Skip to main content
Log in

Transgenic Campanula carpatica plants with reduced ethylene sensitivity

  • Genetic Transformation and Hybridization
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

Fertile transgenic Campanula carpatica Jacq. plants with flowers, which had reduced sensitivity to ethylene were obtained by Agrobacterium tumefaciens that mediated transformation. The construct used for transformation contained the etr1-1 gene from Arabidopsis thaliana under control of the flower specific fbp1-promoter from petunia. More than 100 flowering T0 lines were tested for their ethylene sensitivity using 2 μl l−1 ethylene. The tolerance level to ethylene varied among the lines. While control plants stopped flowering within 3 days of exposure to ethylene, one of the transformed lines flowered for up to 27 days. The presence and the expression pattern of the transgene in various tissues were studied by polymerase chain reaction (PCR) and reverse transcription (RT)-PCR techniques. The expression of etr1-1 was significant in flowers and buds. Transgenic lines did not differ morphologically from control plants. The selected transgenic T0 lines, which were re-established from in vitro cultures showed the same degree of tolerance to exogenous ethylene, confirming the stability of the transgene in in vitro cultures. The rooting ability of the transgenic plants was not affected by the presence of etr1-1. T1 progeny were produced by crossing the transgenic line, which showed the most significant reduction in ethylene sensitivity with a control plant, and the analysis of the T1 plants showed 1:1 segregation in terms of ethylene sensitivity and the presence of the transgene.

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

NAA:

α-Naphthalene acetic acid

RT-PCR:

Reverse transcription PCR

TDZ:

Thidiazuron

References

  • Abeles FB, Morgan PW, Saltveit ME (1992) Ethylene in Plant Biology, 2nd edn. Academic Press, San Diego

    Google Scholar 

  • Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: A new generation of protein database search programs. Nuc Acids Res 25:3389–3402

    Article  CAS  Google Scholar 

  • Angenent GC, Busscher M, Franken J, Mol JNM, van Tunen AJ (1992) Differential expression of two MADS box genes in wild-type and mutant petunia flowers. Plant Cell 4:983–989

    Article  PubMed  CAS  Google Scholar 

  • Bentzen P, Cook D, Denti D, Harris AS, Hofman J, Wright JM (1990) One tube DNA extraction procedure for molecular fingerprinting. Fingerprint News 2:17–21

    Google Scholar 

  • Bovy AG, Angenent GC, Dons HJM, van Altvorst AC (1999) Heterologous expression of the Arabidopsis etr1-1 allele inhibits the senescence of carnation flowers. Mol Breed 5:301–308

    Article  CAS  Google Scholar 

  • Chang C, Kwok SF, Bleecker AB, Meyerowitz EM (1993) Arabidopsis ethylene-response gene ETR1: Similarity of product to two-component regulators. Science 262:539–544

    Article  PubMed  CAS  Google Scholar 

  • Chandler SF, Lu C (2005) Biotechnology in ornamental horticulture. In Vitro Cell Dev Biol-Plant 41:591–601

    Article  Google Scholar 

  • Chen LFO, Huang JY, Wang YH, Chen YT, Shaw JF (2004) Ethylene insensitive and post-harvest yellowing retardation in mutant ethylene response sensor (boers) gene transformed broccoli (Brassica oleracea var: italica). Mol Breed 14:199–213

    Article  Google Scholar 

  • Clark DG, Gubrium EK, Barret JE, Nell TA, Klee HJ (1999) Root formation in ethylene insensitive plants. Plant Physiol 121:53–59

    Article  PubMed  CAS  Google Scholar 

  • Cui ML, Takada K, Ma B, Ezura H (2004) Overexpression of a mutated melon ethylene receptor gene Cm-ETR1/H69A confers reduced ethylene sensitivity in a heterologous plant, Nemesia strumosa. Plant Sci 167:253–258

    Article  CAS  Google Scholar 

  • Engelke T, Tatlioglu T (2000) Mitochondrial genome diversity in connection with male sterility in Allium schoenoprasum L. Theor Appl Genet 100:942–948

    Article  CAS  Google Scholar 

  • Gao Z, Xie X, Ling Y, Muthukrishnan S, Liang GH (2005) Agrobacterium tumefaciens-mediated sorghum transformation using a mannose selection system. Plant Biotechn J 3:591–599

    Article  CAS  Google Scholar 

  • Gubrium EK, Clark DG, Barrett JE, Nell TA (2000) Horticultural performance of transgenic ethylene insensitive Petunias. J Amer Soc Hort Sci 125:277–281

    Google Scholar 

  • Haines MM, Shiel PJ, Fellman JK, Berger PH (2003) Abnormalities in growth, development and physiological responses to biotic and abiotic stress in potato (Solanum tuberosum) transformed with Arabidopsis ETR1. J Agric Sci 141:333–347

    Article  CAS  Google Scholar 

  • Higgens D, Thompson J, Gibson T, Thompson JD, Higgens DG, Gibson TJ (1994) CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nuc Acids Res 22:4673–4680

    Article  Google Scholar 

  • Hoffmann T, Schmidt JS, Zheng X, Bent AF (1999) Isolation of ethylene insensitive soybean mutants that are altered in pathogen susceptibility and gene for gene resistance. Plant Physiol 119:935–949

    Article  Google Scholar 

  • Kim JH, Botella JR (2004) etr1-1 gene expression alters regeneration patterns in transgenic lettuce stimulating root formation. Plant Cell Tissu Organ Cult 78:69–73

    Article  CAS  Google Scholar 

  • Knoester M, van Loon LC, van der Heuvel J, Henning J, Bol JF, Linthorst JM (1998) Ethylene-insensitive tobacco lacks nonhost resistance against soil-borne fungi. Proc Natl Acad Sci USA 95:1933–1937

    Article  PubMed  CAS  Google Scholar 

  • Kumlehn J, Serazetdinova L, Hensel G, Becker D, Loerz H (2006) Genetic transformation of barley (Hordeum vulgare L.) via infection of androgenetic pollen cultures with Agrobacterium tumefaciens. Plant Biotech J 4(2):251–261

    Article  CAS  Google Scholar 

  • Lanahan MB, Yen H, Giovannoni JJ, Klee HJ (1994) The never ripe mutation blocks ethylene perception in tomato. Plant Cell 6:521–530

    Article  PubMed  CAS  Google Scholar 

  • Langston BJ, Bai S, Jones ML (2005) Increases in DNA fragmentation and induction of a senescence-specific nuclease are delayed during corolla senescence in ethylene-insensitive (etr1-1) transgenic petunias. J Exp Bot 56:15–23

    Article  PubMed  CAS  Google Scholar 

  • McAlvin CB, Stacey G (2005) Transgenic expression of the soybean apyrase in Lotus japonicus enhances nodulation. Plant Physiol 137(4):1456–1462

    Article  PubMed  CAS  Google Scholar 

  • Mudge KW (1998) Effect of ethylene on rooting. In: Davis TD, Haissig BE, Sankhla N (eds) Adventitious root formation in cuttings, advances in plant sciences series, vol 2, Dioscorides Press, Portland, pp 150–161

    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 

  • Nakagawa T, Takane K, Sugimoto T, Izui K, Kouchi H, Hata S (2003) Regulatory regions and nuclear factors involved in nodule-enhanced expression of soybean phosphoenolpyruvate carboxylase gene: implications for molecular evolution. Mol Gen Genomics 269:163–172

    Article  CAS  Google Scholar 

  • Ohtsubo N, Mitsuhara I, Koga M, Seo S, Ohashi Y (1999) Ethylene promotes the necrotic lesion formation and basic PR gene expression in TMR-infected tobacco. Plant Cell Physiol 40:808–817

    CAS  Google Scholar 

  • Serek M (1990) Effects of pre-harvest fertilization on the flower longevity of potted Campanula carpatica ‘Karl Foerster’. Sci Hort 44:119–126

    Article  Google Scholar 

  • Serek M, Tamari G, Sisler EC, Borochov A (1995a) Inhibition of ethylene-induced cellular senescence symptoms by 1-methylcyclopropene, a new inhibitor of ethylene action. Physiol Plant 94:229–232

    Article  CAS  Google Scholar 

  • Serek M, Sisler EC, Reid MS (1995b) Effects of 1-MCP on the vase life and ethylene response of cut flowers. Plant Growth Regul 16:93–97

    Article  CAS  Google Scholar 

  • Serek M, Sisler EC (2001) Efficacy of inhibitors of ethylene binding in improvement of the postharvest characteristics of potted flowering plants. Postharvest Biol Technol 23:161–166

    Article  CAS  Google Scholar 

  • Shaw JF, Chen HH, Tsai MF, Kuo CI, Huang LC (2002) Extended flower longevity of Petunia hybrida plants transformed with boers, a mutated ERS gene of Brassica oleracea. Mol Breed 9:211–216

    Article  CAS  Google Scholar 

  • Sisler EC, Serek M (1997) Inhibitors of ethylene responses in plants at the receptor level: Recent developments. Physiol Plant 100:577–582

    Article  CAS  Google Scholar 

  • Sisler EC, Serek M (2003) Compounds interacting with the ethylene receptor in plants. Plant Biol 5:473–480

    Article  Google Scholar 

  • Sriskandarajah S, Frello S, Jørgensen K, Serek M (2004) Agrobacterium tumefaciens—mediated transformation of Campanula carpatica: Factors affecting transformation and regeneration of transgenic shoots. Plant Cell Rep 23:59–63

    Article  PubMed  CAS  Google Scholar 

  • Srivastava V, Anderson OD, Ow DW (1999) Single-copy transgenic wheat generated through the resolution of complex integration patterns. Proc Natl Acad Sci USA 96:11117–11121

    Article  PubMed  CAS  Google Scholar 

  • Staba JE (1969) Plant tissue culture as a technique for the phytochemist. Recent Adv Phytochm 2:80

    Google Scholar 

  • Stepanova AN, Ecker JR (2000) Ethylene signaling: from mutants to molecules. Curr Opin Plant Biol 3(5):353–360

    Article  PubMed  Google Scholar 

  • Tanaka Y, Katsumoto Y, Brugliera F, Mason J (2005) Genetic engineering in floriculture. Plant Cell Tissue Organ Cult 80:1–24

    Article  CAS  Google Scholar 

  • Takada K, Ishimaru K, Kamada, H, Ezura H (2006) Anther-specific expression of mutated melon ethylene receptor gene Cm-ERS1/H70A affected tapetum degeneration and pollen grain production in transgenic tobacco plants. Plant Cell Rep 25:936–941

    Article  PubMed  CAS  Google Scholar 

  • Veen H (1983) Silver thiosulfate—an experimental tool in plant-science. Sci Hort 20:211–224

    Article  CAS  Google Scholar 

  • Wang Y, Kumar PP (2004) Heterologous expression of Arabidopsis ERS1 causes delayed senescence in coriander. Plant Cell Rep 22:678–683

    Article  PubMed  CAS  Google Scholar 

  • Wilkinson JQ, Lanahan MB, Clark DG, Bleeker AB, Chang C, Meyerowitz EM, Klee HJ (1997) A dominant mutant receptor from Arabidopsis confers ethylene insensitivity in heterologous plants. Nat Biotechnol 15:444–447

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

The project was funded by the Directorate for Food and Agricultural Business in Denmark (grant No. 3412-0401001) and Danish Campanula Growers: Gartneriet Thoruplund A/S (Odense) and Gartneriet Elmegaard ApS (Skælskør). The authors would like to thank Professor Gerco Angenent of Plant Research International, The Netherlands, for providing the bacterial strain and the growers Jørgen Andersen and Per Elmegård for the support and cooperation. The authors would also like to thank Dr. Bjarne Mejnert Stummann for critical reading of the manuscript and Annette Steding for her excellent technical assistance.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Sridevy Sriskandarajah.

Additional information

Communicated by P. P. Kumar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sriskandarajah, S., Mibus, H. & Serek, M. Transgenic Campanula carpatica plants with reduced ethylene sensitivity. Plant Cell Rep 26, 805–813 (2007). https://doi.org/10.1007/s00299-006-0291-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-006-0291-6

Keywords

Navigation