山西长治番茄枯萎病病原鉴定及其拮抗菌鉴选

doi:10.16420/j.issn.0513-353x.2024-0811

摘要/Abstract

摘要：

以山西省长治市壶关县番茄枯萎病茎秆处样品为试验材料，采用组织分离培养法对该病病原进行分离且培养，泡根接种法测定分离病原菌的致病性。通过常规形态学、ITS序列分析法鉴定该番茄枯萎病病原菌的种类，并对其生物学特性进行研究。同时，从番茄西葫芦轮作两轮后种植西葫芦的土壤中分离到哈茨木霉菌（Trichoderma harzianum）M408，采用平板对峙法测定哈茨木霉M408菌丝及无菌发酵液对该番茄枯萎病原菌的生防能力。经鉴定，山西省长治市壶关县番茄枯萎病的病原菌为层出镰刀菌（Fusarium proliferatum），命名为K11。其最适生长温度28 ℃，最适pH 6.0，24 h全光照下生长速率最快，可利用多种碳源生长，其中碳源为蔗糖时菌丝生长速率最快。用菌丝平板对峙培养法测定，第7天时，菌株哈茨木霉M408对K11菌丝的抑菌率为68.38%，其无菌发酵液对K11菌丝的抑菌率为93.17%。说明M408对K11具有较好的抑制作用，生防机制可能为竞争作用。菌株哈茨木霉M408具有开发成番茄枯萎病抑菌剂的能力。

关键词: 番茄, 枯萎病, 病原鉴定, 层出镰刀菌, 生物学特性, 拮抗菌

Abstract:

Tomato Fusarium wilt samples from the stems in Huguan County，Changzhi City，Shanxi Province，were used as experimental materials. The pathogen was isolated and cultured using tissue separation methods，and its pathogenicity was determined by root-dipping inoculation. The pathogen species responsible for the tomato wilt disease was identified using conventional morphological analysis and ITS sequence analysis. Additionally，the biological characteristics of the pathogen were studied. Furthermore，after two rounds of tomato-zucchini crop rotation，Trichoderma harzianum strain M408 was isolated from the soil planted with zucchini. The antagonistic activity of Trichoderma harzianum M408 mycelium and its sterile fermentation liquid against the tomato wilt pathogen was measured using the dual-culture plate method. The results revealed that the pathogen causing tomato wilt disease in Huguan County，Changzhi City，Shanxi Province，was identified as Fusarium proliferatum，named K11. The suitable growth temperature for K11 was 28 ℃，and 6.0 was the most suitable pH value. Under 24-hour full light conditions，the growth rate of K11 was fastest. K11 was able to grow using various carbon sources，with the fastest mycelial growth observed when sucrose was used as the carbon source. Using the mycelial confrontation method on plates，after 7 days，Trichoderma harzianum M408 showed a 68.38% inhibition rate on the K11 mycelium，and its sterile fermentation liquid exhibited 93.17% inhibition rate against K11 mycelium.

Key words: tomato, Fusarium wilt, pathogen identification, Fusarium proliferatum, biological characteristics, antagonistic fungus

苏秀敏, 王佼, 韩文清, 李鹏, 王秋兰, 李万星. 山西长治番茄枯萎病病原鉴定及其拮抗菌鉴选[J]. 园艺学报, 2025, 52(9): 2532-2544.

SU Xiumin, WANG Jiao, HAN Wenqing, LI Peng, WANG Qiulan, LI Wanxing. Isolation，Biological Characteristics of the Pathogen Causing Fusarium Wilt on Tomato and Antagonistic Fungus Screening[J]. Acta Horticulturae Sinica, 2025, 52(9): 2532-2544.

图/表 8

图1 田间症状及K11菌丝形态 A：初步分离的菌落形态；B：纯化后的菌落形态；C：番茄枯萎病茎部剖开图；D：番茄枯萎病后期田间症状

Fig. 1 Symptoms of Fusarium and colony morphology A：Colony morphology of primary isolates；B：Colony morphology after purification；C：Dissection section symptom of stem；D：Symptoms at the later period of infection

图2 孢子形态及PDA培养基上K11菌落形态 A：孢子形态；B，C：PDA培养基上K11菌落形态

Fig. 2 Microconidia and colony morphology on PDA medium A：Macroconidia；B，C：Colony morphology on PDA medium

图3 供试菌株K11电泳检测结果

Fig. 3 The electrophoresis analysis result of strain K11

图4 基于 ITS 序列构建的菌株K11系统发育树系统发育树各分支点上的数字代表bootstrap值；标尺代表进化距离

Fig. 4 The phylogenetic tree of the strain K11 based on ITS sequence The number on each branch point represents the bootstrap；The scale bar represents the evolutionary distance

图5 不同生长条件下 K11菌株的菌落直径（第7天）不同小写字母表示0.05水平差异显著

Fig. 5 Colony diameters of the K11 strain under different growth conditions（the 7th day） Different lowercase letters indicate significant differences among different treatments at 0.05 level

图6 菌株哈茨木霉M408菌丝与菌株层出镰刀菌K11菌丝对峙培养的菌落形态（1 ~ 7 d）

Fig. 6 The colony morphology confrontation cultured Trichoderma harzianum strain M408 and Fusarium proliferatum strain K11（1-7 d）

图7 菌株哈茨木霉M408无菌发酵液与层出镰刀菌K11菌株对峙培养的菌落形态（第7天）

Fig. 7 The colony morphology confrontation cultured of Trichoderma harzianum strain M408 and Fusarium proliferatum strain K11（the 7th day）

图8 哈茨木霉M408菌丝及无菌发酵液与层出镰刀菌K11菌丝峙培养的k11菌落直径（1 ~ 7 d）不同小写字母表示0.05水平差异显著

Fig. 8 The colony diameters of K11 confrontation cultured Trichoderma harzianum M408 and Fusarium proliferatum strain K11（1-7 d） Different lowercase letters indicate significant differences among different treatments at 0.05 level

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