Effect of growth regulators on transformed root cultures of Hyoscyamus muticus

Leena Vanhala, Manu Eeva, Seppo Lapinjoki, Raimo Hiltunen, Kirsi-Marja Oksman-Caldentey (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

29 Citations (Scopus)

Abstract

The transformed root cultures of Hyoscyamus muticus are usually cultivated in hormone-free medium in which the production of hyoscyamine is high.
We studied whether the addition of exogenous growth regulators could enhance tropane alkaloid accumulation without affecting the growth and root phenotype.
In order to rapidly screen a large number of hairy root samples a monoclonal antibody based enzyme immunological assay was developed for scopolamine analysis.

The hairy root cultures tolerated exogenous auxins in the concentration range 0.01–5μmol/L well. The growth of the roots was not significantly affected by IAA and NAA, but the alkaloid accumulation was doubled compared with that of the roots growing in the absence of auxin.
At optimum IAA concentration (0.1–1.0 μmol/L) the maximum hyoscyamine content was 117 mg L−1 and scopolamine content 0.2mgL−1. In general, the scopolamine contents were by a factor of 300–500 lower than the hyoscyamine ones.
The phenotype, growth rate and alkaloid accumulation of the roots did not change despite cytokinin addition. Both ABA and GA3 strongly inhibited hyoscyamine accumulation. However, the inhibition was not associated with the changes in root morphology.

We suggest that endogenous auxin production in the roots is not sufficient for maximum tropane alkaloid accumulation.
To our knowledge this is the first time that alkaloid accumulation has been significantly increased by exogenous auxins without influencing growth.

Original languageEnglish
Pages (from-to)475-481
JournalJournal of Plant Physiology
Volume153
Issue number3-4
DOIs
Publication statusPublished - 1998
MoE publication typeA1 Journal article-refereed

Fingerprint

Hyoscyamus
Hyoscyamus muticus
Hyoscyamine
growth regulators
Alkaloids
Indoleacetic Acids
Scopolamine Hydrobromide
Tropanes
atropine
Growth
scopolamine
auxins
tropane alkaloids
alkaloids
Phenotype
Cytokinins
indole acetic acid
Enzyme Assays
phenotype
Monoclonal Antibodies

Cite this

Vanhala, Leena ; Eeva, Manu ; Lapinjoki, Seppo ; Hiltunen, Raimo ; Oksman-Caldentey, Kirsi-Marja. / Effect of growth regulators on transformed root cultures of Hyoscyamus muticus. In: Journal of Plant Physiology. 1998 ; Vol. 153, No. 3-4. pp. 475-481.
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abstract = "The transformed root cultures of Hyoscyamus muticus are usually cultivated in hormone-free medium in which the production of hyoscyamine is high. We studied whether the addition of exogenous growth regulators could enhance tropane alkaloid accumulation without affecting the growth and root phenotype. In order to rapidly screen a large number of hairy root samples a monoclonal antibody based enzyme immunological assay was developed for scopolamine analysis.The hairy root cultures tolerated exogenous auxins in the concentration range 0.01–5μmol/L well. The growth of the roots was not significantly affected by IAA and NAA, but the alkaloid accumulation was doubled compared with that of the roots growing in the absence of auxin. At optimum IAA concentration (0.1–1.0 μmol/L) the maximum hyoscyamine content was 117 mg L−1 and scopolamine content 0.2mgL−1. In general, the scopolamine contents were by a factor of 300–500 lower than the hyoscyamine ones. The phenotype, growth rate and alkaloid accumulation of the roots did not change despite cytokinin addition. Both ABA and GA3 strongly inhibited hyoscyamine accumulation. However, the inhibition was not associated with the changes in root morphology.We suggest that endogenous auxin production in the roots is not sufficient for maximum tropane alkaloid accumulation. To our knowledge this is the first time that alkaloid accumulation has been significantly increased by exogenous auxins without influencing growth.",
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Effect of growth regulators on transformed root cultures of Hyoscyamus muticus. / Vanhala, Leena; Eeva, Manu; Lapinjoki, Seppo; Hiltunen, Raimo; Oksman-Caldentey, Kirsi-Marja (Corresponding Author).

In: Journal of Plant Physiology, Vol. 153, No. 3-4, 1998, p. 475-481.

Research output: Contribution to journalArticleScientificpeer-review

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N2 - The transformed root cultures of Hyoscyamus muticus are usually cultivated in hormone-free medium in which the production of hyoscyamine is high. We studied whether the addition of exogenous growth regulators could enhance tropane alkaloid accumulation without affecting the growth and root phenotype. In order to rapidly screen a large number of hairy root samples a monoclonal antibody based enzyme immunological assay was developed for scopolamine analysis.The hairy root cultures tolerated exogenous auxins in the concentration range 0.01–5μmol/L well. The growth of the roots was not significantly affected by IAA and NAA, but the alkaloid accumulation was doubled compared with that of the roots growing in the absence of auxin. At optimum IAA concentration (0.1–1.0 μmol/L) the maximum hyoscyamine content was 117 mg L−1 and scopolamine content 0.2mgL−1. In general, the scopolamine contents were by a factor of 300–500 lower than the hyoscyamine ones. The phenotype, growth rate and alkaloid accumulation of the roots did not change despite cytokinin addition. Both ABA and GA3 strongly inhibited hyoscyamine accumulation. However, the inhibition was not associated with the changes in root morphology.We suggest that endogenous auxin production in the roots is not sufficient for maximum tropane alkaloid accumulation. To our knowledge this is the first time that alkaloid accumulation has been significantly increased by exogenous auxins without influencing growth.

AB - The transformed root cultures of Hyoscyamus muticus are usually cultivated in hormone-free medium in which the production of hyoscyamine is high. We studied whether the addition of exogenous growth regulators could enhance tropane alkaloid accumulation without affecting the growth and root phenotype. In order to rapidly screen a large number of hairy root samples a monoclonal antibody based enzyme immunological assay was developed for scopolamine analysis.The hairy root cultures tolerated exogenous auxins in the concentration range 0.01–5μmol/L well. The growth of the roots was not significantly affected by IAA and NAA, but the alkaloid accumulation was doubled compared with that of the roots growing in the absence of auxin. At optimum IAA concentration (0.1–1.0 μmol/L) the maximum hyoscyamine content was 117 mg L−1 and scopolamine content 0.2mgL−1. In general, the scopolamine contents were by a factor of 300–500 lower than the hyoscyamine ones. The phenotype, growth rate and alkaloid accumulation of the roots did not change despite cytokinin addition. Both ABA and GA3 strongly inhibited hyoscyamine accumulation. However, the inhibition was not associated with the changes in root morphology.We suggest that endogenous auxin production in the roots is not sufficient for maximum tropane alkaloid accumulation. To our knowledge this is the first time that alkaloid accumulation has been significantly increased by exogenous auxins without influencing growth.

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