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

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

Abstract

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

WANG Changmi, LUO Zhiming, LI Yinhu, WANG Xiaoyan, ZHANG Rongyue, LI Jie, YIN Jiong, SHAN Hongli. Isolation and Identification of Pathogenic Fungus Fusarium sacchari of Sugarcane Pokkah Boeng Disease[J]. Chinese Agricultural Science Bulletin, 2025, 41(4): 114-118.

Figures/Tables 5

References 34

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基因名称	引物名称	引物序列	条带大小/bp
EF-1α	EF-1	5'-ATGGGTAAGGARGACAAGAC-3'	700
EF-1α	EF-2	5'-GGARGTACCAGTSATCATGTT-3'	700
TUB2	T21	5'-AACATGCGTGAGATTGTAAGT-3'	1300
TUB2	T22	5'-TCTGGATGTTGTTGGGAATCC-3'	1300
RPB2	RPB2-Sf	5'-GAYGAYMGWGATCAYTTYGG-3'	1200
RPB2	RPB2-7cR	5'-CCCATRGCTTGYTTRCCCAT-3'	1200

基因名称	引物名称	引物序列	条带大小/bp
EF-1α	EF-1	5'-ATGGGTAAGGARGACAAGAC-3'	700
EF-1α	EF-2	5'-GGARGTACCAGTSATCATGTT-3'	700
TUB2	T21	5'-AACATGCGTGAGATTGTAAGT-3'	1300
TUB2	T22	5'-TCTGGATGTTGTTGGGAATCC-3'	1300
RPB2	RPB2-Sf	5'-GAYGAYMGWGATCAYTTYGG-3'	1200
RPB2	RPB2-7cR	5'-CCCATRGCTTGYTTRCCCAT-3'	1200