Figures (3)  Tables (3)

    • Figure 1. 

      Vectors used and infection flow chart in this study. The plasmid profile of (a) pTRV1, (b) pTRV2, (c) pTRV2::PDS410, (d) pTRV2::CeTCP14. Color change of leaves by (e) leaf injection and (f) bulb vacuum injection methods.

    • Figure 2. 

      VIGS system constructing in taro. (a) Empty vector phenotype at 20 d of leaf infection with OD600 = 0.6, (b) VIGS-PDS410 phenotype at 20d of leaf infection OD600 = 0.6, (c) VIGS-PDS410 phenotype at 30 d of leaf infection with OD600 = 0.6, (d) mock phenotype after 20 d infection, (e) RdRp gene expression in TRV1 and CP gene expression in TRV2 were detected by RT-PCR with actin as the internal reference, (f) detection of CePDS expression level for photobleaching phenotype plants, (g) chlorophyll content detection for photobleaching phenotype plants. Scale bars = 3 cm.

    • Figure 3. 

      Detection of taro VIGS system for CeTCP14 in Ganyu No. 2. (a) RdRp gene expression in pTRV1 and fragement expression in pTRV2 were detected by RT-PCR. (b) Detection of CeTCP14 expression at 20 d after bulb vacuum infiltration. (c) Starch content detection for VIGS-CeTCP14 plants.

    • Primer name Primer sequence (5'−3') Length (bp) Purpose
      CePDS-EcoRI-F GGAATTCATGGGCTTTACCAGTTCTCTTTCGG 410 Used for fragments amplified of CePDS and CeTCP14 inserted in TRV2 vector
      CePDS-BamHI-R CGGGATCCTCCAGCAATATAGGCTTATGACCTG
      TRV2-TCP14_F GTGAGTAAGGTTACCGAATTCATGGGGGAGAGCCACCAG 300
      TRV2-TCP14_R CGTGAGCTCGGTACCGGATCCATCGACGGCCTTGCTGGG
      qCePDS-F GGTCGTTGGGGAGGAAGC 140 Used for the qRT-PCR of CePDS
      qCePDS-R TCTAGTCGGGCGTGGTGA
      qCeTCP14_F CCACACCGCCATCCAGTT 110 Used for the qRT-PCR of CeTCP14
      qP_TCP14_R CGAGCTCGTCTATGGCGG
      CeActinF CTAGTGGTCGCACAACAGGT 191 Used for the qRT-PCR of reference genes
      CeActinR TTCACGCTCAGCAGTGGTAG
      TRV1F1 CGTGTTGCATTTCGATGAA 525 Used for the detection of RdRp in TRV1
      TRV1R1 GACAACGCCACGATTAAGT
      TRV2F1 GTTGAAGAAGTTACACAGCA 407 Used for the detection of coat protein in TRV2
      TRV2R1 TCTTCAACTCCATGTTCTCT
      pTRV2_F TGTCAACAAAGATGGACATTGTTAC 198 / 480 Used for the detection of TRV2 and TRV2-CeTCP14 expression
      pTRV2_R ACACGGATCTACTTAAAGAA
      TRV2_F TGTTACTCAAGGAAGCACGATGAGCT − Used for vector construction sequencing and colony PCR detection
      TRV2_R GTACAGACGGGCGTAATAACGCTTA
      Note: underline indicates cleavage sites, bold indicates protective bases.

      Table 1. 

      Primers used in this study

    • OD600 Total number of inoculation plants Number of photobleaching phenotype The percentage of
      photobleaching phenotype
      0 10 0 d 0
      0.6 30 3.00 ± 1.00 c 10%
      0.8 30 5.33 ± 0.58 b 17.77%
      1.0 30 8.33 ± 0.58 a 27.77%
      1.2 30 7.67 ± 0.58 a 25.57%

      Table 2. 

      Bacterial concentrations optimization for the VIGS system.

    • Infection type Total number of inoculation plants Number of photobleaching phenotype The percentage of
      photobleaching phenotype
      Leaf injection 30 8.33 ± 0.58 a 27.77%
      Bulb evacuation 30 8.00 ± 0.58 a 26.67%

      Table 3. 

      Comparison of the VIGS system based on infection type.