Skip to main content
SpringerLink
Log in

Transformation of pollen embryo-derived explants by Agrobacterium tumefaciens in Hyoscyamus niger

  • Published:
Plant Cell, Tissue and Organ Culture Aims and scope Submit manuscript

Abstract

Leaf, root, stem, petiole, hypocotyl, and zygotic embryo explants, as well as pollen embryoids, and redifferentiated tissues from pollen embryoid-derived plantlets of Hyoscyamus niger L. (black henbane) were inoculated with Agrobacterium tumefaciens, harboring binary vectors (pGS Gluc1) and then cultured on media containing kanamycin. Transient β-glucuronidase activity and kanamycin resistant callus formation were influenced by explant origin. Transgenic calluses were obtained at a frequency of up to 30% from all the explants tested. However, transgenic shoots were obtained only from the hypocotyl of plantlets derived from pollen embryoids. Transformation was confirmed by the ability of leaf segments to produce kanamycin resistant calluses, β-glucuronidase histochemical and flurometric assays, polymerase chain reaction and Southern blot analysis. The results show that pollen embryoid-derived explants may be an alternative source for both efficient transformation and regeneration of transgenic plants in recalcitrant species.

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

Similar content being viewed by others

Abbreviations

BA:

6-benzylaminopurine

2,4-D:

dichlorophenoxy acetic acid

GUS:

β-glucuronidase

IAA:

indole-3-acetic acid

2ip:

N6-(2-isopentenyl) adenine

KmR:

kanamycin resistant

MS:

Murashige and Skoog (1962) medium

MUG:

4-methylumbelliferyl-β-D-glucuronide

NAA:

α-naphthaleneacetic acid

NPTII:

neomycin phosphotransferase-II

PCR:

polymerase chain reaction

X-gluc:

5-bromo-4-chloro-3-indolyl glucuronide

References

  • V Babic RS Datla GJ Scoles WA Keller (1998) ArticleTitleDevelopment of an efficient Agrobacterium-mediated transformation system for Brassica carinata Plant Cell Rep. 17 183–188

    Google Scholar 

  • E Beck G Ludwig EA Averswald B Reiss H Schaller (1982) ArticleTitleNucleotide sequence and exact localization of the neomycin phosphotransferase gene from transposon Tn5 Gene 19 327–336

    Google Scholar 

  • MW Bevan (1984) ArticleTitleBinary Agrobacterium vectors for plant transformation Nucleic Acids Res. 12 8711–8721

    Google Scholar 

  • JC Cheng V Raghavan (1985) ArticleTitleSomatic embryogenesis and plant regeneration in Hyoscyamus niger Amer. J. Bot. 72 IssueID4 580–587

    Google Scholar 

  • RN Chibbar KK Kartha RSS Datla N Leung K Caswell CS Mallard L Steinhauer (1993) ArticleTitleThe effect of different promoter-sequences on transient expression of gus reporter gene in cultured barley (Hordeum vulgare L.) cells Plant Cell Rep. 12 506–509

    Google Scholar 

  • G Corduan (1975) ArticleTitleRegeneration of anther-derived plants of Hyoscyamus niger L Planta 127 27–36

    Google Scholar 

  • ML Cui H Ezura (2003) ArticleTitleAgrobacterium-mediated transformation of Nemesia strumosa Benth, a model plant for asymmetric floral development Plant Sci. 165 863–870

    Google Scholar 

  • R Deblaere B Bytebier H Greve ParticleDe F Deboeck J Schell M Van Montagu J Leemans (1985) ArticleTitleEfficient atropine Ti-plasmid derived vectors for Agrobacterium-mediated gene transfer to plants Nucleic Acids Res. 13 4777–4788

    Google Scholar 

  • A De Bondt K Eggermont I Penninckx Goderis WF Broekaert (1996) ArticleTitleAgrobacterium-mediated transformation of apple (Malus x domestica Borkh.): an assessment of factors affecting regeneration of transgenic plants Plant Cell Rep. 15 549–554

    Google Scholar 

  • M De Block De Brouwer P Tenning (1989) ArticleTitleTransformation of Brassica napus and Brassica oleracea using Agrobacterium tumefaciens and the expression of the bar and neo genes in the transgenic plants Plant Physiol. 91 694–701

    Google Scholar 

  • J De Jong MMJ Mertens W Rademaker (1994) ArticleTitleStable expression of the GUS reporter gene in chrysanthemum depends on binary plasmid T-DNA Plant Cell Rep. 14 59–64

    Google Scholar 

  • A Domínguez J Guerri M Cambra L Navarro P Moreno L Peña (2000) ArticleTitleEfficient production of transgenic citrus plants expressing the coat protein gene of citrus tristeza virus Plant Cell Rep. 19 427–433

    Google Scholar 

  • JZ Dong A McHughen (1991) ArticleTitlePatterns of transformation intensity on flax hypocotyls inoculated with A tumefaciens. Plant Cell Rep. 10 555–560

    Google Scholar 

  • JZ Dong A McHughen (1993) ArticleTitleTransgenic flax plants from Agrobacterium mediated transformation: incidence of chimeric regenerants and inheritance of transgenic plants Plant Sci. 91 139–148

    Google Scholar 

  • C Ducrocq RS Sangwan BS Sangwan-Norreel (1994) ArticleTitleProduction of Agrobacterium-mediated transgenic fertile plants by direct somatic embryogenesis from immature zygotic embryos of Datura innoxia Plant Mol. Biol. 25 995–1009

    Google Scholar 

  • M Fari I Nagy M Csanyi J Mityko A Andrasfalvy (1995) ArticleTitleAgrobacterium-mediated genetic transformation and plant regeneration via organogenesis and somatic embryogenesis from cotyledon leaves in egg plant (Solanum melongena L cv. Kecskemet; lila). Plant Cell Rep. 15 82–86

    Google Scholar 

  • E Firoozabady D DeBoer D Merlo E Halk L Amerson KE Rashka EE Murray (1987) ArticleTitleTransformation of cotton (Gossypium hirsutum L.) by Agrobacterium tumefaciens and regeneration of transgenic plants Plant Mol. Biol. 10 105–116

    Google Scholar 

  • Hasunuma T, Fukusaki E & Kobayashi A (2004) Expression of fungal pectin methylesterase in transgenic tobacco leads to alteration in cell wall metabolism a phenotype. 111: 241–251

  • RB Horsch JE Fry NL Hoffmann D Eichholtz SG Rogers RT Fraley (1985) ArticleTitleA simple and general method for transferring genes into plants Science 227 1229–1231 Occurrence Handle1:CAS:528:DyaL2MXhtFSjsLc%3D

    CAS  Google Scholar 

  • Jacq B, Sangwan RS & Sangwan-Norreel BS (1993) Initiation de cellules transgéniques puis régénération de plantes transformées chez la Betterave sucrière (Beta vulgaris L.). Proceedings of the “Biotechnologie’93 Rencontres en Picardie”, Amiens, France, 13–15 Décembre

  • RA Jefferson SM Burgess D Hirsh (1986) ArticleTitleβ-Glucuronidase from Escherichia coli as a gene-fusion marker Proc. Natl. Acad. Sci. USA 83 8447–8451

    Google Scholar 

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

    Google Scholar 

  • E Knopp A Strauss W Wehrli (1988) ArticleTitleRoot induction on several Solanaceae species by Agrobacterium rhizogenes and the determination of root tropane alkaloid content Plant Cell Rep. 7 590–593

    Google Scholar 

  • M Manoharan CS Sree Vidya G Lakshmisita (1998) ArticleTitleAgrobacterium-mediated genetic transformation in hot chilli (Capsicum annuum L var. Pusa jwala). Plant Sci. 131 77–83

    Google Scholar 

  • S McCormick J Niedermeyer J Fry A Barnason R Horsch R Fraley (1986) ArticleTitleLeaf disc transformation of cultivated tomato (L esculentum) using Agrobacterium tumefaciens. Plant Cell Rep. 5 81–84

    Google Scholar 

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

    Google Scholar 

  • R Nagmani V Raghavan (1983) ArticleTitleInduction of embryogenetic divisions in isolated pollen grains of Hyoscyamus niger in a single-step method Z. Pflanzenphysiol. 109 87–90

    Google Scholar 

  • S Nishibayashi H Kaneko T Hayakawa (1996) ArticleTitleTransformation of cucumber (Cucumis sativus L.) plants using Agrobacterium tumefaciens and regeneration from hypocotyl explants Plant Cell Rep. 15 809–814

    Google Scholar 

  • JP Nitsch C Nitsch (1969) ArticleTitleHaploid plants from pollen grains Science 163 85–87

    Google Scholar 

  • A Passey K Barrett D James (2003) ArticleTitleAdventitious shoot regeneration from seven commercial strawberry cultivars (Fragaria × ananassa Duch.) using a range of explant types Plant Cell Rep. 21 397–401

    Google Scholar 

  • J Paszkowski RD Shillito M Saul V Mandak T Hohn B Hohn I Potrykus (1984) ArticleTitleDirect gene transfer to plants EMBO J. 3 2717–2722

    Google Scholar 

  • V Raghavan (1975) ArticleTitleInduction of haploid plants from anther culture of henbane Z. Pflanzenphysiol. 76 89–92

    Google Scholar 

  • J Reinert YPS Bajaj (1977) Anther culture: haploid production and its significance J Reinert YPS Bajaj (Eds) Applied and Fundamental Aspects of Plant Cell, Tissue and Organ Culture Springer Berlin, Heidelberg, New York 251–267

    Google Scholar 

  • Sambrook J, Fritsch EF & Maniatis T (1989) Molecular Cloning (A Laboratory Manual). 2nd edn. Vol. 1, pp. 1.42–1.46, pp. 6.1–6.19, pp. 9.31–9.57, Vol. 2, pp. 14.1–14.35. Cold Spring Harbor Laboratory Press

  • RS Sangwan Y Bourgeois BS Sangwan-Norreel (1991a) ArticleTitleGenetic transformation of Arabidopsis thaliana zygotic embryos and identification of critical parameters influencing transformation efficiency Mol. Gen. Genet. 230 475–485

    Google Scholar 

  • RS Sangwan C Ducrocq BS Sangwan-Norreel (1991b) ArticleTitleEffect of culture conditions on Agrobacterium-mediated transformation in datura Plant Cell Rep. 10 90–93

    Google Scholar 

  • RS Sangwan Y Bourgeois S Brown G Vasseur BS Sangwan-Norreel (1992) ArticleTitleCharacterization of competent cells and early events of Agrobacterium-mediated genetic transformation in Arabidopsis thaliana Planta 188 439–456

    Google Scholar 

  • RS Sangwan C Ducrocq BS Sangwan-Norreel (1993) ArticleTitleAgrobacterium-mediated genetic transformation of pollen embryo in Datura innoxia and Nicotiana tabacum: production of transgenic haploid and fertile homozygous dihaploid plants Plant Sci. 95 99–115

    Google Scholar 

  • R Sarria A Calderón AM Thro E Torres JE Mayer WM Roca (1994) ArticleTitleAgrobacterium-mediated transformation of Stylosanthes guianensis and production of transgenic plants Plant Sci. 96 119–127

    Google Scholar 

  • VV Satyavathi V Prasad B Gita Lakshmi G Sita Lakshmi (2002) ArticleTitleHigh efficiency transformation protocol for three Indian cotton varieties via Agrobacterium tumefaciens Plant Sci. 162 215–223

    Google Scholar 

  • PL Schuerman AM Dandekar (1993) ArticleTitleTransformation of temperate woody plants: progress and potentials Sci. Horticult. 55 101–124

    Google Scholar 

  • Sigareva M, Spivey R, Willits MG, Kramer CM & Chang YF (2004) An efficient mannose selection protocol for tomato that has no adverse effect on the ploidy level of transgenic plants. Plant Cell Rep., on line first: 9 June

  • S Tu (1995) Contribution à l’étude de la régénération et de la transformation par agrobactéries chez la Jusquiame noire (Hyoscyamus niger L.). Thèse de Doctorat Université de Picardie Jules Verne Amiens, France

    Google Scholar 

  • S Tu T Tetu RS Sangwan BS Sangwan-Norreel (1996) ArticleTitleEffect of 2-(N-morpholino)ethanesulfolic acid and myo-inositol on somatic embryogenesis and plant regeneration from zygotic embryos of Hyoscyamus niger L Plant Sci. 114 193–203

    Google Scholar 

  • T Tzfira A Vainstein A Altman (1999) ArticleTitlerol-Gene expression in transgenic aspen (Populus tremula) plants results in accelerated growth and improved stem production index Trees 14 49–54

    Google Scholar 

  • A Vergauwe E Van Geldre D Inzé M Van Montagu V den Eeckhout (1998) ArticleTitleFactors influencing Agrobacterium tumefaciens-mediated transformation of Artemisiannua L Plant Cell Rep. 18 105–110

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratoire Androgenèse et Biotechnologie, Faculté des Sciences, Université de Picardie Jules Verne, 33 rue Saint-Leu, 80039, Amiens Cedex, France

    Shanjun Tu, R.S. Sangwan & B.S. Sangwan-Norreel

  2. Department of Plant Biology, The Ohio State University, Columbus, Ohio, USA

    V. Raghavan

  3. Verma, Biotechnology Centre, The Ohio State University, Columbus, Ohio, USA

    D.P.S. Verma

  4. Institute of Biotechnology, Zhejiang Wanli University, 8 South Qian-hu Road, Ningbo, PRC, 315100

    Shanjun Tu

Authors
  1. Shanjun Tu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. R.S. Sangwan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. V. Raghavan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D.P.S. Verma
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. B.S. Sangwan-Norreel
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shanjun Tu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tu, S., Sangwan, R., Raghavan, V. et al. Transformation of pollen embryo-derived explants by Agrobacterium tumefaciens in Hyoscyamus niger. Plant Cell Tiss Organ Cult 81, 139–148 (2005). https://doi.org/10.1007/s11240-004-4041-6

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-004-4041-6

Keywords

Search

Navigation