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Genetic transformation of Eustoma grandiflorum Griseb. by microprojectile bombardment

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Summary

Foreign DNA was introduced into cell suspension cultures and leaf tissue of Eustoma grandiflorum Griseb. (lisianthus) by microprojectile bombardment. For this purpose a low-cost bombardment device that uses a helium flux to accelerate microprojectiles was built. When cell suspensions were used, an average of 4.1 Kan resistant calli were recovered per shot after 4 months' cultivation on selective medium. Most of the Kan resistant plants regenerated from calli were positive to GUS assay. Both the nptII and gus genes were successfully amplified from alkali-treated leaves of putative transgenic plants by PCR analysis. Transgenic plants were not recovered from bombarded leaves. Considering the host range specificity of Agrobacterium, and the response of the species to plant regeneration from suspension culture, microprojectile bombardment is, at present, the most efficient procedure for genetic transformation of lisianthus.

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Abbreviations

BA:

6-benzyladenine

Cx:

cefotaxime

2,4 D:

(2,4-dichlorophenoxy) acetic acid

FDA:

fluorescein diacetate

gus :

β-glucuronidase

IAA:

indole-3-acetic acid

IBA:

indole-3-butyric acid

2iP:

(2-isopentenyl) adenine

Kan:

kanamycin

nptII :

neomycin phosphotransferase II

PCR:

polymerase chain reaction

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Authors and Affiliations

  1. Istituto Sperimentale per la Floricoltura, C.so Inglesi, 508, 18038, Sanremo, (IM), Italy

    Laura Semeria & Andrea Allavena

  2. Istituto Fitovirologia Applicata, CNR, Strada Delle Cacce, 73, 10135, Torino, Italy

    Anna Maria Vaira & Gian Paolo Accotto

Authors
  1. Laura Semeria
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  2. Anna Maria Vaira
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  3. Gian Paolo Accotto
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  4. Andrea Allavena
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Semeria, L., Vaira, A.M., Accotto, G.P. et al. Genetic transformation of Eustoma grandiflorum Griseb. by microprojectile bombardment. Euphytica 85, 125–130 (1995). https://doi.org/10.1007/BF00023940

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