Inhibitory Effect of Lipopeptide Secreted by Bacillus Strain FJAT-2349 Against Anthracnose Pathogen
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摘要:
目的 探究解淀粉芽胞杆菌FJAT-2349脂肽对枇杷采后炭疽病菌的抑制作用。 方法 通过平板抑菌试验和活体接种试验测定脂肽对枇杷采后炭疽病的抑制效果，通过电镜分析脂肽对枇杷炭疽病菌生长的抑制作用。 结果 芽胞杆菌菌株FJAT-2349能够有效抑制枇杷采后炭疽病菌的生长，抑制率达87.8%。该菌株产生的脂肽由iturin、fengycin和surfactin组成，其中fengycin为主要成分。该脂肽对枇杷炭疽病菌抑菌圈直径可达22 mm，且抑制效果呈剂量效应。电镜结果显示，解淀粉芽胞杆菌菌株FJAT-2349脂肽会导致菌丝肿胀变形、细胞壁变薄或消失、细胞内容物减少，从而影响枇杷炭疽病菌菌的正常生长。活体接种试验表明，该脂肽能够推迟枇杷炭疽病发病时间1 d，降低枇杷炭疽病的发病率，其中治愈性脂肽处理组和预防性脂肽处理组接种枇杷炭疽病菌第4天的相对防效分别为29.2%和72.7%，平均病斑直径为4.67～5.00 mm，低于清水对照组（6.89~7.25 mm），表明先用脂肽预防性处理枇杷，能更有效地降低枇杷采后炭疽病的发生率，减轻枇杷采后炭疽病的发病程度。 结论 解淀粉芽胞杆菌FJAT-2349脂肽能够有效地抑制枇杷采后炭疽病菌。 Abstract:Objective Inhibitory effect of lipopeptides secreted by Bacillus FJAT-2349 on the pathogen of anthracnose was investigated. Method Agar plate diffusion and pin-prick inoculation were used to determine the inhibition effect of the lipopeptides on the pathogen. The fungal morphology was observed under a scanning and a transmission electron microscope. Result The lipopeptide-producing B. amyloliquefaciens FJAT-2349 could effectively inhibit the growth of Colletotrichum acutatum with an inhibition rate of 87.8%. The lipopeptides were composed of mainly fengycin, but also iturin and surfactin. The diameter of inhibition zone on the agar plate by the lipopeptide against C. acutatum was up to 22 mm. The effect was dose-dependent. Under the electron microscopes, the fungal hyphae appeared deformed and cell walls damaged by the lipopeptides. Inoculation of FJAT-2349 on harvested loquats could delay the onset of anthracnose by one day. The in vivo tests showed that on the 4th day after inoculation the relative biocontrol efficacy on a curative-treatment group was 29.2% and 72.7% on a preventive-treatment group; and the average lesion diameter on the treatment groups was 4.67-5.00 mm, as opposed to the larger 6.89-7.25 mm of control. Conclusion The lipopeptides secreted by B. amyloliquefaciens FJAT-2349 could effectively inhibit the growth of Colletotrichum acutatum. -
Key words:
- Bacillus lipopeptide /
- loquat anthracnose /
- antifungal effect /
- disease incidence
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图 1 菌株FJAT-2349对枇杷炭疽病菌FJAT-30256的抑制作用
注：a，处理组；b，对照组。
Figure 1. Inhibition of FJAT-2349 against C. acutatum FJAT-30256
Note: a, treatment group; b, control group.
图 2 枇杷炭疽病菌菌丝形态扫描电镜观察
注：a，对照组；b，脂肽处理组。
Figure 2. Scanning electron micrograph of C. acutatum hyphae
Note: a, control group; b,lipopeptide-treated group.
图 3 枇杷炭疽病菌菌丝形态透射电镜观察
注：a、b：对照组；c、d：脂肽处理组。
Figure 3. Transmission electron micrograph of C. acutatum hyphae
Note: a, b: Control group; c, d: Lipopeptide-treated group.
表 1 不同含量脂肽对真菌的抑制作用
Table 1. Antifungal effect of lipopeptides
脂肽含量
Lipopeptide content/
（mg·mL−1）抑菌圈直径
The inhibition zone diameter/mm枇杷炭疽病菌
Colletotrichum acutatum
FJAT-30256香蕉尖孢镰刀菌
Fusarium oxysporum
FJAT-370甜瓜尖孢镰刀菌
F. oxysporum
FJAT-9230小新壳梭孢菌
Neofusicoccum parvum
FJAT-353130 22.75±0.31 a 22.35±0.48 a 25.46±0.42 a 21.73±0.66 a 20 21.33±0.18 b 22.34±0.63 a 20.01±0.54 b 19.73±1.75 ab 10 20.95±0.99 b 21.48±0.39 a 17.43±0.83 c 17.61±0.11 b 注：同列数据后不同字母表示差异显著（ P <0.05）。
Note: The difference letter in the same column indicated that the difference between the grades is significantly (P< 0.05).
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表 2 脂肽对枇杷炭疽病害治愈性试验结果
Table 2. Effect of lipopeptides in curing anthracnose on postharvest loquats
时间 Date 治愈性清水对照组
Curative water-treated group治愈性脂肽处理组
Curative lipopeptide-treated group发病率
Incidence/%平均病斑直径
Average lesion diameter/mm发病率
Incidence/%平均病斑直径
Average lesion diameter/mm接种第3天 On the third day after inoculation 60.0 4.00±0.51 0 — 接种第4天 On the forth day after inoculation 100.0 7.25±1.37 a 70.8 5.53±1.33 b 接种第5天 On the fifth day after inoculation 100.0 13.15±2.94 a 100.0 10.08±3.80 b 接种第6天 On the sixth day after inoculation 100.0 18.75±3.23 a 100.0 15.33±2.61 b 注：同行数据后不同字母表示差异显著（P<0.05），表3同。
Note: The difference letter in the same row indicated that the difference between the grades is significantly (P < 0.05), the same as table 3.
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表 3 脂肽对枇杷炭疽病害预防性试验结果
Table 3. Effect of lipopeptides in preventing anthracnose on postharvest loquats
时间 Date 预防性清水对照组
Preventive water-treated group预防性脂肽处理组
Preventive lipopeptide-treated group发病率
Incidence/%平均病斑直径
Average lesion diameter/mm发病率
Incidence/%平均病斑直径
Average lesion diameter/mm接种第3天 On the third day after inoculation 27.8 5.00±0.25 0 — 接种第4天 On the forth day after inoculation 100.0 6.89±2.91 a 27.3 4.67±1.21 a 接种第5天 On the fifth day after inoculation 100.0 11.29±4.60 a 59.1 8.00±5.10 a 接种第6天 On the sixth day after inoculation 100.0 16.29±3.70 a 72.7 16.13±3.82 a 接种第7天 On the seventh day after inoculation 100.0 23.28±4.88 a 90.9 19.75±5.69 a
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