Tracking of the CaMV-35S promoter performance in GFP transgenic tobacco, with a special emphasis on flowers and reproductive organs, confirmed its predominant activity in vascular tissues

Hraška, Marek; Rakouský, S.; Čurn, Vladislav

doi:10.1007/s11240-007-9312-6

Cited by 24 publications

(8 citation statements)

References 25 publications

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“…In the mature cotyledon leaves and young leaves which developed subsequently, the intensity of green fluorescence was maintained (Figs 2f-h). Similar green fluorescence pattern was also demonstrated in transgenic grape (Dhekney et al 2008), tobacco (Hraska et al 2008) and other plant species transformed with gfp gene (Hraska et al 2006) during various developmental stages.…”

Section: Somatic Embryogenesissupporting

confidence: 73%

Green fluorescent protein as a visual selection marker for coffee transformation

et al. 2010

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Abstract:The green fluorescent protein (GFP) was used as a visual selectable marker to produce transgenic coffee (Coffea canephora) plants following Agrobacterium-mediated transformation. The binary vector pBECKS 2000.7 containing synthetic gene for GFP (sgfp) S65T and the hygromycin phosphotransferase gene hph both controlled by 35S cauliflower mosaic virus CaMV35S promoters was used for transformation. Embryogenic cultures were initiated from hypocotyls and cotyledon leaves of in vitro grown seedlings and used as target material. Selection of transformed tissue was carried out using GFP visual selection as the sole screen or in combination with a low level of antibiotics (hygromycin 10 mg/L), and the efficiency was compared with antibiotics selection alone (hygromycin 30 mg/L). GFP selection reduced the time for transformed somatic embryos formation from 18 weeks on a hygromycin (30 mg/L) antibiotics containing medium to 8 weeks. Moreover, visual selection of GFP combined with low level of antibiotics selection improved the transformation efficiency and increased the number of transformed coffee plants compared to selection in the presence of antibiotics. Molecular analysis confirmed the presence of the sgfp-S65T coding region in the regenerated plants. Visual screening of transformed cells using GFP by Agrobacterium-mediated transformation techniques was found to be efficient and therefore has the potential for development of selectable marker-free transgenic coffee plants.

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Section: Somatic Embryogenesissupporting

confidence: 73%

Green fluorescent protein as a visual selection marker for coffee transformation

et al. 2010

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“…The CYP78A9 promoter ( pCYP78A9 ) is specifically active in developing Arabidopsis ovules and seeds (Ito and Meyerowitz, ; Sotelo‐Silveira et al ., ), similar to the TaCYP78A3 promoter, which is specifically active in young panicles and immature wheat seeds (Figure S4). However, the 35S promoter showed relatively low activity in the ovules but drove high expression in developing seeds (Jenik and Irish, ; Hraška et al ., ). To further increase TaCYP78A3 expression in ovules only, we also expressed TaCYP78A3 under the control of the INNER NO OUTER promoter ( pINO ) (Villanueva et al ., ).…”

Section: Resultsmentioning

confidence: 97%

Expression of TaCYP78A3, a gene encoding cytochrome P450 CYP78A3 protein in wheat (Triticum aestivum L.), affects seed size

Wang

et al. 2015

The Plant Journal

120

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Several studies have described quantitative trait loci (QTL) for seed size in wheat, but the relevant genes and molecular mechanisms remain largely unknown. Here we report the functional characterization of the wheat TaCYP78A3 gene and its effect on seed size. TaCYP78A3 encoded wheat cytochrome P450 CYP78A3, and was specifically expressed in wheat reproductive organs. TaCYP78A3 activity was positively correlated with the final seed size. Its silencing caused a reduction of cell number in the seed coat, resulting in an 11% decrease in wheat seed size, whereas TaCYP78A3 over-expression induced production of more cells in the seed coat, leading to an 11-48% increase in Arabidopsis seed size. In addition, the cell number in the final seed coat was determined by the TaCYP78A3 expression level, which affected the extent of integument cell proliferation in the developing ovule and seed. Unfortunately, TaCYP78A3 over-expression in Arabidopsis caused a reduced seed set due to an ovule developmental defect. Moreover, TaCYP78A3 over-expression affected embryo development by promoting embryo integument cell proliferation during seed development, which also ultimately affected the final seed size in Arabidopsis. In summary, our results indicated that TaCYP78A3 plays critical roles in influencing seed size by affecting the extent of integument cell proliferation. The present study provides direct evidence that TaCYP78A3 affects seed size in wheat, and contributes to an understanding of the cellular basis of the gene influencing seed development.

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“…The overwhelming majority of pollen grains in the 35S pro :INP1 lines looked normal. Although cauliflower mosaic virus 35S is a strong constitutive promoter commonly used to overexpress genes in plants, its expression in anther tissues is somewhat controversial (McCormick et al, 1991;de Mesa et al, 2004;Hraška et al, 2008;Zhang et al, 2008). The levels of INP1 transcript in stage 9 buds were significantly higher in the 35S pro :INP1 lines than in the wild type (see Supplemental Figure 6 online); however, to prove that the construct is similarly overexpressed in tetrads is more challenging.…”

Section: Abnormalities In Inp1 Expression May Affect Global Exine Patmentioning

confidence: 99%

The Novel Plant Protein INAPERTURATE POLLEN1 Marks Distinct Cellular Domains and Controls Formation of Apertures in the Arabidopsis Pollen Exine

2012

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Pollen grains protect the sperm cells inside them with the help of the unique cell wall, the exine, which exhibits enormous morphological variation across plant taxa, assembling into intricate and diverse species-specific patterns. How this complex extracellular structure is faithfully deposited at precise sites and acquires precise shape within a species is not understood. Here, we describe the isolation and characterization of the novel Arabidopsis thaliana gene INAPERTURATE POLLEN1 (INP1), which is specifically involved in formation of the pollen surface apertures, which arise by restriction of exine deposition at specific sites. Loss of INP1 leads to the loss of all three apertures in Arabidopsis pollen, and INP1 protein exhibits a unique tripartite localization in developing pollen, indicative of its direct involvement in specification of aperture positions. We also show that aperture length appears to be sensitive to INP1 dosage and INP1 misexpression can affect global exine patterning. Phenotypes of some inp1 mutants indicate that Arabidopsis apertures are initiated at three nonrandom positions around the pollen equator. The identification of INP1 opens up new avenues for studies of how formation of distinct cellular domains results in the production of different extracellular morphologies.

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Tracking of the CaMV-35S promoter performance in GFP transgenic tobacco, with a special emphasis on flowers and reproductive organs, confirmed its predominant activity in vascular tissues

Cited by 24 publications

References 25 publications

Green fluorescent protein as a visual selection marker for coffee transformation

Green fluorescent protein as a visual selection marker for coffee transformation

Expression of TaCYP78A3, a gene encoding cytochrome P450 CYP78A3 protein in wheat (Triticum aestivum L.), affects seed size

The Novel Plant Protein INAPERTURATE POLLEN1 Marks Distinct Cellular Domains and Controls Formation of Apertures in the Arabidopsis Pollen Exine

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