telomerase activity in tree species of various life-spans, and with age in the bristlecone pine Pinus longaeva. Biogerontology 6:101–111. CrossRefGoogle Scholar
  12. Flanary BE, Streit WJ (2003) Telomeres shorten with age in rat cerebellum and cortex in vivo. J Anti Aging Med 6:299–308. CrossRefGoogle Scholar
  13. Gentry HS (1982) Agaves of continental North America. The University of Arizona Press, TucsonGoogle Scholar
  14. Göhring J, Fulcher N, Jacak J, Riha K (2014) TeloTool: a new tool for telomere length measurement from terminal restriction fragment analysis with improved probe intensity correction. Nucleic Acids Res 42:1–10. CrossRefGoogle Scholar
  15. Gonçalves S, Martins N, Romano A (2017) Physiological traits and oxidative stress markers during acclimatization of micropropagated plants from two endangered Plantago species: P. algarbiensis Samp. and P. almogravensis Franco. In Vitro Cell Dev Biol Plant 53:249–255. CrossRefGoogle Scholar
  16. Grygoryev D, Zimbrick J (2010) Effect of quadeuplex conformation on radiation-induced formation of 8-hydroxyguanine and unaltered base release in polyguanylic acid. Radiat Res 173:110–118. CrossRefGoogle Scholar
  17. Harley CB, Futcher B, Greider C (1990) Telomeres shorten during ageing of human fibroblasts. Nature 345:458–460CrossRefGoogle Scholar
  18. Heacock M, Spangler E, Riha K et al (2004) Molecular analysis of telomere fusions in Arabidopsis: multiple pathways for chromosome end-joining. EMBO J 23:2304–2313. CrossRefGoogle Scholar
  19. Ijdo JW, Wells RA, Baldini A, Reeders ST (1991) Improved telomere detection using a telomere repeat probe (TTAGGG)n generated by PCR. Nucleic Acids Res 19:4780. CrossRefGoogle Scholar
  20. Kawanishi S, Oikawa S (2004) Mechanism of telomere shortening by oxidative stress. Ann N Y Acad Sci 1019:278–284. CrossRefGoogle Scholar
  21. Kilian A, Stiff C, Kleinhofs A (1995) Barley telomeres shorten during differentiation but grow in callus culture. Proc Natl Acad Sci USA 92:9555–9559. CrossRefGoogle Scholar
  22. Kilian A, Heller K, Kleinhofs A (1998) Development patterns of telomerase activity in barley and maize. Plant Mol Biol 37:621–628. CrossRefGoogle Scholar
  23. Kimura M, Stone RC, Hunt SC et al (2010) Measurement of telomere length by the Southern blot analysis of terminal restriction fragment lengths. Nat Protoc 5:1596–1607. CrossRefGoogle Scholar
  24. Lee H-T, Bose A, Lee C-Y et al (2017) Molecular mechanisms by which oxidative DNA damage promotes telomerase activity. Nucleic Acids Res. Google Scholar
  25. Lewis K, Wuttke DS (2012) Telomerase and telomere-associated proteins: structural insights into mechanism and evolution. Structure 20:28–39. CrossRefGoogle Scholar
  26. Liang J, Jiang C, Peng H et al (2015) Analysis of the age of Panax ginseng based on telomere length and telomerase activity. Sci Rep. Google Scholar
  27. Liu D, Qiao N, Song H et al (2007) Comparative analysis of telomeric restriction fragment lengths in different tissues of Ginkgo biloba trees of different age. J Plant Res 120:523–528. CrossRefGoogle Scholar
  28. McKnight TD, Riha K, Shippen DE (2002) Telomeres, telomerase, and stability of the plant genome. Plant Mol Biol 48:331–337. CrossRefGoogle Scholar
  29. Monja-Mio KM, Robert ML (2013) Direct somatic embryogenesis of Agave fourcroydes Lem. through thin cell layer culture. In Vitro Cell Dev Biol Plant. Google Scholar
  30. Monja-Mio KM, Robert ML (2016) Somatic embryogenesis in Agave: an overview. In: Loyola-Vargas VM, Ochoa-Alejo N (eds) Somatic embryogenesis: fundamental aspects and applications. Springer, Switzerland, pp 283–296CrossRefGoogle Scholar
  31. Monja-Mio KM, Pool FB, Herrera GH et al (2015) Development of the stomatal complex and leaf surface of Agave angustifolia Haw. ‘Bacanora’ plantlets during the in vitro to ex vitro transition process. Sci Hortic 189:32–40. CrossRefGoogle Scholar
  32. Moriguchi R, Kato K, Kanahama K et al (2007) Analysis of telomere lengths in apple and cherry trees. In: Acta horticulturae. International Society for Horticultural Science (ISHS), Leuven, pp 389–395Google Scholar
  33. Mu Y, Ren L, Hu X et al (2015) Season-specific changes in telomere length and telomerase activity in Chinese pine (Pinus tabulaeformis Carr.). Russ J Plant Physiol 62:487–493. CrossRefGoogle Scholar
  34. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  35. Narváez-Zapata J, Sánchez-Teyer LF (2009) Agaves as a raw material: recent technologies and applications. Recent Pat Biotechnol 3:185–191. CrossRefGoogle Scholar
  36. Nava-Cruza NY, Medina-Moralesa MA, Martineza JL et al (2015) Agave biotechnology: an overview. Crit Rev Biotechnol 35:546–559. CrossRefGoogle Scholar
  37. Pérez-Jiménez M, López-Pérez AJ, Otálora-Alcón G et al (2015) A regime of high CO2 concentration improves the acclimatization process and increases plant quality and survival. Plant Cell Tissue Organ Cult 121:547–557. CrossRefGoogle Scholar
  38. Plot V, Criscuolo F, Zahn S, Georges JY (2012) Telomeres, age and reproduction in a long-lived reptile. PLoS ONE 7:1–6. CrossRefGoogle Scholar
  39. Pospíšilová J, Tichá I, Kadleček P et al (1999) Acclimatization of micropropagated plants to ex vitro conditions. Biol Plant 42:481–497CrossRefGoogle Scholar
  40. Rescalvo-Morales A, Monja-Mio KM, Herrera-Herrera G et al (2016) Analysis of telomere length during the organogenesis induction of Agave fourcroydes Lem and Agave tequilana Weber. Plant Cell Tissue Organ Cult. Google Scholar
  41. Rice C, Skordalakes E (2016) Structure and function of the telomeric CST complex. Comput Struct Biotechnol J 14:161–167. CrossRefGoogle Scholar
  42. Richards EJ, Ausubel FM (1988) Isolation of a higher eukaryotic telomere from Arabidopsis thaliana. Cell 53:127–136. CrossRefGoogle Scholar
  43. Riha K, Fajkus J, Siroky J, Vyskot B (1998) Developmental control of telomere lengths and telomerase activity in plants. Plant Cell 10:1691–1698. CrossRefGoogle Scholar
  44. Robert ML, Herrera-Herrera JL, Castillo E et al (2006) An efficient method for the micropropagation of Agave species. In: Loyola-Vargas VM, Vázquez-Flota F (eds) Plant cell culture protocols, 2nd edn. Humana Press Inc., Totowa, pp 165–178Google Scholar
  45. Rubio MA, Davalos AR, Campisi J (2004) Telomere length mediates the effects of telomerase on the cellular response to genotoxic stress. Exp Cell Res 298:17–27. CrossRefGoogle Scholar
  46. Santamaría JM, Herrera JL, Robert ML (1995) Stomatal physiology of a micropropagated CAM plant; Agave tequilana (Weber). Plant Growth Regul 16:211–214. CrossRefGoogle Scholar
  47. Schrumpfová PP, Kuchar M, Palecek J, Fajkus J (2008) Mapping of interaction domains of putative telomere-binding proteins AtTRB1 and AtPOT1b from Arabidopsis thaliana. FEBS Lett 582:1400–1406. CrossRefGoogle Scholar
  48. Serra V, Grune T, Sitte N et al (2000) Telomere length as a marker of oxidative stress in primary human fibroblast cultures. Ann N Y Acad Sci 908:327–330CrossRefGoogle Scholar
  49. Shakirov EV, Salzberg SL, Alam M, Shippen DE (2008) Analysis of Carica papaya telomeres and telomere-associated proteins: insights into the evolution of telomere maintenance in Brassicales. Trop Plant Biol 1:202–215. CrossRefGoogle Scholar
  50. Song H, Liu D, Chen X et al (2010) Change of season-specific telomere lengths in Ginkgo biloba L. Mol Biol Rep 37:819–824. CrossRefGoogle Scholar
  51. Song H, Liu D, Li F, Lu H (2011) Season- and age-associated telomerase activity in Ginkgo biloba L. Mol Biol Rep 38:1799–1805. CrossRefGoogle Scholar
  52. Stout GJ, Blasco MA (2013) Telomere length and telomerase activity impact the UV sensitivity syndrome xeroderma pigmentosum C. Cancer Res 73:1844–1854. CrossRefGoogle Scholar
  53. Tichá I, Radochová B, Kadleček P (1999) Stomatal morphology during acclimatization of tobacco plantlets to ex vitro conditions. Biol Plant 42:469–474CrossRefGoogle Scholar
  54. Uziel O, Reshef H, Ravid A et al (2008) Oxidative stress causes telomere damage in Fanconi anaemia cells—a possible predisposition for malignant transformation. Br J Haematol 142:82–93. CrossRefGoogle Scholar
  55. Watson JM, Riha K (2011) Telomeres, aging, and plants: from weeds to Methuselah—a mini-review. Gerontology 57:129–136. CrossRefGoogle Scholar
  56. Zellinger B, Riha K (2007) Composition of plant telomeres. Biochim Biophys Acta 1769:399–409. CrossRefGoogle Scholar
  57. Zellinger B, Akimcheva S, Puizina J et al (2007) Ku suppresses formation of telomeric circles and alternative telomere lengthening in Arabidopsis. Mol Cell 27:163–169. CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Centro de Investigación Científica de YucatánUnidad de BiotecnologíaMéridaMexico

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