Virus-Induced Gene Silencing Identifies an Important Role of the TaRSR1 Transcription Factor in Starch Synthesis in Bread Wheat
AbstractThe function of a wheat starch regulator 1 (TaRSR1) in regulating the synthesis of grain storage starch was determined using the barley stripe mosaic virus—virus induced gene-silencing (BSMV-VIGS) method in field experiments. Chlorotic stripes appeared on the wheat spikes infected with barley stripe mosaic virus-virus induced gene-silencing- wheat starch regulator 1 (BSMV-VIGS-TaRSR1) at 15 days after anthesis, at which time the transcription levels of the TaRSR1 gene significantly decreased. Quantitative real-time PCR was also used to measure the transcription levels of 26 starch synthesis-related enzyme genes in the grains of BSMV-VIGS-TaRSR1-silenced wheat plants at 20, 27, and 31 days after anthesis. The results showed that the transcription levels of some starch synthesis-related enzyme genes were markedly induced at different sampling time points: TaSSI, TaSSIV, TaBEIII, TaISA1, TaISA3, TaPHOL, and TaDPE1 genes were induced at each of the three sampling time points and TaAGPS1-b, TaAGPL1, TaAGPL2, TaSSIIb, TaSSIIc, TaSSIIIb, TaBEI, TaBEIIa, TaBEIIb, TaISA2, TaPHOH, and TaDPE2 genes were induced at one sampling time point. Moreover, both the grain starch contents, one thousand kernel weights, grain length and width of BSMV-VIGS-TaRSR1-infected wheat plants significantly increased. These results suggest that TaRSR1 acts as a negative regulator and plays an important role in starch synthesis in wheat grains by temporally regulating the expression of specific starch synthesis-related enzyme genes. View Full-Text
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Description: Figure S1. cDNA sequences for three copies of f TaRSR1 gene. The fragment used to VIGS silencing has been underlined. Figure S2. Transcription levels of the TaRSR1 gene in the BSMV-TaRSR1-infected and BSMV-GFP-infected wheat plants. Figure S3. Transcription levels of the 26 starch synthesis enzyme genes in the grains of BSMV-TaRSR1-infected and BSMV-GFP-infected wheat plants. Figure S4. Schematic construction of the recombinant BSMV-TaRSR1 vector. Figure S5. BSMV-TaRSR1 and BSMV-GFP virus inoculation under field conditions. Table S1. Target genes for analyzing transcription levels and their primer sequences.
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Liu, G.; Wu, Y.; Xu, M.; Gao, T.; Wang, P.; Wang, L.; Guo, T.; Kang, G. Virus-Induced Gene Silencing Identifies an Important Role of the TaRSR1 Transcription Factor in Starch Synthesis in Bread Wheat. Int. J. Mol. Sci. 2016, 17, 1557.
Liu G, Wu Y, Xu M, Gao T, Wang P, Wang L, Guo T, Kang G. Virus-Induced Gene Silencing Identifies an Important Role of the TaRSR1 Transcription Factor in Starch Synthesis in Bread Wheat. International Journal of Molecular Sciences. 2016; 17(10):1557.Chicago/Turabian Style
Liu, Guoyu; Wu, Yufang; Xu, Mengjun; Gao, Tian; Wang, Pengfei; Wang, Lina; Guo, Tiancai; Kang, Guozhang. 2016. "Virus-Induced Gene Silencing Identifies an Important Role of the TaRSR1 Transcription Factor in Starch Synthesis in Bread Wheat." Int. J. Mol. Sci. 17, no. 10: 1557.
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