Cloning and characterization of a Verticillium wilt resistance gene from Gossypium barbadense and functional analysis in Arabidopsis thaliana

Verticillium wilt causes enormous loss to yield or quality in many crops. In an effort to help controlling this disease through genetic engineering, we first cloned and characterized a Verticillium wilt resistance gene (GbVe) from cotton (Gossypium barbadense) and analyzed its function in Arabidopsis thaliana. Its nucleotide sequence is 3,819 bp long, with an open reading frame of 3,387 bp, and encoding an 1,128-aa protein precursor. Sequence analysis shows that GbVe produces a leucine-rich repeat receptor-like protein. It shares identities of 55.9% and 57.4% with tomato Ve1 and Ve2, respectively. Quantitative real-time PCR indicated that the Ve gene expression pattern was different between the resistant and susceptible cultivars. In the resistant Pima90–53, GbVe was quickly induced and reached to a peak at 2 h after inoculation, two-fold higher than that of control. We localized the GbVe–GFP fusion protein to the cytomembrane in onion epidermal cells. By inserting GbVe into Arabidopsis via Agrobacterium-mediated transformation, T3 transgenic lines were obtained. Compared with the wild-type control, GbVe-overexpressing plants had greater levels of resistance to V. dahliae. This suggests that GbVe is a useful gene for improving the plant resistance against fungal diseases.

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