Isolation and Identification of Pathogenic Fungus Fusarium sacchari of Sugarcane Pokkah Boeng Disease

WANG Changmi; LUO Zhiming; LI Yinhu; WANG Xiaoyan; ZHANG Rongyue; LI Jie; YIN Jiong; SHAN Hongli

doi:10.11924/j.issn.1000-6850.casb2024-0277

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Chin Agric Sci Bull ›› 2025, Vol. 41 ›› Issue (4) : 114-118. DOI: 10.11924/j.issn.1000-6850.casb2024-0277

Isolation and Identification of Pathogenic Fungus Fusarium sacchari of Sugarcane Pokkah Boeng Disease

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Abstract

Sugarcane pokkah boeng disease is a fungal disease caused by multiple species of Fusarium. In order to clarify the pathogen species of sugarcane pokkah boeng disease in Yunnan, thirty-three samples of sugarcane pokkah boeng disease were collected from Kaiyuan and Menglian in Yunnan, and strains FS1 and FS2 were isolated and purified, and their DNA was extracted; the primers of transcription elongation factor (EF-1α), tubulin gene (TUB2), polymerase gene (RPB2) were used for PCR amplification of the isolated strains. The result was that a clear and bright band consistent with the target band was obtained, and the products were sequenced and analyzed by BLAST on the NCBI website, the EF-1α, TUB2 and RPB2 sequences of strains FS1 and FS2 were compared with EF-1α (accession number: MK609907.1), TUB2 (accession number: MT011039.1) and RPB2 (accession number: MW238849.1) of Fusarium sacchari with 100% similarity and coverage. Phylogenetic trees of strains FS1 and FS2 were constructed by maximum likelihood method with MEGA6.0 software, and the results showed that FS1 and FS2 clustered on the same branch with F. sacchari. Therefore, based on the morphological characteristics of strains FS1 and FS2 and the results of molecular biological identification, the pathogen of sugarcane pokkah boeng disease was identified as F. sacchari. F. sacchari spores were inoculated with ‘Yunzhe 08-1609’, and the symptoms of leaf regression and leaf yellowing appeared on the 7^th day of inoculation, the infected leaves were isolated and sequenced again. The sequence similarity and coverage between EF-1α of the isolated strain and EF-1α of F. sacchari (accession number: MK609907.1) were 100%. F. sacchari was shown to be the causal agent of sugarcane pokkah boeng disease in Kaiyuan and Menglian.

Key words

sugarcane / pokkah boeng disease / Fusarium sacchari / isolation / identification / PCR amplification / phylogenetic analysis

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WANG Changmi , LUO Zhiming , LI Yinhu , et al . Isolation and Identification of Pathogenic Fungus Fusarium sacchari of Sugarcane Pokkah Boeng Disease[J]. Chinese Agricultural Science Bulletin. 2025, 41(4): 114-118 https://doi.org/10.11924/j.issn.1000-6850.casb2024-0277

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根腐病是危害浙江省铁皮石斛的重要病害之一，鉴定根腐病的病原菌是有效防治该病害的前提条件。为进一步明确前期经柯赫法则验证为病原菌的镰刀菌（编号为GF-14）分类地位，本研究通过形态学鉴定，并结合内源转录间隔区（rDNA-ITS）、翻译延伸因子（TEF-1α）和RNA聚合酶基因（RPB1和RPB2）等序列的同源性比对及系统发育分析，确认该致病菌属于变红-木贼镰刀菌复合种群Fusarium incarnatum-equiseti species complex。进一步采用菌丝生长速率法，测定了该病原菌对多种杀菌剂的敏感性，并选取对菌丝生长抑制效果最佳的杀菌剂进行田间防效的测定。结果表明：本实验室分离获得的生防菌淀粉酶产色链霉菌Streptomyces diastatochramogenes 1628发酵液对该致病菌的抑制效果最佳，发酵液稀释10倍后抑制率高达98.22%；而宁南霉素、井冈霉素则对该致病菌无显著抑制作用。菌株对峙试验发现淀粉酶产色链霉菌1628对菌株GF-14的菌丝生长抑制率为32.65%。田间防效试验表明其发酵液对铁皮石斛根腐病病株具有防治效果，为34.90%。本研究明确了铁皮石斛镰刀菌根腐病病原菌的分类地位，并筛选到能有效抑制该病原菌的微生物杀菌剂，为后续铁皮石斛根腐病的防治提供依据。

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