Abstract：Cotton verticillium wilt has always been an important obstacle to the sustainable development of China's cotton (Gossypium hirsutum) industry. It has been shown that plant elicitor peptide (Pep) can promote immune responses in multiple plant families against many kinds of pathogens. The purpose of this study was to construct an engineering strain of plant immune-inducing peptide for cotton verticillium wilt, and explore the antibacterial effect of protein-inducing peptide after activating plant immunity, then screen the engineering strain with good control effect on cotton verticillium wilt. Firstly, the plant immune-inducing protein genes PEL1 (pectate lyase 1) and PevD1 (protein elicitor 1) in Verticillium dahliae, a pathogenic bacterium of cotton wilt disease, were cloned, and the recombinant plasmids pHT43-PEL1 and pHT43-PevD1 were constructed. After being verified by enzyme digestion and sequencing, the plasmid was introduced into Bacillus sp. T6, which was screened in the early study, to construct engineering strain. The heterologous expression of the immune-inducing peptide was verified by SDS-PAGE, and the activity of the pot experiment was verified, so as to further confirm the engineering strain with efficient control of cotton verticillium wilt. The results of SDS-PAGE showed that the 2 inducible peptides were successfully overexpressed in T6 strain, and the engineering strain V3-T with PEL1 overexpression and the engineering strain P18-T with PevD1 overexpression were obtained respectively. The engineering strain P18-T was injected into the leaves of cotton plants for 15 min, and the active oxygen content increased significantly (P<0.01). In the cotton pot experiment, the engineering strain P18-T acted on the susceptible cotton plants for 30 d. Compared with the control wild strain T6 group, the incidence of cotton verticillium wilt in the P18-T treatment group decreased significantly (P<0.05), and the control effect reached to 78.85% (P<0.05). In this study, an engineering strain P18-T with high efficiency against cotton verticillium wilt was obtained, which provides basic data for broadening the comprehensive prevention and control technology of cotton verticillium wilt.
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