Skip to main content
SpringerLink
Log in

Agrobacterium-mediated transformation of Eupatorium adenophorum

  • Research Note
  • Published:
Plant Cell, Tissue and Organ Culture (PCTOC) Aims and scope Submit manuscript

Abstract

A simple and efficient protocol for Agrobacterium tumefaciens-mediated genetic transformation of Eupatorium adenophorum has been developed. Stem segments were pre-cultured on Murashige and Skoog (MS) medium for 24 h and then co-cultured with A. tumefaciens strain LBA4404 harboring the pBI121 plasmid on the same medium in darkness for 3 days. After co-cultivation, explants were transferred to MS medium supplemented with 1.0 mg l−1 naphthaleneacetic acid (NAA), 1.0 mg l−1 6-benzyladenine (BA), 150 mg l−1 kanamycin (Kan), and 200 mg l−1 cefotaxime (Cef), and incubated at 25 ± 1°C under a 16-h photoperiod. Callus formation was induced for 3–4 weeks. Induced calluses were transferred to fresh selection medium and subcultured every 15 days. After 6–8 weeks, regenerated plantlets from embryogenic callus could be observed. Buds of 1 cm in length were transferred to half-strength MS medium containing 150 mg l−1 Kan and 200 mg l−1 Cef, and rooted plantlets were obtained after 4 weeks. Putative transgenic plants were confirmed using GUS histochemical assay, polymerase chain reaction (PCR), and reverse transcriptase (RT)-PCR. A transformation efficiency of 2% was obtained.

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
Fig. 6
Fig. 7

Abbreviations

Kan:

Kanamycin

KanR :

Kanamycin-resistant

Str:

Streptomycin

Cef:

Cefotaxime

BA:

6-benzyladenine

NAA:

Naphthaleneacetic acid

MS:

Murashige and Skoog basal medium

YEB:

Yeast extract medium

PGR:

Plant growth regulator

References

  • Arunachalam K, Arunachalam A, Melkania NP (1999) Influence of soil properties on microbial populations, activity and biomass in humid subtropical mountainous ecosystems of India. Biol Fertil Soils 30:217–223

    Article  Google Scholar 

  • Auld BA (1970) Eupatorium weed species in Australia. Pest Articles News Summ 16:82–86

    Google Scholar 

  • Bevan M (1984) Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res 12:8711–8721

    Article  CAS  PubMed  Google Scholar 

  • Borthakur M, Dutta K, Nath SC, Singh RS (2000) Micropropagation of Eclipta alba and Eupatorium adenophorum using a single-step nodal cutting technique. Plant Cell Tiss Organ Cult 62:239–242

    Article  Google Scholar 

  • Dey S, Sinha B, Kalita J (2005) Effect of Eupatorium adenophorum Spreng leaf extracts on the mustard aphid, Lipaphis erysimi Kalt: a scanning electron microscope study. Microsc Res Tech 66(1):31–36

    Article  PubMed  Google Scholar 

  • Duan H, Qiang S, Su XH, Wu HR, Zhu YZ, Liu LL (2004) Genetic diversity of Eupatorium adenophorum determined by AFLP marker. Acta Ecol Sin 25(8):2109–2114 (in Chinese)

    Google Scholar 

  • Feng YL, Wang YH, Liu YY, Ding KY (2006) Karyotype of Chromolaena odorata and Ageratina adenophora. Bull Bot Res 26(3):356–360 (in Chinese)

    Google Scholar 

  • Flores-Benítez S, Jiménez-Bremont JF, Rosales-Mendoza S, Argüello-Astorga GR, Castillo-Collazo R, Alpuche-Solís ÁG (2007) Genetic transformation of Agave salmiana by Agrobacterium tumefaciens and particle bombardment. Plant Cell Tiss Organ Cult 91:215–224

    Article  Google Scholar 

  • Hassanein A, Chevreau E, Borion N (2005) Highly efficient transformation of zonal (Pelargonium × hortorum) and scented (P. capitatum) geraniums via Agrobacterium tumefaciens using leaf discs. Plant Sci 169:532–541

    Article  CAS  Google Scholar 

  • Herman I, Jacobs A, Van Montagu M, Depicker A (1990) Plant chromosome/marker gene fusion assay for study of normal and truncated T-DNA integration events. Mol Gen Genet 224:248–256

    Article  CAS  PubMed  Google Scholar 

  • Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405

    Article  CAS  Google Scholar 

  • Keil DJ, Ludkow MA, Pinkava DJ (1988) Chromosome studies in Asteraceae from the United States, Mexico, the West Indies, and South America. Am J Bot 75:652–668

    Article  Google Scholar 

  • Li H, Qiang S, Cui J (2005) Eupatroium adenophorum micro-propagation by bud and callus culture. Acta Bot Boreali-occident Sin 25(7):1458–1462 (in Chinese)

    CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Papes M, Peterson AT (2003) Predicting the potential invasive distribution for Eupatorium adenophorum Spreng. J Wuhan Bot Res 21:137–142 (in Chinese)

    Google Scholar 

  • Qiang S (1998) The history and status of the study on Crofton weed (Eupatorium adenophorum Spreng.) a worst worldwide weed. J Wuhan Bot Res 16(4):366–372 (in Chinese)

    Google Scholar 

  • Song QS, Fu Y, Tang JW, Feng ZL, Yang CR (2000) Allelopathic potential of Eupatorium adenophorum. Acta Phytoecol Sin 24(3):362–365 (in Chinese)

    Google Scholar 

  • Song GQ, Loskutov AV, Sink KC (2007) Highly efficient Agrobacterium tumefaciens-mediated transformation of celery (Apium graveolens L.) through somatic embryogenesis. Plant Cell Tiss Organ Cult 88:193–200

    Article  CAS  Google Scholar 

  • Tel-Zur N, Abbo S, Mylaboski D, Mizrahi Y (1999) Modified CTAB procedure for DNA isolation from epiphytic cacti of the genera Hylocereus and Selenicereus (Cacataceae). Plant Mol Biol Rep 17:249–254

    Article  CAS  Google Scholar 

  • Wang HJ, He P, Ma JL (1994) An Investigation and research report on the dissemination of Ageratina adenophora on rangeland areas in Liangshan District of Sichuan Province. Grassl China 87:62–64 (in Chinese)

    Google Scholar 

  • Yan YP, Wang ZZ (2007) Genetic transformation of the medicinal plant Salvia miltiorrhiza by Agrobacterium tumefaciens-mediated method. Plant Cell Tiss Organ Cult 88:175–184

    Article  CAS  Google Scholar 

  • Yin Z, Wang GL (2000) Evidence of multiple complex patterns of T-DNA integration into the rice genome. Theor Appl Genet 100:461–470

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This research was supported by the National Natural Science Foundation of China (30771422) and a 973 Project grant (2009CB119200) from the Ministry of Science and Technology of China.

Author information

Authors and Affiliations

  1. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

    Huiming Guo, Yongqiang Zhang & Hongmei Cheng

  2. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100094, China

    Fanghao Wan

Authors
  1. Huiming Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yongqiang Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Fanghao Wan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hongmei Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hongmei Cheng.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Guo, H., Zhang, Y., Wan, F. et al. Agrobacterium-mediated transformation of Eupatorium adenophorum. Plant Cell Tiss Organ Cult 103, 417–422 (2010). https://doi.org/10.1007/s11240-010-9784-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-010-9784-7

Keywords

Search

Navigation