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Tissue‐specific and pathogen‐inducible expression of a fusion protein containing a Fusarium‐specific antibody and a fungal chitinase protects wheat against Fusarium pathogens and mycotoxins
Authors:Wei Cheng  He‐Ping Li  Jing‐Bo Zhang  Hong‐Jie Du  Qi‐Yong Wei  Tao Huang  Peng Yang  Xian‐Wei Kong  Yu‐Cai Liao
Affiliation:1. Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan, China;2. College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, China;3. College of Life Science and Technology, Huazhong Agricultural University, Wuhan, China;4. National Center of Plant Gene Research (Wuhan), Huazhong Agricultural University, Wuhan, China
Abstract:Fusarium head blight (FHB) in wheat and other small grain cereals is a globally devastating disease caused by toxigenic Fusarium pathogens. Controlling FHB is a challenge because germplasm that is naturally resistant against these pathogens is inadequate. Current control measures rely on fungicides. Here, an antibody fusion comprised of the Fusarium spp.‐specific recombinant antibody gene CWP2 derived from chicken, and the endochitinase gene Ech42 from the biocontrol fungus Trichoderma atroviride was introduced into the elite wheat cultivar Zhengmai9023 by particle bombardment. Expression of this fusion gene was regulated by the lemma/palea‐specific promoter Lem2 derived from barley; its expression was confirmed as lemma/palea‐specific in transgenic wheat. Single‐floret inoculation of independent transgenic wheat lines of the T3 to T6 generations revealed significant resistance (type II) to fungal spreading, and natural infection assays in the field showed significant resistance (type I) to initial infection. Gas chromatography–mass spectrometry analysis revealed marked reduction of mycotoxins in the grains of the transgenic wheat lines. Progenies of crosses between the transgenic lines and the FHB‐susceptible cultivar Huamai13 also showed significantly enhanced FHB resistance. Quantitative real‐time PCR analysis revealed that the tissue‐specific expression of the antibody fusion was induced by salicylic acid drenching and induced to a greater extent by F. graminearum infection. Histochemical analysis showed substantial restriction of mycelial growth in the lemma tissues of the transgenic plants. Thus, the combined tissue‐specific and pathogen‐inducible expression of this Fusarium‐specific antibody fusion can effectively protect wheat against Fusarium pathogens and reduce mycotoxin content in grain.
Keywords:antibody fusion  chitinase  Fusarium head blight  mycotoxins  pathogen‐inducible expression  tissue‐specific promoter  transgenic wheat
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