Abstract:

【Objective】 The objective of this study is to establish high efficient REMI mutagenesis system, which is able to construct mutants that can be used to clone genes related to growth, development and pathogenicity of S. turcica.【Method】 REMI mutagenesis system was established through seeking the optimal different ages of this fungus, cell wall digest enzymes and its condition and collection method which affect protoplast yield, and seeking hygromycin B concentration for transformants screening, and seeking plasmid for transformation, which was used to achieve transformants. 【Result】 When lyallzyme, snailase, and drislase were mixed and at concentration of 1.25%, respectively, and when mycelium was cultured for 20 h, and 3 000 r/min for collection, the protoplast production was the maximum. Hygromycin B at 50 μg?mL-1 was determined as the optimal concentration for transformants screening, in which conidial germination and colony growth of wild type was completely inhibited. The plasmid pAN7-1 had higher transformation efficiency. The optimal system was used to induce mutagenesis and lots of transformants were obtained, from which some conidia production-changed and pathogencity-changed transformants were found. 【Conclusion】 The REMI mutagenesis of S. turcica was established, which had a great significance in construction of mutant library and in cloning the pathogencity-related genes.

Key words: Setosphaeria turcica, REMI, protoplast, transformantion