Transgenic rapeseed plants obtained by the microinjection of DNA into microspore-derived embryoids

Neuhaus, Gunther; Spangenberg, Germán; Scheid, Ortrun Mittelsten; Schweiger, Hans−Georg

doi:10.1007/bf00249138

Cited by 200 publications

(42 citation statements)

References 44 publications

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“…By employing an ultrafine needle, the DNA was injected into the nucleus of cells of multicellular structures having a high potential for plant regeneration. Microsporederived embryos were chosen as targets for these experiments and transgenic plants were regenerated (Neuhaus et al 1987). Following the use of multicellular explants, the resulting plants were chimeric.…”

Section: Vectorless Transformationmentioning

confidence: 99%

Genetic transformation of Brassica

Poulsen

1996

Plant Breeding

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Tissue culture regeneration of plants from Brassica species can be achieved from various explant types. Their performance, however, varies with the genotype and age of the explant and is sensitive to medium supplements and Agrobacterium cocultivation. Most Brassica transformations were carried out using Agrobacterium and it was established that nopaline and agropine strains of .4. tumefaciens and A. rhizogenes, respectively, are the most efficient. Application of virulence gene induction is ambiguous. Binary vectors are mostly apphed in spite of indications that cointegrate vectors are more efficient. Cotransformation is successful with two different bacteria or with one bacterium carrying both plasmids, however, it has proved problematic to integrate the two transgenes into separate loci. Transformations with wild type rol genes develop hairy roots, from which transgenic plants can be regenerated. Vectorless transformation is feasible with microprojectile bombardment of microspore derived embryos being the most promising.Antibiotic markers providing resistance to kanamycin, hygromycin, and spectinomycin are applicable as selectable markers, while chloramphenicol is not suitable. Among herbicides, phosphinotricin gives good results while chlorsulfuron is unsuitable for in vitro selection. Major therapeutic antibiotics being successfully applied are carbenicillin, cefotaxime, amoxycillin, and ticarcillin. Standard methods for evaluating transgenic plants also have been applied to Brassica species. Some breeding objectives obtained through genetic transformation are presented.

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Section: Vectorless Transformationmentioning

confidence: 99%

Genetic transformation of Brassica

Poulsen

1996

Plant Breeding

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“…Similar time periods are required for DNA uptake and regeneration of protoplasts to produce stably transformed plants. Microinjection of DNA has been tested successfully in isolated cells cultured in vitro (3,12), but this method may not be applicable to other cell types or cells located below the surface of certain tissues and organs.…”

Section: Plasmidsmentioning

confidence: 99%

Transient Gene Expression in Maize, Rice, and Wheat Cells Using an Airgun Apparatus

Oard

Paige²,

Simmonds³

et al. 1990

Plant Physiol.

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An airgun apparatus has been constructed for transient gene expression studies of monocots. This device utilizes compressed air from a commercial airgun to propel macroprojectile and DNAcoated tungsten particles. The ,-glucuronidase (GUS) reporter gene was used to monitor transient expression in three distinct cell types of maize (Zea mays), rice (Oryza sativa), and wheat (Triticum aestivum). The highest level of GUS activity in cultured maize cells was observed when distance between stopping plate and target cells was adjusted to 4.3 centimeters. Efficiency of transformation was estimated to be 4.4 x 10-3. In a partial vacuum of 700 millimeters Hg, velocity of macroprojectile was measured at 520 meters per second with a 6% reduction in velocity at atmospheric pressure. A polyethylene film placed in the breech before firing contributed to a 12% increase in muzzle velocity. A 700 millimeters Hg level of vacuum was necessary for maximum number of transfornants. GUS expression was also detected in wheat leaf base tissue of microdissected shoot apices. High levels of transient gene expression were also observed in hard, compact embryogenic callus of rice. These results show that the airgun apparatus is a convenient, safe, and low-cost device for rapid transient gene expression studies in cereals.

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“…is lower (0.08 %). It has been reported by Neuhaus et al (1987) that transformation efficiency of plant cells can be significantly increased when the DNA is directly injected into the nucleus. Moreover it has been also reported (Kohler et al, 1989) that the transfer of interspecific genes is enhanced by irradiating the donor DNA with low doses of X-rays.…”

Section: Discussionmentioning

confidence: 99%

Introgression of G. barbadense genes into G. hirsutum through DNA-mediated Embryo Transformation approach

Aslam¹,

.²,

Iqbal³

1997

Pakistan J. of Biological Sciences

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Attempts were made to incorporate intraploid interpsecific gene(s) from Gossyium barbadense into G. hirsutum through DNA-mediated embryo transformation approach. G. barbadense DNA was injected into the styles/ovaries of G. hirsutum, 24 hours after self pollination. Two phenotypic types of plants i.e. transformed and G. hirsutum like were observed in D population. The transformed plants had clearcut variations in boll weight, yield and other quality traits confirmed to recipient parent. D transformations were also heritable and were transmitted to D generation. The transformed plants had better boll weight (4.9 gm), better fibre fineness (3.9 µg/in) and better fibre strength (101.0 TPPSI) compared to the boll weight (3.0 gms), fibre fineness (4.5 µg/in) and fibre strength (91.0 TPPSI) of G. hirsutum. The transformation efficiency for quantitative traits was about 20 % where as low for qualitative traits like flower petals with red spot (0.08 %). In the present studies transformed genotypes illustrated higher yield potential and better fibre quality than the recipient (G. hirsutum) parent.

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Transgenic rapeseed plants obtained by the microinjection of DNA into microspore-derived embryoids

Cited by 200 publications

References 44 publications

Genetic transformation of Brassica

Genetic transformation of Brassica

Transient Gene Expression in Maize, Rice, and Wheat Cells Using an Airgun Apparatus

Introgression of G. barbadense genes into G. hirsutum through DNA-mediated Embryo Transformation approach

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