Efficient plant regeneration from hairy root-derived protoplasts of Hyoscyamus muticus

Nina Sevón, Kirsi Marja Oksman-Caldentey, Raimo Hiltunen

Research output: Contribution to journalArticleScientificpeer-review

23 Citations (Scopus)

Abstract

Successful plant regeneration was achieved for the first time from hairy root-derived protoplasts of Hyoscyamus muticus. High yields (7 × 106 / g fresh weight) of protoplasts were isolated directly from the transformed roots of Hyoscyamus muticus using an enzyme mixture comprising 1 % macerozyme and 2 % cellulase in an osmoticum consisting of 0.2 M CaCl2 and 0.6 M mannitol. Protoplasts were first cultured in liquid NT/PRO I medium and further on semi-solid NT/PRO II agar medium. The procedure permits highly efficient formation of colonies. The plating efficiency varied from 1-9 %. The small individual colonies regenerated easily into shoots and roots at frequencies of 18 % and 70 %, respectively. The time required for the development of small plantlets from protoplasts was 8-11 weeks. The regenerated plants contained rolB from Ri-T-DNA and exhibited an altered phenotype compared to the control plants.

Original languageEnglish
Pages (from-to)738-742
Number of pages5
JournalPlant Cell Reports
Volume14
Issue number11
DOIs
Publication statusPublished - 1 Aug 1995
MoE publication typeA1 Journal article-refereed

Fingerprint

Hyoscyamus muticus
protoplasts
mannitol
endo-1,4-beta-glucanase
plantlets
agar
phenotype
shoots
liquids
DNA
enzymes

Keywords

  • Hyoscyamus muticus
  • plant regeneration
  • protoplasts
  • transformed roots

Cite this

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title = "Efficient plant regeneration from hairy root-derived protoplasts of Hyoscyamus muticus",
abstract = "Successful plant regeneration was achieved for the first time from hairy root-derived protoplasts of Hyoscyamus muticus. High yields (7 × 106 / g fresh weight) of protoplasts were isolated directly from the transformed roots of Hyoscyamus muticus using an enzyme mixture comprising 1 {\%} macerozyme and 2 {\%} cellulase in an osmoticum consisting of 0.2 M CaCl2 and 0.6 M mannitol. Protoplasts were first cultured in liquid NT/PRO I medium and further on semi-solid NT/PRO II agar medium. The procedure permits highly efficient formation of colonies. The plating efficiency varied from 1-9 {\%}. The small individual colonies regenerated easily into shoots and roots at frequencies of 18 {\%} and 70 {\%}, respectively. The time required for the development of small plantlets from protoplasts was 8-11 weeks. The regenerated plants contained rolB from Ri-T-DNA and exhibited an altered phenotype compared to the control plants.",
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Efficient plant regeneration from hairy root-derived protoplasts of Hyoscyamus muticus. / Sevón, Nina; Oksman-Caldentey, Kirsi Marja; Hiltunen, Raimo.

In: Plant Cell Reports, Vol. 14, No. 11, 01.08.1995, p. 738-742.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Sevón, Nina

AU - Oksman-Caldentey, Kirsi Marja

AU - Hiltunen, Raimo

PY - 1995/8/1

Y1 - 1995/8/1

N2 - Successful plant regeneration was achieved for the first time from hairy root-derived protoplasts of Hyoscyamus muticus. High yields (7 × 106 / g fresh weight) of protoplasts were isolated directly from the transformed roots of Hyoscyamus muticus using an enzyme mixture comprising 1 % macerozyme and 2 % cellulase in an osmoticum consisting of 0.2 M CaCl2 and 0.6 M mannitol. Protoplasts were first cultured in liquid NT/PRO I medium and further on semi-solid NT/PRO II agar medium. The procedure permits highly efficient formation of colonies. The plating efficiency varied from 1-9 %. The small individual colonies regenerated easily into shoots and roots at frequencies of 18 % and 70 %, respectively. The time required for the development of small plantlets from protoplasts was 8-11 weeks. The regenerated plants contained rolB from Ri-T-DNA and exhibited an altered phenotype compared to the control plants.

AB - Successful plant regeneration was achieved for the first time from hairy root-derived protoplasts of Hyoscyamus muticus. High yields (7 × 106 / g fresh weight) of protoplasts were isolated directly from the transformed roots of Hyoscyamus muticus using an enzyme mixture comprising 1 % macerozyme and 2 % cellulase in an osmoticum consisting of 0.2 M CaCl2 and 0.6 M mannitol. Protoplasts were first cultured in liquid NT/PRO I medium and further on semi-solid NT/PRO II agar medium. The procedure permits highly efficient formation of colonies. The plating efficiency varied from 1-9 %. The small individual colonies regenerated easily into shoots and roots at frequencies of 18 % and 70 %, respectively. The time required for the development of small plantlets from protoplasts was 8-11 weeks. The regenerated plants contained rolB from Ri-T-DNA and exhibited an altered phenotype compared to the control plants.

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