Abstract
Key message
Over-expression of the potato Gibberellin Stimulated-Like 2 ( GSL2 ) gene in transgenic potato confers resistance to blackleg disease incited by Pectobacterium atrosepticum and confirms a role for GSL2 in plant defence.
Abstract
The Gibberellin Stimulated-Like 2 (GSL2) gene (also known as Snakin 2) encodes a cysteine-rich, low-molecular weight antimicrobial peptide produced in potato plants. This protein is thought to play important roles in the innate defence against invading microbes. Over-expression of the GSL2 gene in potato (cultivar Iwa) was achieved using Agrobacterium-mediated gene transfer of a plant expression vector with the potato GSL2 gene under the regulatory control elements of the potato light-inducible Lhca3 gene. The resulting plants were confirmed as being transgenic by PCR, and subsequently analysed for transcriptional expression of the Lhca3-GSL2-Lhca3 chimeric potato gene. Quantitative RT-PCR analysis demonstrated that the majority of the transgenic potato lines over-expressed the GSL2 gene at the mRNA level. Based on qRT-PCR results and evaluation of phenotypic appearance, eight lines were selected for further characterisation and evaluated in bioassays for resistance to Pectobacterium atrosepticum (formerly Erwinia carotovora subsp. atroseptica), the causal agent of blackleg in potato. Three independent pathogenicity bioassays showed that transgenic lines with significantly increased transcriptional expression of the GSL2 gene exhibit resistance to blackleg disease. This establishes a functional role for GSL2 in plant defence against pathogens in potato.
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Acknowledgments
We thank David Lewis, Ian King and Stuart Larsen for providing plant growth facilities, Ruth Butler for help with analysis of the qPCR data to determine the density of P. atrosepticum and Alasdair Noble for assistance with analysis of variance. This work was supported by AGMARDT Grant 20791 and Ministry of Business, Innovation and Employment (New Zealand) contract C02X0202 to The New Zealand Institute for Plant & Food Research Ltd.
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The authors declare that they have no conflict of interest.
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The authors declare that the experiments comply with the laws of New Zealand.
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Communicated by H. J. van Eck.
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Mohan, S., Meiyalaghan, S., Latimer, J.M. et al. GSL2 over-expression confers resistance to Pectobacterium atrosepticum in potato. Theor Appl Genet 127, 677–689 (2014). https://doi.org/10.1007/s00122-013-2250-2
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DOI: https://doi.org/10.1007/s00122-013-2250-2