Efficient transformation of scutellar tissue of immature maize embryos

Brettschneider, Reinhold; Becker, Dirk; Lörz, Horst

doi:10.1007/s001220050473

Cited by 96 publications

(59 citation statements)

References 31 publications

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“…Thus, this strategy may make it unnecessary to use herbicide selection of T0 plants to eliminate untransformed escapes. Selection pressures of 5.0 and 10.0 mg/l of bialaphos have been reported in transformation of barley (Wan and Lemaux 1994), rice (Toki et al 1992), and maize (Brettschneider et al 1997). Our selection approach is similar to that reported by Wan and Lemaux (1994) for barley transformation.…”

Section: Molecular Analyses (Pcr Southern and Western) Of 33supporting

confidence: 74%

An efficient wheat transformation procedure: transformed calli with long-term morphogenic potential for plant regeneration

Zhang

Rybczyński

Langenberg³

et al. 2000

Plant Cell Reports

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Langenberg, W.G.; Mitra, Amitava; and French, Roy C., "An efficient wheat transformation procedure: transformed calli with long-term morphogenic potential for plant regeneration" (2000). Abstract A method for producing large numbers of transgenic wheat plants has been developed. With this approach, an average of 9.7% of immature embryo explants were transformed and generated multiple selffertile, independently transformed plants. No untransformed plants, or escapes, were regenerated. This transformation procedure uses morphogenic calli derived from scutellum tissue of immature embryos of Triticum aestivum cv. Bobwhite co-bombarded with separate plasmids carrying a selectable marker gene (bar) and a gene of interest, respectively. Transformed wheat calli with a vigorous growth phenotype were obtained by extended culture on media containing 5.0 mg/l bialaphos. These calli retained morphogenic potential and were competent for plant regeneration for as long as 11 months. The bar gene and the gene of interest were co-expressed in T0 progeny plants. This wheat transformation protocol may facilitate quantitative production of multiple transgenic plants and significantly reduce the cost and labor otherwise required for screening out untransformed escapes.

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Section: Molecular Analyses (Pcr Southern and Western) Of 33supporting

confidence: 74%

An efficient wheat transformation procedure: transformed calli with long-term morphogenic potential for plant regeneration

Zhang

Rybczyński

Langenberg³

et al. 2000

Plant Cell Reports

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“…A glufosinate leaf-spray test (Brettschneider et al, 1997) was used to establish segregation ratios for expression of the bar gene in progeny. The herbicide Liberty (Aventis, Strasbourg, France) was dissolved in water (1.25 mL L Ϫ1 ) along with 0.1% (v/v) Tween 20 for a final glufosinate concentration of 250 mg L Ϫ1 .…”

Section: Progeny Segregation Analysis For Bar Gene Expressionmentioning

confidence: 99%

Agrobacterium tumefaciens-Mediated Transformation of Maize Embryos Using a Standard Binary Vector System

Frame¹,

Shou²,

Chikwamba³

et al. 2002

Plant Physiology

412

323

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We have achieved routine transformation of maize (Zea mays) using an Agrobacterium tumefaciens standard binary (non-super binary) vector system. Immature zygotic embryos of the hybrid line Hi II were infected with A. tumefaciens strain EHA101 harboring a standard binary vector and cocultivated in the presence of 400 mg L Ϫ1 l-cysteine. Inclusion of l-cysteine in cocultivation medium lead to an improvement in transient ␤-glucuronidase expression observed in targeted cells and a significant increase in stable transformation efficiency, but was associated with a decrease in embryo response after cocultivation. The average stable transformation efficiency (no. of bialaphos-resistant events recovered per 100 embryos infected) of the present protocol was 5.5%. Southern-blot and progeny analyses confirmed the integration, expression, and inheritance of the bar and gus transgenes in R 0 , R 1 , and R 2 generations of transgenic events. To our knowledge, this represents the first report in which fertile, stable transgenic maize has been routinely produced using an A. tumefaciens standard binary vector system.

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“…O emprego de embriões imaturos de milho para indução de embriogênese somática e regeneração de plantas de milho submetidas à transformação por biobalística é frequente na literatura (GORDON-KAMM et al, 1990;SONGSTAD et al, 1996;BRETTSCHNEIDER et al, 1997;ZHANG et al, 2012). De acordo com SONGSTAD et al (1996), esse tipo de explante permite a obtenção de plantas transgênicas com elevada fertilidade em comparação a outros tipos de explantes.…”

Section: Resultsunclassified

“…Os tecidos primários mais efi cientes na regeneração de plantas por embriogênese somática são os embriões imaturos obtidos de genótipos capazes de produzir calos embriogênicos do tipo II (BRETTSCHNEIDER et al, 1997;KOMARI, et al, 1998;SAHRAWAT et al, 2003;WANG et al, 2009). Estes calos são friáveis, amarelados, de consistência macia, podem ser mantidos em cultivo in vitro por maior tempo (GREEN et al, 1983).…”

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Regeneração de plantas a partir de dois tipos de explantes de milho submetidos à transformação genética por biobalística

Vidigal

Schuster

Texeira³

et al. 2014

Cienc. Rural

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O uso da biobalística na transformação genética de plantas requer a otimização de diversos parâmetros, entre eles, o desenvolvimento de protocolos para regeneração de plantas a partir de células transformadas. Este trabalho teve como objetivo avaliar a eficiência de regeneração de plantas de milho a partir de calos embriogênicos e de explantes sementes divididas, submetidos à transformação por biobalística, utilizando o gene bar como marcador de seleção. Foram submetidos 2000 agrupamentos de calos embriogênicos de milho e 2000 explantes sementes divididas à transformação por biobalística. A seleção dos eventos de transformação foi realizada em meios de cultivo suplementados com glifosinato de amônio. As plântulas selecionadas foram aclimatadas e transplantadas para vasos em casa de vegetação. Com a utilização de embriogênese somática, a eficiência de regeneração foi de 0,2%, enquanto que, com a utilização de sementes divididas, a eficiência de regeneração foi de 1,3%. Embora ambos os protocolos possibilitem a regeneração de plantas de milho submetidas à transformação por biobalística, há a necessidade de aumentar sua eficiência.

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Efficient transformation of scutellar tissue of immature maize embryos

Cited by 96 publications

References 31 publications

An efficient wheat transformation procedure: transformed calli with long-term morphogenic potential for plant regeneration

An efficient wheat transformation procedure: transformed calli with long-term morphogenic potential for plant regeneration

Agrobacterium tumefaciens-Mediated Transformation of Maize Embryos Using a Standard Binary Vector System

Regeneração de plantas a partir de dois tipos de explantes de milho submetidos à transformação genética por biobalística

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