Coordinated Expression of Cytosolic and Chloroplastic Glutamine Synthetase During Reproductive Stage and Its Impact in GS1 RNAi Transgenic Rice

doi:10.1016/j.rsci.2018.08.001

摘要/Abstract

. [J]. Rice Science, 2018, 25(5): 250-260.

Fig. 1. Expression level of glutamine synthetase (GS) genes in various organs of rice plants. A, GS mRNA expression. RNA was extracted from the root (R), leaf sheath (LS), and leaf (L) of 4-week-old seedling, while panicle (P) was collected at the pre-flowering stage. B, GS protein accumulation. C, Immunolocalization of GS protein in flag leaf. D, Immunolocalization of GS protein in leaf sheath. E, Immunolocalization of GS protein in anther. F, Immunolocalization of GS protein in carpel.Arrows indicate vascular bundles. An, Anther; Ca, Carpel; Fu, Funiculus; Ld, Lodicule; Lo, Locule.

Fig. 2. Glutamine synthetase (GS) expression in wild type (WT) and GS1 RNAi transgenic (GS1-RNAi) rice plants. A, GS mRNA expression. reverse-transcription PCR analysis was performed using the leaves of 4-week-old seedlings. B, GS protein accumulation in the leaves of transgenic plants. C, Immunolocalization of GS protein in the leaf sheath of transgenic rice. D, Immunolocalization of GS protein in the leaf sheath of the WT. Arrows indicate vascular bundles.

Fig. 3. Expression of glutamine synthetase (GS) mRNAs in flag leaf (A), leaf sheath (B) and panicle (C) during reproductive stages from pre- to post-flowering.WT, Wild type; T, Transgenic rice; I, Pre-flowering stage (7 d before flowering); II, Flowering stage; III, Post-flowering stage (7 d after flowering).

Fig. 4. Accumulation of GS protein in flag leaf, leaf sheath and panicle from pre- to post-flowering stage. A, GS1 and GS2 protein analysis in the RNAi transgenic rice. B, Immunoblot of 2D electrophoresis profile. Total protein (40 μg) was separated by two-dimensional gel electrophoresis. WT, Wild type; T, GS1;1 RNAi transgenic rice; I, Pre-flowering stage (7 d before flowering); II, Flowering stage; III, Post-flowering stage (7 d after flowering).

Fig. 5. Ammonium (NH4+) contents in the flag leaves and panicles between GS1 RNAi transgenic and wild-type rice during reproductive stages.A, Ammonium content in leaves; B, Ammonium content in panicles; C, Total soluble protein in leaves; D, Total soluble protein in panicles.WT, Wild type; T, Transgenic rice; I, Pre-flowering stage (7 d before flowering); II, Flowering stage; III, Post-flowering stage (7 d after flowering). Data are mean ± SE from three independent experiments using three randomly selected plants. Asterisks represent significant differences (two-way ANOVA and Bonferroni post-test, *, P < 0.05; **, P < 0.01; ***, P < 0.001).

Fig. 6. Amino acid concentrations in flag leaves and panicles of GS1 RNAi transgenic and wild-type rice during the reproductive stages. T, Transgenic rice; WT, Wild type; Asp, Aspartate; Glu, Glutamic acid; Gln, Glutamine; Asn, Asparagine; Lys, Lycine; Thr, Threonine; Met, Methionine; Ile, Isoleucine.Data are mean ± SE of measurements from three independent experiments using three randomly selected plants. Asterisks represent significant differences (two-way ANOVA and Bonferroni post-test, *, P < 0.05; **, P < 0.01; ***, P < 0.001).

Fig. 7. Phenotype of GS1 RNAi transgenic (T) and wild-type (WT) rice at 16th week after transplanting. A, Panicles; B, Ears; C, Number of panicles per plant; D, Number of seeds per panicle. Data are mean ± SE of measurements from 12 individual plants in three replicates. Asterisk indicates that the value significantly differed from the wild type (one-way ANOVA; ***, P < 0.001).

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