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Biolistic transformation of Eucalyptus grandis x E. urophylla callus.
Sartoretto, Laudete Maria; Cid, Luis Pedro Barrueto; Brasileiro, Ana Cristina Miranda.
Afiliação
  • Sartoretto LM; Laboratório de Transferência de Genes, Embrapa Recursos Genéticos e Biotecnologia, Parque Estação Biológica (PqEB), 70.849-970, Brazil.
  • Cid LPB; Laboratório de Transferência de Genes, Embrapa Recursos Genéticos e Biotecnologia, Parque Estação Biológica (PqEB), 70.849-970, Brazil.
  • Brasileiro ACM; Laboratório de Transferência de Genes, Embrapa Recursos Genéticos e Biotecnologia, Parque Estação Biológica (PqEB), 70.849-970, Brazil. Current address: Embrapa Labex - France / Agropolis International, Avenue Agropolis, 34934 Montpellier Cedex 5, France.Corresponding author; email: anacmb@cenargen.embrapa.br.
Funct Plant Biol ; 29(8): 917-924, 2002 Aug.
Article em En | MEDLINE | ID: mdl-32689542
A procedure for genetic transformation of the hybrid Eucalyptus grandis × E. urophylla using particle bombardment is described. Cotyledon- and hypocotyl-derived calli growing on SP medium supplemented with 2mthidiazuron or on MS modified (MSM) medium supplemented with 10 m 2,4-dichlorophenoxyacetic acid (2,4-D) and 2.5m6-benzylaminopurine (BAP), were used as target material for bombardment assays. Multiple preincubation and bombardment conditions were tested. Tungsten particles were coated with the plasmid pBI426 harbouring a ß-glucuronidase (gus) and neomycin phosphotransferase II (npt II) gene fusion controlled by a double 35S cauliflower mosaic virus (CaMV) promoter. Four days after bombardment, the transient transformation efficiency was determined by expression of the gus gene. Fully GUS-positive calli were then obtained after 105 d in MSM medium supplemented with 2,4-D, BAP, and the selective agent kanamycin at 200 mg L-1. The presence of the gus gene in these kanamycin-resistant calli was confirmed by polymerase chain reaction analysis. Extensive experiments were performed aiming to identify conditions for the regeneration of these GUS-expressing calli. However, they were unable to regenerate transgenic shoots, suggesting that conditions suitable for regeneration are unsuitable for transformation and vice versa.

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Funct Plant Biol Ano de publicação: 2002 Tipo de documento: Article País de afiliação: Brasil País de publicação: Austrália

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Funct Plant Biol Ano de publicação: 2002 Tipo de documento: Article País de afiliação: Brasil País de publicação: Austrália