Abstract
Ceratocystis manginecans causes mango wilt with significant economic losses. In the infection court, cerato-platanin (CP) family proteins (CPPs) are believed to involve in pathogenesis but has not been determined in C. manginecans. To confirm this function, a CP protein (CmCP) of C. manginecans was characterized in this study. A protoplast of C. manginecans was prepared by treating its mycelia with driselase and lysing enzymes. The cmcp gene was edited using CRISPR/Cas-U6-1 expression vectors in 60% PEG and 50 μg/mL hygromycin B in the medium, resulting in mutants with cmcp deletion (Δcmcp). A complemented mutant (Δcmcp-C) was obtained by transforming cmcp to Δcmcp. Both Δcmcp and Δcmcp-C were characterized by comparing them with a wild-type strain on morphology, mycelial growth, conidial production and pathogenicity. Additionally, cmcp was transformed and expressed in Pichia pastoris, and the derived recombinant protein CmCP caused a severe necrosis on Nicotiana tabacum leaves. CmCP-treated plant leaves showed symptoms of hypersensitive response including electrolyte leakage, reactive oxygen species generation and overexpression of defense-related genes PR-1, PAD3, ERF1, HSR203J, and HIN1. All those results suggested that cmcp gene was required for the growth development of C. manginecans and functioned as a major pathogenicity factor in mango infection.
Highlights
Ceratocystis spp. attack a wide range of economically important plants, causing cankers, lethal, wilt-type diseases, and black rot of storage roots on many plants
The purpose of this study was to establish a system of mutation in C. manginecans using clustered regularly interspaced short palindromic repeat (CRISPR)/Cas technology and gene knockout method and characterize the function of cmcp gene based on this system
Hypersensitive response (HR) comprises a series of characteristic symptoms that include the induction of reactive oxygen species (ROS) and electrolyte leakage (Frías et al, 2011)
Summary
Ceratocystis spp. attack a wide range of economically important plants, causing cankers, lethal, wilt-type diseases, and black rot of storage roots on many plants. Ceratocystis fimbriata was firstly reported on mango in Brazil (Viégas, 1960). Several other pathogenic species were reported in the world, including C. manginecans, C. acaciivora, and C. omanensis (Tarigan et al, 2011; Van Wyk et al, 2011). C. manginecans is considered as an important pathogen of mango tree and several other plants (Fourie et al, 2016). Mango wilt caused by C. manginecans is a serious vascular disease and has caused significantly economic losses to mango industry (Van Wyk et al, 2007; Al Adawi et al, 2013; Zhang et al, 2017)
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
