陕西猕猴桃常见病害生物防治技术及田间应用
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摘要
陕西作为我国猕猴桃的主要产区,也是猕猴桃病害的重灾区。各种病害特别是猕猴桃细菌性溃疡病和猕猴桃细菌性叶枯病,发病面积广、发病程度重,严重影响着果实的数量和质量,阻碍了整个猕猴桃产业的发展。
本研究从不同来源的样品中进行猕猴桃病害拮抗菌的筛选鉴定,并尝试将拮抗作用好且具有亲和性的两株生防菌株进行复配,进而进行室内盆栽试验比较复合和单一菌剂的生防效果,以观察复合菌剂防效的提高情况,最后进行田间防治试验,以期能够获得在田间实际应用中较单一拮抗菌株仿效更高、稳定性好的复合拮抗菌剂。
本实验分别采用抑菌圈法和滤纸片法初筛和复筛,从猕猴桃根际土壤等七个不同来源的样品中分离得到的96株菌株中,筛选出2株对猕猴桃细菌性溃疡病病原菌(Pseudomonas syringae Pv. Actinidiae)和猕猴桃细菌性叶枯病病原菌(Pseudomonas syringaepv. syringae)均具有良好拮抗作用的菌株ZM12和SF6；菌株ZM12和SF6对甜瓜枯萎病、白菜软腐病等果几种蔬常见真菌和细菌性病害病原菌均表现出较强的抑制作用,是两株具有广谱拮抗作用的菌株。
依据两菌株的形态及生理生化特征,初步推断菌株ZM12和SF6均为芽孢杆菌属(Bacillus),16S rDNA序列分析结果表明菌株ZM12为枯草芽抱杆菌(Bacillus subtilis),菌株SF6为巨大芽孢杆菌(Bacillus megaterium)
平板交互划线培养结果表明菌株ZM12和SF6能够在同一平板上互无拮抗作用共存,两者之间表现出良好的亲和性；混合液体培养发酵液中的活菌数高于两菌单独培养的,表明混合培养有利于菌的生长和增殖；盆栽试验中混合发酵液对猕猴桃细菌性溃疡病和猕猴桃细菌性叶枯病的防治效果分别高达92.4%和93.5%,明显高于菌株ZM12和SF6。
田间防治试验结果表明复合拮抗菌剂三个不同浓度的处理均对猕猴桃病害表现出较好的防治效果,其原液、50倍和100倍稀释液对猕猴桃细菌性溃疡病的防治效果分别为89.8%、85.6%和83.7%,病斑治愈率分别为94.0%、88.0%和86.0%,对猕猴桃细菌性叶枯病的防治效果分别为90.4%、87.0%和85.9%,均显著高于95%CT 1000倍稀释液(p<0.05)。
复合拮抗菌剂处理组猕猴桃果实经农业部食品质量监督检验测试中心进行感官评价、果实质量检测、农药及重金属残留检测,表明果实生长健康完好,无任何农药残留,无重金属残留或远低于国家标准,符合国家无公害食品的要求；Vc含量糖度及可溶性固形物含量均均得到提升,总酸含量下降,果实质量比对照更为优质。同时复合拮抗菌剂对果树树势的生长表现出正效应。
As the main kiwifruit-growing areas, Shaanxi also is a worst-hit area of kiwifruit disease, especially kiwifruit bacterial canker and kiwifruit bacterial leaf blight disease, decreasing not only the yield and quality, but also the whole industrialization of kiwifruit. Because of the overuse of chemical pesticides inevitably led to pesticide residues in fruit and environmental pollution, it is necessary to seek an approach that healthy, green and environment protected, and antagonistic strains is proved to be the most effectively and feasiblely.
96 strains were isolated from rhizosphere soil of kiwifruit and other different source, in primary and further screening, strains ZM12 and SF6 revealed obviously antibacterial activities against the pathogen of kiwifruit bacterial canker(Pseudomonas syringae Pv. Actinidiae) and kiwifruit bacterial leaf blight disease(Pseudomonas syringae Pv. Actinidiae). And the two strains also revealed good antagonism to common diseases pathogens of fruits and vagetables, Fusarim axysporium, Erwinia carotovora subsp.carotovora and so on, they are broad-spectrum antagonistic strains.
Based on the morphological characteristics and physiological-biochemical characteristics of stains ZM12 and SF6, they are regarded as Bacillus preliminarily, and the analysis of 16S rDNA sequence showed stain ZM12 is Bacillus subtilis, SF6 is Bacillus megaterium.
Culture ZM12 and SF6 in one NA plate, it showed normal growth, did not express any antibacterial activities to each other; Do mix culture of ZM12 and SF6 in liquid medium, the viable counts of mixed fermentation broth are evidently more than the other two when strains ZM12 and SF6 were cultured individually, display that mix culture can enhance the growth of strains. In pot experiments, mixed fermentation broth showed excellent control effect to kiwifruit bacterial canker and kiwifruit bacterial leaf blight disease, can reach 92.4% and 93.5% respectively.
Field experiment showed that all of the three concentration of mixed antagonistic inoculum have achieved the desirable control efficacy towards kiwifruit diseases. The control effect to kiwifruit bacterial canker were 89.8%,85.6%,83.7%, and spots cure effect were 94.0%,88.0%,86.0%, respectively, the control effect to kiwifruit bacterial leaf blight disease reached to 90.4%,87.0% and 85.9%, significantly higher than those of chemical bactericide 95% CT (p<0.05).
The kiwifruit of mixed antagonistic inoculum treatment no pesticide residues, no heavy metals or extra-low than national request, accord with national standard of non-pollution food, and the quality of kiwifruit has been improved. Mixed antagonistic inoculum benefit the growth of kiwifruit tree.
本研究从不同来源的样品中进行猕猴桃病害拮抗菌的筛选鉴定,并尝试将拮抗作用好且具有亲和性的两株生防菌株进行复配,进而进行室内盆栽试验比较复合和单一菌剂的生防效果,以观察复合菌剂防效的提高情况,最后进行田间防治试验,以期能够获得在田间实际应用中较单一拮抗菌株仿效更高、稳定性好的复合拮抗菌剂。
本实验分别采用抑菌圈法和滤纸片法初筛和复筛,从猕猴桃根际土壤等七个不同来源的样品中分离得到的96株菌株中,筛选出2株对猕猴桃细菌性溃疡病病原菌(Pseudomonas syringae Pv. Actinidiae)和猕猴桃细菌性叶枯病病原菌(Pseudomonas syringaepv. syringae)均具有良好拮抗作用的菌株ZM12和SF6；菌株ZM12和SF6对甜瓜枯萎病、白菜软腐病等果几种蔬常见真菌和细菌性病害病原菌均表现出较强的抑制作用,是两株具有广谱拮抗作用的菌株。
依据两菌株的形态及生理生化特征,初步推断菌株ZM12和SF6均为芽孢杆菌属(Bacillus),16S rDNA序列分析结果表明菌株ZM12为枯草芽抱杆菌(Bacillus subtilis),菌株SF6为巨大芽孢杆菌(Bacillus megaterium)
平板交互划线培养结果表明菌株ZM12和SF6能够在同一平板上互无拮抗作用共存,两者之间表现出良好的亲和性；混合液体培养发酵液中的活菌数高于两菌单独培养的,表明混合培养有利于菌的生长和增殖；盆栽试验中混合发酵液对猕猴桃细菌性溃疡病和猕猴桃细菌性叶枯病的防治效果分别高达92.4%和93.5%,明显高于菌株ZM12和SF6。
田间防治试验结果表明复合拮抗菌剂三个不同浓度的处理均对猕猴桃病害表现出较好的防治效果,其原液、50倍和100倍稀释液对猕猴桃细菌性溃疡病的防治效果分别为89.8%、85.6%和83.7%,病斑治愈率分别为94.0%、88.0%和86.0%,对猕猴桃细菌性叶枯病的防治效果分别为90.4%、87.0%和85.9%,均显著高于95%CT 1000倍稀释液(p<0.05)。
复合拮抗菌剂处理组猕猴桃果实经农业部食品质量监督检验测试中心进行感官评价、果实质量检测、农药及重金属残留检测,表明果实生长健康完好,无任何农药残留,无重金属残留或远低于国家标准,符合国家无公害食品的要求；Vc含量糖度及可溶性固形物含量均均得到提升,总酸含量下降,果实质量比对照更为优质。同时复合拮抗菌剂对果树树势的生长表现出正效应。
As the main kiwifruit-growing areas, Shaanxi also is a worst-hit area of kiwifruit disease, especially kiwifruit bacterial canker and kiwifruit bacterial leaf blight disease, decreasing not only the yield and quality, but also the whole industrialization of kiwifruit. Because of the overuse of chemical pesticides inevitably led to pesticide residues in fruit and environmental pollution, it is necessary to seek an approach that healthy, green and environment protected, and antagonistic strains is proved to be the most effectively and feasiblely.
96 strains were isolated from rhizosphere soil of kiwifruit and other different source, in primary and further screening, strains ZM12 and SF6 revealed obviously antibacterial activities against the pathogen of kiwifruit bacterial canker(Pseudomonas syringae Pv. Actinidiae) and kiwifruit bacterial leaf blight disease(Pseudomonas syringae Pv. Actinidiae). And the two strains also revealed good antagonism to common diseases pathogens of fruits and vagetables, Fusarim axysporium, Erwinia carotovora subsp.carotovora and so on, they are broad-spectrum antagonistic strains.
Based on the morphological characteristics and physiological-biochemical characteristics of stains ZM12 and SF6, they are regarded as Bacillus preliminarily, and the analysis of 16S rDNA sequence showed stain ZM12 is Bacillus subtilis, SF6 is Bacillus megaterium.
Culture ZM12 and SF6 in one NA plate, it showed normal growth, did not express any antibacterial activities to each other; Do mix culture of ZM12 and SF6 in liquid medium, the viable counts of mixed fermentation broth are evidently more than the other two when strains ZM12 and SF6 were cultured individually, display that mix culture can enhance the growth of strains. In pot experiments, mixed fermentation broth showed excellent control effect to kiwifruit bacterial canker and kiwifruit bacterial leaf blight disease, can reach 92.4% and 93.5% respectively.
Field experiment showed that all of the three concentration of mixed antagonistic inoculum have achieved the desirable control efficacy towards kiwifruit diseases. The control effect to kiwifruit bacterial canker were 89.8%,85.6%,83.7%, and spots cure effect were 94.0%,88.0%,86.0%, respectively, the control effect to kiwifruit bacterial leaf blight disease reached to 90.4%,87.0% and 85.9%, significantly higher than those of chemical bactericide 95% CT (p<0.05).
The kiwifruit of mixed antagonistic inoculum treatment no pesticide residues, no heavy metals or extra-low than national request, accord with national standard of non-pollution food, and the quality of kiwifruit has been improved. Mixed antagonistic inoculum benefit the growth of kiwifruit tree.
引文
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