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Journal of Biotechnology and Biodiversity
Gessiel Newton Scheidt
ISSN: 2179-4804
Vol. 2, No. 1, 2011, pp. 1-6
Bioline Code: jy11001
Full paper language: English
Document type: Research Article
Document available free of charge

Journal of Biotechnology and Biodiversity, Vol. 2, No. 1, 2011, pp. 1-6

 en Preliminary results for genetic transformation of shoot tip of Eucalyptus saligna check for this species in other resources Sm. via Agrobacterium tumefaciens check for this species in other resources
da Silva, ALL; de Oliveira, Y; Costa, JL; Mudry, CS; Procopiuk, M; Scheidt, GN & Brondani, GE

Abstract

The regeneration of transgenic plants of Eucalypt is the largest difficulty for the genetic transformation of this genus; in addition a low rate of transformed plants is reached. The aim of this research was to evaluate acetosyringone (3',5'-Dimethoxy-4'-hydroxyacetophenone) on the co-culture medium during genetic transformation of shoot tips of Eucalyptus saligna check for this species in other resources via Agrobacterium tumefaciens check for this species in other resources and to promote the explants selection supposedly transformed. Shoot tip from multiple shoots was used as explants. These explants were pre-cultured during two days before the transformation. Strain EHA105 of A. tumefaciens harboring the plasmid pBI121 was used. The treatments were: 0 and 100 μM acetosyringone added to the co-culture, after co-culture the explants were cultured in multiplication medium supplemented with 250 mg.L-1 Cefotaxime® and each sub-culture the kanamycin levels were increased from 50 to 150 mg.L-1. The transient expression of the uidA gene in shoot tips was evaluated after the end of the co-culture (fifth day) and after seven days of culture on medium containing kanamycin. The presence of 100 µM acetosyringone at the co-culture of shoot tips of Eucalyptus saligna promoted higher transient expression of the uidA gene and retards toxic effects caused by kanamycin.

Keywords
Strain EHA105; GUS gene; Eucalypt; B-glucuronidase

 
 pt
da Silva, ALL; de Oliveira, Y; Costa, JL; Mudry, CS; Procopiuk, M; Scheidt, GN & Brondani, GE

Resumo

A regeneração de plantas transgênicas de eucalipto representa a maior dificuldade para a transformação genética de plantas do gênero Eucalyptus, além de que taxas baixas de plantas transformadas são alcançadas. O objetivo desse trabalho foi avaliar a acetosiringona no meio de co-cultura durante a transformação genética de ápices caulinares de Eucalyptus saligna check for this species in other resources via Agrobacterium tumefaciens check for this species in other resources e promover a seleção dos supostos explantes transformados. Ápices caulinares originados de brotações múltiplas foram usados como explantes. Estes explantes foram pré-cultivados por dois dias antes da transformação. A linhagem EHA105 de A. tumefaciens contendo o plasmídeo pBI120 foi utilizada. Os tratamentos foram: 0 e 100 μM de acetosiringona adicionada ao meio de co-cultura, após a co-cultura os explantes foram cultivados em meio de multiplicação suplementado com 250 mg.L-1 de cefotaxima e a cada subcultivo a concentração de canamicina foi aumentada, de 50 até 150 mg.L-1. A expressão transiente do gene uidA nos ápices caulinares foi avaliada no final da co-cultura e após sete dias em meio com canamicina (agente seletivo). A presença de 100 μM de acetosiringona no meio de co-cultura dos ápices caulinares de Eucalyptus saligna promoveu maior expressão transiente do gene uidA e retardou a ação tóxica causada pela canamicina.

Palavras-chave
Linhagem EHA105; gene GUS; Eucalipto; B-glucuronidase

 
© Journal of Biotechnology and Biodiversity
Alternative site location: http://revista.uft.edu.br/index.php/jbb/index

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