Scientia Agricultura Sinica ›› 2018, Vol. 51 ›› Issue (6): 1079-1088.doi: 10.3864/j.issn.0578-1752.2018.06.007

• PLANT PROTECTION • Previous Articles     Next Articles

Identification and Analysis of Magnaporthe oryzae of Foxtail Millet Avirulence Genes

REN ShiLong1,2, BAI Hui1, WANG yongFang1, QUAN JianZhang1, DONG ZhiPing1,  LI ZhiYong1, XING JiHong2   

  1. 1Millet Institute, Hebei Academy of Agricultural and Forestry Sciences/Minor Cereal Crops Laboratory of Hebei Province/National Foxtail Millet Improvement Center, Shijiazhuang 050035; 2Hebei Agricultural University/Key Laboratory of Hebei Province for Plant Physiology and Molecular Pathology, Baoding 071001, Hebei
  • Received:2017-09-26 Online:2018-03-16 Published:2018-03-16

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Abstract: 【Objective】The objective of this study is to identify avirulence genes (Avr-genes) of Magnaporthe oryzae, understand the distribution and variation of Avr-genes in the epidemic strains of different regions, and to provide a reference for further study of the mechanism of Avr-genes variation in M. oryzae. 【Method】 A total of 76 strains of M. oryzae were isolated from different regions of millet growing area in northern China. The genomic DNA of 76 M. oryzae strains was extracted and used as a template for PCR amplification using the specific primers designed according to the 7 Avr-genes cloned from rice blast. AGE (agarose gel electrophoresis) analysis and sequencing of the PCR products were used to analyze the distribution of Avr-genes in M. oryzae strains of different regions from foxtail millet. 【Result】 The amplification rate of ACE1, Avr-pita, Avr1-CO39 and AvrPiz-t genes was 100% in 76 strains of M. oryzae and the amplification rate of avirulent genes Avr-pik, Avr-pia and Avr-pii was 63.2%, 42.1% and 21.1%, respectively. The amplification of Avr1-CO39 in the strains P11 and P34 was about 490 bp larger than the expected fragments. Sequencing of the Avr1-CO39 in the strains P11 and P34 showed that the Avr1-CO39 sequences in strains P11 and P34 were identical and the promoter region contained a 490 bp inserted fragment which shared 99.16% similarity with non-LTR retrotransposon: Mg-SINE. A high level of Avr-pita polymorphisms were found in M. oryzae and the major variation was single nucleotide variation, including single base insertion, deletion, and SNP of multiple loci. The main sequence variation of Avr-pia was the absence of the whole Avr-gene, and their sequencing verification results showed that there were 4 genotypes of Avr-pia allele in the strains. The Avr-pia-A was identical to the reference sequence (AB498873.1) and contained 10 strains. Avr-pia-B contained 20 strains, and there were C/T, G/T and C/A mutations at -116, -109, and -16 bp sites, but the CDS region of Avr-pia-B was the same as the reference sequence. The Avr-pia-C type contained only the strain P10, and there was a T/G mutation at +150 bp, but it is a synonymous mutation. The Avr-pia-D contained only the strain P18, and there was a C/T mutation at +212 bp site, which resulted in the mutation of coding amino acid from threonine to isoleucine. The Avr-pii of M. oryzae contained 3 alleles. The Avr-pii-A was the same as the reference sequence (AB498874.1) and contained 14 strains. An A/G mutation was found in Avr-pii-B and Avr-pii-C at +139 and +64 bp sites, respectively, which caused the amino sequence in the site to mutate from threonine to alanine. Avr-pii-B and Avr-pii-C were both reported for the first time. The haplotype analysis showed that AG2 containing 23 strains was the dominant type, which accounted for 30.2% of the tested strains. 【Conclusion】 Avr-genes ACE1, Avr-pita, Avr1-CO39 and AvrPiz-t had no geographical origin difference and the frequency distribution of Avr-genes Avr-pik, Avr-pia and Avr-pii varied in different regions. The AG2 haplotype was predominant in M. oryzae from foxtail millet, followed by AG1 and AG5 haplotypes.

Key words: foxtail millet, Magnaporthe oryzae, avirulence gene, mutation

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