Plant regeneration from different explant types of Bituminaria bituminosa and furanocoumarin content along plant regeneration stages

Maria Pazos-Navarro, J.A. Del Río, A. Ortuño, P. Romero-Espinar, E. Correal, M. Dabauza

    Research output: Contribution to journalArticle

    9 Citations (Scopus)

    Abstract

    The first protocol for in vitro plant regeneration from different explants of Bituminaria bituminosa, a pasture and medicinal species, has been established. Three explant types (petiole, leaflet and petiole-leaflet attachment "PLA") cultured on media with different combinations of benzylaminopurine (BA; 5.0, 10.0 or 20.0 μM) and naphthalene acetic acid (NAA) or indole acetic acid (IAA; 0.5 or 5.0 μM) were tested for calli induction, and with 5 μM BA + 0. 5 μM NAA or IAA for shoot development. The average number of shoots (≥5 mm) per callus depended on the explant type and the calli induction medium. The highest average number of shoots per callus was achieved by culturing leaflet and PLA explants on 5 μM IAA + 10 μM BA for calli induction and on 0. 5 μM IAA + 5 μM BA for shoot development, and by culturing petiole explants on 0. 5 μM NAA + 10 μM BA followed by a second culture on 0. 5 μM NAA + 5 μM BA. The highest frequency of shoot rooting was achieved with 10. 0 μM NAA and 1. 0 μM gibberellic acid (GA3). Rooted plants were acclimatised in a culture chamber, reaching 96 % survival. Acclimatised plants were transferred to a greenhouse and finally to the field, reaching 100 % survival. The furanocoumarin (FC) accumulation was evaluated in organogenic calli, in vitro shoots, ex vitro plants in the greenhouse and in ex vitro plants in the field (after 1 and 4 months of acclimatisation). The content of FCs depended on the plant material evaluated, being higher in ex vitro plants in the field (up to 9,824 μg g-1 DW total FC) and lowest in organogenic calli (up to 50 μg g-1 DW total FC). This effect may be due to cell organization, longer exposure to environmental factors and the developmental stage. © 2013 Springer Science+Business Media Dordrecht.
    Original languageEnglish
    Pages (from-to)123-129
    JournalPlant Growth Regulation
    Volume70
    Issue number2
    DOIs
    Publication statusPublished - 2013

    Fingerprint

    Bituminaria bituminosa
    psoralens
    Bony Callus
    Regeneration
    explants
    callus
    naphthaleneacetic acid
    Acetic Acid
    indole acetic acid
    shoots
    greenhouses
    Acclimatization
    Environmental Exposure
    Furocoumarins
    gibberellic acid
    rooting
    acclimation
    pastures
    In Vitro Techniques
    naphthalene

    Cite this

    Pazos-Navarro, Maria ; Del Río, J.A. ; Ortuño, A. ; Romero-Espinar, P. ; Correal, E. ; Dabauza, M. / Plant regeneration from different explant types of Bituminaria bituminosa and furanocoumarin content along plant regeneration stages. In: Plant Growth Regulation. 2013 ; Vol. 70, No. 2. pp. 123-129.
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    abstract = "The first protocol for in vitro plant regeneration from different explants of Bituminaria bituminosa, a pasture and medicinal species, has been established. Three explant types (petiole, leaflet and petiole-leaflet attachment {"}PLA{"}) cultured on media with different combinations of benzylaminopurine (BA; 5.0, 10.0 or 20.0 μM) and naphthalene acetic acid (NAA) or indole acetic acid (IAA; 0.5 or 5.0 μM) were tested for calli induction, and with 5 μM BA + 0. 5 μM NAA or IAA for shoot development. The average number of shoots (≥5 mm) per callus depended on the explant type and the calli induction medium. The highest average number of shoots per callus was achieved by culturing leaflet and PLA explants on 5 μM IAA + 10 μM BA for calli induction and on 0. 5 μM IAA + 5 μM BA for shoot development, and by culturing petiole explants on 0. 5 μM NAA + 10 μM BA followed by a second culture on 0. 5 μM NAA + 5 μM BA. The highest frequency of shoot rooting was achieved with 10. 0 μM NAA and 1. 0 μM gibberellic acid (GA3). Rooted plants were acclimatised in a culture chamber, reaching 96 {\%} survival. Acclimatised plants were transferred to a greenhouse and finally to the field, reaching 100 {\%} survival. The furanocoumarin (FC) accumulation was evaluated in organogenic calli, in vitro shoots, ex vitro plants in the greenhouse and in ex vitro plants in the field (after 1 and 4 months of acclimatisation). The content of FCs depended on the plant material evaluated, being higher in ex vitro plants in the field (up to 9,824 μg g-1 DW total FC) and lowest in organogenic calli (up to 50 μg g-1 DW total FC). This effect may be due to cell organization, longer exposure to environmental factors and the developmental stage. {\circledC} 2013 Springer Science+Business Media Dordrecht.",
    author = "Maria Pazos-Navarro and {Del R{\'i}o}, J.A. and A. Ortu{\~n}o and P. Romero-Espinar and E. Correal and M. Dabauza",
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    Plant regeneration from different explant types of Bituminaria bituminosa and furanocoumarin content along plant regeneration stages. / Pazos-Navarro, Maria; Del Río, J.A.; Ortuño, A.; Romero-Espinar, P.; Correal, E.; Dabauza, M.

    In: Plant Growth Regulation, Vol. 70, No. 2, 2013, p. 123-129.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Plant regeneration from different explant types of Bituminaria bituminosa and furanocoumarin content along plant regeneration stages

    AU - Pazos-Navarro, Maria

    AU - Del Río, J.A.

    AU - Ortuño, A.

    AU - Romero-Espinar, P.

    AU - Correal, E.

    AU - Dabauza, M.

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    AB - The first protocol for in vitro plant regeneration from different explants of Bituminaria bituminosa, a pasture and medicinal species, has been established. Three explant types (petiole, leaflet and petiole-leaflet attachment "PLA") cultured on media with different combinations of benzylaminopurine (BA; 5.0, 10.0 or 20.0 μM) and naphthalene acetic acid (NAA) or indole acetic acid (IAA; 0.5 or 5.0 μM) were tested for calli induction, and with 5 μM BA + 0. 5 μM NAA or IAA for shoot development. The average number of shoots (≥5 mm) per callus depended on the explant type and the calli induction medium. The highest average number of shoots per callus was achieved by culturing leaflet and PLA explants on 5 μM IAA + 10 μM BA for calli induction and on 0. 5 μM IAA + 5 μM BA for shoot development, and by culturing petiole explants on 0. 5 μM NAA + 10 μM BA followed by a second culture on 0. 5 μM NAA + 5 μM BA. The highest frequency of shoot rooting was achieved with 10. 0 μM NAA and 1. 0 μM gibberellic acid (GA3). Rooted plants were acclimatised in a culture chamber, reaching 96 % survival. Acclimatised plants were transferred to a greenhouse and finally to the field, reaching 100 % survival. The furanocoumarin (FC) accumulation was evaluated in organogenic calli, in vitro shoots, ex vitro plants in the greenhouse and in ex vitro plants in the field (after 1 and 4 months of acclimatisation). The content of FCs depended on the plant material evaluated, being higher in ex vitro plants in the field (up to 9,824 μg g-1 DW total FC) and lowest in organogenic calli (up to 50 μg g-1 DW total FC). This effect may be due to cell organization, longer exposure to environmental factors and the developmental stage. © 2013 Springer Science+Business Media Dordrecht.

    U2 - 10.1007/s10725-013-9784-9

    DO - 10.1007/s10725-013-9784-9

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