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Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 131, Issue 3, pp 499–512 | Cite as

Micropropagation of mature Quercus ilex L. trees by axillary budding

  • M. T. Martínez
  • E. Corredoira
  • A. M. Vieitez
  • M. J. Cernadas
  • R. Montenegro
  • A. Ballester
  • F. J. Vieitez
  • M. C. San JoséEmail author
Original Article

Abstract

This paper reports the successful micropropagation of mature Quercus ilex trees known as reluctant to in vitro propagation. Crown branch segments collected from 30 to 100 year-old trees were forced in order to promote the production of sprouting shoots that were used as a source of explants for initiating the cultures. Sterilization was critical and required low-level disinfestation protocols. Six out of the eight mature genotypes attempted were successfully inoculated and then maintained in culture with varying responses. Shoot proliferation of holm oak was influenced by BA concentration, with improved multiplication and shoot appearance when the BA concentration was sequentially reduced over the culture period. Micropropagation by axillary budding was achieved by culturing shoots on a sequence of cytokinin-enriched Lloyd and McCown (WPM) media alternating 2 week-long subcultures on 0.44 µM benzyadenine (BA) first, followed by 0.22 µM BA, then 0.044 µM BA plus 0.46 µM zeatin. Sucrose concentration and agar brand affected shoot proliferation, and the best results were obtained on WPM medium supplemented with 8 g L−1 Sigma agar (A-1296; Sigma-Aldrich) and 30 g L−1 sucrose. Addition of 20 µM silver thiosulphate had a significant positive effect on the appearance and development of shoots with a higher number of shoots being healthy and showing reduced shoot tip necrosis and early senescence of leaves. The 18.8% of the microshoots obtained for one clone could be rooted within 15 days on a half-strength Murashige and Skoog medium containing 14.8 µM or 24.6 µM indole-3-butyric acid and 0.54 µM α-naphthalene acetic acid.

Keywords

Holm oak Ethylene inhibitors Gelling agents Rejuvenation Rooting Shoot culture initiation 

Notes

Acknowledgements

We thank Dr M Toribio for kindly providing the plant material used in this study. We also thank JC Suárez San Martín for technical support.

Funding

This research has been partly funded by Ministerio de Economía y Competitividad, MINECO, Spain (AGL 2013-47400-C4-3R and AGL2016-76143-C4-4-R).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11240_2017_1300_MOESM1_ESM.docx (17 kb)
Supplementary material 1 (DOCX 17 KB)
11240_2017_1300_MOESM2_ESM.docx (19 kb)
Supplementary material 2 (DOCX 18 KB)
11240_2017_1300_MOESM3_ESM.docx (16 kb)
Supplementary material 3 (DOCX 15 KB)

References

  1. Alaska-Kennedy Y, Yoshida H, Takahata Y (2005) Efficient plant regeneration from leaves of rapeseed (Brassica napus L.): the influence of AgNO3 and genotype. Plant Cell Rep 24:649–654CrossRefGoogle Scholar
  2. Bairu MW, Stirk WA, Dolezal K, Van Staden JV (2007) Optimizing the micropropagation protocol for the endangered Aloe polyphylla: can meta-polin and its derivates serve as replacement for benzyladenine and zeatin? Plant Cell Tissue Organ Cult 90:15–23CrossRefGoogle Scholar
  3. Ballester A, Vidal N, Vieitez AM (2009) Developmental stages during in vitro rooting of hardwood trees from material with juvenile and mature characteristics. In: Niemi K, Scagel C (eds) Adventitious root formation of forest trees and horticultural plants-from genes to applications. Research Signpost, Thiruvananthapuram, pp 277–296Google Scholar
  4. Ballester A, Corredoira E, Vieitez AM (2016) Limitations of somatic embryogenesis in hardwood trees. In: Park Y-S, Bonga JM, Moon H-K (eds) Vegetative propagation of forest trees. National Institute of Forest Science (Nifos), Seoul, pp 56–74Google Scholar
  5. Benson E (2000) In vitro plant recalcitrance: an introduction. In Vitro Cell Dev Biol Plant 36:141–148CrossRefGoogle Scholar
  6. Blasco M, Barra A, Brisa C, Corredoira E, Segura J, Toribio M, Arrillaga I (2013) Somatic embryogenesis in holm oak male catkins. Plant Growth Regul 71:261–270CrossRefGoogle Scholar
  7. Bon MC, Riccardi F, Monteuuis O (1994) Influence of phase change within a 90-year-old Sequoia sempervirens on its in vitro organogenic capacity and protein patterns. Trees 8:283–287CrossRefGoogle Scholar
  8. Bonga JM (2017) Can explant choice help resolve recalcitrance problems in in vitro propagation, a problem still acute especially for adult conifers? Trees 31:781–789CrossRefGoogle Scholar
  9. Bonga JM, von Aderkas P (1992) In vitro culture of trees. Kluwer Academic Publishers, DordrechtCrossRefGoogle Scholar
  10. Bonga JM, Klimaszewska KK, von Aderkas P (2010) Recalcitrance in clonal propagation, in particular of conifers. Plant Cell Tiss Organ Cult 100:241–254CrossRefGoogle Scholar
  11. Burgos L, Alburquerque N (2003) Ethylene inhibitors and low kanamycin concentrations improve adventitious regeneration from apricot leaves. Plant Cell Rep 31:1167–1174CrossRefGoogle Scholar
  12. Corcobado T, Cubera E, Moreno G, Solla A (2013) Quercus ilex forests are influenced by annual variations in water table, soil water deficit and fine root loss caused by Phythophtora cinnamomi. Agric For Meteorol 169:92–99CrossRefGoogle Scholar
  13. Corredoira E, Ballester A, Vieitez AM (2008) Thidiazuron-induced high frequency plant regeneration from leaf explants of Paulownia tomentosa mature trees. Plant Cell Tiss Organ Cult 95:197–208CrossRefGoogle Scholar
  14. Corredoira E, San-José MC, Vieitez AM (2012) Induction of somatic embryogenesis from different explants of shoot cultures derived from young Quercus alba trees. Trees 26:881–891CrossRefGoogle Scholar
  15. Corredoira E, Ballester A, Ibarra M, Vieitez AM (2015) Induction of somatic embryogenesis in leaf and shoot apex explants of shoot cultures derived from adult Eucalyptus globulus and Eucalyptus saligna × E. maidenii trees. Tree Physiol 35:663–677CrossRefGoogle Scholar
  16. Correia S, Lopes ML, Canhoto JM (2011) Somatic embryogenesis induction system for cloning an adult Cyphomandra betacea (Cav.) Sendt. (tamarillo). Trees 25:1009–1020CrossRefGoogle Scholar
  17. Council of Europe, UNEP & ECNC (1996) The Pan-European biological and landscape diversity strategy: a vision for Europe’s natural heritage. European Centre for Nature Conservation, TilburgGoogle Scholar
  18. Dal Vesco LL, Guerra MP (2001) The effectiveness of nitrogen sources in Feijoa somatic embryogenesis. Plant Cell Tiss Org Cult 64:19–25CrossRefGoogle Scholar
  19. Ďurkovič J, Mišalová A (2008) Micropropagation of temperate hardwoods: an overview. Funct Plant Sci 2:1–19Google Scholar
  20. El Kbiach ML, Lamarti A, Abdali A, Badoc A (2004) Micropropagation du Chêne-liège (Quercus suber L.) par bourgeonnement axillaire. Acta Bot Gallica 151:415–427CrossRefGoogle Scholar
  21. Gaspar T, Kevers C, Penel C, Greppin H, Reid DM, Thorpe TA (1996) Plant hormones and plant growth regulators in plant tissue culture. In Vitro Cell Dev Biol Plant 32:272–289CrossRefGoogle Scholar
  22. George EF, Debergh PC (2008) Micropropagation: uses and methods. In: George EF, Hall MA, de Klerk GJ (eds) Plant propagation by tissue culture, 3rd edn. Springer, Dordrecht, pp 29–64Google Scholar
  23. Gomes F, Canhoto JM (2009) Micropropagation of strawberry tree (Arbustus nedo L.) from adult plants. In Vitro Cell Dev Biol Plant 45:72–82CrossRefGoogle Scholar
  24. Gomes F, Simỡes M, Lopes ML, Canhoto JM (2010) Effect of plant growth regulators and genotype on the micropropagation of Arbutus unedo L. (strawberry tree). New Biotechnol 27:882–892CrossRefGoogle Scholar
  25. Gresshoff PM, Doy CH (1972) Development and differentiation of haploid Lycopersicon esculentum. Planta 107:161–170CrossRefPubMedGoogle Scholar
  26. Howell SH, Lall S, Che P (2003) Cytokinins and shoot development. Trends Plant Sci 8:453–459CrossRefPubMedGoogle Scholar
  27. Huntsinger L, Campos P, Starrs PF, Oviedo JL, Díaz M, Standiford RB, Montero G (2013) Working landscapes of the spanish dehesa and the California oak woodlands: an introduction. In: Campos P, Huntsinger L, Oviedo JL et al (eds) Mediterranean oak woodland working landscapes. Springer, Dordrecht, pp 3–23CrossRefGoogle Scholar
  28. L´Helgoual’ch M, Espagnac H (1987) First observations on the adventitious rhizogenic capacity of holm oak (Quercus ilex L.). Ann Sci For 44:325–334CrossRefGoogle Scholar
  29. Liñan J, Cantos M, Troncoso J, García JL, Fernández A, Troncoso A (2011) Some propagation methods for cloning holm oak (Quercus ilex L.) plants. Cent Eur J Biol 6:359–364Google Scholar
  30. Liu X, Pijut P (2008) Plant regeneration from in vitro leaves of mature black cherry (Prunus serotina). Plant Cell Tiss Organ Cult 94:113–123CrossRefGoogle Scholar
  31. Lloyd G, McCown B (1980) Commercially-feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot-tip culture. Comb Proc Int Plant Prop Soc 30:421–427Google Scholar
  32. Martínez MT, Vidal N, Ballester A, Vieitez AM (2012) Improved organogenic capacity of shot cultures from mature pedunculate oak trees through somatic embryogenesis as rejuvenation technique. Trees 26:321–330CrossRefGoogle Scholar
  33. Martínez MT, Vieitez AM, Corredoira E (2015) Improved secondary embryo production in Quercus alba and Quercus rubra by activated charcoal, silver thiosulphate and sucrose: influence of embryogenic explant used for subculture. Plant Cell Tiss Organ Cult 212:531–546CrossRefGoogle Scholar
  34. Martínez MT, Vieitez AM, Corredoira E, Cernadas MJ, Montenegro R, Ballester A, Vieitez FJ, San José MC (2017) Vegetative propagation of adult Quercus ilex L. (holm oak) trees. I. Micropropagation by axillary budding. Plant Cell Tiss Organ Cult (in press)Google Scholar
  35. Mauri PV, Manzanera JA (2005) Protocol of somatic embryogenesis: holm oak (Quercus ilex L.). In: Jain SM, Gupta PK (eds) Protocol for somatic embryogenesis in woody plants. Springer, Dordrecht, pp 469–482CrossRefGoogle Scholar
  36. McCown BH (2000) Recalcitrance of woody and herbaceous plants: dealing with genetic predeterminism. In vitro Cell Dev Biol Plant 36:149–154CrossRefGoogle Scholar
  37. Mesías FJ, Pulido F, Gaspar P, Escribano M, Pulido AF (2010) Management of Spanish rangelands (Dehesas): an example of sustainable use. In: Veress B, Szigethy J (eds) Horizons in earth science research, Vol 1. Nova Science Publishers, New York, pp 163–186Google Scholar
  38. Millán-Orozco L, Corredoira E, San José MC (2011) In vitro rhizogenesis: histoanatomy of Cedrela odorata (Meliaceae) microcuttings. Rev Biol Trop 59:447–453PubMedGoogle Scholar
  39. Monteuuis O (2016) Micropropagation and production of forest trees. In: Park Y-S, Bonga JM, Moon H-K (eds) Vegetative propagation of forest trees. National Institute of Forest Science (NIFOS), Seoul, pp 32–55Google Scholar
  40. Monteuuis O, Doulbeau S, Verdeil JL (2008) DNA methylation in different origin clonal offspring from mature Sequoiadendron giganteum genotype. Trees 22:779–784CrossRefGoogle Scholar
  41. Moreno G, Pulido F (2009) The functioning, management, and persistence of dehesas. In: Rigueiro-Rodríguez A, McAdam J, Mosquera-Losada MR (eds) Agroforestry in Europe. Advances in agroforestry. Springer, Dordrecht, pp 127–160Google Scholar
  42. Moshkov IE, Novikova GV, Hall MA, George EF (2008) Plant growth regulators III: ethylene. In: George EF, Hall MA, de Klerk GJ (eds) Plant propagation by tissue culture, Vol 1, 3rd edn. Springer, Dordrecht, pp 239–248Google Scholar
  43. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497CrossRefGoogle Scholar
  44. Ostrolucká MG, Gajdosová A, Libiaková G (2007) Protocol for micropropagation of Quercus spp. In: Jain SM, Häggman H (eds) Protocols for micropropagation of woody trees and fruits. Springer, The Netherlands, pp 85–91CrossRefGoogle Scholar
  45. Park JS, Naing AH, Kim CK (2016) Effects of ethylene on shoot initiation, leaf yellowing, and shoot tip necrosis in rose. Plant Cell Tissue Organ Cult 127:425–431CrossRefGoogle Scholar
  46. Pereira-Netto AB, Petkowic CLO, Cruz-Silva TA, Gazzoni MT, Mello AFP, Silveira JLM (2007) Differential performance of marabakaido apple rootstock shoots grown in culture media containing different agar brands: dynamic rheological analysis. In vitro Cell Dev Biol Plant 43:356–363CrossRefGoogle Scholar
  47. Plieninger T, Pulido FJ, Schaich H (2004) Effects of land-use and landscape structure on holm oak recruitment and regeneration at farm level in Quercus ilex L. dehesas. J Arid Environ 57:345–364CrossRefGoogle Scholar
  48. Poothong S, Reed BM (2014) Modeling the effects of mineral nutrition for improving growth and development of micropropagated red raspberries. Sci Hort 165:132–141CrossRefGoogle Scholar
  49. Preece J (2008) Stock plant physiological factors affecting growth and morphogenesis. In: George EF, Hall MA, de Klerk GJ (eds) Plant propagation by tissue culture, 3rd edn. Springer, Dordrecht, pp 403–422Google Scholar
  50. Qin YH, Zhang SL, Zhang LX, Zhu DY, Syed A (2005) Response to in vitro strawberry to silver nitrate (AgNO3). HortScience 40:747–751Google Scholar
  51. Ramage CM, Williams RR (2002) Mineral nutrition and plant morphogenesis. In vitro Cell Dev Biol Plant 38:116–124CrossRefGoogle Scholar
  52. Reis LB, PaivaNeto VB, Toledo Picoli EA, Costa NGC, Rêgo MM, Carvalho CR, Finger FL, Otoni WC (2003) Axillary bud development of passion fruit as affected by ethylene precursor and inhibitors. In Vitro Cell Dev Biol Plant 39:618–622CrossRefGoogle Scholar
  53. Ruíz de la Torre J (2006) Flora mayor. Organismo Autónomo de Parques Nacionales. Dirección General para la Biodiversidad, MadridGoogle Scholar
  54. San José MC, Vieitez AM, Ballester A (1990) Clonal propagation of juvenile and adult trees of sessile oak by tissue culture. Silvae Genet 39:50–55Google Scholar
  55. San José MC, Corredoira E, Martínez MT, Vidal N, Valladares S, Mallón R, Vieitez AM (2010) Shoot apex explants for induction of somatic embryogenesis in mature Quercus robur L. trees. Plant Cell Rep 29:661–67CrossRefPubMedGoogle Scholar
  56. San José MC, Janeiro LV, Corredoira E (2013) Micropropagation of threatened black alder. Silva Fenn 47:1–12CrossRefGoogle Scholar
  57. Sánchez MC, Vieitez AM (1991) In vitro morphogenetic competence of basal sprouts and crown branches of mature chestnut. Tree Physiol 8:59–70CrossRefPubMedGoogle Scholar
  58. Sánchez ME, Caetano P, Ferraz J, Trapero A (2002) Phytophthora diseases of Quercus ilex in south-western Spain. For Pathol 32:5–18CrossRefGoogle Scholar
  59. Schenk RU, Hildebrand AC (1972) Medium and techniques for induction of growth of monocotyledonous and dicotyledonous plant cell culture. Can J Bot 50:199–204CrossRefGoogle Scholar
  60. Scholten HJ, Pierik RLM (1998) Agar as a gelling: differential biological effects in vitro. Sci Hort 77:1009–1116Google Scholar
  61. SPSS for Windows (2010) Guía Breve de IBM SPSS Statistics 19.SPSS Inc., IBM Company, Chicago, IL, p. 171Google Scholar
  62. Steinitz B, Barr N, Tabib Y, Vaknin Y, Bernstein N (2010) Control of in vitro rooting and plant development in Corymbia maculate by silver nitrate, silver thiosulphate and thiosulphate ion. Plant Cell Rep 29:1315–1323CrossRefPubMedGoogle Scholar
  63. Thorpe T, Stasolla C, Yeung EC, de Klerk G-J, Roberts A, George EF (2008) The components of plant tissue culture media II: organic additions, osmotic and pH effects, and support systems. In: George EF, Hall MA, de Klerk GJ (eds) Plant propagation by tissue culture, 3rd edn. Springer, Dordrecht, pp 115–174Google Scholar
  64. Van Staden J, Zazimalova E, George EF (2008) Plant growth regulators II: Cytokinins, their analogues and antagonists. In: George EF, Hall MA, de Klerk GJ (eds) Plant propagation by tissue culture, 3rd edn. Springer, Dordrecht, pp 205–226Google Scholar
  65. Vengadesan G, Pijut PM (2009) Somatic embryogenesis and plant regeneration of northern red oak (Quercus rubra L.). Plant Cell Tissue Organ Cult 97:141–149CrossRefGoogle Scholar
  66. Vidal N (2002) Relación entre el contenido endógeno de reguladores de crecimiento, el estado ontogenético y la capacidad de enraizamiento de brotes de castaño y roble cultivados in vitro. Tesis Doctoral. Universidad de Santiago de Compostela, SpainGoogle Scholar
  67. Vieitez AM, Ferro EM, Ballester A (1993a) Micropropagation of Fagus sylvatica L. In Vitro Cell Dev Biol Plant 29:183–188CrossRefGoogle Scholar
  68. Vieitez AM, Pintos F, San José MC, Ballester A (1993b) In vitro shoot proliferation determined by explant orientation of juvenile and mature Quercus rubra L. Tree Physiol 12:107–117CrossRefPubMedGoogle Scholar
  69. Vieitez AM, Sánchez MC, Amomarco JB, Ballester A (1994) Forced flushing of branch segments as a method for obtaining reactive explants of mature Quercus robur trees for micropropagation. Plant Cell Tissue Organ Cult 37:287–295Google Scholar
  70. Vieitez AM, San José MC, Sánchez MC, Ballester A (2003) Micropropagation of Fagus spp. In: Jain SM, Ishii K (eds) Micropropagation of woody trees and fruits. Kluwer Academic Publishers, Netherlands, pp 181–215CrossRefGoogle Scholar
  71. Vieitez AM, Sánchez MC, García-Nimo ML, Ballester A (2007) Protocol for micropropagation of Castanea sativa Mill. In: Jain SM, Häggman H (eds) Protocols for micropropagation of woody trees and fruits. Springer, Heidelberg, pp 299–312CrossRefGoogle Scholar
  72. Vieitez AM, Corredoira E, Ballester A, Muñoz F, Durán J, Ibarra M (2009) In vitro regeneration of important North American oak species Quercus alba, Quercus bicolor and Quercus rubra. PlantCellTissueOrganCult 98:135–145Google Scholar
  73. Vieitez AM, Corredoira E, Martínez MT, San José MC, Sánchez C, Valladares S, Vidal N, Ballester A (2012) Application of biotechnological tools to Quercus improvement. Eur J Forest Res 131:519–539CrossRefGoogle Scholar
  74. Wendling I, Trueman SJ, Xavier A (2014) Maturation and related aspects in clonal forestry-part II: reinvigoration, rejuvenation and juvenility maintenance. New For 45:473–486CrossRefGoogle Scholar

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© Springer Science+Business Media B.V. 2017

Authors and Affiliations

  1. 1.Group of Biotechnology and Forestry Improvement, Department of Plant PhysiologyInstituto de Investigaciones Agrobiológicas de Galicia (IIAG-CSIC)Santiago de CompostelaSpain

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