Enhancement of resistance against fungal pathogens in peanut (Arachis hypogaea L.) cultivar L14 by heterologous expression of gene encoding chitinase 42 kDa from Trichoderma asperellum SH16

Abstract

This study aims to extracellularly express the 42 kDa chitinase from Trichoderma asperellum SH16 in peanut (Arachis hypogaea) roots under the regulation of a tissue-specific Asy promoter and the driving of the signal peptide of the rice amylase 3D gene. The 42 kDa chitinase genes were employed in this study, which included one wild-type sequence (Chi42) and two synthetic sequences (syncodChi42–1 and syncodChi42–2) optimized for codon usage for plant expression. From 102 42 kDa chitinase transgenic peanut lines, seven elite lines were discovered, among them two for syncodChi42–1, three for syncodChi42–2, and two for Chi42. The study also revealed chitinase from two synthetic genes had higher expression levels than chitinase from the wild-type gene, with the highest specific activities of approximately 901 U/mg and 1124 U/mg, respectively, compared to 739 U/mg. All seven elite transgenic peanut lines exhibited strong extracellular chitinase activity, and some of them were able to fight the soil-borne pathogenic fungus Sclerotium rolfsii in both in vitro and in vivo assays. The significant chitinase activity of seven elite transgenic peanut lines, particularly five lines carrying one of two synthetic genes, suggests that transgenic crops might be used to prevent phytopathogenic fungi.