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Protoplasma

, Volume 250, Issue 6, pp 1381–1392 | Cite as

The sub-cellular localisation of the potato (Solanum tuberosum L.) carotenoid biosynthetic enzymes, CrtRb2 and PSY2

  • Stefania Pasare
  • Kathryn Wright
  • Raymond Campbell
  • Wayne Morris
  • Laurence Ducreux
  • Sean Chapman
  • Peter Bramley
  • Paul Fraser
  • Alison Roberts
  • Mark TaylorEmail author
Original Article

Abstract

Carotenoids are isoprenoids with important biological roles both for plants and animals. The yellow flesh colour of potato (Solanum tuberosum L.) tubers is a quality trait dependent on the types and levels of carotenoids that accumulate. The carotenoid biosynthetic pathway is well characterised, facilitating the successful engineering of carotenoid content in numerous crops including potato. However, a clear understanding concerning the factors regulating carotenoid accumulation and localisation in plant storage organs, such as tubers, is lacking. In the present study, the localisation of key carotenoid biosynthetic enzymes was investigated, as one of the unexplored factors that could influence the accumulation of carotenoids in potato tubers. Stable transgenic potato plants were generated by over-expressing β-CAROTENE HYDROXYLASE 2 (CrtRb2) and PHYTOENE SYNTHASE 2 (PSY2) genes, fused to red fluorescent protein (RFP). Gene expression and carotenoid levels were both significantly increased, confirming functionality of the fluorescently tagged proteins. Confocal microscopy studies revealed different sub-organellar localisations of CrtRb2-RFP and PSY2-RFP within amyloplasts. CrtRb2 was detected in small vesicular structures, inside amyloplasts, whereas PSY2 was localised in the stroma of amyloplasts. We conclude that it is important to consider the location of biosynthetic enzymes when engineering the carotenoid metabolic pathway in storage organs such as tubers.

Keywords

Amyloplast Carotenoid Localisation Plastid Potato Tuber 

Abbreviations

CrtRb

β-Carotene hydroxylase

DiOC6

3,3′-Dihexyloxacarbocyanine iodide

GFP

Green fluorescent protein

HPLC

High-performance liquid chromatography

PG

Plastoglobuli

PSY

PHYTOENE SYNTHASE

RFP

Red fluorescent protein

Notes

Acknowledgments

This work was funded by the Scottish Government Rural and Environment Science and Analytical Services Division and EU-FP7 METAPRO 244348.

Conflict of interest

None.

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Copyright information

© Springer-Verlag Wien 2013

Authors and Affiliations

  • Stefania Pasare
    • 1
    • 2
  • Kathryn Wright
    • 1
  • Raymond Campbell
    • 1
  • Wayne Morris
    • 1
  • Laurence Ducreux
    • 1
  • Sean Chapman
    • 1
  • Peter Bramley
    • 2
  • Paul Fraser
    • 2
  • Alison Roberts
    • 1
  • Mark Taylor
    • 1
    Email author
  1. 1.Cell and Molecular Sciences, The James Hutton InstituteDundeeUK
  2. 2.School of Biological Sciences, Royal HollowayUniversity of LondonEghamUK

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