生防细菌HK11-9对黄瓜棒孢叶斑病的防病能力及其鉴定

doi:10.13560/j.cnki.biotech.bull.1985.2023-0407

摘要/Abstract

摘要：

为探讨黄瓜棒孢叶斑病生防细菌HK11-9的防病能力，通过皿内试验测定HK11-9对黄瓜棒孢叶斑病菌多主棒孢霉的抑菌活性、通过离体和活体接种试验评价其对黄瓜棒孢叶斑病的防治效果，进行盆栽试验分析HK11-9在黄瓜叶部的定殖能力及其对黄瓜叶部防御酶活性的影响，同时，测定HK11-9的生物学功能并对其进行鉴定。结果显示，HK11-9显著抑制多主棒孢霉的菌丝生长和孢子萌发；在离体接种和活体接种条件下，HK11-9均对黄瓜棒孢叶斑病具有明显的防治效果，其防治效果分别达到62.80%和67.73%；同时，HK11-9在黄瓜叶部具有稳定的定殖能力，在黄瓜叶部能保持10⁴CFU/g的定殖密度，能够显著提高黄瓜叶部PAL、POD和PPO活性。HK11-9菌株具有分解蛋白、分解纤维素、分解淀粉和产嗜铁素作用，对多种植物病原真菌具有抑菌活性；并根据形态特征、生理生化特性和16S rRNA序列同源性分析，将HK11-9菌株鉴定为多黏类芽孢杆菌（Paenibacillus polymyxa）。综上所述，生防细菌HK11-9防治黄瓜棒孢叶斑病具有极大的潜力。

关键词: 黄瓜棒孢叶斑病, 多黏类芽孢杆菌HK11-9, 定殖量, 防御酶活性, 鉴定, 生物防治

Abstract:

In order to explore the biocontrol ability of biocontrol agent HK11-9 against Corynespora leaf spot of cucumber, the antifungal activity of HK11-9 against Corynespora cassiicola were detected by in petri dishes experiments, the control effect of HK11-9 against Corynespora leaf spot of cucumber were detected by in vitro and in vivo inoculation experiments, the colonization ability of HK11-9 on cucumber leaves and its effect on the activities of defense enzymes were conducted by pot experiment, at the same time, the biological functions of HK11-9 was determined. The results showed that, HK11-9 significantly inhibited the mycelia growth and conidia germination of Corynespora cassiicola in the petri dishes experiment, HK11-9 presented significant control effect against Corynespora leaf spot of cucumber at the in vitro and in vivo inoculation experiment, the control efficiencies were 62.80% and 67.73% respectively. HK11-9 colonized stably in cucumber leaves, maintained a density of 10⁴CFU/g in cucumber leaves, and HK11-9 significantly improved the activities of defense enzymes in cucumber leaves. In addition, HK11-9 decomposed protein, cellulose and starch, produced siderophore, and HK11-9 demonstrated antifungal activity against a variety plant pathogenic fungi. Based on the morphological, physiological characteristics and 16S rRNA sequence analysis, the strain HK11-9 was identified as Paenibacillus polymyxa. In summary, the biocontrol agent HK11-9 has great potential to control Corynespora leaf spot of cucumber.

Key words: Corynespora leaf spot of cucumber, Paenibacillus polymyxa HK11-9, colonization density, activity of defense enzyme, identification, biological control

张林林, 沈虎生, 杨冰, 何梦菡, 朴凤植, 申顺善. 生防细菌HK11-9对黄瓜棒孢叶斑病的防病能力及其鉴定[J]. 生物技术通报, 2023, 39(12): 209-218.

ZHANG Lin-lin, SHEN Hu-sheng, YANG Bing, HE Meng-han, PIAO Feng-zhi, SHEN Shun-shan. Control Ability and Identification of Biocontrol Agent HK11-9 Against Corynespora Leaf Spot of Cucumber[J]. Biotechnology Bulletin, 2023, 39(12): 209-218.

图/表 14

参考文献 38

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处理 Treatment	离体接种 In vitro inoculation		活体接种In vivo inoculation
处理 Treatment	病情指数Disease index	防治效果Control effect/%	病情指数Disease index	防治效果Control effect/%
HK11-9	22.03±1.13a	62.80	21.17±1.67a	67.73
化学药剂Pesticides	24.21±0.97a	59.12	25.25±2.33a	61.52
对照Control	59.23±1.33b		65.61±2.21b

处理 Treatment	离体接种 In vitro inoculation		活体接种In vivo inoculation
处理 Treatment	病情指数Disease index	防治效果Control effect/%	病情指数Disease index	防治效果Control effect/%
HK11-9	22.03±1.13a	62.80	21.17±1.67a	67.73
化学药剂Pesticides	24.21±0.97a	59.12	25.25±2.33a	61.52
对照Control	59.23±1.33b		65.61±2.21b

处理 Treatment	病情指数 Disease index	防治效果 Control effect/%
菌悬液Bacteria suspension	24.38±0.87a	66.35
发酵液Fermentation liquid	25.50±1.12a	64.80
上清液Fermentation solution	69.96±2.04b	3.44
对照 Control	72.45±1.67b

处理 Treatment	病情指数 Disease index	防治效果 Control effect/%
菌悬液Bacteria suspension	24.38±0.87a	66.35
发酵液Fermentation liquid	25.50±1.12a	64.80
上清液Fermentation solution	69.96±2.04b	3.44
对照 Control	72.45±1.67b

鉴定指标Identification index	结果 Result
菌体形态Cell morphology	直杆状Straight rod
菌体大小Cell size	（0.5-0.8）μm×（2.4-2.8）μm
鞭毛Flagellum	周生Multi flagella
革兰氏染色Gram reaction	G+
耐盐度Salt concentration	≤4%
生长温度Growth temperature	4-50℃
最适生长温度 Optimal growth temperature	30℃
运动性Motility	+
生长所需营养 Nutrition
蔗糖Sucrose	+
葡萄糖Glucose	+
甘油Glycerol	+
麦芽糖Maltose	+
半乳糖Galactose	-
单宁酸Tannin	-
甘露醇Mannitol	+
柠檬酸Citric acid	-
硝酸盐还原Nitrate reduction	-
亚硝酸还原nitrous acid	+
硫化氢Hydrogen sulfide	-
明胶液化Gelatin liquefaction	-
淀粉水解Amylolysis	+
葡萄糖氧化Glucose oxidative	发酵型 Fermentation