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A simple and reliable method for the screening of transgenic tobacco plants PAB
Freitas-Astua,Juliana; Astua-Monge,Gustavo; Polston,Jane Elisabeth; Hiebert,Ernest.
Even though much improvement has been made in plant transformation methods, the screening of transgenic plants is often a laborious work. Most approaches for detecting the transgene in transformed plants are still timeconsuming, and can be quite expensive. The objective of this study was to search for a simpler method to screen for transgenic plants. The infiltration of kanamycin (100 mg/mL) into tobacco leaves resulted in conspicuous chlorotic spots on the non-transgenic plant leaves, while no spots were seen on the leaves of transformed plants. This reaction occurred regardless of age of the tested plants, and the method has proven to be simple, fast, non-destructive, relatively cheap, and reliable. These results were comparable to those obtained by the...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Kanamycin; Genetically modified organisms; Transgenic plants; Testing.
Ano: 2003 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-204X2003000700015
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Agrobacterium-mediated transformation of white poplar (Populus alba L.): (with 3 figures & 1 table) Phyton
Sánchez,N; Manzanera,JA; Bueno,MA.
A simple method is presented for the genetic transformation of white poplar (Populus alba L.). Root suckers and buds from branches of adult trees were established in vitro. Callus was induced on injured leaves cultured on medium supplemented with 0.45 µM thidiazuron and 0.1 µM a-naphthalene acetic acid. Non-transformed leaves and calli did not survive concentrations higher than 85 µM kanamycin or 18 mM hygromycin. The leaf explants were inoculated with the disarmed Agrobacterium tumefaciens LBA4404/p35S GUS INT/pCAMBIA 2301 strain. Twenty % of the cultured callus explants was kanamycin-resistant two months after the co-inoculation experiment. The rate of adventitious bud induction on resistant calli was 17 %. The analysis of b-glucuronidase expression two...
Tipo: Info:eu-repo/semantics/article Palavras-chave: BAP benzil aminopurine TDZ; Thidiazuron IBA; Indole butyric acid NAA; Naphthalene acetic acid KAN; Kanamycin.
Ano: 2004 URL: http://www.scielo.org.ar/scielo.php?script=sci_arttext&pid=S1851-56572004000100015
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Rooting in Km selective media as efficient in vitro selection method for sunflower genetic transformation Electron. J. Biotechnol.
Radonic,Laura Mabel; Zimmermann,Julián Marcos; Zavallo,Diego; López,Nilda; López Bilbao,Marisa.
Despite of numerous publications in sunflower genetic transformation, there is no efficient or reproducible protocol with low number of escapes. The latter would indicate that the selection method is not effective. In this work we used Km as selective agent, Agrobacterium tumefaciens EHA105 strain and a vector with the nptII gene under the nos promoter and uidA gene under 35S promoter. The response of agroinfected (A) and control (C) explants during the in vitro culture was studied and in both cases in presence or absence of Km in order to assign a differential morphologic response between transformed and non-transformed plants. The characteristics analyzed were: height, colour/aspect of the plantlets, in vitro rooting and in vitro bud-flower development....
Tipo: Journal article Palavras-chave: Genetic transformation; Kanamycin; Selection; Sunflower.
Ano: 2006 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000300027
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Selective agent and A. tumefaciens overgrowth-control antibiotics in Eucalyptus camaldulensis Cotiledonary culture BABT
Quisen,Regina; Oliveira,Yohana de; Pileggi,Marcos; Cuquel,Francine; Quoirin,Marguerite.
The objectives of the present work were to establish the minimal lethal dose of the selective agent to determine the type and concentration of appropriate antibiotics for the elimination of Agrobacterium tumefaciens inoculated explants, without interfering with the regenerative potential of the E. camaldulensis cotyledonary explants. Non-transformed explants were cultivated in medium supplemented with kanamycin. The results showed that the antibiotic was suitable for the selection of transformed cells in the concentration of 9 mg L-1 as it inhibited the growth of non-transformed cells. Cotyledons infected with A. tumefaciens were cultivated in MS N/2 medium supplemented with BAP, ANA, Km and cefotaxime or AugmentinÒ . The highest average of regenerated...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Morphogenesis; Kanamycin; Cefotaxime; Augmentin; Agrobacterium tumefaciens; Eucalypt.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132009000600020
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