Visualisation of plastids in endosperm, pollen and roots of transgenic wheat expressing modified GFP fused to transit peptides from wheat SSU RubisCO, rice FtsZ and maize ferredoxin III proteins

Lucia F. Primavesi, Huixia Wu, Elisabeth A. Mudd, Anil Day, Huw D. Jones*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

30 Citations (SciVal)

Abstract

The ability to target marker proteins to specific subcellular compartments is a powerful research tool to study the structure and development of organelles. Here transit sequences from nuclear-encoded, plastid proteins, namely rice FtsZ, maize non-photosynthetic ferredoxin III (FdIII) and the small subunit of RubisCO were used to target a modified synthetic GFP (S65G, S72A) to plastids. The localisations of the fusion proteins expressed in transgenic wheat plants and under the control of the rice actin promoter were compared to an untargeted GFP control. GFP fluorescence was localised to non-green plastids in pollen, roots and seed endosperm and detected in isolated leaf chloroplasts using a GFP-specific antibody. Transit peptides appeared to influence the relative fluorescence intensities of plastids in different tissues. This is consistent with differential targeting and/or turnover of GFP fusion proteins in different plastid types. Replacement of GFP sequences with alternative coding regions enables immediate applications of our vectors for academic research and commercial applications.

Original languageEnglish
Pages (from-to)529-543
Number of pages15
JournalTransgenic Research
Volume17
Issue number4
Early online date21 Aug 2007
DOIs
Publication statusPublished - 01 Aug 2008
Externally publishedYes

Keywords

  • GFP
  • wheat
  • confocal microscopy
  • non-green plastids
  • protein targeting
  • stromule
  • GREEN FLUORESCENT PROTEIN
  • HIGH-LEVEL EXPRESSION
  • CHLOROPLAST DIVISION
  • CEREAL ENDOSPERM
  • STARCH SYNTHESIS
  • GENE FAMILIES
  • TRANSFORMATION
  • PLANTS
  • PROMOTER
  • IMPORT

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