Histochemical analysis of CaMV 35S promoter-?-glucuronidase gene expression in transgenic rice plants

Battraw, Michael J.; Hall, Timothy C.

doi:10.1007/bf00017828

Cited by 192 publications

(93 citation statements)

References 24 publications

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“…For this purpose, transient expression of the uidA reporter gene was evaluated under the control of a number of promoters. Although the CaMV 35S promoter has been shown to be active in most tissues of several plant species (26), in this research it was found to be the weakest of the promoters tested in sugarcane. While differences in transient gene expression level were observed among the monocot promoters, the highest activity was obtained with the Act1 promoter, which is in agreement with previous reports on the Act1-D promoter activity in other monocots such as date palm (16) and rice (27).…”

Section: Discussionmentioning

confidence: 52%

Sugarcane (NCo310) Transient Transformation Using uidA Reporter Gene

et al. 2013

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Background: Sugarcane is a monocotyledonous crop that is cultivated in the tropical and subtropical regions of the world. One of the most important criteria, influencing the efficiency of the sugarcane transformation is known to be related to physical and biological factors during the transformation procedure. Objectives: The objective of this research was to study the response of callus induction and embryogenic callus production and to identify the major parameters controlling DNA delivery by particle bombardment into sugarcane (Saccharum officinarum L.) cv. NCo310. Materials and Methods: For callus induction and embryogenic callus production, leaf base segments were subjected to in vitro culture medium supplemented with two plant growth regulators (2,4-D and Dicamba). Results showed that 1 mg.L -1 2,4-D was significantly influential in callus induction and embryogenic callus production. Considering both physical and biological factors, the efficiency of DNA (uidA gene) delivery was assessed by scoring transient GUS (gene (β-glucuronidase) ) expression in bombarded tissues. Results: The highest transient GUS expression was obtained when callus was bombarded with the construct harboring rice Act1 promoter, and having 9 cm target distance, 25 inHg vacuum pressure, 1 µm gold particles, 12.5 µg DNA per bombardment and one day pre-culture prior to the bombardment. Conclusions: A bombardment procedure suitable for elite sugarcane varieties was developed, which allowed high-efficiency DNA delivery combined with reduced damage to target tissues.

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Section: Discussionmentioning

confidence: 52%

Sugarcane (NCo310) Transient Transformation Using uidA Reporter Gene

et al. 2013

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“…As the somatic embryos matured into plantlets, there was preferential expression of GUS in leaves over roots. Higher levels of GUS activity were found in leaves than in roots of rice (2) but not tobacco (3, 1 1) transformed with CaMV 35S-GUS constructions. The CaMV 35S promoter possesses domains that confer tissue-specific and developmentally regulated expression (3) that may be recognized differently in plants evolutionarily divergent from tobacco, such as rice and yellow-poplar, a member of the Magnoliaceae.…”

Section: Discussionmentioning

confidence: 88%

Expression of Foreign Genes in Transgenic Yellow-Poplar Plants

Wilde

Meagher²,

Merkle³

1992

Plant Physiol.

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Cells of yellow-poplar (Liriodendron tulipifera L.) were transformed by direct gene transfer and regenerated into plants by somatic embryogenesis. Plasmid DNA bearing marker genes encoding,8-glucuronidase (GUS) and neomycin phosphotransferase (NPT II) were introduced by microprojectile bombardment into single cells and small cell clusters isolated from embryogenic suspension cultures. The number of full-length copies of the GUS gene in independently transformed callus lines ranged from approximately 3 to 30. An enzyme-linked immunosorbent assay for NPT II and a fluorometric assay for GUS showed that the expression of both enzymes varied by less than fourfold among callus lines. A histochemical assay for GUS activity revealed a heterogeneous pattern of staining with the substrate 5-bromo-4-chloro-3-indoyl-O-D-glucuronic acid in some transformed cell cultures. However, cell clusters reacting positively (blue) or negatively (white) with 5-bromo-4-chloro-3-indoyl-,8-D-glucuronic acid demonstrated both GUS activity and NPT II expression in quantitative assays. Somatic embryos induced from transformed cell cultures were found to be uniformly GUS positive by histochemical analysis. All transgenic plants sampled expressed the two marker genes in both root and shoot tissues. GUS activity was found to be higher in leaves than roots by fluorometric and histochemical assays. Conversely, roots expressed higher levels of NPT II than leaves.

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“…GUS histochemical staining was performed essentially as described previously (1,19). GUS activity was localized histochemically using a staining solution containing 1 mM X-Gluc (Sigma, Aldrich, MO), 50 mM sodium phosphate (pH 7.0), 10 mM sodium EDTA (pH 8.0), 0.5 mM potassium ferrocyanide, 0.5 mM potassium ferricyanide, 20% (vol/vol) methanol, and 0.05% Triton X-100.…”

Section: Methodsmentioning

confidence: 99%

Genetic Determinants of Symptoms on Viral DNA Satellites

Ding

Qing

et al. 2009

Appl Environ Microbiol

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Begomovirus-DNA-␤ disease complexes induce different symptom phenotypes in their hosts. To investigate the genetic determinants of the phenotypic differences, Nicotiana spp. and tomato plants were inoculated with infectious clones of Tobacco curly shoot virus (TbCSV)/TbCSV DNA-␤ (TbCSB) and Tomato yellow leaf curl China virus (TYLCCNV)/TYLCCNV DNA-␤ (TYLCCNB) pseudorecombinants and showed that TYLCCNB induced characteristic vein-thickening and enation symptoms, while TbCSB only slightly exacerbated the leaf-curling symptoms, regardless of the helper virus being used. The roles of DNA-␤-encoded ␤C1 and a 430-nucleotide fragment containing the A-rich region and the putative ␤C1 promoter region of the ␤C1 gene (referred to as AP) in symptom development were further investigated by constructing hybrid satellites in which the ␤C1 coding region or AP was exchanged between the two satellite molecules. A TYLCCNB hybrid with TbCSB ␤C1 lost the ability to elicit the vein-thickening and enation phenotypes. TbCSB hybrids containing the TYLCCNB ␤C1 or AP fragment failed to induce the characteristic vein thickening and enations. A TYLCCNB hybrid having the TbCSB AP fragment produced the enations, but the number of enations was less and their sizes were reduced. Differently from the phloem-specific pattern of the TYLCCNB promoter, a full-length fragment upstream of the TbCSB ␤C1 gene confers a constitutive ␤-glucuronidase expression pattern in transgenic tobacco plants. The above results indicate that the DNA-␤-encoded ␤C1 protein is the symptom determinant, but the promoter of the ␤C1 gene has influence on symptom production.Geminiviruses are small plant viruses with circular singlestranded DNA (ssDNA) genomes that are encapsidated in unique twinned (geminate) particles. Members of the genus Begomovirus are transmitted by whiteflies (Bemisia tabaci) and infect dicotyledonous plants (42). Begomoviruses have either one or two circular ssDNA genomic components (DNA-A and DNA-B). The DNA-A component is capable of autonomous replication and encapsidation, whereas the DNA-B component encodes two proteins (BC1 and BV1) involved in movement (14). Recently, some monopartite begomoviruses have been found in association with a novel satellite DNA molecule, referred to as DNA-␤ and now known as a betasatellite (2,5,20,22,38,45). DNA-␤ is approximately half the size of the viral genomic DNA, and apart from a nonanucleotide sequence (TA ATATTAC), it has little sequence identity with viral genomic DNA. DNA-␤ depends on the helper virus for replication and encapsidation and, in turn, is required for the induction of bona fide disease symptoms. DNA-␤ bears a ␤C1 open reading frame (ORF) on the complementary-sense strand, which is conserved among distinct betasatellites in terms of position and size. Mutational analyses and constitutive expression have shown that ␤C1 is a strong pathogenicity/symptom determinant (7, 34, 39).Begomovirus-DNA-␤ disease complexes are associated with a wide range of plant species and induce different sets of sympto...

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Histochemical analysis of CaMV 35S promoter-?-glucuronidase gene expression in transgenic rice plants

Cited by 192 publications

References 24 publications

Sugarcane (NCo310) Transient Transformation Using uidA Reporter Gene

Sugarcane (NCo310) Transient Transformation Using uidA Reporter Gene

Expression of Foreign Genes in Transgenic Yellow-Poplar Plants

Genetic Determinants of Symptoms on Viral DNA Satellites

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