Figure 1.
Phylogenetic and structural analysis of Ve-like genes.
Domain A is the predicted signal peptide. Domain B that constitutes the main body of the protein is mainly composed of LRRs. The membrane-associated hydrophobic domain D is flanked by a neutral/acidic amino acids rich domain C and by a neutral/basic amino acids rich domain E. Domain F contains a PEST sequence, while domain G contains an endocytosis signal. The detail information about the sequences and domains of GbVe1 protein are shown in Figure S1. Other genes could be found in the GenBank database using following accession numbers: GbVe (EU855795), GbVd2 (GU299534), SlVe1 (AY262016), StVe (AY311527), Ve1 (AF272367), Ve2 (AF365929), and the two A. thaliana genes, AT1G47890 and AT1G45616, could be found in www.arabidopsis.org.
Figure 2.
Expression patterns of the Gbve1 and Ghve1 genes.
(A). Expression patterns of Gbve1/Ghve1 genes in H7124 (Gb) and Yumian 1 (Gh) under infection by V. dahliae isolate V991 and BP2. dpi means days post inoculation. (B). Expression patterns of the Gbve1 gene by different phytohormones.
Figure 3.
GUS activities in transgenic Arabidopsis plants containing the PGbve1:GUS construct.
(A). GUS activities in the Arabidopsis plantlet. (B). Magnified view of vascular bundles in roots and stems.
Figure 4.
The Gbve1 gene endowed Verticillium wilt resistance in cotton.
(A). The cotton chlI gene was used as a positive control with a phenotype of yellow-colored leave after VIGS in cotton (Left: 3 weeks; Right: blossom stage). (B). The phenotypes of H7124 under infection by V. dahliae isolates V991 and BP2 after VIGS with CLCrV containing a fragment of Gbve1 gene. Photos were taken at 42 d after V. dahliae inoculation (55 d after VIGS). (C). The disease indices Gbve1 gene-silence lines. The results were presented as means±SE from three replications with at least 25 plants per replication. (D). Expression levels of the Gbve1 gene and its homolog (the Gbve gene) in the silenced lines.
Figure 5.
Gbve1 confers Verticillium wilt resistance in transgenic Arabidopsis.
(A). The resistant levels of transgenic Arabidopsis (left panel) to V. dahliae was related to the expression levels of the Gbve1 gene in the T3 independent transgenic lines (right panel). The disease indices were presented means±SE from three replications with at least 16 plants per replication. (B). Phenotype comparison of transgenic line R5 and WT infected with V. dahliae isolates V991 and BP2. (C).The Gbve1 transgenic Arabidopsis were resistance to V. dahliae through cell death response. Bar = 200 µm. (D). Expressional levels of genes relative to pathogen resistance in transgenic line R5.
Figure 6.
Gbve1 confers Verticillium wilt resistance in transgenic upland cotton.
(A). Relative expression level of Gbve1 genes in T1 transgenic cotton lines. (B). The disease indices of T1 transgenic cottons with infection by V. dahliae defoliating isolate V991 (upper panel) and non-defoliating isolate BP2 (down panel). Results were presented as means±SE from three replications with at least 10 plants per replication. (C). The phenotypes of transgenic cotton lines with infection by V. dahliae isolates.