Factors Influencing Gene Delivery into Zea Mays Cells by High–Velocity Microprojectiles

Klein, Theodore M.; Gradziel, Thomas M.; Fromm, Michael; Sanford, Jay P.

doi:10.1038/nbt0588-559

Cited by 232 publications

(141 citation statements)

References 17 publications

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“…DNA (1 &shot final concentration) was precipitated onto gold or tungsten (1-3 pm diameter) particles and used for bombardment, according to the method of Klein et al (1988). Tissue to be used for particle bombardment was excised and placed on nutritive agar plates (Horsch et al, 1988).…”

Section: Particle Gun Bombardmentsmentioning

confidence: 99%

The S-Ribonuclease Gene of Petunia hybrida Is Expressed in Nonstylar Tissue, Including Immature Anthers

Clark

Sims

1994

Plant Physiol.

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To determine the ability of isolated S-locus promoter sequences to direct organ-specific gene expression, we used microprojectile bombardment to introduce chimeric S-allele/&glucuronidase genes into different tissues of Petunia hybrida for transient expression. Histochemical staining showed that S-locus/B-glucuronidase fusions were expressed in pistil, ovary, and petal tissue. No expression of the chimeric genes was detected in leaves or in mature pollen, either by histochemical staining or by fluorescence assays. RNA blot hybridization confirmed that low levels of S-locus mRNA accumulate in petals and ovaries in vivo. Analysis of the expression pattern of S-locus promoter deletions showed that sequences in the immediate vicinity of the TATA box were sufficient to confer qualitatively correct organ-specific expression of &glucuronidase. To further investigate the potential for S-ribonuclease expression in pollen, we used the polymerase chain reaction to amplify RNA accumulated in developing anthers. These assays demonstrated that mRNA for the S-ribonuclease accumulates to low levels in developing anthers several days prior to corolla opening and pollen anthesis. We discuss these results in light of current models of selfincompatibility.

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Section: Particle Gun Bombardmentsmentioning

confidence: 99%

The S-Ribonuclease Gene of Petunia hybrida Is Expressed in Nonstylar Tissue, Including Immature Anthers

Clark

Sims

1994

Plant Physiol.

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“…Plasmid DNA was delivered to intact tissues by high-velocity microprojectiles using the Biolistics device as described (Klein et al 1988(Klein et al , 1989. A total of 8 ^lg of plasmid DNA (at 1.0 mg/ml) was precipitated onto microprojectiles by the addition of 25 yA of 50 mg/ml 1.0 ji-m tungsten, followed by the addition of 25 (xl of 1.0 M CaCli and 10 p-l 100 mM spermidine free base.…”

Section: High-velocity Microprojectile Bombardmentmentioning

confidence: 99%

Identification of functional domains in the maize transcriptional activator C1: comparison of wild-type and dominant inhibitor proteins.

1991

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Genes encoding fusions between the maize regulatory protein CI and the yeast transcriptional activator GAL4 and mutant CI proteins were assayed for their ability to trans-activate anthocyanin biosynthetic genes in intact maize tissues. The putative DNA-binding region of CI fused to the transcriptional activation domain of GAL4 activated transcription of anthocyanin structural gene promoters in cl aleurones, cl Rscm2 embryos, and cl r embryogenic callus. Cells receiving these constructs accumulated purple anthocyanin pigments. The Cl acidic region fused to the GAL4 DNA-binding domain activated transcription of a GAL4-regulated promoter. An internal deletion of Cl also induced pigmentation; however, frameshifts in either the amino-terminal basic or carboxy-terminal acidic region blocked tra/is-activation, and the latter generated a dominant inhibitory protein. Fusion constructs between the wild-type Cl cDNA and the dominant inhibitor allele Cl-I cDNA were used to identify the amino acid changes in Cl responsible for the Cl-I inhibitory phenotype. Results from these studies establish that amino acids within the myb-homologous domain are critical for transcriptional activation.

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“…The speed of the microprojectiles was controlled by pushing the macroprojectiles to a specific depth in the gun barrel. The depth (in 0.5-cm increments) that resulted in the highest GUS activity in tobacco suspension culture cells was used for a11 the experiments (Klein et al, 1988). The procedures for precipitating plasmid DNA on the microprojectiles were as described by Klein et al (1988).…”

Section: Particle-gun Experimentsmentioning

confidence: 99%

“…The depth (in 0.5-cm increments) that resulted in the highest GUS activity in tobacco suspension culture cells was used for a11 the experiments (Klein et al, 1988). The procedures for precipitating plasmid DNA on the microprojectiles were as described by Klein et al (1988). After bombardment, the plant material was incubated at 25°C overnight on the same p1,ates.…”

Section: Particle-gun Experimentsmentioning

confidence: 99%

“…Although others have found such transient assays to be reliable methods to measure gene activities, we have relied on the data obtained from the analyses of transgenic plants to study the isocitrate lyase 5' flanking region because of the potential problems in interpreting the results of the bombardment experiments (Klein et al, 1988(Klein et al, , 1989Bruce et al, 1989;Goff et al, 1990;Rolfe and Tobin, 1991;Twell et al, 1991).…”

Section: Comparison Of Promoter Activities In a Transient Expression mentioning

confidence: 99%

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DNA Sequences That Activate Isocitrate Lyase Gene Expression during Late Embryogenesis and during Postgerminative Growth

et al. 1996

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We analyzed DNA sequences that regulate the expression of an isocitrate lyase gene from Brasska napus L. during late embryogenesis and during postgerminative growth to determine whether glyoxysomal function is induced by a common mechanism at different developmental stages. P-Clucuronidase constructs were used both in transient expression assays in 6. napus and in transgenic Arabidopsis thaliana to identify the segments of the isocitrate lyase 5' flanking region that influence promoter activity. DNA sequences that play the principal role in activating the promoter during postgerminative growth are located more than 1200 bp upstream of the gene. Distinct DNA sequences that were sufficient for high-leve1 expression during late embryogenesis but only low-leve1 expression during postgerminative growth were also identified. Other parts of the 5' flanking region increased promoter activity both in developing seed and in seedlings. We conclude that a combination of elements is involved in regulating the isocitrate lyase gene and that distinct DNA sequences play primary roles in activating the gene in embryos and in seedlings. These findings suggest that different signals contribute to the induction of glyoxysomal function during these two developmental stages. We also showed that some of the constructs were expressed differently in transient expression assays and in transgenic plants.Glyoxysomes are peroxisomes with a specialized function in lipid mobilization. They contain enzymes of the P-oxidation pathway and the glyoxylate cycle that largely account for the ability of plants to utilize lipids as a carbon source (Trelease and Doman, 1984). These organelles play a prominent role in the metabolism of postgerminative seedlings by converting storage lipids into carbohydrates that serve as carbon and energy sources for the growing seedling (Bridenbach et al., 1967;Huang et al., 1983;Trelease, 1984). Functional glyoxysomes are also present in senescing organs, where they are thought to play a similar role in transforming membrane lipids into carbohydrates that can This work was supported by grants from the National Science Foundation. C.M.S. was supported in part by a fellowship from the North Atlantic Treaty Organization. Bellis and Nishimura, 1991;Wanner et al., 1991;Graham et al., 1992). The findings that genes encoding the two glyoxysome-specific enzymes, isocitrate lyase and malate synthase, are active during pollen and seed development as well as during postgerminative growth opens the possibility that peroxisomes present at these stages of the life cycle may have a glyoxysomal function (Pais and Feijo, 1987;Charzynska et al., 1989;Comai et al., 1989;Turley and Trelease, 1990;Zhang et al., 1994). Thus, glyoxysomal enzymes are present at severa1 stages of the plant life cycle.Although genes encoding the key glyoxylate-cycle enzymes are activated in developing pollen and in maturing seed, the pathway's precise metabolic role(s) at these developmental stages is not known. Understanding the mechanisms that induce these...

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Factors Influencing Gene Delivery into Zea Mays Cells by High–Velocity Microprojectiles

Cited by 232 publications

References 17 publications

The S-Ribonuclease Gene of Petunia hybrida Is Expressed in Nonstylar Tissue, Including Immature Anthers

The S-Ribonuclease Gene of Petunia hybrida Is Expressed in Nonstylar Tissue, Including Immature Anthers

Identification of functional domains in the maize transcriptional activator C1: comparison of wild-type and dominant inhibitor proteins.

DNA Sequences That Activate Isocitrate Lyase Gene Expression during Late Embryogenesis and during Postgerminative Growth

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