Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (5): 881-888.doi: 10.3864/j.issn.0578-1752.2013.05.002

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Cloning and Functional Analysis of StAC Gene in Setosphaeria turcica

 SHEN  Shen, WANG  Jing-Jing, TONG  Ya-Meng, LI  Po, HAO  Zhi-Min, DONG  Jin-Gao   

  1. 1.College of Life Sciences, Agricultural University of Hebei/Mycotoxin and Plant Molecular Pathology Laboratory, Baoding 071001, Hebei
    2.Plant Protection Institute, Hebei Academy of Agricultural and Forestry Sciences, Baoding 071001, Hebei
  • Received:2012-11-02 Online:2013-03-01 Published:2013-01-23

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Abstract: 【Objective】 To understand the function of adenylate cyclase (AC) during regulating the fungal pathogenicity, the gene encoding AC in S. turcica was cloned and knocked out.【Method】Degenerated primer-PCR and genome walking were used to obtain the full length DNA of StAC in S. turcica. The structure and homology sequence alignment of StAC were analyzed by bioinformatics methods and its copy number was verified by Southern blotting. Furthermore, the function of StAC was analyzed by gene knockout technology.【Result】StAC, encoding a protein of 2 005 amino acid residues and including 5 exons and 4 introns, was 6 816 bp of DNA and 6 018 bp of ORF. The nucleotide sequence of StAC gene showed 96% identity with its homology in Pyrenophora tritici-repentis. Southern blotting showed that there was only single copy of StAC in genome of S. turcica. The phenotypic analysis showed that the aerial hyphae of StAC knockout mutant named Δstac were gray. The mutant failed to sporulate, and the toxin activity and pathogenicity on leaves of susceptible host was significantly reduced, but the resistance against hyperosmotic stress was enhanced.【Conclusion】It is suggested that StAC is involved in many procedures of S. turcica, including conidial formation, pathogenicity, hyperosmotic stress response, HT-toxin activity and the biosynthesis metabolism of pigmen.

Key words: Setosphaeria turcica , adenylate cyclase , pathogenicity , hyperosmotic stress

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