Welcome to Chinese Agricultural Science Bulletin,

Chinese Agricultural Science Bulletin ›› 2022, Vol. 38 ›› Issue (4): 81-85.doi: 10.11924/j.issn.1000-6850.casb2021-0031

Special Issue: 生物技术 植物保护

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Isolation and Identification of the Pathogen Causing Leaf Spot on Trifolium repens

SHANG Na1,2(), REN Aizhi1, LIU Bing1, ZHAO Peibao1()   

  1. 1College of Agronomy, Liaocheng University, Liaocheng, Shandong 252000
    2Liaocheng Academy of Agricultural Sciences, Liaocheng, Shandong 252000
  • Received:2021-01-13 Revised:2021-11-26 Online:2022-02-05 Published:2022-03-16
  • Contact: ZHAO Peibao E-mail:shangna126@126.com;zhaopeibao@163.com

Abstract:

An investigation in Liaocheng of Shandong Province shows that a pathogen causing leaf spot was found on Trifolium repens. Therefore, it is of great significance to find out the pathogen species and provide reference for the prevention and control of the disease. The leaves of T. repens with spot disease collected from Liaocheng were used as materials. The pathogenic bacteria were separated by tissue separation method, and the pathogenicity was verified by Koch's rule. Based on morphological observation, molecular biological method was used to identify pathogenic bacteria. The phylogenetic tree was constructed to analyze the fungus genetic diversity. The pathogenic bacteria (labelled SY1) isolated from diseased leaves were pathogenic and consistent with the field symptoms. To further classify the taxonomic status of the pathogen, the generic fungal primers ITS4/ITS5 were used for PCR amplification and sequencing. Sequence analysis showed that the SY1 was 100% similar to Alternaria arborescens (serial number: KC464334.1), and gathered together on the phylogenetic tree. According to the morphological characteristics and gene sequence identification results, the pathogen of T. repens leaf spot was identified as A. arborescens, which was the first report of A. arborescens causing T. repens leaf spot in China.

Key words: Trifolium repens, Alternaria arborescens, molecular biological identification, morphological identification, leaf spot, pathogen

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