Skip to main content

Advertisement

SpringerLink
Log in

Controlling the barriers of cloning mature Picea abies (L.) H. Karst. via tissue culture and co-cultivation with Agrobacterium rhizogenes

  • Original Article
  • Published:
Trees Aims and scope Submit manuscript

Abstract

Key message

Providing an improved protocol for cloning of Picea abies (L.) H. Karst. mature trees via Agrobacterium rhizogenes.

Abstract

In reforestation programs, rejuvenation of superior genotypes is of great importance in helping juvenile rootstock production. In this regard, the use of plant tissue culture to produce axillary and/or adventitious shoots from explants of mature trees has been the objective of many researchers. There are a few reports about the successful in vitro rooting of the regenerated shoots from explants of mature trees. There is a very low rate of natural rooting in conifers, between 1 and 5%. Furthermore, they are still hard to root even when auxins are applied. In this study, in vitro propagation of Norway spruce explants and its rooting improvement by A. rhizogenes with the aim of receiving an efficient clonal propagation method was investigated. The effects of BA (0.5, 1, 1.5, and 2 μM) and TDZ (0.0025, 0.025, 0.25, and 2.5 μM) were examined on axillary shoot development from shoot-tip explants of the Norway spruce. The effects of three strains of Agrobacterium rhizogenes (K599, AR15834 and GMI9534) were also examined on enhancement of the root induction. According to the results, after the treatment of 1.5 μM BA, 87.5% of shoot-tip explants produced axillary shoots (the highest percentage among treatments), and also the maximum mean number of regenerated shoots per explant (6.5) was achieved in this treatment. Furthermore, the induction of rooting was performed using IBA (7.5 µM) and IAA (0.5 or 1 μM) in the WPM medium along with keeping the explants in the darkness at 21 °C, which yielded 37.5% and 12.5% rooting, respectively. Moreover, the application of coumarin along with IAA caused an increase of 22% in rooting, compared to the use of IAA alone. Two strains, K599 and AR15834, succeeded in improving the percentage of rooting, with the highest being 54.2% as a result of using the K599 strain alongside 7.5 μM IBA. The proposed treatment can be used efficiently for the propagation of Norway spruce in landscaping and/or reforestation programs.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

BA:

Benzyl adenine

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

MES:

2-(N-morpholino) ethanesulfonic acid

MS:

Murashige and Skoog (1962) medium

NAA:

Naphthalene acetic acid

TDZ:

Thidiazuron

WPM:

Woody plant medium (Lloyd and McCown 1981)

YEP:

Yeast extract peptone medium

Z:

Zeatin

References

  • Alonso P, Moncaleán P, Fernández B, Rodríguez A, Centeno ML, Ordás RJ (2006) An improved micropropagation protocol for stone pine (Pinus pinea L.). Ann For Sci 63:879–885

    CAS  Google Scholar 

  • Bonga J (1991) In vitro propagation of conifers: fidelity of the clonal offspring. Woody Plant Biotechnol. Springer, Berlin, pp 13–21

    Google Scholar 

  • Bonga J (2014) A comparative evaluation of the application of somatic embryogenesis, rooting of cuttings, and organogenesis of conifers. Can J For Res 45:379–383

    Google Scholar 

  • Bonga J, Pond S (1991) Adventitious shoot formation in cultures of 30-year-old Larix decidua, L. leptolepis, L. eurolepis, and L. laricina trees. Plant Cell Tiss Org 26:45–51

    Google Scholar 

  • Chalupa V (1989) Micropropagation of Larix decidua Mill. and Pinus sylvestris L. Biol Plant 31:400–407

    Google Scholar 

  • Cortizo M, de Diego N, Moncaleán P, Ordás RJ (2009) Micropropagation of adult stone pine (Pinus pinea L.). Trees 23:835–842

    Google Scholar 

  • De Diego N, Montalbán I, Fernández de Larrinoa E, Moncaleán P (2008) In vitro regeneration of Pinus pinaster adult trees. Can J For Res 38:2607–2615

    Google Scholar 

  • Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15

    Google Scholar 

  • Drake P, John A, Power J, Davey M (1997) Cytokinin pulse-mediated shoot organogenesis from cotyledons of Sitka spruce [Picea sitchensis (Bong.) Carr.] and high frequency in vitro rooting of shoots. Plant Cell Tiss Org 50:147–151

    CAS  Google Scholar 

  • Druart P, Kevers C, Boxus P, Gaspar T (1982) vitro promotion of root formation by apple shoots through darkness effect on endogenous phenols and peroxidases. Z Pflanzenphysiol 108:429–436

    CAS  Google Scholar 

  • Ellenberg H (1988) Vegetation ecology of central Europe. Cambridge University Press, Cambridge

    Google Scholar 

  • Escobar MA, Dandekar AM (2003) Agrobacterium tumefaciens as an agent of disease. Trends Plant Sci 8:380–386

    CAS  PubMed  Google Scholar 

  • Faye M, Ourry A, Saidali-Savi C, Dargent R, Boucaud J, David A (1989) Effects of glutamine and K-glutamate on assimilation of [15 N]-nitrate during auxin treatment for root formation in vitro (Pinus pinaster). Physiol Plant 76:277–282

    CAS  Google Scholar 

  • Gahan P (2007) Totipotency and the cell cycle. Protocols for micropropagation of woody trees and fruits. Springer, Berlin, pp 3–14

    Google Scholar 

  • Gomez M, Segura J (1994) Factors controlling adventitious bud induction and plant regeneration in mature Juniperus oxycedrus leaves cultured in vitro. In Vitro Cell Dev Biol-Plant 30:210–218

    Google Scholar 

  • Gould JH, Zhou Y, Padmanabhan V, Magallanes-Cedeno ME, Newton RJ (2002) Transformation and regeneration of loblolly pine: shoot apex inoculation with Agrobacterium. Mol Breed 10:131–141

    CAS  Google Scholar 

  • Gupta PK, Durzan DJ (1985) Shoot multiplication from mature trees of Douglas-fir (Pseudotsuga menziesii) and sugar pine (Pinus lambertiana). Plant Cell Rep 4:177–179

    CAS  PubMed  Google Scholar 

  • Hansen G (1994) Constitutive expression of the virulence genes improves the efficiency of plant transformation by Agrobacterium. PNAS 91:7603–7607

    CAS  PubMed  Google Scholar 

  • Hartmann HT, Kester DE, Davies J (2002) Plant propagation: principles and practices. Prentice-Hall, New Jersey

    Google Scholar 

  • Harvengt L, Trontin J-F, Reymond I, Canlet F, Paques M (2001) Molecular evidence of true-to-type propagation of a 3-year-old Norway spruce through somatic embryogenesis. Planta 213:828–832

    CAS  PubMed  Google Scholar 

  • Hatta M, Beyl CA, Garton S, Diner A (1996) Induction of roots on jujube softwood cuttings using Agrobacterium rhizogenes. J Hortic Sci 71:881–886

    Google Scholar 

  • James D, Thurbon IJ (1979) Rapid in vitro rooting of the apple rootstock M. 9 J Hortic Sci 54:309–311

    Google Scholar 

  • Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. The EMBO J 6:3901–3907

    CAS  PubMed  Google Scholar 

  • Källman T (2009) Adaptive evolution and demographic history of Norway spruce (Picea abies). Acta Universitatis Upsaliensis

  • Khoshhal Sarmast M, Salehi H, Khosh-Khui M (2012) In vitro rooting of Araucaria excelsa R. Br. var. glauca using Agrobacterium rhizogenes. J Cent Eur Agric 13:1

    Google Scholar 

  • Klimaszewska K, Hargreaves C, Lelu-Walter M-A, Trontin J-F (2016) Advances in conifer somatic embryogenesis since year 2000. In vitro embryogenesis in higher plants. Springer, Berlin, pp 131–166

    Google Scholar 

  • Kling G, Meyer Jr M (1983) Effects of phenolic compounds and indoleacetic acid on adventitious root initiation in cuttings of Phaseolus aureus, Acer saccharinum, and Acer griseum [Mung bean bioassay, silver maple, paperbark maple, propagation, woody plants] HortScience

  • Kupidlowska E, Kowalec M, Sulkowski G, Zobel A (1994) The effect of coumarins on root elongation and ultrastructure of meristematic cell protoplast. Ann Bot 73:525–530

    CAS  Google Scholar 

  • Li M, Leung DW (2003) Root induction in radiata pine using Agrobacterium rhizogenes. Electron J Biotechnol 6:254–261

    Google Scholar 

  • Liu C, Zhu J, Liu Z, Li L, Pan R, Jin L (2002) Exogenous auxin effects on growth and phenotype of normal and hairy roots of Pueraria lobata (Willd.). Ohwi Plant Growth Regul 38:37–43

    CAS  Google Scholar 

  • Lloyd G, McCown B (1981) Commercially-feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot-tip culture. Proc Intl Plant Prop Soc 30:421–427

    Google Scholar 

  • López-Escamilla AL, Olguín-Santos LP, Márquez J, Chávez VM, Bye R (2000) Adventitious bud formation from mature embryos of Picea chihuahuana Martínez, an endangered Mexican spruce tree. Ann Bot 86:921–927

    Google Scholar 

  • Magnussen D, Clapham D, Grönroos R, von Arnold S (1994) Induction of hairy and normal roots on Picea abies, Pinus sylvestris and Pinus cortorta by Agrobacterium rhizogenes. cand J Forest Res 9:46–51

    Google Scholar 

  • McAfee B, White E, Pelcher L, Lapp M (1993) Root induction in pine (Pinus) and larch (Larix) spp. using Agrobacterium rhizogenes. Plant Cell Tiss Org 34:53–62

    Google Scholar 

  • Mihaljević S, Stipković S, Jelaska S (1996) Increase of root induction in Pinus nigra explants using agrobacteria. Plant Cell Rep 15:610–614

    PubMed  Google Scholar 

  • Mihaljević S, Katavić V, Jelaska S (1999) Root formation in micropropagated shoots of Sequoia sempervirens using Agrobacterium. Plant Sci 141:73–80

    Google Scholar 

  • Mohammed GH, Vidaver WE (1988) Root production and plantlet development in tissue-cultured conifers Plant Cell. Tiss Org Cult 14:137–160

    Google Scholar 

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

    CAS  Google Scholar 

  • Olhoft PM et al (2007) A novel Agrobacterium rhizogenes-mediated transformation method of soybean [Glycine max (L.) Merrill] using primary-node explants from seedlings. In Vitro Cell Dev Biol-Plant 43:536–549

    CAS  Google Scholar 

  • Ordás R, Alonso P, Cuesta C, Cortizo M, Rodríguez A, Fernández B (2007) Micropropagation of Pinus pinea L. Protocols for micropropagation of woody trees and fruits. Springer, Berlin, pp 33–39

    Google Scholar 

  • Park Y-S, Klimaszewska K (2003) Achievements and challenges in conifer somatic embryogenesis for clonal forestry. In: Paper presented at the XII world forestry congress, Quebec City, Quebec, Canada

  • Prehn D, Serrano C, Mercado A, Stange C, Barrales L, Arce-Johnson P (2003) Regeneration of whole plants from apical meristems of Pinus radiata. Plant Cell Tiss Org Cult 73:91–94

    CAS  Google Scholar 

  • Pulido CM, Harry IS, Thorpe TA (1990) In vitro regeneration of plantlets of Canary Island pine (Pinus canariensis). Can J For Res 20:1200–1211

    Google Scholar 

  • Ragonezi C, Klimaszewska K, Castro MR, Lima M, de Oliveira P, Zavattieri MA (2010) Adventitious rooting of conifers: influence of physical and chemical factors. Trees 24:975–992

    CAS  Google Scholar 

  • Renau-Morata B, Nebauer SG, Sales E, Allainguillaume J, Caligari P, Segura J (2005) Genetic diversity and structure of natural and managed populations of Cedrus atlantica (Pinaceae) assessed using random amplified polymorphic DNA. Am J Bot 92:875–884

    CAS  PubMed  Google Scholar 

  • Rumary C, Thorpe TA (1984) Plantlet formation in black and white spruce. I. In vitro techniques. Can J For Res 14:10–16

    Google Scholar 

  • Saborio F, Dvorak WS, Donahue JK, Thorpe TA (1997) vitro regeneration of plantlets from mature embryos of Pinus ayacahuite. Tree Physiol 17:787–796

    CAS  PubMed  Google Scholar 

  • Sarmast MK (2016) Genetic transformation and somaclonal variation in conifers Plant. Biotechnol Rep 10:309–325

    Google Scholar 

  • Sarmast MK (2018) In vitro propagation of conifers using mature shoots. J For Res 29:565–574

    Google Scholar 

  • Sarmast MK, Salehi H, Khosh-Khui M (2012) Micropropagation of Araucaria excelsa R. Br. var. glauca Carrière from orthotropic stem explants. Physiol Mol Biol Plant 18:265–271

    Google Scholar 

  • Sen S, Magallanes-Cedeno M, Kamps R, McKinley C, Newton R (1994) In vitro micropropagation of Afghan pine. Can J For Res 24:1248–1252

    Google Scholar 

  • Sheikholeslam SN, Weeks DP (1987) Acetosyringone promotes high efficiency transformation of Arabidopsis thaliana explants by Agrobacterium tumefaciens. Plant Mol Biol 8:291–298

    CAS  PubMed  Google Scholar 

  • Silen RR (1978) Genetics of Douglas-fir. USDA For Serv 35:22

    Google Scholar 

  • Slightom JL, Durand-Tardif M, Jouanin L, Tepfer D (1986) Nucleotide sequence analysis of TL-DNA of Agrobacterium rhizogenes agropine type plasmid. Identification of open reading frames. J Biol Chem 261:108–121

    CAS  PubMed  Google Scholar 

  • Smith D et al (1986) Forest and nut trees. Trees I. Springer, Berlin, pp 274–450

    Google Scholar 

  • Stojicić D, Budimir S, Ćulafić L (1999) Micropropagation of Pinus heldreichii. Plant Cell Tiss Org Cult 59:147–150

    Google Scholar 

  • Sunilkumar G, Vijayachandra K, Veluthambi K (1999) Preincubation of cut tobacco leaf explants promotes Agrobacterium-mediated transformation by increasing vir gene induction. Plant Sci 141:51–58

    CAS  Google Scholar 

  • Tang W, Newton R (2007) Micropropagation via organogenesis in slash pine. Protocols for micropropagation of woody trees and fruits. Springer, Berlin, pp 15–22

    Google Scholar 

  • Varis S, Klimaszewska K, Aronen T (2018) Somatic embryogenesis and plant regeneration from primordial shoot explants of Picea abies (L.) H. Karst. somatic trees. Front Plant Sci 9:1551

    PubMed  PubMed Central  Google Scholar 

  • Veena V, Taylor CG (2007) Agrobacterium rhizogenes: recent developments and promising applications. In Vitro Cell Dev Biol-Plant 43:383–403

    CAS  Google Scholar 

  • Veluthambi K, Krishnan M, Gould JH, Smith RH, Gelvin SB (1989) Opines stimulate induction of the vir genes of the Agrobacterium tumefaciens Ti plasmid. J Bacteriol 171:3696–3703

    CAS  PubMed  PubMed Central  Google Scholar 

  • Villalobos-Amador E, Rodríguez-Hernández G, Pérez-Molphe-Balch E (2002) Organogenesis and Agrobacterium rhizogenes-induced rooting in Pinus maximartinezii Rzedowsky and P. pinceana Gordon. Plant Cell Rep 20:779–785

    CAS  Google Scholar 

  • Von Arnold S (1982) Factors influencing formation, development and rooting of adventitious shoots from embryos of Picea abies (L.) Karst. Plant Sci Lett 27:275–287

    Google Scholar 

  • Von Arnold S, Eriksson T (1984) Effect of agar concentration on growth and anatomy of adventitious shoots of Picea abies (L.) Karst. Plant Cell Tiss Org Cult 3:257–264

    Google Scholar 

  • Zarei M, Salehi H, Jowkar A (2017) Effects of temperature and season on in vitro establishment and shoot multiplication of Picea abies (L.) H. Karst Int J Hortic Sci Technol 4:51–56

    CAS  Google Scholar 

  • Zavattieri A, Lima M, Sobral V, Oliveira P, Costa A (2007) Effects of carbon source, carbon concentration and culture conditions on in vitro rooting of Pinus pinea L. microshoots. In: III International symposium on acclimatization and establishment of micropropagated plants 812. pp 173–180

  • Zavattieri MA, Ragonezi C, Klimaszewska K (2016) Adventitious rooting of conifers: influence of biological factors. Trees 30:1021–1032

    CAS  Google Scholar 

  • Žel J, Gogala N, Camloh M (1988) Micropropagation of Pinus sylvestris. Plant Cell Tiss Org Cult 14:169–175

    Google Scholar 

  • Zhihua L, Li L, Yuxi L, Lizhi Q (1991) Micropropagation in tissue culture of Dahurian Larch. J For Res 2:1–8

    Google Scholar 

Download references

Funding

This study was funded by Shiraz University.

Author information

Authors and Affiliations

  1. Department of Horticultural Science, School of Agriculture, Shiraz University, Shiraz, Iran

    Mehrnaz Zarei, Hassan Salehi & Abolfazl Jowkar

Authors
  1. Mehrnaz Zarei
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hassan Salehi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Abolfazl Jowkar
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hassan Salehi.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Communicated by K. Klimaszewska.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zarei, M., Salehi, H. & Jowkar, A. Controlling the barriers of cloning mature Picea abies (L.) H. Karst. via tissue culture and co-cultivation with Agrobacterium rhizogenes. Trees 34, 637–647 (2020). https://doi.org/10.1007/s00468-019-01945-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00468-019-01945-z

Keywords

Search

Navigation