Transformation and inheritance of a hygromycin phosphotransferase gene in maize plants

Walters, David A.; Vetsch, Clayton S.; Potts, Diane E.; Lundquist, Ronald C.

doi:10.1007/bf00034948

Cited by 158 publications

(75 citation statements)

References 36 publications

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“…For monocots, there has been a progression from particle gun transformation to that mediated by Agrobacterium tumefaciens, as well as refinements in tissue culture protocols and selection strategies (Shrawat and Lörz, 2006). Since the earliest reports of maize protoplast transformation (Rhodes et al, 1988;Shillito et al, 1989;Golovkin et al, 1993), the preferred target cells for transformation have gone from maize cells in liquid suspension (Fromm et al, 1990;Gordon-Kamm et al, 1990;Frame et al, 1994), through embryogenic callus (Walters et al, 1992;Wan et al, 1995), and finally to the transformation of scutellar cells of freshly isolated immature embryos (Ishida et al, 1996;Songstad et al, 1996;Frame et al, 2002). For other monocots such as barley (Hordeum vulgare), wheat, rice, and sorghum, immature embryos remain the predominant transformation target, despite reports over the years of successfully initiating tissue culture responses from explants other than immature embryos.…”

Section: Introductionmentioning

confidence: 99%

Morphogenic Regulators Baby boom and Wuschel Improve Monocot Transformation

et al. 2016

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While transformation of the major monocot crops is currently possible, the process typically remains confined to one or two genotypes per species, often with poor agronomics, and efficiencies that place these methods beyond the reach of most academic laboratories. Here, we report a transformation approach involving overexpression of the maize (Zea mays) Baby boom (Bbm) and maize Wuschel2 (Wus2) genes, which produced high transformation frequencies in numerous previously nontransformable maize inbred lines. For example, the Pioneer inbred PHH5G is recalcitrant to biolistic and Agrobacterium tumefaciens transformation. However, when Bbm and Wus2 were expressed, transgenic calli were recovered from over 40% of the starting explants, with most producing healthy, fertile plants. Another limitation for many monocots is the intensive labor and greenhouse space required to supply immature embryos for transformation. This problem could be alleviated using alternative target tissues that could be supplied consistently with automated preparation. As a major step toward this objective, we transformed Bbm and Wus2 directly into either embryo slices from mature seed or leaf segments from seedlings in a variety of Pioneer inbred lines, routinely recovering healthy, fertile T0 plants. Finally, we demonstrated that the maize Bbm and Wus2 genes stimulate transformation in sorghum (Sorghum bicolor) immature embryos, sugarcane (Saccharum officinarum) callus, and indica rice (Oryza sativa ssp indica) callus.

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Section: Introductionmentioning

confidence: 99%

Morphogenic Regulators Baby boom and Wuschel Improve Monocot Transformation

et al. 2016

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“…Transgene expression levels and inheritance show wide variation among independently transformed plants carrying the same construct (Peach and Velten 1991;Spencer et al 1992;Walters et al 1992). Many factors can contribute to the variation in transgene expression.…”

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Gene dosage and genetic background affect miraculin accumulation in transgenic tomato fruits

Kim

Hirai

Kato

et al. 2010

Plant Biotechnology

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Gene dosage and genetic background are factors that influence transgene expression in transgenic plants. In our previous studies, we produced transgenic tomato plants that accumulate miraculin, a taste-modifying protein, in a genetically stable manner. To elucidate the effects of gene dosage and genetic background on miraculin accumulation in transgenic tomato fruits, we generated hybrid tomato lines between the homozygous transgenic line 56B (background cultivar 'Moneymaker') and the pure cultivars 'Micro-Tom,' 'Moneymaker,' 'Ailsa Craig,' 'M82,' 'Rutgers' and 'Aichi-first' and analyzed them for miraculin mRNA expression and miraculin protein accumulation. The hybrid lines exhibited variation in their fruit structures. Among the hybrid lines heterozygous for the miraculin gene, miraculin accumulation in the fruits varied from 111.0 mg g Ϫ1 fresh weight (FW) to 159.4 mg g Ϫ1 FW. Furthermore, the homozygous line 56B showed higher miraculin accumulation and miraculin mRNA expression than the heterozygous line 56Bϫ'Moneymaker.' These results demonstrate the profound effects of gene dosage and genetic background on miraculin accumulation in transgenic tomato fruits.Key words: Gene dosage, genetic background, miraculin, taste-modifying protein.Plant Biotechnology 27, 333-338 (2010) Original PaperThis article can be found at http://www.jspcmb.jp/ allelic composition and genetic background affect transgene expression and inheritance in white clover. Another study in rice showed that the expression pattern of Xa3/Xa26, which is a rice disease resistance gene, was affected by the genetic background (Cao et al. 2007). The full effects of gene dosage and genetic background on transgene expression and translation remain to be elucidated.In this study, we generated hybrid lines between transgenic line 56B (background cultivar 'Moneymaker'), which is homozygous for the miraculin gene, and the pure line cultivars 'Micro-Tom,' 'Moneymaker,' 'Ailsa Craig,' 'M82,' 'Rutgers' and 'Aichi-first.' Characterization of the hybrids' miraculin mRNA expression and miraculin protein accumulation, revealed the effects of gene dosage and genetic background on miraculin accumulation in transgenic tomato fruits, and in addition we discuss the significance of combining molecular breeding with cross-breeding to produce transgenic tomatoes that accumulate miraculin. Materials and methods Plant material and growth conditionsThe transgenic tomato line 56B was produced in our previous work (Sun et al. 2007). In brief miraculin encoding 660-bp DNA fragment was inserted into the XbaI/SacI sites of the plant transformation vectors pBI121 (Sun et al. 2006). Line 56B possesses a single copy of the miraculin gene driven by the CaMV 35S promoter.In the present study, the miraculin-accumulating transgenic tomato line 56B (background cultivars 'Moneymaker') was crossed with six other cultivars: 'Micro-Tom (accession number TOMJPF00001), ' 'Moneymaker (accession number TOMJPF00002), ' 'Aichi-first (accession number TOMJPF00003), ' 'Ailsa Cra...

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“…Concurrent with gene delivery methods, selectable marker development has been integral in developing efficient maize transformation. Kanamycin (Fromm et al, 1986;Rhodes et al, 1988;Lyznik et al, 1989) and hygromycin (Walters et al, 1992) were two of the earliest antibiotics used as selection agents in corn. The first herbicide used as a selection agent for maize transformation was the Glu analog phosphinothricin or, more commonly, the tripeptide bialaphos (lphosphinothricyl-l-alanyl-l-ananine), which contains phosphinothricin as the active ingredient (Fromm et al, 1990;Gordon-Kamm et al, 1990).…”

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Development of Protoporphyrinogen Oxidase as an Efficient Selection Marker forAgrobacterium tumefaciens-Mediated Transformation of Maize

Volrath

Nicholl

et al. 2003

Plant Physiology

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In this article, we report the isolation of plant protoporphyrinogen oxidase (PPO) genes and the isolation of herbicide-tolerant mutants. Subsequently, an Arabidopsis double mutant (Y426M + S305L) was used to develop a selectable marker system for Agrobacterium tumefaciens-mediated transformation of maize (Zea mays) and to obtain multiple events tolerant to the PPO family of herbicides. Maize transformants were produced via butafenacil selection using a flexible light regime to increase selection pressure. Butafenacil selection per se did not change transgene copy number distribution relative to other selectable marker systems, but the most tolerant events identified in the greenhouse were more likely to contain multiple copies of the introduced mutant PPO gene. To date, more than 2,500 independent transgenic maize events have been produced using butafenacil selection. The high frequency of A. tumefaciens-mediated transformation via PPO selection enabled us to obtain single-copy transgenic maize lines tolerant to field levels of butafenacil

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Transformation and inheritance of a hygromycin phosphotransferase gene in maize plants

Cited by 158 publications

References 36 publications

Morphogenic Regulators Baby boom and Wuschel Improve Monocot Transformation

Morphogenic Regulators Baby boom and Wuschel Improve Monocot Transformation

Gene dosage and genetic background affect miraculin accumulation in transgenic tomato fruits

Development of Protoporphyrinogen Oxidase as an Efficient Selection Marker forAgrobacterium tumefaciens-Mediated Transformation of Maize

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