Transcription profiling of wheat treated with organic and inorganic fertlizers (N source), dose comparison
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ABSTRACT: Effect of nitrogen supply and nitrogen supply form on the transcriptome of wheat grain. Differences reflecting the use of inorganic versus organic fertiliser regimes.
Project description:B1355-4-2 expresses five HMW subunits encoded by Glu-B1 (14, 15) and Glu D1 (5, 10) and transgene Glu-A1 (Ax1).Cadenza does not express Glu-A1 (Ax1. Line B1355-4-2(18) was generated by co-transformation with the ?clean? fragments of the HMW-GS 1Ax1 transgene (Halford, N.G. et al. Analysis of HMW glutenin subunits encoded by chromosome 1A of bread wheat (Triticum aestivum L.) indicates quantitative effects on grain quality. Theor Appl Genet 83, 373-378 (1992).)and the bar gene sequence. We compared the transcriptome of transgenic B1355-4-2(18) wheat line with their its background control-Cadenza bread wheat line. Transcriptomes comparisons were performed in endosperm tissue (14 and 28 days post anthesis-dpa) and in leaf tissue (8 days post germination ?dpg). Each of the transcriptome comparisons analaysis was performed using three biological replicates (i.e. per line/tissue/developmental stage selected). Hybridisations were performed in reverse dye labelling.
Project description:The high molecular weight (HMW) subunits of wheat glutenin are synthesised only in the starchy endosperm tissue of the developing wheat grain. We studied the effect of introducing transgenes on the global gene expression profiles of selected transgenic wheat lines, particularly during wheat seed development. For these particular set of experiments a direct comparison between the hexaploid bread transgenic line B102,1-1 (Rooke, L., Steele, S.H., Barcelo, P.,Shewry, P.R. & Lazzeri,P. Transgene inheritance, segregation and expression in bread wheat. Euphytica 129, 301-309 (2003)) and it background, non transformed L88-31 wheat line (Lawrence,G.J., Macritchie, F. & Wrigley, C.W. Dough and baking quality of wheat lines in glutenin subunits controlled by Glu-A1, Glu-B1 and Glu-D1 loci. J. Cereal. Sci. 7,109-112 (1988)) was performed. Transcriptome comparison analysis was performed in endosperm tissue (14 and 28 days post anthesis-dpa) and in leaf tissue (8 days post germination ?dpg). The transcriptome comparisons analysis was performed using three biological replicates (i.e. per line/tissue /developmental stage selected). Hybridisations were performed in reverse dye labelling.
Project description:Transgenic cadenza lines express five HMW subunits encoded by:Glu-B1 (14, 15) and Glu D1 (5, 10) and transgene Glu-A1 (Ax1). Transcriptomes of the wheat line B1118-8-4(6) (produced by co transformation with two plasmids: one carried the transgene HMW-GS 1Ax1and other the bar and gus gene sequences) and wheat line B1355-4-2(18) (generated by co transformation with the clean fragments of the HMW-GS 1Ax1 transgene and the bar gene sequence) were compared. Transcriptomes analysis was performed in endosperm tissue (14 and 28 days post anthesis-dpa) and in leaf tissue (8 days post germination dpg). Each of the transcriptome comparisons was performed using three biological replicates (i.e. per line/tissue /developmental stage selected). Hybridisations were performed in reverse dye labelling
Project description:The high molecular weight (HMW) subunits of wheat glutenin are synthesised only in the starchy endosperm tissue of the developing wheat grain. We compared the expressed genomes of the transgenic wheat line B102,1-1 (Rooke et al. Transgene inheritance, segregation and expression in bread wheat. Euphytica 129, 301-309 (2003)). Both lines were shown to express the HMW-GS Ax1 gene (Halford, N.G. et al. Analysis of HMW glutenin subunits encoded bychromosome 1A of bread wheat (Triticum aestivum L.) indicates quantitative effects on grain quality. Theor Appl Genet 83, 373-378 (1992).) to the expressed genome of conventionally bred wheat line L88-18 (Lawrence et al. Dough and baking quality of wheat lines in glutenin subunits controlled by Glu-A1, Glu-B1 and Glu-D1 loci. J. Cereal. Sci. 7,109-112 (1988)) which results in the same effects on traits. Transcriptomes comparison analysis was performed in endosperm tissue (14 and 28 days post anthesis-dpa) and in leaf tissue (8 days post germination dpg), respectively. Each of the transcriptome comparisons was performed using three biological replicates (i.e. per line/tissue /developmental stage selected). Hybridisations were performed in reverse dye labelling.Exceptionally, biological replica 2 was only performed for B102,1-1 (green)/L88-18 (red) labelling and not swap
Project description:The high molecular weight (HMW) subunits of wheat glutenin are synthesised only in the starchy endosperm tissue of the developing wheat grain. The transcriptomes of the lines L88-18 and L88-31 (Lawrence, G.J., Macritchie, F. & Wrigley, C.W. Dough and baking quality of wheat lines in glutenin subunits controlled by Glu-A1, Glu-B1 and Glu-D1 loci. J. Cereal. Sci. 7,109-112 (1988)) coming from the same cross were compared. Transcriptomes analysis was performed in endosperm tissue (14 and 28 days post anthesis-dpa) and in leaf tissue (8 days post germination dpg). Each of the transcriptome comparisons was performed using three biological replicates (i.e. per line/tissue /developmental stage selected). Hybridisations were performed in reverse dye labelling.
Project description:B1118-8-4 expresses five HMW subunits encoded by Glu-B1 (14, 15) and Glu D1 (5, 10) and transgene Glu-A1 (Ax1).Cadenza does not express Glu-A1 (Ax1. B111-8-8-4 (6)transgenic wheat line was produced by co-transformation with two plasmids: one of the plasmids carring the transgene HMW-GS 1Ax1 (Halford, N.G. et al. Analysis of HMW glutenin subunits encoded bychromosome 1A of bread wheat (Triticum aestivum L.) and the other plasmid harbour the marker genes bar and gus indicates quantitative effects on grain quality. Theor Appl Genet 83, 373-378 (1992).) and the other the bar gus gene sequences. We compared the transcriptome of transgenic B1118-8 4(6) wheat line with its background control-Cadenza bread wheat line. Transcriptomes analysis was performed in endosperm tissue (14 and 28 days post anthesis-dpa) and in leaf tissue (8 days post germination dpg). Each of the transcriptome comparisons was performed using three biological replicates (i.e. per line/tissue/developmental stage selected). Hybridisations were performed in reverse dye labelling.
Project description:The high molecular weight (HMW) subunits of wheat glutenin are synthesised only in the starchy endosperm tissue of the developing wheat grain. To place the differences observed between the endosperms of the transgenic and non-transgenic lines in a wider developmental context, the transcriptomes of endosperm at 14 dpa and leaf at 8 dpg of the transgenic line B102,1-1 were also compared.The experiment was performed with three biological replicates and hybridisations were performed in reverse dye labelling.
Project description:Isofemale line ED321 Heterogonic was used. Fresh faeces were collected from S. ratti-infected rats at days 5, 6, 7 and 8 p.i. and L1s prepared with a Baermann funnel held for 6 h at 19oC. The larvae were concentrated by centrifugation and then cleaned by flotation on 60% v/v sucrose. Infective L3s were harvested from 14 day-old faecal cultures that had been maintained at 19oC. The iL3s were cleaned by sucrose flotation as for the L1s, above. In excess of 150,000 larvae of either stage were routinely isolated from 6 infected hosts. The experimental design used, was to have at least three biological replicates for each sample (i.e. three independent preparations of the relevant worm samples and their RNA) and to have at least three technical replicates (i.e. independent, separate cDNA synthesis, amplification and hybridization etc.) for each biological replicate. For each hybridisation a dye-swap was used i.e. each sample to be used in a hybridisation was labelled, separately, with each of the two dyes.
Project description:S. ratti Isofemale line ED321 Heterogonic predominantly undergoes indirect development; isofemale line ED5 Homogonic predominantly undergoes direct development. Therefore, L2 stages of ED321 Heterogonic and of ED5 Homogonic are destined for indirect (i.e. L2 indirect) and direct (i.e. L2 direct) development, respectively; these sources of material were used in this comparison. To do this, for both isofemale lines, rats were infected with ED321 Heterogonic or ED5 Homogonic and faeces collected on days 5, 6, 7 and 8 p.i. and cultured for 24 h at 19oC, after which larvae were prepared with a Baermann funnel held for 6 h at 19oC, larvae were concentrated and cleaned by sucrose flotation, as above. In excess of 75,000 worms were routinely isolated from three infected hosts. The experimental design used, was to have at least three biological replicates for each sample (i.e. three independent preparations of the relevant worm samples and their RNA) and to have at least three technical replicates (i.e. independent, separate cDNA synthesis, amplification and hybridization etc.) for each biological replicate. For each hybridisation a dye-swap was used i.e. each sample to be used in a hybridisation was labelled, separately, with each of the two dyes.
Project description:During grain filling in barley reserves are remobilized from vegetative organs like glumes. In this expression analysis values from glumes and endosperm material were compared from 0 - 24 day after pollination (DAP). This study showed that glumes metabolism and development is adjusted to changing grain demands. Candidate genes are potentially involved in assimilate conversion and translocation.