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Plant defense gene promoter enhances the reliability of shiva-1 gene-induced resistance to soft rot disease in potato

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Abstract

PAL5, a tomato (Lycopersicon esculentum Mill.) plant defense gene that encodes phenylalanine ammonia-lyase, is known to respond to a variety of environmental stresses including pathogen infection and wounding. A shiva-1 gene recombinant that encodes a small synthetic antibacterial peptide under the PAL5 gene promoter was transformed into potato (Solanum tuberosum L.) and its ability to induce resistance to Erwinia carotovora was compared with a construct under the control of the constitutive and widely used cauliflower mosaic virus (CaMV) 35S promoter. The shiva-1 peptide, an analog of natural cecropin B, was shown previously to have high bactericidal activity in vitro, but when expressed in vivo under the control of the CaMV 35S promoter, the effects were very inconsistent. As observed previously, in the present studies a few transformants with the CaMV 35S promoter were highly resistant when assayed for susceptibility to soft rot disease. In marked contrast the majority of transformants with the PAL5 gene promoter were highly resistant. More-detailed analyses of the incorporated DNA indicated that most of the transformants with the CaMV 35S promoter contained multiple copies of the transforming DNA while all of the PAL5 recombinants contained single copies. The highly resistant CaMV 35S recombinant also was present as a single copy. The results indicate that, at least in this instance, a constitutive promoter may not be ideal for the effective expression of a foreign gene and suggest that multiple insertions may have negative consequences.

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Abbreviations

CaMV:

Cauliflower mosaic virus

GUS:

β-Glucuronidase

NOS:

Nopaline synthase

NPTII:

Neomycin phosphotransferase II

PAL:

Phenylalanine ammonia-lyase

PCR:

Polymerase chain reaction

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Acknowledgements

This study was supported by the BioGreen 21 Project, RDA, Korea and the Natural Sciences and Engineering Research Council of Canada.

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Authors and Affiliations

  1. Department of Crop Biotechnology, Jinju National University, 660-758, Jinju, Korea

    Shin Woo Lee

  2. National Institute of Alpine Agriculture, RDA, 232-955, Pyeongchang, Korea

    Jung Yoon Yi & Hyo Won Seo

  3. Biological Sciences, Chonbuk National University, 561-756, Chonju, Korea

    Moon Sik Yang

  4. Department of Molecular Biology & Genetics, University of Guelph, Guelph, Ontario, N1G 2W1, Canada

    E. Jane Robb & Ross N. Nazar

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  1. Jung Yoon Yi
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  2. Hyo Won Seo
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  3. Moon Sik Yang
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  4. E. Jane Robb
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  6. Shin Woo Lee
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Correspondence to Shin Woo Lee.

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Yi, J.Y., Seo, H.W., Yang, M.S. et al. Plant defense gene promoter enhances the reliability of shiva-1 gene-induced resistance to soft rot disease in potato. Planta 220, 165–171 (2004). https://doi.org/10.1007/s00425-004-1346-y

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