Plant Cell Reports

, Volume 11, Issue 9, pp 452–456

Endophytic bacteria expressing β-glucuronidase cause false positives in transformation of Dioscorea species

Authors

  • Mahmut Tör
    • Wye College, University of London
  • Sinclair H. Mantell
    • Wye College, University of London
  • Charles Ainsworth
    • Wye College, University of London
Article

DOI: 10.1007/BF00232689

Cite this article as:
Tör, M., Mantell, S.H. & Ainsworth, C. Plant Cell Reports (1992) 11: 452. doi:10.1007/BF00232689

Summary

False positive transformants obtained during plant transformation experiments on species of the monocotyledonous genus Dioscorea (yam) are described. The false positive results were found to be due to endophytic bacteria which exist within aseptically micropropagated shoot cultures and which express β-glucuronidase (GUS). The bacteria were isolated and identified as two species of Curtobacterium. The expression of GUS in these organisms was found to be induced by a variety of glucuronide substrates. The induction of GUS activity in the bacteria can be inhibited by chloramphenicol, tetracycline, ticarcillin and sodium azide. Implications of these results for use of the gus gene in plant transformation work are discussed.

Keywords

β-GlucuronidaseDioscoreaendophytic bacteriafalse positive transformants

Abbreviations

DTT

Dithiothreitol

EDTA

Ethylenediamine tetra-acetic acid

ELISA

Enzyme-linked immuno-sorbent assay

GUS

β-glucuronidase

LB

Luria Bertani

MUG

4-Methylumbelliferyl-β-D-glucuronide

PNPG

p-nitrophenyl-β-D-glucuronide

PVP

Polyvinylpyrrolidone

SDS

Sodium dodecyl sulphate

TAE

Tris-acetate-EDTA buffer

X-Gluc

5-Bromo-4-chloro-3-indolyl-β-D-glucuronide

Copyright information

© Springer-Verlag 1992