Abstract
Background TRR14 protein is a small protein, a member of a multigene family in Arabidopsis which was found as the fi rst protein during screening seedlings for their resistant to the trehalose sugar. Objectives A number of TRR14-overexpressing plants were subjected to the characterization in the present research, among which, the associated morphological features and changes accompany growth pattern and photosynthesis related parameters. Materials and Methods TRR14 gene was isolated from Arabidopsis Thaliana and cloned into the pBin-35S vector. Recombinant vector was transferred to the Arabidopsis (Col-0) via Agrobacterium tumefaciens using the Floral Dipping method. Seeds from the TRR14 overexpressed (TRR14) and the Col-0 wild-type (WT) plants were shown on soil under long day conditions. Several measurements were then performed including determination of the fresh and dry weights, leaf area, chlorophyll a and b (Chl a and Chl b) content, Chl a/b ratio, total chlorophyll and carotenoids content, soluble and insoluble sugars content, total and soluble protein content, the Hill reaction rate, chlorophyll fl uorescence, as well as photorespiration rate. Meanwhile, the chloroplastic proteins were investigated by SDS-PAGE analysis. Results TRR14 plants showed a signifi cant increase in fresh and dry weights, leaf area, and total and soluble protein content along with a signifi cant decrease in the insoluble sugar contents was observed in comparison to the WT plants. Chl a, Chl b, total chlorophyll content, Chl a/b ratio, carotenoids content, Hill reaction rate, and chlorophyll fl uorescence didn’t show a signifi cant diff erence between TRR14 and WT plants. The SDS-PAGE gel electrophoresis of the chloroplastic proteins showed a thick band with a molecular mass of 25 kDa in TRR14-overexpressed plants, compared to the WT plants. Remarkably, photorespiration rate was decreased in TRR14 plants compared to WT plants. Conclusion The increased biomass of TRR14 transformed plants might be due to its ability in reducing photorespiration through concentrating CO2 in the leaf’s intercellular spaces.
Highlights
TRR14 protein is a small protein, a member of a multigene family in Arabidopsis which was found as the first protein during screening seedlings for their resistant to the trehalose sugar
Overexpression of TRR14 To investigate the physiologic role of TRR14, the corresponding cDNA was integrated into a binary vector pBin19 under the control of CaMV 35S promoter and transformed into the WT (Colombia-0 ecotype) seedlings
Three independent transgenic lines were selected for TRR14 gene expression analysis (T7, T12, and T17)
Summary
TRR14 protein is a small protein, a member of a multigene family in Arabidopsis which was found as the first protein during screening seedlings for their resistant to the trehalose sugar. Several measurements were performed including determination of the fresh and dry weights, leaf area, chlorophyll a and b (Chl a and Chl b) content, Chl a/b ratio, total chlorophyll and carotenoids content, soluble and insoluble sugars content, total and soluble protein content, the Hill reaction rate, chlorophyll fluorescence, as well as photorespiration rate. Results: TRR14 plants showed a significant increase in fresh and dry weights, leaf area, and total and soluble protein content along with a significant decrease in the insoluble sugar contents was observed in comparison to the WT plants. Proteins containing conserved Cupin domain play diverse functions in the metabolism and signaling. Enzymes such as: phosphomanoseisomerase, polyketide synthase, dioxygenase, oxalate oxidase (germins), and auxin binding protein belong to this group [2, 3]. This protein has been localized in the chloroplast and its expression is ubiquitous [4]
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