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Phosphorylation of TGB1 by protein kinase CK2 promotes barley stripe mosaic virus movement in monocots and dicots.

Hu Y, Li Z, Yuan C, Jin X, Yan L, Zhao X, Zhang Y, Jackson AO, Wang X, Han C, Yu J, Li D - J. Exp. Bot. (2015)

Bottom Line: Substitution of Thr-395 or Thr-401 with aspartic acid interfered with monocot and dicot cell-to-cell movement and the plants failed to develop systemic infections.The mutant XJTGB1T395A/T401A weakened in vitro interactions between XJTGB1 and XJTGB3 proteins but had little effect on XJTGB1 RNA-binding ability.Taken together, our results support a critical role of CK2 phosphorylation in the movement of BSMV in monocots and dicots, and provide new insights into the roles of phosphorylation in TGB protein functions.

View Article: PubMed Central - PubMed

Affiliation: State Key laboratory of Agro-Biotechnology and Ministry of Agriculture Key Laboratory of Soil Microbiology, College of Biological Sciences, China Agricultural University, Beijing 100193, PR China.

No MeSH data available.


Related in: MedlinePlus

Diagram of XJBSMV strain infectious clones and cereal and dicot host infectivity test results. (A) Illustration of XJBSMV infectious clones under the T7 promoter or double cauliflower mosaic virus 35S promoter as described previously (Yuan et al., 2011; Lee et al., 2012). (B) Infectivity assays with in vitro-synthesized gRNAs of barley, wheat, and B. distachyon Bd21. (C) Agroinfiltration was used to initiate infections of N. benthamiana. Typical chlorotic stripes and mosaic symptoms (top) of BSMV appeared on emerging uninoculated leaves by 7–9 dpi. Upper uninoculated leaf tissue was harvested at 12 dpi, and the relative BSMV RNA and CP amounts were evaluated by RT-PCR (middle) and Western blotting with the antibody against BSMV CP (bottom). BSMV RNAγ was detected by RT-PCR with the primer pair BS-10/BS-32 (Supplementary Table S1). (This figure is available in colour at JXB online.)
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Figure 1: Diagram of XJBSMV strain infectious clones and cereal and dicot host infectivity test results. (A) Illustration of XJBSMV infectious clones under the T7 promoter or double cauliflower mosaic virus 35S promoter as described previously (Yuan et al., 2011; Lee et al., 2012). (B) Infectivity assays with in vitro-synthesized gRNAs of barley, wheat, and B. distachyon Bd21. (C) Agroinfiltration was used to initiate infections of N. benthamiana. Typical chlorotic stripes and mosaic symptoms (top) of BSMV appeared on emerging uninoculated leaves by 7–9 dpi. Upper uninoculated leaf tissue was harvested at 12 dpi, and the relative BSMV RNA and CP amounts were evaluated by RT-PCR (middle) and Western blotting with the antibody against BSMV CP (bottom). BSMV RNAγ was detected by RT-PCR with the primer pair BS-10/BS-32 (Supplementary Table S1). (This figure is available in colour at JXB online.)

Mentions: Several BSMV field strains from China collected in our laboratory have broader host ranges than the more extensively studied NDBSMV and Type BSMV (TYBSMV) strains. To evaluate the diversity of the more virulent BSMV strains (Lee et al., 2012), we constructed infectious clones of the XJBSMV strain (Xie et al., 1981) under the bacteriophage T7 or double cauliflower mosaic virus 35S promoters (Petty et al., 1989; Yuan et al., 2011) (Fig. 1A). The infectivity of in vitro transcripts synthesized from linearized pT7-αXJ, pT7-βXJ, and pT7-γXJ plasmids was tested by mechanical inoculation to barley, wheat, and B. distachyon Bd21. Inoculated plants consistently developed chlorotic stripes and mosaic symptoms typical of BSMV infections on upper uninoculated leaves by 6–7 dpi (Fig. 1B) and the efficiency of infectivity in barley, wheat, and B. distachyon Bd21 was 70–80, 80–90, and 50–60%, respectively (also see Supplementary Table S6). Agroinfiltration was used to initiate infections of N. benthamiana because only 10–30% of the plants became infected when using in vitro transcripts as inocula. Agrobacterium harbouring the plasmids pCa-αXJ, pCa-βXJ, and pCa-γXJ were infiltrated into the basal sides of the leaves. Newly emerging leaves developed mild mottling symptoms at 7–9 d after agroinfiltration (Fig. 1C), and the efficiency of infectivity was increased to approximate 90%. RT-PCR and Western blot analysis verified the infectivity of the XJBSMV infectious clones in the monocot hosts (Fig. 1B) and in N. benthamiana (Fig. 1C).


Phosphorylation of TGB1 by protein kinase CK2 promotes barley stripe mosaic virus movement in monocots and dicots.

Hu Y, Li Z, Yuan C, Jin X, Yan L, Zhao X, Zhang Y, Jackson AO, Wang X, Han C, Yu J, Li D - J. Exp. Bot. (2015)

Diagram of XJBSMV strain infectious clones and cereal and dicot host infectivity test results. (A) Illustration of XJBSMV infectious clones under the T7 promoter or double cauliflower mosaic virus 35S promoter as described previously (Yuan et al., 2011; Lee et al., 2012). (B) Infectivity assays with in vitro-synthesized gRNAs of barley, wheat, and B. distachyon Bd21. (C) Agroinfiltration was used to initiate infections of N. benthamiana. Typical chlorotic stripes and mosaic symptoms (top) of BSMV appeared on emerging uninoculated leaves by 7–9 dpi. Upper uninoculated leaf tissue was harvested at 12 dpi, and the relative BSMV RNA and CP amounts were evaluated by RT-PCR (middle) and Western blotting with the antibody against BSMV CP (bottom). BSMV RNAγ was detected by RT-PCR with the primer pair BS-10/BS-32 (Supplementary Table S1). (This figure is available in colour at JXB online.)
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Figure 1: Diagram of XJBSMV strain infectious clones and cereal and dicot host infectivity test results. (A) Illustration of XJBSMV infectious clones under the T7 promoter or double cauliflower mosaic virus 35S promoter as described previously (Yuan et al., 2011; Lee et al., 2012). (B) Infectivity assays with in vitro-synthesized gRNAs of barley, wheat, and B. distachyon Bd21. (C) Agroinfiltration was used to initiate infections of N. benthamiana. Typical chlorotic stripes and mosaic symptoms (top) of BSMV appeared on emerging uninoculated leaves by 7–9 dpi. Upper uninoculated leaf tissue was harvested at 12 dpi, and the relative BSMV RNA and CP amounts were evaluated by RT-PCR (middle) and Western blotting with the antibody against BSMV CP (bottom). BSMV RNAγ was detected by RT-PCR with the primer pair BS-10/BS-32 (Supplementary Table S1). (This figure is available in colour at JXB online.)
Mentions: Several BSMV field strains from China collected in our laboratory have broader host ranges than the more extensively studied NDBSMV and Type BSMV (TYBSMV) strains. To evaluate the diversity of the more virulent BSMV strains (Lee et al., 2012), we constructed infectious clones of the XJBSMV strain (Xie et al., 1981) under the bacteriophage T7 or double cauliflower mosaic virus 35S promoters (Petty et al., 1989; Yuan et al., 2011) (Fig. 1A). The infectivity of in vitro transcripts synthesized from linearized pT7-αXJ, pT7-βXJ, and pT7-γXJ plasmids was tested by mechanical inoculation to barley, wheat, and B. distachyon Bd21. Inoculated plants consistently developed chlorotic stripes and mosaic symptoms typical of BSMV infections on upper uninoculated leaves by 6–7 dpi (Fig. 1B) and the efficiency of infectivity in barley, wheat, and B. distachyon Bd21 was 70–80, 80–90, and 50–60%, respectively (also see Supplementary Table S6). Agroinfiltration was used to initiate infections of N. benthamiana because only 10–30% of the plants became infected when using in vitro transcripts as inocula. Agrobacterium harbouring the plasmids pCa-αXJ, pCa-βXJ, and pCa-γXJ were infiltrated into the basal sides of the leaves. Newly emerging leaves developed mild mottling symptoms at 7–9 d after agroinfiltration (Fig. 1C), and the efficiency of infectivity was increased to approximate 90%. RT-PCR and Western blot analysis verified the infectivity of the XJBSMV infectious clones in the monocot hosts (Fig. 1B) and in N. benthamiana (Fig. 1C).

Bottom Line: Substitution of Thr-395 or Thr-401 with aspartic acid interfered with monocot and dicot cell-to-cell movement and the plants failed to develop systemic infections.The mutant XJTGB1T395A/T401A weakened in vitro interactions between XJTGB1 and XJTGB3 proteins but had little effect on XJTGB1 RNA-binding ability.Taken together, our results support a critical role of CK2 phosphorylation in the movement of BSMV in monocots and dicots, and provide new insights into the roles of phosphorylation in TGB protein functions.

View Article: PubMed Central - PubMed

Affiliation: State Key laboratory of Agro-Biotechnology and Ministry of Agriculture Key Laboratory of Soil Microbiology, College of Biological Sciences, China Agricultural University, Beijing 100193, PR China.

No MeSH data available.


Related in: MedlinePlus