An effective and reproducible transformation protocol for the model resurrection plant Craterostigma plantagineum Hochst.

O. Toldi, S. Tóth, T. Pónyi, P. Scott

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Procedures previously established for plant regeneration and Agrobacterium tumefaciens-mediated genetic transformation of the desiccation-tolerant plant, Craterostigma plantagineum, have been further developed. A highly effective tissue culture system was established based on the integrated optimisation of antioxidant and growth regulator composition and the stabilisation of the pH of the culture media by means of a potassium phosphate buffer. The undesirable hyperhydricity of Craterostigma tissue in tissue culture was also circumvented by these modifications, which serve as an alternative to the previously described procedures. The high efficiency of plant regeneration from the callus phase provided the basis for optimising genetic transformation in Craterostigma. For gene delivery, both a standard (method A) and a modified protocol (method B), the latter having previously resulted in successful Agrobacterium-mediated transformation of monocot cereals, were applied. Physical and biochemical key variables in transformation were evaluated, such as gene gun-mediated microwounding of plant explants, infiltration of Agrobacterium suspension cultures into target tissues and the influence of in vitro pre-induction of vir genes. While the physical enhancement of Agrobacterium infection (microwounding, infiltration) had no positive effect, the in vitro pre-induction of vir genes (biochemical enhancement) resulted in a twofold increase in the transformation frequency as compared to the conventional protocol (method A).

Original languageEnglish
Pages (from-to)63-69
Number of pages7
JournalPlant Cell Reports
Volume21
Issue number1
DOIs
Publication statusPublished - 2002

Fingerprint

Agrobacterium
gene induction
genetic transformation
tissue culture
hyperhydricity
potassium phosphates
Liliopsida
Agrobacterium radiobacter
growth regulators
gene transfer
explants
callus
buffers
culture media
methodology
antioxidants
Craterostigma plantagineum
infection
genes
Craterostigma

Keywords

  • Desiccation tolerance
  • Genetic transformation
  • Hyperhydricity

ASJC Scopus subject areas

  • Plant Science

Cite this

An effective and reproducible transformation protocol for the model resurrection plant Craterostigma plantagineum Hochst. / Toldi, O.; Tóth, S.; Pónyi, T.; Scott, P.

In: Plant Cell Reports, Vol. 21, No. 1, 2002, p. 63-69.

Research output: Contribution to journalArticle

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