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Protoplasts to plants of Gentianaceae. Regeneration of lisianthus (Eustoma grandiflorum) is affected by calcium ion preconditioning, osmolality and pH of the culture media

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Abstract

A reliable method has been developed for regeneration of whole plants from isolated protoplasts of five cultivars of lisianthus,Eustoma grandiflorum (Griseb.) Schinners (Gentianaceae). Protoplasts were isolated from either cotyledons or leaves and cultured in agarose beads surrounded by liquid V-KM media containing 5.37 µM 1-naphthyleneacetic acid (NAA) and 2.28 µM zeatin. When microcalli were approximately 1 mm in diameter, the agarose beads were transferred to shoot regeneration media containing 0.1 µM indolebutyric acid (IBA) and 4.44 µM 6-benzylaminopurine (BAP). Shoots were produced from the calli during several sub-culture periods. Protoplast viability and the subsequent regeneration of plants were dependent on calcium levels and growth regulator presence in thein vitro seed germination media, on the osmolality of the protoplast purification solution, and osmolality increase and pH of the culture media. Shoots were rooted in Murashige & Skoog (1962) media containing 5.71 µM indole-3-acetic acid (IAA). Plantlets derived from protoplasts of five lisianthus cultivars (Fresh White, Hakusen, Miss Lilac, Fresh Purple and Doremi Wine Red) have been successfully transferred to the glasshouse.

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Abbreviations

B5:

Gamborg et al. (1968)

BAP:

6-benzylaminopurine

IAA:

indole-3-acetic acid

IBA:

indolebutyric acid

GA3 :

gibberellic acid

mOsm:

(negative) milli Osmoles per kilogram water

MES:

2[N-morpholino]ethane sulfonic acid

MS:

Murashige & Skoog (1962)

NAA:

α-naphthaleneacetic acid

pfd:

photonfluence density

V-KM:

Binding & Nehls (1977)

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Author notes

  1. Iona E. W. O'Brien

    Present address: HortResearch, Mt Albert Research Centre, Private Bag 92169, Auckland, New Zealand

Authors and Affiliations

  1. MAF Technology, Horticultural Research Centre, Private Bag, Levin, New Zealand

    Iona E. W. O'Brien & Graeme C. Lindsay

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  1. Iona E. W. O'Brien
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  2. Graeme C. Lindsay
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O'Brien, I.E.W., Lindsay, G.C. Protoplasts to plants of Gentianaceae. Regeneration of lisianthus (Eustoma grandiflorum) is affected by calcium ion preconditioning, osmolality and pH of the culture media. Plant Cell Tiss Organ Cult 33, 31–37 (1993). https://doi.org/10.1007/BF01997595

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  • DOI: https://doi.org/10.1007/BF01997595

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