玉米大斑病菌ATMT突变体库的构建及其分析

doi:10.3864/j.issn.0578-1752.2012.12.006

Abstract

Abstract: 【Objective】 The objective of this study is to use Agrobacterium tumefaciens-mediated transformation (ATMT) to mutate the genes of Setosphaeria turcica and construct ATMT mutant library, which will lay a foundation for illuminating the pathogenesis of S. turcica at molecular level. 【Method】A. tumefacien with binary recombination vector was transformed into S. turcica, and the mutants were screened by hygromycin B and PCR technology. Based on the methods, a preliminary ATMT mutant library was constructed. Some mutants were selected randomly and analysed about the colony morphologies, mycelium and conidium development, and pathogenicity.【Result】A total of 1 265 T-DNA insertion mutants were obtained in this experiment. Hygromycin B resistance screening and PCR technology were used to confirm 36 mutants, which were selected randomly from the mutant library. The results showed that hygromycin phosphotransferase genes had been integrated into wild-type genome of S. turcica and the mutants were stable in their characteristics of genetics. Compared with wild-type strains, most of the mutants did not changed their colony morphologies and growth rates, only minorities of the mutants varied in their characteristics, in which 13.8% strains slowed down their growth rate obviously, 16.7% strains accelerated their growth rate distinctly, about 5.6% strains altered their shapes of conidia, 13.5% strains reduced the number of conidia, 16.6% strains changed their germination rate evidently, and 1 strain reinforced its pathogenicity, accounting 2.8% in all selected strains.【Conclusion】 ATMT mutant library of S. turcica was constructed and some mutants were analysed primarily. The study will lay a foundation for cloning genes related to growth, development and pathogenicity of S. turcica in the future.

Key words: Setosphaeria turcica, ATMT, mutant library, T-DNA insertion mutant

WANG Mei-Juan, LI Po, WU Min, FAN Yong-Shan, GU Shou-Qin, DONG Jin-Gao. Constuction and Evaluation of ATMT Mutant Library of Setosphaeria turcica[J].Scientia Agricultura Sinica, 2012, 45(12): 2384-2392.

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