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Fig. 3. | BMC Biology

Fig. 3.

From: Comparative gene retention analysis in barley, wild emmer, and bread wheat pangenome lines reveals factors affecting gene retention following gene duplication

Fig. 3.

Gene expression pattern, subcellular localization of HvHPT2, and tocopherol levels in HvHPT2 overexpression transgenic lines. A Semi-quantitative RT-PCR of HvHPT1 and HvHPT2 in different tissues of barley. B Quantitative RT-PCR was applied to detect the expression level of HvHGGT, HvHPT1, and HvHPT2 in embryo, endosperm, and husk of barley grain (16 days after pollination, DAP), respectively. Cβ-Glucuronidase (GUS) reporter gene expression in transgenic rice plants expressing the HvHPT2 promoter-GUS constructs. The spike of heading stage and the grains of 14 DAP were excised and stained with GUS solution. Bar: 2mm. D Transient expression of proteins with GFP-tag in protoplast cells of barely. GFP fluorescence, chlorophyll autofluorescence, merged signals, and bright field are indicated above the panels. Bar: 10 μm. E The expression level of HvHPT2 in transgenic barley leaves and grains. F, G HPLC profiles of tocopherol isomers in the leaf and grain of wild-type barley and its transgenic lines (35S:HPT2_2#, 16# and 27#), respectively. H The content of four tocopherol isomers in leaf. I The content of four tocopherol isomers in grain. J The total content of tocopherols in leaf and grain. The leaves of 2-month-old T1 transgenic lines and their mature grains were used for HPLC analysis. Fifty-milligrams of leaf or grain powder was used in each replicate. The means and standard errors of at least three replicates are presented. Asterisks (* or **) indicate a significant difference between the wild-type and transgenic lines at P < 0.05 or P < 0.01, respectively, as determined by Student’s t tests. D, day; DW, dry weight; T, tocopherol

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