Transient expression of foreign genes in rice, wheat and soybean cells following particle bombardment

Wang, Yi Chang; Klein, Theodore M.; Fromm, Michael; Cao, Jun; Sanford, Jay P.; Wü, Ray

doi:10.1007/bf00039024

Cited by 177 publications

(51 citation statements)

References 28 publications

(32 reference statements)

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“…Previous work based on the analysis of transient gene expression has shown that microprojectiles can be used for the delivery of DNA to a wide range of intact plant cells and tissues including those from onion, maize, tobacco (13)(14)(15)(16), rice, wheat, and soybean (22). Stable transformation by particle bombardment has been shown in tobacco (16) and soybean (18).…”

mentioning

confidence: 99%

Genetic Transformation of Maize Cells by Particle Bombardment

Klein¹,

Kornstein²,

Sanford³

et al. 1989

Plant Physiol.

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142

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Intact maize cells were bombarded with microprojectiles bearing plasmid DNA coding for selectable (neomycin phosphotransferase [NPT II]) and screenable (,l-glucuronidase [GUS]) marker genes. Kanamycin-resistant calli were selected from bombarded cells, and these calli carried copies of the NPT II and GUS genes as determined by Southern blot analysis. All MATERIALS AND METHODS PlasmidsThe plasmid pNGI (8.2 kb) contains both a neomycin phosphotransferase II (NPT II) and a 3-glucuronidase (GUS) gene cloned in pUC8 (Fig. 1). The GUS gene comprises the promoter and intron 1 (nucleotides 1-1775) from the Alcohol dehydrogenase 1 (Adh 1) gene of maize (6), a GUS coding region consisting of the PstI fragment from pRAJ260 (10), and a modified 3' end from the nopaline synthase gene (8). The NPT II gene in pNGI was derived from pCaMVNEO (9) and is composed ofthe 35S promoter from cauliflower mosaic 440 virus, the NPT II coding region, and the nopaline synthase 3' end. pCaMVINEO (2) is similar to pCaMVNEO except it contains the Adhl intron 1 between the 35S promoter and NPT II coding region. Bombardment of CellsThe suspension culture of Z. mays cv Black Mexican Sweet (BMS; ATCC No 54022) was maintained in MS media as previously described (8). The bombardment (15) and subsequent selection (9) of kanamycin-resistant clones was performed as detailed previously. Briefly, 2 mL of suspension culture (about 2x I05 cells) was distributed over the surface of a filter paper (Whatman No. 4, 5.5 cm in diameter). The filter paper bearing the cells was placed over three layers of filter paper to which 2.5 mL of MS media had been added. The cells were then bombarded under a partial vacuum with tungsten particles (average diameter 1.2 ,tm) to which DNA was adsorbed using a calcium-spermidine precipitation procedure (15). Following bombardment, the cells were washed from the filter with 6 mL of MS medium into a 10 cm Petri dish. The Petri dish was sealed and then incubated in the dark at 26°C. After 2 d, 6 mL of fresh medium containing 300 ,ug of kanamycin per mL was added to each Petri dish and half of the volume was transferred to a fresh dish. After 1 week the culture was again diluted twofold with fresh medium containing 150 /Ag of kanamycin per mL and half the culture transferred to a new Petri dish. Subsequent additions of fresh medium containing kanamycin were made at 2 week intervals until the majority ofcells in the culture clearly ceased to grow. This generally occurred after 4 to 6 weeks of selection. The cells were then transferred from the liquid medium to a membrane filter supported on a cellulose adsorbent pad (Gelman) that was on agarose-solidified MS medium containing 100 ,ug/mL kanamycin. Calli that developed on the filter were transferred to agarose-solidified medium supplemented with kanamycin after they had reached a diameter of about 0.5 cm. GUS AssaysThe presence of the GUS enzyme in the transformants was visualized using a histochemical reaction (1 1). A small amount of a kanamycin-resistant callus was plac...

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mentioning

confidence: 99%

Genetic Transformation of Maize Cells by Particle Bombardment

Klein¹,

Kornstein²,

Sanford³

et al. 1989

Plant Physiol.

Self Cite

142

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“…Reggiardo et al (1991) found that more than one bombardment per plate had deleterious effects on barley and maize cells. However, in wheat, Wang et al (1988) observed a direct relationship between an increase in the number of cells transiently expressing GUS and an increase in the number of shots per plate. …”

Section: Resultsmentioning

confidence: 94%

Optimization of particle bombardment parameters for the genetic transformation of Brazilian maize inbred lines

Petrillo

Carneiro

Purcino

et al. 2008

Pesq. agropec. bras.

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-The objective of this work was to develop a genetic transformation system for tropical maize genotypes via particle bombardment of immature zygotic embryos. Particle bombardment was carried out using a genetic construct with bar and uidA genes under control of CaMV35S promoter. The best conditions to transform maize tropical inbred lines L3 and L1345 were obtained when immature embryos were cultivated, prior to the bombardment, in higher osmolarity during 4 hours and bombarded at an acceleration helium gas pressure of 1,100 psi, two shots per plate, and a microcarrier flying distance of 6.6 cm. Transformation frequencies obtained using these conditions ranged from 0.9 to 2.31%. Integration of foreign genes into the genome of maize plants was confirmed by Southern blot analysis as well as bar and uidA gene expressions. The maize genetic transformation protocol developed in this work will possibly improve the efficiency to produce new transgenic tropical maize lines expressing desirable agronomic characteristics.

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“…Best results were obtained at pressure of 1100 psi, target distance of 6cm and 28 Hg in vacuum. Bombarding the explants two times increased the number of cells giving transient gene expression and this could be due to the fact that multiple bombardments allow better coverage of the target areas and compensate for misfires from faulty and poorly set rupture discs as was the case in rice (Wang et al, 1988). By using the same range of sizes reported for maize , gene transfer in wheat was shown to be most efficient with microprojectiles 1.2 µm in diameter.…”

Section: Optimization Of Conditions For Dna Deliverymentioning

confidence: 97%

Stable gene transformation in cowpea (Vigna unguiculata L. walp.) using particle gun method

Ikea¹,

Ingelbrecht²,

Uwaifo³

et al. 2003

Afr. J. Biotechnol.

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We investigated the possibility of transforming and obtaining transgenic cowpea (Vigna unguiculata L Walp) plants using the particle bombardment process. Meristematic explants that could give rise to whole fertile plants were used in transformation experiments with reporter and selectable marker genes driven by a 35S CaMV promoter. Conditions for optimal delivery of DNA to explants were established based on transient gus expression assays two days after bombardment. The size of microcarriers, microflight distance and helium pressure significantly affected transient expression of reporter genes. A total of 1692 explants were bombarded with DNA-coated particles and placed on 3 mg/l bialaphos selective medium. Only 12 regenerated shoots produced seeds eventually, and all were Gus negative even though

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Transient expression of foreign genes in rice, wheat and soybean cells following particle bombardment

Cited by 177 publications

References 28 publications

Genetic Transformation of Maize Cells by Particle Bombardment

Genetic Transformation of Maize Cells by Particle Bombardment

Optimization of particle bombardment parameters for the genetic transformation of Brazilian maize inbred lines

Stable gene transformation in cowpea (Vigna unguiculata L. walp.) using particle gun method

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