| Verticillium wilt is a disastrous plant vascular disease causing severe yield and quality losses in many crops, including tomato, eggplant, cotton, potato and etc. It is of great importance to clone Verticillium wilt resistance-related genes in wild eggplant for plant Verticillium wilt-resistance breeding.The stem segments with axillary bud of Solarium torvum were cultured on MS medium plus 6-BA to induce plantlet regeneration. At the early period, the high concentration of 6-BA benefited bud inducement while inhibiting further growth of buds. After transfering to MS medium with lower concentration of 6-BA, the buds elongated and developed into complete plantlets.Primers used for RT-PCR and 3'-RACE were designed based on conserved sequences of Ve1 and SaVe1 cDNA sequences respectively isolated from tomato and Solarium aethiopicum. We isolated a cDNA sequence relating to Verticillium wilt and denominated as StoVe1. The nucleotide sequence of StoVe1 is 3400 bp long encoding a protein of 1051 amino acids which has a calculated molecular weight of 117 kDa with an isoelectric point of 5.74. The protein StoVe1 possesses a hydrophobic N-terminal signal peptide containing a leucine zipper-like sequence, 23 predicted leucine-rich repeats containing 28 potential N-glycosylation sites, a hydrophobic membrane-spanning domain and a C-terminal domain with the receptor-mediated endocytosis signal. StoVe1 has high sequence similarity to Ve proteins from tomato, Solarium aethiopicum and Solarium Lycopersicoides, and has lower sequence similarity to some resistant and putative resistant proteins from rice, tomato and other plants.Phylogenetic analysis showed StoVe1 has higher homology to tomato Ve2 compared with tomato Ve1, and it revealed great variation of Ve1 genes between Solarium torvum and tomato. Semi-quantitive RT-PCR results displayed higher expression level of StoVe1in stems and in roots than in leaves . which may correlated with the root-infectig speciality of Verticillium species. |
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