Genetic transformation of the C<sub>4</sub> plant, <i>Flaveria bidentis</i>

Chitty, Julie A.; Furbank, Robert T.; Marshall, Jerry S.; Chen, Zhenghua; Taylor, William C.

doi:10.1046/j.1365-313x.1994.6060949.x

Cited by 64 publications

(53 citation statements)

References 33 publications

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“…2A) and examined its expression behavior in transgenic F. bidentis plants. The C 4 species F. bidentis is a close relative to F. trinervia, but unlike F. trinervia it is suitable for transformation by Agrobacterium tumefaciens-mediated gene transfer (Chitty et al, 1994).…”

Section: In Situ Rna Hybridizationmentioning

confidence: 99%

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The Gene for the P-Subunit of Glycine Decarboxylase from the C4 SpeciesFlaveria trinervia: Analysis of Transcriptional Control in TransgenicFlaveria bidentis(C4) and Arabidopsis (C3)

et al. 2008

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Glycine decarboxylase (GDC) plays an important role in the photorespiratory metabolism of plants. GDC is composed of four subunits (P, H, L, and T) with the P-subunit (GLDP) serving as the actual decarboxylating unit. In C3 plants, GDC can be found in all photosynthetic cells, whereas in leaves of C3-C4 intermediate and C4 species its occurrence is restricted to bundle-sheath cells. The specific expression of GLDP in bundle-sheath cells might have constituted a biochemical starting point for the evolution of C4 photosynthesis. To understand the molecular mechanisms responsible for restricting GLDP expression to bundle-sheath cells, we performed a functional analysis of the GLDPA promoter from the C4 species Flaveria trinervia. Expression of a promoter-reporter gene fusion in transgenic plants of the transformable C4 species Flaveria bidentis (C4) showed that 1,571 bp of the GLDPA 5′ flanking region contain all the necessary information for the specific expression in bundle-sheath cells and vascular bundles. Interestingly, we found that the GLDPA promoter of F. trinervia exhibits a C4-like spatial activity also in the C3 plant Arabidopsis (Arabidopsis thaliana), indicating that a mechanism for bundle-sheath-specific expression is also present in this C3 species. Using transgenic Arabidopsis, promoter deletion studies identified two regions in the GLDPA promoter, one conferring repression of gene expression in mesophyll cells and one functioning as a general transcriptional enhancer. Subsequent analyses in transgenic F. bidentis confirmed that these two segments fulfill the same function also in the C4 context.

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Section: In Situ Rna Hybridizationmentioning

confidence: 99%

“…Arabidopsis plants were transformed via the floral dip method according to Clough and Bent (1998). The transformation of Flaveria bidentis was performed as described by Chitty et al (1994). The integration of the transgenes into the genome of regenerated F. bidentis or T 1 Arabidopsis plants was proved by PCR analyses.…”

Section: Plant Transformationmentioning

confidence: 99%

The Gene for the P-Subunit of Glycine Decarboxylase from the C4 SpeciesFlaveria trinervia: Analysis of Transcriptional Control in TransgenicFlaveria bidentis(C4) and Arabidopsis (C3)

et al. 2008

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“…F. bidentis and Arabidopsis were transformed as described (Chitty et al, 1994;Clough and Bent, 1998). Integration of the chimerical genes into the genome was confirmed by PCR.…”

Section: Generation Of Transgenic F Bidentis and Arabidopsis Thalianamentioning

confidence: 99%

Evolution and Function of acis-Regulatory Module for Mesophyll-Specific Gene Expression in the C4 DicotFlaveria trinervia

et al. 2007

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C4 photosynthesis presents a sophisticated integration of two complementary cell types, mesophyll and bundle sheath cells. It relies on the differential expression of the genes encoding the component enzymes and transporters of this pathway. The entry enzyme of C4 photosynthesis, phosphoenolpyruvate carboxylase (PEPC), is found exclusively in mesophyll cells, and the expression of the corresponding gene is regulated at the transcriptional level. In the C4 dicot Flaveria trinervia, the mesophyll-specific expression of the C4 PEPC gene (ppcA) depends on a 41-bp segment in the distal promoter region referred to as MEM1 (for mesophyll expression module1). Here, we show that a MEM1 sequence found in the orthologous ppcA gene from the C3 species Flaveria pringlei is not able to direct mesophyll-specific gene expression. The two orthologous MEM1 sequences of F. pringlei and F. trinervia differ at two positions, a G-to-A exchange and the insertion of the tetranucleotide CACT. Changes at these two positions in the C3 MEM1 sequence were necessary and sufficient to create a mesophyll-specificity element during C4 evolution. The MEM1 of F. trinervia enhances mesophyll expression and concomitantly represses expression in bundle sheath cells and vascular bundles.

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“…For production of transgenic plants, the AhRbcS1 UTR-derived expression constructs and the control construct described above were inserted into the binary vector pGA482 (An, 1987) and used for transformation of F. bidentis hypocotyls (Chitty et al 1994). Although all three of the AhRbcS1 UTR-derived expression constructs described above were used for transformation of F. bidentis hypocotyls, only the binary vector containing the CaMV/AhRbcS1-5#-UTRgusA-3#-UTR expression cassette (from pGG53) led to the recovery of regenerated C 4 plants.…”

Section: Ahrbcs1-5#-utr-gusa-3#-utr Expression Levels In Leaves Of Trmentioning

confidence: 99%

Untranslated Regions from C4 Amaranth AhRbcS1 mRNAs Confer Translational Enhancement and Preferential Bundle Sheath Cell Expression in Transgenic C4Flaveria bidentis

et al. 2004

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Many aspects of photosynthetic gene expression are posttranscriptionally regulated in C4 plants. To determine if RbcS mRNA untranslated regions (UTRs) in themselves could confer any characteristic C4 expression patterns, 5′- and 3′-UTRs of AhRbcS1 mRNA from the C4 dicot amaranth were linked to a gusA reporter gene. These were constitutively transcribed from a cauliflower mosaic virus promoter and assayed for posttranscriptional expression patterns in transgenic lines of the C4 dicot Flaveria bidentis. Three characteristic C4 expression patterns were conferred by heterologous AhRbcS1 UTRs in transgenic F. bidentis. First, the AhRbcS1 UTRs conferred strong translational enhancement of gusA expression, relative to control constructs lacking these UTRs. Second, while the UTRs did not appear to confer tissue-specific expression when analyzed by β-glucuronidase activity assays, differences in gusA mRNA accumulation were observed in leaves, stems, and roots. Third, the AhRbcS1 UTRs conferred preferential gusA expression (enzyme activity and gusA mRNA accumulation) in leaf bundle sheath cells. AhRbcS1 UTR-mediated translational enhancement was also observed in transgenic C3 plants (tobacco [Nicotiana tabacum]) and in in vitro translation extracts. These mRNAs appear to be translated with different efficiencies in C4 versus C3 plants, indicating that processes determining overall translational efficiency may vary between these two categories of higher plants. Our findings suggest that the AhRbcS1 5′-UTR functions as a strong translational enhancer in leaves and other tissues, and may work synergistically with the 3′-UTR to modulate overall levels of Rubisco gene expression in different tissues and cell types of C4 plants.

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Genetic transformation of the C₄ plant, Flaveria bidentis

Cited by 64 publications

References 33 publications

The Gene for the P-Subunit of Glycine Decarboxylase from the C4 SpeciesFlaveria trinervia: Analysis of Transcriptional Control in TransgenicFlaveria bidentis(C4) and Arabidopsis (C3)

The Gene for the P-Subunit of Glycine Decarboxylase from the C4 SpeciesFlaveria trinervia: Analysis of Transcriptional Control in TransgenicFlaveria bidentis(C4) and Arabidopsis (C3)

Evolution and Function of acis-Regulatory Module for Mesophyll-Specific Gene Expression in the C4 DicotFlaveria trinervia

Untranslated Regions from C4 Amaranth AhRbcS1 mRNAs Confer Translational Enhancement and Preferential Bundle Sheath Cell Expression in Transgenic C4Flaveria bidentis

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Genetic transformation of the C4 plant, Flaveria bidentis

Cited by 64 publications

References 33 publications

The Gene for the P-Subunit of Glycine Decarboxylase from the C4 SpeciesFlaveria trinervia: Analysis of Transcriptional Control in TransgenicFlaveria bidentis(C4) and Arabidopsis (C3)

The Gene for the P-Subunit of Glycine Decarboxylase from the C4 SpeciesFlaveria trinervia: Analysis of Transcriptional Control in TransgenicFlaveria bidentis(C4) and Arabidopsis (C3)

Evolution and Function of acis-Regulatory Module for Mesophyll-Specific Gene Expression in the C4 DicotFlaveria trinervia

Untranslated Regions from C4 Amaranth AhRbcS1 mRNAs Confer Translational Enhancement and Preferential Bundle Sheath Cell Expression in Transgenic C4Flaveria bidentis

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Genetic transformation of the C₄ plant, Flaveria bidentis