title: Disruption and molecular characterization of calpains-related (MoCAPN1, MoCAPN3 and MoCAPN4) genes in Magnaporthe oryzae.
abstract: Calpains are intracellular, cysteine proteases found in plants, animals and fungi functioning as signal transduction components in different cellular pathways including sporulation and alkaline adaptation in fungi. Calpains-related MoCAPN1 (MGG_14872), MoCAPN3 (MGG_15810) and MoCAPN4 (MGG_04818) genes from Magnaporthe oryzae genome which are 2604, 3513 and 771-bp in length and encoding identical proteins of 867, 1170 and 256 amino acids were functionally characterized for different phenotypes through gene disruption method. All the mutants except those for MoCAPN1 showed normal phenotypes. In pathogenicity test, the mutants did not lead to any visible changes in phenotypes causing similar blast lesions on blast susceptible rice and barley leaves as those of the Guy-11 strain suggesting no major role in pathogenicity. Germ tubes formation, appressorium formation, mycelium radial growth and mating with 2539 strain were indistinguishable among the mutants and Guy-11 strains. Cell wall integrity (congo red) test, stress response under chemical pressure (ZnSO4, CuSO4 and CdCl2), osmotic and oxidative (NaCl and H2O2) stress response, growth response on glucose and nitrogen deficient media resulted in similar results in the mutants and Guy-11 strains. However, mutants for ΔMoCAPN1 gene produced reduced (0.57±0.15B and 0.54±0.05B) conidia compared to that (1.69±0.13A) of the Guy-11 strain showing its involvement in conidiation.
authors.affiliation: State Key Laboratory for Rice Biology, Biotechnology Institute, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, China; Department of Plant Pathology, Faculty of Crop Protection Sciences, The University of Agriculture, Peshawar, Pakistan.
authors.forename: Irshad Ali
authors.initials: IA
authors.lastname: Khan
authors.affiliation: State Key Laboratory for Rice Biology, Biotechnology Institute, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Wang
authors.affiliation: State Key Laboratory for Rice Biology, Biotechnology Institute, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, China.
authors.forename: Hai-Jiao
authors.initials: HJ
authors.lastname: Li
authors.affiliation: School of Life Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, China.
authors.forename: Jian-Ping
authors.initials: JP
authors.lastname: Lu
authors.affiliation: State Key Laboratory for Rice Biology, Biotechnology Institute, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, China.
authors.forename: Xiao-Hong
authors.initials: XH
authors.lastname: Liu
authors.affiliation: State Key Laboratory for Rice Biology, Biotechnology Institute, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, China; China Tobacco Gene Research Center, Zhengzhou Tobacco Institute of CNTC, Zhengzhou 450001, China. Electronic address: fuchenglin@zju.edu.cn.
authors.forename: Fu-Cheng
authors.initials: FC
authors.lastname: Lin
mesh_terms: D002154:Calpain / Q000235:genetics* / Q000378:metabolism; D002473:Cell Wall / Q000235:genetics / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D001467:Hordeum / Q000382:microbiology; D020082:Magnaporthe / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D025282:Mycelium / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D012275:Oryza / Q000382:microbiology; D010935:Plant Diseases / Q000382:microbiology*; D013172:Spores, Fungal / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D014774:Virulence
chemical_list: D005656:Fungal Proteins; D002154:Calpain
keywords: Calpains; Conidiation; Growth; Magnaporthe oryzae; Pathogenicity