Molecular detection and beta-glucuronidase expression of gus-marked Bacillus subtilis L-form bacteria in developing Chinese cabbage seedlings

E Tsomlexoglou, P W H K P Daulagala, G W Gooday, L A Glover, B Seddon, E J Allan

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Aim: To detect L-form bacteria in developing Chinese cabbage seedlings.

Methods and Results: Stable Bacillus subtilis L-forms were genetically modified to express the gus gene (encoding beta -glucuronidase). Germinated seeds of Chinese cabbage were soaked in mannitol based suspensions of the L-form bacteria or with mannitol alone and after washing were grown in aseptic conditions on plant growth medium. Histochemical staining of beta-glucuronidase activity (X-gluc) and Polymerase Chain Reaction (PCR) detection of the gus gene were achieved in the L-form associated seedlings. beta-Glucuronidase was localized in discrete spots, mainly in the roots with staining, and was also observed in the cotyledons and base of stems. Correlation was observed between PCR detection of the gus gene and histochemical staining with detection in similar tissues. Stable L-form bacteria were non-culturable after their association with plant material.

Conclusions: The gus reporter gene system with its associated histological staining for enzyme activity was used successfully for detecting B. subtilis L-form bacteria in plant material.

Significance and Impact of the Study: These molecular marked L-forms should provide a specific and sensitive technique for detecting L-form bacteria in planta and offer a method for further understanding the L-form/plant association.

Original languageEnglish
Pages (from-to)218-224
Number of pages7
JournalJournal of Applied Microbiology
Volume95
Publication statusPublished - 2003

Keywords

  • beta-glucuronidase
  • Bacillus subtilis
  • gus gene
  • L-forms
  • Polymerase Chain Reaction
  • symbioses
  • SYRINGAE PV PHASEOLICOLA
  • GENE
  • PLANTS
  • TRANSFORMATION
  • ASSOCIATIONS
  • INDUCTION
  • SYMBIOSIS
  • DISEASE
  • GROWTH

Cite this

Tsomlexoglou, E., Daulagala, P. W. H. K. P., Gooday, G. W., Glover, L. A., Seddon, B., & Allan, E. J. (2003). Molecular detection and beta-glucuronidase expression of gus-marked Bacillus subtilis L-form bacteria in developing Chinese cabbage seedlings. Journal of Applied Microbiology, 95, 218-224.

Molecular detection and beta-glucuronidase expression of gus-marked Bacillus subtilis L-form bacteria in developing Chinese cabbage seedlings. / Tsomlexoglou, E ; Daulagala, P W H K P ; Gooday, G W ; Glover, L A ; Seddon, B ; Allan, E J .

In: Journal of Applied Microbiology, Vol. 95, 2003, p. 218-224.

Research output: Contribution to journalArticle

Tsomlexoglou, E ; Daulagala, P W H K P ; Gooday, G W ; Glover, L A ; Seddon, B ; Allan, E J . / Molecular detection and beta-glucuronidase expression of gus-marked Bacillus subtilis L-form bacteria in developing Chinese cabbage seedlings. In: Journal of Applied Microbiology. 2003 ; Vol. 95. pp. 218-224.
@article{8f68ed560c464cd489e3986315cb7375,
title = "Molecular detection and beta-glucuronidase expression of gus-marked Bacillus subtilis L-form bacteria in developing Chinese cabbage seedlings",
abstract = "Aim: To detect L-form bacteria in developing Chinese cabbage seedlings.Methods and Results: Stable Bacillus subtilis L-forms were genetically modified to express the gus gene (encoding beta -glucuronidase). Germinated seeds of Chinese cabbage were soaked in mannitol based suspensions of the L-form bacteria or with mannitol alone and after washing were grown in aseptic conditions on plant growth medium. Histochemical staining of beta-glucuronidase activity (X-gluc) and Polymerase Chain Reaction (PCR) detection of the gus gene were achieved in the L-form associated seedlings. beta-Glucuronidase was localized in discrete spots, mainly in the roots with staining, and was also observed in the cotyledons and base of stems. Correlation was observed between PCR detection of the gus gene and histochemical staining with detection in similar tissues. Stable L-form bacteria were non-culturable after their association with plant material.Conclusions: The gus reporter gene system with its associated histological staining for enzyme activity was used successfully for detecting B. subtilis L-form bacteria in plant material.Significance and Impact of the Study: These molecular marked L-forms should provide a specific and sensitive technique for detecting L-form bacteria in planta and offer a method for further understanding the L-form/plant association.",
keywords = "beta-glucuronidase, Bacillus subtilis, gus gene, L-forms, Polymerase Chain Reaction, symbioses, SYRINGAE PV PHASEOLICOLA, GENE, PLANTS, TRANSFORMATION, ASSOCIATIONS, INDUCTION, SYMBIOSIS, DISEASE, GROWTH",
author = "E Tsomlexoglou and Daulagala, {P W H K P} and Gooday, {G W} and Glover, {L A} and B Seddon and Allan, {E J}",
year = "2003",
language = "English",
volume = "95",
pages = "218--224",
journal = "Journal of Applied Microbiology",
issn = "1364-5072",
publisher = "Wiley-Blackwell",

}

TY - JOUR

T1 - Molecular detection and beta-glucuronidase expression of gus-marked Bacillus subtilis L-form bacteria in developing Chinese cabbage seedlings

AU - Tsomlexoglou, E

AU - Daulagala, P W H K P

AU - Gooday, G W

AU - Glover, L A

AU - Seddon, B

AU - Allan, E J

PY - 2003

Y1 - 2003

N2 - Aim: To detect L-form bacteria in developing Chinese cabbage seedlings.Methods and Results: Stable Bacillus subtilis L-forms were genetically modified to express the gus gene (encoding beta -glucuronidase). Germinated seeds of Chinese cabbage were soaked in mannitol based suspensions of the L-form bacteria or with mannitol alone and after washing were grown in aseptic conditions on plant growth medium. Histochemical staining of beta-glucuronidase activity (X-gluc) and Polymerase Chain Reaction (PCR) detection of the gus gene were achieved in the L-form associated seedlings. beta-Glucuronidase was localized in discrete spots, mainly in the roots with staining, and was also observed in the cotyledons and base of stems. Correlation was observed between PCR detection of the gus gene and histochemical staining with detection in similar tissues. Stable L-form bacteria were non-culturable after their association with plant material.Conclusions: The gus reporter gene system with its associated histological staining for enzyme activity was used successfully for detecting B. subtilis L-form bacteria in plant material.Significance and Impact of the Study: These molecular marked L-forms should provide a specific and sensitive technique for detecting L-form bacteria in planta and offer a method for further understanding the L-form/plant association.

AB - Aim: To detect L-form bacteria in developing Chinese cabbage seedlings.Methods and Results: Stable Bacillus subtilis L-forms were genetically modified to express the gus gene (encoding beta -glucuronidase). Germinated seeds of Chinese cabbage were soaked in mannitol based suspensions of the L-form bacteria or with mannitol alone and after washing were grown in aseptic conditions on plant growth medium. Histochemical staining of beta-glucuronidase activity (X-gluc) and Polymerase Chain Reaction (PCR) detection of the gus gene were achieved in the L-form associated seedlings. beta-Glucuronidase was localized in discrete spots, mainly in the roots with staining, and was also observed in the cotyledons and base of stems. Correlation was observed between PCR detection of the gus gene and histochemical staining with detection in similar tissues. Stable L-form bacteria were non-culturable after their association with plant material.Conclusions: The gus reporter gene system with its associated histological staining for enzyme activity was used successfully for detecting B. subtilis L-form bacteria in plant material.Significance and Impact of the Study: These molecular marked L-forms should provide a specific and sensitive technique for detecting L-form bacteria in planta and offer a method for further understanding the L-form/plant association.

KW - beta-glucuronidase

KW - Bacillus subtilis

KW - gus gene

KW - L-forms

KW - Polymerase Chain Reaction

KW - symbioses

KW - SYRINGAE PV PHASEOLICOLA

KW - GENE

KW - PLANTS

KW - TRANSFORMATION

KW - ASSOCIATIONS

KW - INDUCTION

KW - SYMBIOSIS

KW - DISEASE

KW - GROWTH

M3 - Article

VL - 95

SP - 218

EP - 224

JO - Journal of Applied Microbiology

JF - Journal of Applied Microbiology

SN - 1364-5072

ER -