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中国农学通报

中国农学通报 ›› 2025, Vol. 41 ›› Issue (4): 114-118.doi: 10.11924/j.issn.1000-6850.casb2024-0277

• 植物保护·农药 • 上一篇    下一篇

甘蔗梢腐病致病菌甘蔗镰刀菌的分离及鉴定

王长秘(), 罗志明, 李银煳, 王晓燕, 张荣跃, 李婕, 尹炯, 单红丽()   

  1. 云南省农业科学院甘蔗研究所/云南省甘蔗遗传改良重点实验室/云南省蔗糖产业工程研究中心，云南开远 661699
  • 收稿日期:2024-04-28 修回日期:2024-11-29 出版日期:2025-01-23 发布日期:2025-01-23
  • 通讯作者:
    单红丽，女，1984年出生，云南保山人，副研究员，硕士，主要从事甘蔗病害研究。E-mail：、
  • 作者简介:

    王长秘，男，1991年出生，云南永善人，助理研究员，硕士，主要从事甘蔗病害研究。E-mail：、

  • 基金资助:
    财政部和农业农村部国家现代农业产业技术体系专项资金(CARS-170303); 云南省科技厅“云南省现代农业产业技术体系建设专项资金”(YNGZTX-4-92); 云南省科技厅“云南省技术创新人才培养对象项目”(202405AD350069); 中央引导地方科技发展基金“丘陵山地甘蔗新品种及绿色低碳高效生产技术产业化应用”(202307AD110002)

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()   

  1. Sugarcane Research Institute, Yunnan Province Academy of Agricultural Sciences/ Yunnan Key Laboratory of Sugarcane Genetic Improvement/ Yunnan Engineering Research Center of Sugarcane Industry, Kaiyuan, Yunnan 661699
  • Received:2024-04-28 Revised:2024-11-29 Published:2025-01-23 Online:2025-01-23

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摘要：

甘蔗梢腐病是由多种镰刀菌引起的真菌性病害。为明确云南甘蔗梢腐病病原种类，从云南开远和孟连采集甘蔗梢腐病样品33份，分离纯化得到菌株FS1和FS2，提取其DNA；利用转录延长因子基因(EF-1α)、微管蛋白基因(TUB2)、聚合酶基因(RPB2)引物序列对分离到菌株进行PCR扩增，获得了与目的条带大小一致的清晰明亮条带；将产物测序后在NCBI网站上进行BLAST比对分析，菌株FS1和FS2的EF-1α序列、TUB2序列和RPB2序列与Fusarium sacchari的EF-1α（登录号MK609907.1）、TUB2（登录号MT011039.1）和RPB2（登录号MW238849.1）序列的相似性和覆盖性均为100%；同时通过MEGA6.0软件以最大似然法建立菌株FS1和FS2的系统发育树，发现FS1、FS2与F. sacchari均聚在同一分支上；因此，根据菌株FS1和FS2的形态学特征以及分子生物学鉴定结果，甘蔗梢腐病的病原菌鉴定为F. sacchari。用F. sacchari孢子接种‘云蔗08-1609’，接种7 d叶片出现退绿和叶片黄化症状；将接种发病叶片进行再次分离和测序，分离到菌株EF-1α序列与F. sacchari的EF-1α（登录号MK609907.1）序列的相似性和覆盖性均为100%。研究表明，F. sacchari为开远和孟连引起梢腐病的致病菌。

关键词: 甘蔗, 梢腐病, 镰刀菌, 分离, 鉴定, PCR扩增, 系统发育分析

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 7th 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