Green fluorescent protein (GFP) as a reporter gene for the plant pathogenic oomycete Phytophthora palmivora

P van West, B Reid, T A Campbell, R W Sandrock, W E Fry, S Kamoun, N A R Gow

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

Transgenic Phytophthora palmivora strains that produce green fluorescent protein (GFP) or beta-glucuronidase (GUS) constitutively were obtained after stable DNA integration using a polyethylene-glycol and CaCl2-based transformation protocol. GFP and GUS production were monitored during several stages of the life cycle of P. palmivora to evaluate their use in molecular and physiological studies, 40% of the GFP transformants produced the GFP to a level detectable by a confocal laser scanning microscope, whereas 75% of the GUS transformants produced GUS. GFP could be visualised readily in swimming zoospores and other developmental stages of P. palmivora cells. For high magnification microscopic studies, GFP is better visualised and was superior to GUS. In contrast, for macroscopic examination, GUS was superior. Our findings indicate that both GFP and GUS call be used successfully as reporter genes in P. palmivora. (C) 1999 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

Original languageEnglish
Pages (from-to)71-80
Number of pages10
JournalFEMS Microbiology Letters
Volume178
Publication statusPublished - 1999

Keywords

  • beta-glucuronidase
  • green fluorescent protein
  • oomycete
  • Phytophthora palmivora
  • REGULATORY SEQUENCES
  • INFESTANS
  • EXPRESSION
  • FUNGI
  • ZOOSPORES
  • PYTHIUM

Cite this

Green fluorescent protein (GFP) as a reporter gene for the plant pathogenic oomycete Phytophthora palmivora. / van West, P ; Reid, B ; Campbell, T A ; Sandrock, R W ; Fry, W E ; Kamoun, S ; Gow, N A R .

In: FEMS Microbiology Letters, Vol. 178, 1999, p. 71-80.

Research output: Contribution to journalArticle

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abstract = "Transgenic Phytophthora palmivora strains that produce green fluorescent protein (GFP) or beta-glucuronidase (GUS) constitutively were obtained after stable DNA integration using a polyethylene-glycol and CaCl2-based transformation protocol. GFP and GUS production were monitored during several stages of the life cycle of P. palmivora to evaluate their use in molecular and physiological studies, 40{\%} of the GFP transformants produced the GFP to a level detectable by a confocal laser scanning microscope, whereas 75{\%} of the GUS transformants produced GUS. GFP could be visualised readily in swimming zoospores and other developmental stages of P. palmivora cells. For high magnification microscopic studies, GFP is better visualised and was superior to GUS. In contrast, for macroscopic examination, GUS was superior. Our findings indicate that both GFP and GUS call be used successfully as reporter genes in P. palmivora. (C) 1999 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.",
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T1 - Green fluorescent protein (GFP) as a reporter gene for the plant pathogenic oomycete Phytophthora palmivora

AU - van West, P

AU - Reid, B

AU - Campbell, T A

AU - Sandrock, R W

AU - Fry, W E

AU - Kamoun, S

AU - Gow, N A R

PY - 1999

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N2 - Transgenic Phytophthora palmivora strains that produce green fluorescent protein (GFP) or beta-glucuronidase (GUS) constitutively were obtained after stable DNA integration using a polyethylene-glycol and CaCl2-based transformation protocol. GFP and GUS production were monitored during several stages of the life cycle of P. palmivora to evaluate their use in molecular and physiological studies, 40% of the GFP transformants produced the GFP to a level detectable by a confocal laser scanning microscope, whereas 75% of the GUS transformants produced GUS. GFP could be visualised readily in swimming zoospores and other developmental stages of P. palmivora cells. For high magnification microscopic studies, GFP is better visualised and was superior to GUS. In contrast, for macroscopic examination, GUS was superior. Our findings indicate that both GFP and GUS call be used successfully as reporter genes in P. palmivora. (C) 1999 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

AB - Transgenic Phytophthora palmivora strains that produce green fluorescent protein (GFP) or beta-glucuronidase (GUS) constitutively were obtained after stable DNA integration using a polyethylene-glycol and CaCl2-based transformation protocol. GFP and GUS production were monitored during several stages of the life cycle of P. palmivora to evaluate their use in molecular and physiological studies, 40% of the GFP transformants produced the GFP to a level detectable by a confocal laser scanning microscope, whereas 75% of the GUS transformants produced GUS. GFP could be visualised readily in swimming zoospores and other developmental stages of P. palmivora cells. For high magnification microscopic studies, GFP is better visualised and was superior to GUS. In contrast, for macroscopic examination, GUS was superior. Our findings indicate that both GFP and GUS call be used successfully as reporter genes in P. palmivora. (C) 1999 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

KW - beta-glucuronidase

KW - green fluorescent protein

KW - oomycete

KW - Phytophthora palmivora

KW - REGULATORY SEQUENCES

KW - INFESTANS

KW - EXPRESSION

KW - FUNGI

KW - ZOOSPORES

KW - PYTHIUM

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JO - FEMS Microbiology Letters

JF - FEMS Microbiology Letters

SN - 0378-1097

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