Abstract
Ubiquitination is a post-translational regulatory mechanism that controls a variety of biological processes in plants. The E3 ligases confer specificity by recognizing target proteins for ubiquitination. Here, we identified SEVEN IN ABSENTIA (SINA) ubiquitin ligases, which belong to the RING-type E3 ligase family, in upland cotton (Gossypium hirsutum). Twenty-four GhSINAs were characterized, and the expression levels of GhSINA7, GhSINA8, and GhSINA9 were upregulated at 24 h after inoculation with Verticillium dahliae. In vitro ubiquitination assays indicated that the three GhSINAs possessed E3 ubiquitin ligase activities. Transient expression in Nicotiana benthamiana leaves showed that they localized to the nucleus. And yeast two-hybrid (Y2H) screening revealed that they could interact with each other. The ectopic overexpression of GhSINA7, GhSINA8, and GhSINA9 independently in Arabidopsis thaliana resulted in increased tolerance to V. dahliae, while individual knockdowns of GhSINA7, GhSINA8, and GhSINA9 compromised cotton resistance to the pathogen. Thus, GhSINA7, GhSINA8, and GhSINA9 act as positive regulators of defense responses against V. dahliae in cotton plants.
Highlights
Verticillium dahliae, a soil-borne fungal pathogen, causes Verticillium wilt, which is a destructive vascular disease affecting more than 200 plant species, including agro-economically important cotton (BejaranoAlcazar et al, 1996; Fradin and Thomma, 2006; Cai et al, 2009)
To investigate the functions of E3 ubiquitin ligase genes in response to V. dahliae in upland cotton (Gossypium hirsutum), we identified three SEVEN IN ABSENTIA (SINA) E3 ligase genes, GhSINA7, GhSINA8, and GhSINA9, which are induced by V. dahliae infections
All the identified SINA proteins were divided into two groups, with 32 cotton SINA genes (8, 8, and 16 from G. raimondii, G. arboreum, and G. hirsutum, respectively) and 3 Arabidopsis SINA genes in Group I, and 16 cotton SINA genes (4, 4, and 8 from G. raimondii, G. arboreum, and G. hirsutum, respectively) and 2 Arabidopsis SINA genes in Group II (Figure 1C)
Summary
Verticillium dahliae, a soil-borne fungal pathogen, causes Verticillium wilt, which is a destructive vascular disease affecting more than 200 plant species, including agro-economically important cotton (BejaranoAlcazar et al, 1996; Fradin and Thomma, 2006; Cai et al, 2009). V. dahliae is notoriously difficult to control because of its strongly invasive pathogenicity and its ability to persist in soil, allowing it to penetrate host root xylem vessels and vascular tissues, which severely block. The breeding of disease-resistant cultivars is an effective and practical management strategy to control the V. dahliae threat. Genetic engineering has become a promosing and environmentally friendly strategy to cope with V. dahliae, based on the characterization of cotton resistant candidate genes, like GhMYB1, GbSOBIR1, GhCRR1, GbTSA1, and GhWAK7A (Cheng et al, 2016; Han et al, 2019; Miao et al, 2019; Zhou et al, 2019; Wang et al, 2020)
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