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 Taylor (Lead / Corresponding author)

    Research output: Contribution to journalArticle

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    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 ß-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.
    Original languageEnglish
    Pages (from-to)1381-1392
    Number of pages12
    JournalProtoplasma
    Volume250
    Issue number6
    DOIs
    Publication statusPublished - Dec 2013

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    Carotenoids
    Solanum tuberosum
    carotenoids
    potatoes
    Enzymes
    enzymes
    amyloplasts
    Plastids
    tubers
    storage organs
    biochemical pathways
    engineering
    Genetically Modified Plants
    isoprenoids
    Biosynthetic Pathways
    Terpenes
    Metabolic Networks and Pathways
    Confocal Microscopy
    Color
    genetically modified organisms

    Cite this

    Pasare, S., Wright, K., Campbell, R., Morris, W., Ducreux, L., Chapman, S., ... Taylor, M. (2013). The sub-cellular localisation of the potato (Solanum tuberosum L.) carotenoid biosynthetic enzymes, CrtRb2 and PSY2. Protoplasma, 250(6), 1381-1392. https://doi.org/10.1007/s00709-013-0521-z
    Pasare, Stefania ; Wright, Kathryn ; Campbell, Raymond ; Morris, Wayne ; Ducreux, Laurence ; Chapman, Sean ; Bramley, Peter ; Fraser, Paul ; Roberts, Alison ; Taylor, Mark. / The sub-cellular localisation of the potato (Solanum tuberosum L.) carotenoid biosynthetic enzymes, CrtRb2 and PSY2. In: Protoplasma. 2013 ; Vol. 250, No. 6. pp. 1381-1392.
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    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 {\ss}-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.",
    author = "Stefania Pasare and Kathryn Wright and Raymond Campbell and Wayne Morris and Laurence Ducreux and Sean Chapman and Peter Bramley and Paul Fraser and Alison Roberts and Mark Taylor",
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    Pasare, S, Wright, K, Campbell, R, Morris, W, Ducreux, L, Chapman, S, Bramley, P, Fraser, P, Roberts, A & Taylor, M 2013, 'The sub-cellular localisation of the potato (Solanum tuberosum L.) carotenoid biosynthetic enzymes, CrtRb2 and PSY2', Protoplasma, vol. 250, no. 6, pp. 1381-1392. https://doi.org/10.1007/s00709-013-0521-z

    The sub-cellular localisation of the potato (Solanum tuberosum L.) carotenoid biosynthetic enzymes, CrtRb2 and PSY2. / Pasare, Stefania; Wright, Kathryn; Campbell, Raymond; Morris, Wayne; Ducreux, Laurence; Chapman, Sean; Bramley, Peter; Fraser, Paul; Roberts, Alison; Taylor, Mark (Lead / Corresponding author).

    In: Protoplasma, Vol. 250, No. 6, 12.2013, p. 1381-1392.

    Research output: Contribution to journalArticle

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    AU - Pasare, Stefania

    AU - Wright, Kathryn

    AU - Campbell, Raymond

    AU - Morris, Wayne

    AU - Ducreux, Laurence

    AU - Chapman, Sean

    AU - Bramley, Peter

    AU - Fraser, Paul

    AU - Roberts, Alison

    AU - Taylor, Mark

    PY - 2013/12

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    N2 - 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 ß-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.

    AB - 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 ß-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.

    U2 - 10.1007/s00709-013-0521-z

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