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In vitro propagation of conifers using mature shoots

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Abstract

Micropropagation mostly leads to the production of innumerable true-to-type plants. However, establishing pathogen-free explants through in vitro culture requires a precise management of time for the exposure of explants to antimicrobial chemicals. The application of antimicrobial chemicals must also be managed to impose the least injury on explants. This review discusses the contributions of micropropagation procedures, explant types, subculture duration, media ingredients and plant growth regulators to the in vitro response of conifer explants. Even though regeneration from mature conifer explants such as mature shoots are laborious, the chances of variation, induced in vitro, are unlikely.

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References

  • Aitken-Christie J, Singh A, Davies H (1988) Multiplication of meristematic tissue: a new tissue culture system for radiata pine. In: Hanover JW, Keathley DE (eds) Genetic manipulation of woody plants, vol 44. Plenum Publishing Corporation, New York, pp 413–432

  • Alonso P, Cortizo M, Cantón FR, Fernández B, Rodríguez A, Centeno ML, Cánovas FM, Ordás RJ (2007) Identification of genes differentially expressed during adventitious shoot induction in Pinus pinea cotyledons by subtractive hybridization and quantitative PCR. Tree Physiol 27:1721–1730

    Article  CAS  PubMed  Google Scholar 

  • Arya S, Kalia RK, Arya ID (2000) Induction of somatic embryogenesis in Pinus roxburghii Sarg. Plant Cell Rep 19:775–780

    Article  CAS  Google Scholar 

  • Bastol B, Jasik J, Mantell S (2000) In vitro propagation of a Himalayan pine P. wallichiana A. B. Jacks. Curr Sci 78:338–341

    Google Scholar 

  • Becwar M, Nagmani R, Wann S (1990) Initiation of embryogenic cultures and somatic embryo development in loblolly pine (Pinus taeda). Can J For Res 20:810–817

    Article  Google Scholar 

  • Bigot C, Engelmann F (1987) Vegetative propagation in vitro of Cunninghamia lanceolata (Lamb.) Hook. In: Bonga JM, Durzan DJ (eds) Cell and tissue culture in forestry, case histories, gymnosperms, angiosperms and palms, vol 3. Martinus Nijhoff, The Hague, pp 114–127

    Chapter  Google Scholar 

  • Bonga JM, Klimaszewska KK, von Aderkas P (2010) Recalcitrance in clonal propagation, in particular of conifers. Plant Cell Tissue Organ Cult 100:241–254

    Article  Google Scholar 

  • Boulay M (1979) Multiplication et clonage rapide du Sequoia sempervirens par la culture in vitro. Annales de Recherche Sylvicole, AFOCEL, pp 49–55

    Google Scholar 

  • Bourgin JP, Nitsch JP (1967) Obtention de Nicotiana haploides à partir d’étamines cultivées in vitro. Ann Physiol Vég 9:337–338

    Google Scholar 

  • Buckley PM, DeWild TN, Reed BM (1995) Characterization and identification of bacteria isolated from micropropagated mint plant. In Vitro Cell Dev Biol Plant 31:58–64

    Article  Google Scholar 

  • Burrows GE, Doley DD, Haines RJ, Nikles DG (1988) In vitro propagation of Araucaria cunninghamii and other species of the Araucariaceae via axillary meristems. Aust J Bot 36:665–676

    Article  Google Scholar 

  • Cassells AC (1991) Problems in tissue culture: culture contamination. In: Debergh PC, Zimmerm RH (eds) Micropropagation technology and application. Kluwer Academic Publishers, Dordrecht, pp 31–44

    Chapter  Google Scholar 

  • Chanway C (1998) Bacterial endophytes: ecological and practical implication. Sydowia 50:149–170

    Google Scholar 

  • Cortizo M, Cuestam C, Centeno ML, Rodriguez A, Fernandez B, Ordas R (2009a) Benzyladenine metabolism and temporal compedence of Pinus pinea cotyledons to form buds in vitro. J Plant Physiol 166:1069–1076

    Article  CAS  PubMed  Google Scholar 

  • Cortizo M, De Diego N, Ordás R, Moncaleán P (2009b) Micropropagation of adult Stone Pine (Pinus pinea L.). Trees 23:835–842

    Article  Google Scholar 

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

    Article  Google Scholar 

  • De Diego N, Montalbán IA, Moncaleán P (2010) In vitro regeneration of adult Pinus sylvestris L. trees. S Afr J Bot 76:158–162

    Article  Google Scholar 

  • Debergh PC, Vanderschaeghe AM (1988) Some symptoms indicating the presence of bacterial contaminants in plant tissue culture. Acta Hortic 255:77–81

    Article  Google Scholar 

  • Dodds JH, Roberts WL (1981) Some inhibitory effectors on gentamicin on plant tissue culture. In Vitro 17:467–470

    Article  CAS  Google Scholar 

  • Dos Santos ALW, Silveria V, Steinr N, Vidor M, Guerra MP (2002) Somatic embryogenesis in Parana Pine (Araucaria angustifolia (Bert.) O. Kuntze). Braz Arch Biol Technol 45:97–106

    Article  Google Scholar 

  • Drake PMW, John A, Power JB, Davey MR (1997) Expression of the gus A gene in embryogenic cell lines of Sitka spruce following Agrobacterium-mediated transformation. J Exp Bot 48:151–155

    Article  CAS  Google Scholar 

  • Driver JA, Kuniyuki AH (1984) In vitro propagation of paradox walnut root stocks. Hortic Sci 19:507–509

    Google Scholar 

  • Ellis DD, Lazaroff WR, Roberts DR, Flinn BS, Webb DT (1989) The effect of antibiotics on elongation and callus and bud formation from embryogenic tissue of Picea glauca. Can J For Res 19:1343–1346

    Article  CAS  Google Scholar 

  • Ewald D, Suss R (1993) A system for repetable formation of elongating adventitious buds in Norway spruce tissue cultures. Silvae Genetica 42:169–175

    Google Scholar 

  • Ewald D, Kretzschmar U, Chen Y (1997) Continuous micropropagation of juvenile larch from different species via adventitious bud formation. Biol Plant 39:321–329

    Article  Google Scholar 

  • Ewald D, Zaspel I, Naujoks G, Behrendt U (2000) Endogenous bacteria in tissue cultures of conifers—appearance and action. Acta Hortic 530:137–144

    Article  Google Scholar 

  • Falkiner, FR (1990) The criteria for choosing an antibiotic for control of bacteria in plant tissue culture in TCL. Assoc Plant Tiss Cult Newsl 60:13–23

    Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    Article  CAS  PubMed  Google Scholar 

  • Gresshoff PM, Doy CH (1972) Development and differentiation of haploid Lycopersicon esculentum (tomato). Planta 107:161–170

    Article  CAS  PubMed  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

    Article  CAS  PubMed  Google Scholar 

  • Haines RJ, de Fossard RA (1977) Propagation of Hoop pine (Araucaria cunninghamii) by organ culture. Acta Hortic 78:297–302

    Article  Google Scholar 

  • Harry IS, Thompson MR, Lu CY, Thorpe TA (1987) In vitro plantlet formation from embryonic explants of eastern white cedar (Thuja occidentalis L.). Tree Physiol 3:273–283

    Article  CAS  PubMed  Google Scholar 

  • Hohtola A (1988) Seasonal changes in explant viability and contamination of Scots pine tissue cultures from mature Scots pine. Plant Cell Tissue Organ Cult 15:211–222

    Article  Google Scholar 

  • Holford P, Newbury HJ (1992) The effects of antibiotics and their breakdown products on the in vitro growth of Antirrhinum majus. Plant Cell Rep 11:93–96

    CAS  PubMed  Google Scholar 

  • Holland L, Gemmell JE, Chanty JA, Walter C (1997) Foreign gene transfer into Pinus radiata cotyledons by Agrobacterium tumefaciens. NZ J Forest Sci 27:289–304

    CAS  Google Scholar 

  • Hood EE, Clapham DH, Ekberg I, Johanson T (1990) T-DNA presence and opine production in tumors of Picea abies (L.) Karst induced by Agrobacterium tumefaciens A281. Plant Mol Biol 14:111–117

    Article  CAS  PubMed  Google Scholar 

  • Humara JM, Ordas RJ (1999) The toxicity of antibiotics and herbicides on in vitro adventitious shoot formation on Pius pinea L. cotyledone. In Vitro Cell Dev Biol Plant 35:339–343

    Article  CAS  Google Scholar 

  • Keathley DE (1984) In: Proceedings of the international symposium of recent advances in forest biotechnology, Michigan Biotechnology Institute, Traverse City, Michigan, pp 58–63, 10–13 June 1984

  • Kolevska-Pletikapic B, Buturovic-Deric Z (1995) Regeneration of Picea omarika plants via organogenesis. Plant Cell Tissue Organ Cult 41:189–192

    Article  CAS  Google Scholar 

  • Laukkanen H, Soini H, Kontunen-Soppela S, Hohtola A, Viljanen M (2000) A mycobacterium isolated from tissue cultures of mature Pinus sylvestris interferes with growth of Scots pine seedlings. Tree Physiol 20:915–920

    Article  PubMed  Google Scholar 

  • Le-Feuvre R, Triviño C, Sabja AM, Bernier-Cardou M, Moynihan MR, Klimaszewska K (2013) Organic nitrogen composition of the tissue culture medium influences Agrobacterium tumefaciens growth and the recovery of transformed Pinus radiata embryonal masses after cocultivation. In Vitro Cell Dev Biol Plant 49:30–40

    Article  CAS  Google Scholar 

  • Leifert C, Waites WM, Nicholas LR (1989) Bacterial contamination of micropropagated plant tissue cultures. J Appl Bacteriol 67:353–361

    Article  Google Scholar 

  • Leifert C, Camotla H, Wright SM, Waites B, Cheyne VA, Waites WM (1991) Elimination of Lactobacillus plantarum, Corynebacterium spp. Staphylococcus saprophyticus and Pseudomonas paucimobilis from micropropagated Hemerocallis, Choisya and Delphinium cultures using antibiotics. J Appl Bacteriol 71:307–330

    Article  Google Scholar 

  • Leifert C, Camotla H, Wailes WM (1992) Effect of combinations of antibiotics on micropropagated Clematis, Delphinium, Hosta, Iris and Photinia. Plant Cell Tissue Organ Cult 29:153–160

    Article  CAS  Google Scholar 

  • Levee V, Lelu MA, Jouanin L, Cornu D, Pilati G (1997) Agrobacterium tumefaciens-mediated transformation of hybrid larch (Larix kaempferi × L. decidua) and transgenic plant regeneration. Plant Cell Rep 16:680–685

    Article  CAS  Google Scholar 

  • López M, Humara JM, Rodríguez R, Ordás RJ (2000) Factors involved in Agrobacterium tumefaciens-mediated gene transfer into Pinus nigra Arn. ssp. Salzmannii (Dunal) Franco. Euphytica 114:195–203

    Article  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

    Article  Google Scholar 

  • Loureiro J, Capelo A, Brito G, Rodriguez E, Silva S, Pinto G, Santos C (2007) Micropropagation of Juniperus phoenicea from adult plant explants and analysis of ploidy stability using flow cytometry. Biol Plant 51:7–14

    Article  CAS  Google Scholar 

  • Malá J, Pavingerová D, Cvrčková H, Bříza J, Dostál J, Šíma P (2009) Tolerance of Norway spruce (Picea abies [L.] Karst.) embryogenic tissue to penicillin, carbapenem and aminoglycoside antibiotics. J For Sci 55:156–161

    Article  Google Scholar 

  • Malabadi RB, Van Staden J (2005) Somatic embryogenesis from vegetative shoot apices of mature trees of Pinus patula. Tree Physiol 25:11–16

    Article  PubMed  Google Scholar 

  • McCown BH, Lloyd G (1981) Woody plant medium (WPM)—a mineral nutrient formulation for microculture for woody plant species. Hortic Sci 16:453

    Google Scholar 

  • Mendes AFS, Cidade LC, de Oliveira MLP, Otoni WC, Soares-Filho WS, Costa MGC (2009) Evaluation of novel beta-lactam antibiotics in comparison to cefotaxime on plant regeneration of Citrus sinensis L. Osb. Plant Cell Tissue Organ Cult 97:331–336

    Article  Google Scholar 

  • Moncaleán P, Alonso P, Centeno ML, Cortizo M, Rodríguez A, Fernández B, Ordás RJ (2005) Organogenic responses of Pinus pinea cotyledons to hormonal contents: BA metabolism and cytokinin content. Tree Physiol 25:1–19

    Article  PubMed  Google Scholar 

  • Montalbán IA, De Diego N, Igartua EA, Setién A, Moncaleán P (2011) A combined pathway of somatic embryogenesis and organogenesis to regenerate radiata pine plants. Plant Biotechnol Rep 5:177–186

    Article  Google Scholar 

  • Murashige T (1974) Plant propagation through tissue culture. Ann Rev Plant Physiol 25:135–166

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Nauerby B, Billing K, Wyndaele R (1997) Influence of the antibiotic timentin on plant regeneration compared to carbenicillin and cefotaxime in concentrations suitable for elimination of Agrobacterium tumefaciens. Plant Sci 123:169–177

    Article  CAS  Google Scholar 

  • Ogawa Y, Mii M (2005) Evaluation of 12 b-lactam antibiotics for Agrobacterium-mediated transformation through in planta antibacterial activities and phytotoxicities. Plant Cell Rep 23:736–743

    Article  CAS  PubMed  Google Scholar 

  • Piola F, Rohr R, Heizmann P (1999) Rapid detection of genetic variation within and among in vitro propagated cedar (Cedrus libani Loudon) clones. Plant Sci 141:159–163

    Article  CAS  Google Scholar 

  • 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 Tissue Organ Cult 73:91–94

    Article  CAS  Google Scholar 

  • Pulido CM, Harry IS, Thorpe TA (1992) Optimization of bud induction in cotyledonary explants of Pinus canariensis. Plant Cell Tissue Organ Cult 29:247–255

    Article  Google Scholar 

  • Quoirin M, Lepoivre P (1977) Elude de milieu adaptes aux cultures in vitro de Prunus. Acta Hortic 78:437–442

    Article  Google Scholar 

  • Reed BM, Tanprasert P (1995) Detection and control of bacterial contaminations of plant tissue culture: a review of recent literature. Plant Tissue Cult Biotechnol 1:137–142

    Google Scholar 

  • Renau-Morata B, Ollero J, Arrillagam I, Segura J (2005) Factor influencing axillary proliferation and adventitious budding in cedar. Tree Physiol 25:477–486

    Article  PubMed  Google Scholar 

  • Rugini E (1984) In vitro propagation of some olive (Olea europaea L. var. sativa) cultivars with different root-ability, and medium development using analytical data from developing shoots and embryos. Sci Hortic 24:123–134

    Article  CAS  Google Scholar 

  • Saravitz CH, Blazich FA, Amerson HV (1991) In vitro propagation of virginia pine from cotyledons. J Am Soc Hortic Sci 116:362–365

    Google Scholar 

  • Sarma KS, Evans NE, Selby CH (1995) Effect of carbenicillin and cefotaxime on somatic embryogenesis of Sitka spruce (Picea sitchensis (Bong.) Carr.). J Exp Bot 46:1779–1781

    Article  CAS  Google Scholar 

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

    Article  Google Scholar 

  • Sarmast MK, Salehi H, Khosh-Khui M, Niazi A, Bastani R (2010) Nano silver functionalization of Agrobacterium mediated transformation with companionship of nanobiotechnology. Symposium on nanotechnologies applied to biosystems engineering and the environment. CIGR. Québec City, 13–17 June, Canada

  • Sarmast MK, Salehi H, Khosh-Khui M (2011) Nano silver treatment is effective in reducing bacterial contamination of Araucaria excelsa R. Br. var. glauca explants. Acta Biol Hung 62:477–484

    Article  CAS  PubMed  Google Scholar 

  • Sarmast MK, Salehi H, Ramzani Abolimoghadam AA, Niazi A, Khosh-Khui M (2012a) RAPD fingerprint to appraise the genetic fidelity of in vitro propagated Araucaria excelsa R. Br. var. glauca plantlets. Mol Biotechnol 50:181–188

    Article  CAS  PubMed  Google Scholar 

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

    Article  PubMed Central  PubMed  Google Scholar 

  • Sarmast MK, Niazi A, Salehi H, Abolimoghadam A (2015) Silver nanoparticles affect ACS expression in Tecomella undulata in vitro culture. Plant Cell Tiss Organ Cult 121:227–236

    Article  CAS  Google Scholar 

  • Sehgal L, Sehgal OP, Khosla PK (1989) Micropropagation of Araucaria columnaris Hook. Ann Sci For 46:158–160

    Article  Google Scholar 

  • Selby C, Watson S, Harvey BMR (2005) Morphogenesis in Sitka spruce (Picea sitchensis (Bong.) Carr.) bud cultures—tree maturation and explants from epicormic shoots. Plant Cell Tissue Organ Cult 83:279–285

    Article  Google Scholar 

  • Skoog F (1944) Growth and organ formation in tobacco tissue cultures. Am J Bot 31:19–24

    Article  Google Scholar 

  • Stojicic D, Budimir S (2004) Cytokinin-mediated axillary shoots formation in Pinus heldreichii. Biol Plant 48:477–479

    Article  CAS  Google Scholar 

  • Stojicic D, Budimir S, Culafic L (1999) Micropropagation of Pinus heldrechii. Plant Cell Tissue Organ Cult 59:147–150

    Article  Google Scholar 

  • Tamta S, Palni LMS (2004) Studies of in vitro propagation of Himalayan cedar (Cedrus deodara) using zygotic embryos and stem segments. Indian J Biotechnol 3:209–215

    Google Scholar 

  • Tang W, Newton RJ (2005) Plant regeneration from callus cultures derived from mature zygotic embryos in white pine (Pinus strobus L.). Plant Cell Rep 24:1–9

    Article  CAS  PubMed  Google Scholar 

  • Tang W, Sederoff R, Whetten R (2001) Regeneration of transgenic loblolly pine (Pinus taeda L.) from zygotic embryos transformed with Agrobacterium tumefaciencs. Planta 213:981–989

    Article  CAS  PubMed  Google Scholar 

  • Tang W, Newton RJ, Charles TM (2006) Plant regeneration through multiple adventitious shoot differentiation from callus cultures of slash pine (Pinus elliottii). J Plant Physiol 163:98–101

    Article  CAS  PubMed  Google Scholar 

  • Tang W, Harris LC, Outhavong V, Newton RJ (2004) The effect of different plant growth regulators on adventitious shoot formation from Virginia pine (Pinus virginiana) zygotic embryo explants. Plant Cell Tiss Org Cult 78:237–240

    Article  CAS  Google Scholar 

  • Thorpe TA (1985) Application of tissue culture to forest tree improvement. For Chron 61:436–438

    Article  Google Scholar 

  • Thorpe TA (2007) History of plant tissue culture. Mol Biotech 37:169–180

    Article  CAS  Google Scholar 

  • Timmis R, Ritchie GA (1984) In: Proceedings of the international symposium of recent advances in forest biotechnology, Michigan Biotechnology Institute, Traverse City, Michigan, pp 37–46, 10–13 June 1984

  • Traore A, Xing Z, Bonser A, Carlson J (2005) Optimizing a protocol for sterilization and in vitro establishment of vegetative bud from mature Douglas fir trees. HortScience 40:1464–1468

    Google Scholar 

  • Valdés AE, Ordás RJ, Fernández B, Centeno ML (2001) Relationships between hormonal content sand the organogenic response in Pinus pinea cotyledons. Plant Physiol Biochem 39:377–384

    Article  Google Scholar 

  • Vidal N, Arellano G, San-Jose MC, Vieitez AM, Ballester A (2003) Developmental stages during the rooting of in vitro cultured Quercus robur shoots from material of juvenile and mature origin. Tree Physiol 23:1247–1254

    Article  CAS  PubMed  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

    Article  CAS  Google Scholar 

  • Viss PR, Brooks EM, Driver JA (1991) A simplified method for the control of bacterial contamination in woody plant tissue culture. In vitro Cell Dev Biol Plant 27:42

    Article  Google Scholar 

  • Webb KJ, Street HE (1977) Morphogenesis in vitro of Pinus and Picea. Acta Horticult 78:259–267

    Article  Google Scholar 

  • Yepes LM, Aldwinckle HS (1994) Factors that affect leaf regeneration efficiency in apple and effect of antibiotics in morphogenesis. Plant Cell Tissue Organ Cult 37:257–269

    CAS  Google Scholar 

  • Zhang Y, Wei Z, Xi M, Shi J (2006) Direct organogenesis and plantlet regeneration from mature zygotic embryos of masson pine (Pinus massoniana L.). Plant Cell Tissue Organ Cult 84:119–123

    Article  CAS  Google Scholar 

  • Zhu LH, Wu XQ, Qu HY, Ji J, Ye J (2010) Micropropagation of Pinus massoniana and mycorrhiza formation in vitro. Plant Cell Tissue Organ Cult 102:121–128

    Article  Google Scholar 

Download references

Acknowledgements

I thank Dr. Jenna E. Gallegos (Department of Molecular and Cellular Biology, University of California, Davis) for critically reading the manuscript. Mohsen Hamedpour-Darabi is thanked for editing the English.

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  1. Department of Horticultural Science, Faculty of Plant Production, Gorgan University of Agricultural Sciences and Natural Resources (GUASNR), Gorgan, Golestan, 49138-43464, Iran

    Mostafa K. Sarmast

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Correspondence to Mostafa K. Sarmast.

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Project funding: This work was supported by grant from Gorgan University of Agricultural Sciences and Natural Resources (GUASNR) for multiyear project on micropropagation of Juniperus species.

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Corresponding editor: Chai Ruihai

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Sarmast, M.K. In vitro propagation of conifers using mature shoots. J. For. Res. 29, 565–574 (2018). https://doi.org/10.1007/s11676-018-0608-7

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