Scientia Agricultura Sinica ›› 2019, Vol. 52 ›› Issue (11): 1895-1907.doi: 10.3864/j.issn.0578-1752.2019.11.005

• PLANT PROTECTION • Previous Articles     Next Articles

Composition of Fusarium Species Causing Maize Ear Rot and Analysis of Toxigenic Chemotype in Guangxi

DU Qing1,TANG ZhaoLei1,LI ShiChu1,SHANGGUAN LingLing2,LI HuaJiao2,DUAN CanXing3()   

  1. 1 Institute of Maize Research, Guangxi Academy of Agricultural Sciences, Nanning 530007
    2 Guangxi Agricultural Vocational and Technical College, Nanning 530007
    3 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences/National Key Facility of Crop Gene Resources and Genetic Improvement, Beijing 100081
  • Received:2019-02-19 Accepted:2019-03-21 Online:2019-06-01 Published:2019-06-11
  • Contact: CanXing DUAN E-mail:duancanxing@caas.cn

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Abstract:

【Objective】 The objective of this study is to clarify the composition and toxigenic chemotype of Fusarium species causing maize ear rot in Guangxi, and to provide important guidance and theoretical basis for comprehensive control of maize ear rot and reasonable distribution of varieties and resistance breeding.【Method】 The diseased ear samples were collected from main maize producing areas in Guangxi between 2016 and 2018, a total of 138 isolates from 21 counties (districts) were obtained by tissue separation and single-spore purification. Fusarium species were identified and determined according to morphological characteristics and molecular methods. The phylogenetic tree was constructed based on TEF-1α gene sequences, and specific primers were used to detect toxigenic chemotype.【Result】 A total of 10 Fusarium species were identified and confirmed among 138 isolates, including F. verticillioides, F. proliferatum, F. kyushuense, F. meridionale, F. sacchari, F. fujikuroi, F. asiaticum, F. concentricum, F. incarnatum and F. graminearum, with the isolation frequencies of 50.72%, 12.32%, 10.87%, 8.70%, 6.52%, 3.62%, 3.62%, 1.45%, 1.45% and 0.72%, respectively. F. graminearum species complex (FGSC) contained three independent species, i.e. F. meridionale, F. asiaticum and F. graminearum. F. verticillioides was the predominant pathogen, FGSC, F. proliferatum and F. kyushuense were the secondary predominant pathogens. F. sacchari and F. concentricum were the first reported to be the pathogen of maize ear rot in China. The key gene FUM1 responsible for the biosynthesis of fumonisins was detected among 67 F. verticillioides, 13 F. proliferatum, 5 F. sacchari and 3 F. fujikuroi strains, respectively, which indicated the potential ability of producing fumonisins. FUM1 was not detected in the F. concentricum strains. Four toxigenic chemotypes including NIV, 15-ADON, NIV+15-ADON and DON+15-ADON were detected among FGSC, F. kyushuense and F. incarnatum strains. Strains containing NIV chemotype included 8 F. kyushuense, 2 F. asiaticum, 2 F. meridionale, and 1 F. incarnatum. Strains containing 15-ADON chemotype included 2 F. meridionale. The NIV+15-ADON chemotype was detected among 8 F. meridionale, 2 F. kyushuense, 1 F. asiaticum and 1 F. incarnatum strains. Only 1 F. graminearum strain carried DON+15-ADON chemotype. The 3-ADON chemotype was not detected among these strains.【Conclusion】 F. verticillioides is the predominant pathogen of maize ear rot in Guangxi. FGSC, F. proliferatum and F. kyushuense are the secondary predominant species. FUM1 was detected among F. verticillioides, F. proliferatum, F. sacchari and F. fujikuroi. The main toxigenic chemtype of FGSC in Guangxi is NIV+15-ADON, while NIV is the main toxigenic chemotype of F. incarnatum and some F. kyushuense. The population composition of Fusarium species causing maize ear rot in Guangxi is different from equivalent study of temperate zone in China, which may be due to the fact that Fusarium species adapt to the high temperature and humidity growth environment in tropical and subtropical Guangxi and thus leads to the differences of toxigenic chemotypes.

Key words: maize, ear rot, Fusarium spp., isolation frequency, toxigenic chemotype

Table 1

Specific primers for Fusarium spp. identification"

检测真菌
Detected fungus		 引物
Primer		 引物序列
Primer sequence (5′-3′)		 扩增片段
Target fragment (bp)		 退火温度
Tm (℃)
镰孢菌 ItsF AACTCCCAAACCCCTGTGAACATA 431 58
Fusarium spp. ItsR TTTAACGGCGTGGCCGC
禾谷镰孢复合种 Fg16NF ACAGATGACAAGATTCAGGCACA 280 57
F. graminearum Fg16NR TTCTTTGACATCTGTTCAACCCA
species complex
拟轮枝镰孢 VER1 CTTCCTGCGATGTTTCTCC 578 56
F. verticillioides VER2 AATTGGCCATTGGTATTATATATCTA
层出镰孢 PRO1 CTTTCCGCCAAGTTTCTTC 585 56
F. proliferatum PRO2 TGTCAGTAACTCGACGTTGTTG

Table 2

Primers used for identification of toxigenic chemotype"

引物
Primer		 引物序列
Primer sequence (5′-3′)		 毒素化学型
Toxigenic chemotype		 扩增片段
Product size (bp)		 退火温度
Tm (℃)
Tri13F TACGTGAAACATTGTTGGC DON, NIV 234, 415 57
Tri13R GGTGTCCCAGGATCTGCG
Tri303F GATGGCCGCAAGTGGA 3-DON 586 56
Tri303R GCCGGACTGCCCTATTG
Tri315F CTCGCTGAAGTTGGACGTAA 15-ADON 864 58
Tri315R GTCTATGCTCTCAACGGACAAC

Table 3

Occurrence of Fusarium spp. in 21 counties (districts) in Guangxi"

镰孢菌
Fusarium spp.		 拟轮枝镰孢
F. v		 层出镰孢
F. p		 甘蔗镰孢
F. s		 藤仓镰孢
F. f		 轮纹镰孢
F. c		 亚洲镰孢
F. a		 禾谷镰孢
F. g		 南方镰孢
F. m		 九州镰孢
F. k		 变红镰孢
F. i		 总计
Total
巴马 Bama 2 4 - - - 1 - - 3 - 10
大化 Dahua 2 - 1 1 - - - - - - 4
东兰 Donglan 9 - - - - - - - - - 9
都安 Duan 4 - 1 - - 1 - 2 3 - 11
凤山Fengshan 5 - - - - - - 3 - - 8
金城江Jinchengjiang - - 2 - - - - - - - 2
乐业 Leye 3 - - - - 2 - - - - 5
凌云 Lingyun 9 - - 2 - - - - - - 11
平果 Pingguo 1 1 - - - - 1 1 3 1 8
靖西 Jingxi - 6 - - 2 - - - - - 8
田东Tiandong 6 - - - - - - 1 - - 7
田林 Tianlin 2 - - - - - - - - - 2
田阳 Tianyang 2 - - - - - - - - - 2
德保 Debao 1 2 - 1 - - - 2 2 - 8
西林 Xilin - 1 - - - - - - - - 1
隆安 Longan - - - - - - - - - 1 1
江南 Jiangnan 7 - - - - - - - - - 7
马山 Mashan 7 - - - - - - - - - 7
武宣 Wuxuan - 2 2 1 - - - - 2 - 7
大新 Daxin 3 - 1 - - 1 - - 1 - 6
天等 Tiandeng 7 1 2 - - - - 3 1 - 14
总计 Total 70 17 9 5 2 5 1 12 15 2 138

Fig. 1

Morphological characteristic and maize symptom after inoculation of F. sacchari and F. concentricum"

Fig. 2

Construction of phylogenetic tree of some Fusarium spp. strains based on TEF-1α gene sequence"

Fig. 3

PCR amplification of toxigenic gene and chemotypes"

Table 4

Toxigenic chemotype detection of Fusarium spp. strains"

菌株编号
Strain number		 镰孢菌
Fusarium spp.		 毒素化学型 Toxigenic chemotype
NIV DON 3-ADON 15-ADON
Bama7 F. asiaticum - - - -
Daxin1 F. asiaticum + - - -
Duan1 F. asiaticum - - - -
Leye2 F. asiaticum + - - -
Leye4 F. asiaticum + - - +
Pingguo4 F. graminearum - + - +
Longan1 F. incarnatum + - - -
Pingguo8 F. incarnatum + - - +
Bama2 F. kyushuense + - - +
Bama3 F. kyushuense + - - +
Bama9 F. kyushuense - - - -
Daxin5 F. kyushuense - - - -
Debao3 F. kyushuense + - - -
Debao5 F. kyushuense - - - -
Duan3 F. kyushuense + - - -
Duan4 F. kyushuense + - - -
Duan5 F. kyushuense + - - -
Pingguo3 F. kyushuense + - - -
Pingguo5 F. kyushuense - - - -
Pingguo6 F. kyushuense + - - -
Tiandeng12 F. kyushuense + - - -
Wuxuan6 F. kyushuense + - - -
Wuxuan7 F. kyushuense - - - -
Debao6 F. meridionale + - - +
Debao7 F. meridionale + - - -
Duan2 F. meridionale - - - +
Duan8 F. meridionale + - - +
FengShan2 F. meridionale - - - +
FengShan3 F. meridionale + - - +
FengShan7 F. meridionale + - - +
Pingguo2 F. meridionale + - - +
Tiandeng1 F. meridionale + - - +
Tiandeng2 F. meridionale + - - +
Tiandeng8 F. meridionale + - - +
Tiandong4 F. meridionale + - - -
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