Skip to main content
SpringerLink
Log in

The same treatment for transgenic shoot regeneration elicits the opposite effect in mature explants from two closely related sweet orange (Citrus sinensis (L.) Osb.) genotypes

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

Abstract

In citrus, production of mature transgenic plants belonging to different genotypes is an important biotechnological objective. In the present study, we tried to genetically transform and regenerate mature plants from the economically important Navelina sweet orange cultivar by using the procedure previously established for the genetically close Pineapple sweet orange variety. The use of BAP at 3 mg l−1 promoted efficient shoot organogenesis in Pineapple as expected, but not in Navelina. Furthermore, different effects were observed when the auxin α-naphtalene acetic acid (NAA) was added to BAP-containing regeneration media. Although NAA addition at 0.5 mg l−1 enhanced cambial callus formation, number of shoots and their elongation in Navelina, the contrary effect was observed in Pineapple. Moreover, transformation efficiency in Navelina rose from 0 to 3% but declined from 6 to 0% in Pineapple, indicating that BAP and BAP + NAA exerted the opposite effect in transgenic shoot regeneration from two closely related cultivars. This suggests that small changes in the procedure could induce drastic alterations in regeneration and even increase the likelihood of obtaining transformants from non-responsive genotypes. Moreover, the vigour of the starting plant material and the addition of kanamycin as selective agent were determining for the generation of mature sweet orange transgenic plants.

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

BAP:

6-Benzylaminopurine

CM:

Co-cultivation medium

IM:

Inoculation medium

NAA:

α-Naphtalene acetic acid

SRM:

Shoot regeneration medium

References

  • Almeida WAB, Mourao Filho FAA, Pino LE et al (2003) Genetic transformation and plant recovery from mature tissues of Citrus sinensis L. Osbeck Plant Sci 164:203–211

    Article  CAS  Google Scholar 

  • Bordón Y, Guardiola JL, García-Luis A (2000) Genotype affects the morphogenic response in vitro of epicotyl segments of Citrus rootstocks. Ann Bot 86:159–166

    Article  Google Scholar 

  • Cervera M, Pina JA, Juárez J et al (1998a) Agrobacterium-mediated transformation of citrange: factors affecting transformation and regeneration. Plant Cell Rep 18:271–278

    Article  CAS  Google Scholar 

  • Cervera M, Juárez J, Navarro A et al (1998b) Genetic transformation and regeneration of mature tissues of woody fruit plants bypassing the juvenile stage. Transgenic Res 7:51–59

    Article  CAS  Google Scholar 

  • Cervera M, López MM, Navarro L et al (1998c) Virulence and supervirulence of Agrobacterium tumefaciens in woody fruit plants. Physiol Mol Plant Pathol 52:67–78

    Article  Google Scholar 

  • Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: Version II. Plant Mol Biol Rep 4:19–21

    Google Scholar 

  • Domínguez A, Cervera M, Pérez R et al (2004) Characterisation of regenerants obtained under selective conditions after Agrobacterium-mediated transformation of citrus explants reveals production of silenced and chimeric plants at unexpected high frequencies. Mol Breed 14:171–183

    Article  Google Scholar 

  • Duran-Vila N, Ortega V, Navarro L (1989) Morphogenesis and tissue cultures of three citrus species. Plant Cell Tiss Organ Cult 16:123–133

    Article  Google Scholar 

  • Durzan D (1990) Adult vs. juvenile explants: directed totipotency. In: Rodríguez R, Sánchez-Tamés R, Durzan DJ (eds) Plant aging. Basic and applied approaches, NATO ASI Series. Series A: Life Sciences, vol 186, pp 19–25

  • Edriss MH, Burger DW (1984) In vitro propagation of troyer citrange from epicotyl segments. Scientia Horticulturae 23:159–162

    Article  CAS  Google Scholar 

  • Fang DQ, Roose ML (1997) Identification of closely related citrus cultivars with inter-simple sequence repeat markers. Theor Appl Genet 95:408–417

    Article  CAS  Google Scholar 

  • García-Luis A, Bordón Y, Moreira-Dias JM et al (1999) Explant orientation and polarity determine the morphogenic response of epicotyl segments of Troyer citrange. Ann Bot 84:715–723

    Article  Google Scholar 

  • Ghorbel R, Juárez J, Navarro L et al (1999) Green fluorescent protein as a screenable marker to increase the efficiency of generating transgenic woody fruit plants. Theor Appl Genet 99:350–358

    Article  Google Scholar 

  • Ghorbel R, Domínguez A, Navarro L et al (2000) High efficiency genetic transformation of sour orange (Citrus aurantium L.) and production of transgenic trees containing the coat protein gene of Citrus Tristeza Virus. Tree Physiol 20:1183–1189

    PubMed  Google Scholar 

  • Grosser JW, Ollitrault P, Olivares-Fuster O (2000) Somatic hybridization in citrus: an effective tool to facilitate variety improvement. In vitro Cell Dev Biol Plant 36:439–449

    Article  Google Scholar 

  • Hood EE, Gelvin SB, Melchers LS et al (1993) New Agrobacterium helper plasmids for gene transfer to plants. Transgenic Res 2:208–218

    Article  CAS  Google Scholar 

  • Kaneyoshi J, Kobayashi S, Nakamura Y et al (1994) A simple and efficient gene transfer system of trifoliate orange (Poncirus trifoliata Raf.). Plant Cell Rep 13:541–545

    Article  CAS  Google Scholar 

  • McGarvey PB, Kaper JM (1991) A simple and rapid method for screening transgenic plants using the PCR. Biotechniques 11:428–432

    PubMed  CAS  Google Scholar 

  • Moore GA (1986) In vitro propagation of citrus rootstocks. HortScience 21:300–301

    CAS  Google Scholar 

  • Moreira-Dias JM, Molina RV, Bordón Y et al (2000) Direct and indirect shoot organogenic pathways in epicotyl cuttings of Troyer citrange differ in hormone requirements and in their response to light. Ann Bot 85:103–110

    Article  CAS  Google Scholar 

  • Nicolosi E (2007) Origin and Taxonomy. In: Khan I (ed) Citrus genetics, breeding and biotechnology. CABI Publishing, CAB International, Wallington, UK, pp 19–43

  • Pedrosa A, Schweizer D, Guerra M (2000) Cytological heterozygosity and the hybrid origin of sweet orange [Citrus sinensis (L.) Osbeck]. Theor Appl Genet 100:361–367

    Article  Google Scholar 

  • Peña L, Cervera M, Juárez J et al (1995) Agrobacterium-mediated transformation of sweet orange and regeneration of transgenic plants. Plant Cell Rep 14:616–619

    Article  Google Scholar 

  • Peña L, Cervera M, Juárez J et al (1997) Genetic transformation of lime (Citrus aurantifolia Swing.): factors affecting transformation and regeneration. Plant Cell Rep 16:731–737

    Article  Google Scholar 

  • Peña L, Cervera M, Fagoaga C et al (2004a) Agrobacterium-mediated transformation of citrus. In: Curtis IS (ed) Transgenic crops of the world. Kluwer Academic Publishers, Dordrecht, The Netherlands, pp 145–157

    Google Scholar 

  • Peña L, Perez R, Cervera M et al (2004b) Early events in Agrobacterium-mediated genetic transformation of citrus explants. Ann Bot 94:67–74

    Article  PubMed  CAS  Google Scholar 

  • Pérez-Molphe-Balch E, Ochoa-Alejo N (1997) In vitro plant regeneration of Mexican lime and mandarin by direct organogenesis. HortScience 32:931–934

    Google Scholar 

  • Roose ML (1988) Isozymes and DNA restriction fragment length polymorphisms in citrus breeding and systematics. In: Goren R, Mendel K (eds) Proceedings of the 6th International Citrus Congress. Balaban Publishers, Rehovot, Israel, vol 1, pp 155–165

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, USA

  • Yu C, Huang S, Chen C et al (2002) Factors affecting Agrobacterium-mediated transformation and regeneration of sweet orange and citrange. Plant Cell Tiss Organ Cult 71:147–155

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank J. A. Pina for technical assistance, and Dr. E. Carbonell and J. Pérez for statistical analyses. This research was supported by grant AGL2006-03673. A. Rodríguez and M. Cervera were recipients of a Generalitat Valenciana fellowship and a “Ramón y Cajal” MEC postdoctoral contract, respectively. English text revised by F. Barraclough.

Author information

Authors and Affiliations

  1. Centre of Plant Protection and Biotechnology, Instituto Valenciano de Investigaciones Agrarias (IVIA), Ctra. Moncada-Náquera, km. 5, 46113, Moncada, Valencia, Spain

    Ana Rodríguez, Magdalena Cervera, Josep Enric Peris & Leandro Peña

Authors
  1. Ana Rodríguez
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Magdalena Cervera
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Josep Enric Peris
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Leandro Peña
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Leandro Peña.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rodríguez, A., Cervera, M., Peris, J.E. et al. The same treatment for transgenic shoot regeneration elicits the opposite effect in mature explants from two closely related sweet orange (Citrus sinensis (L.) Osb.) genotypes. Plant Cell Tiss Organ Cult 93, 97–106 (2008). https://doi.org/10.1007/s11240-008-9347-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-008-9347-3

Keywords

Search

Navigation