CN114831120B - 一种含有脂肽的组合物及其在害虫防治中的应用 - Google Patents

一种含有脂肽的组合物及其在害虫防治中的应用 Download PDF

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CN114831120B
CN114831120B CN202210681601.8A CN202210681601A CN114831120B CN 114831120 B CN114831120 B CN 114831120B CN 202210681601 A CN202210681601 A CN 202210681601A CN 114831120 B CN114831120 B CN 114831120B
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pests
bacillus
lipopeptide
composition
pest control
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CN114831120A (zh
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刘洪伟
赵雯雅
刘永锋
姚彩苗
张丽萍
张飞燕
王雅娜
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Institute of Biology of Hebei Academy of Sciences
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Abstract

本发明公开了一种含有脂肽的组合物及其在害虫防治中的应用，属于害虫防治技术领域。所述组合物包括丰原素家族脂肽、伊枯草素家族脂肽和表面活性素家族脂肽。将所述组合物和不同农药复配，能够用于防治半翅目害虫、鞘翅目害虫、直翅目害虫、蜚蠊目害虫以及鳞翅目害虫，尤其是对半翅目昆虫或鞘翅目昆虫的杀虫活性非常明显，24小时在低浓度时杀虫效果可达90％以上，因此，本发明公开的组合物可以应用于农业害虫的防治中，为害虫防治新药物开发奠定基础。

Description

一种含有脂肽的组合物及其在害虫防治中的应用
技术领域
本发明涉及害虫防治技术领域，特别是涉及一种含有脂肽的组合物及其在害虫防治中的应用。
背景技术
丰原素，也叫芬荠素，包括丰原素(fengycin)A、B和制磷脂菌素(plipastatin)A、B。1986年，德国和日本团队同时发现了丰原素：枯草芽孢杆菌产生的丰原素和蜡状芽孢杆菌的制磷脂菌素。丰原素被确定为抗真菌剂，制磷脂菌素被确定为磷脂酶A2的抑制剂。它们之间仅存在着微小的结构差异(它们的差异在于第9位氨基酸-酪氨酸的构象，丰原素为L型，而制磷脂菌素则为D型)。而后Kim等人证实苏云金芽孢杆菌也产生丰原素。Tosato等人在枯草芽孢杆菌168中发现编码丰原素或制磷脂菌素合成酶的操纵子，而后在枯草芽孢杆菌b213以及解淀粉芽孢杆菌FZB42中也陆续发现。
伊枯草素(Iturin)家族化合物是Delcambe和Devignat在1957年从枯草芽孢杆菌(Bacillus subtilis)中分离得到，至今已发现了很多属于Iturins家族中的脂肽，比如Iturin A、B、C、D、E，芽孢菌霉素(bacillomycin)D、F、L以及抗霉枯草菌素(mycosubtilin)等。近期研究发现产生iturins家族脂肽的其他菌种还有类芽孢杆菌、解淀粉芽孢杆菌和短小芽孢杆菌等。
表面活性素(Surfactin)是1968年Arima等人从枯草芽孢杆菌胞外次级代谢产物中分离出具有特殊生物表面活性的化合物，经鉴定为脂肽结构。表面活性素由于其卓越的表面活性剂性质最初由Arima等鉴定和命名，其化学结构由Kakinuma等确定其含有7个α-氨基酸和具有13、14或15个碳原子的β-羟基脂肪酸构成的大环内脂型脂肽类化合物，被称为标准表面活性素(Standard surfactin)。此后表面活性素家族的成员不断增加，分别来源于天然的、生物合成的或基因工程产生的肽链或脂肪酸链的变异体。
害虫防治是农业生产中非常重要的组成部分，随着农业生产方式的转变以及科学技术的进步，害虫防治策略先后经历了农业防治、化学防治、综合防治和绿色防治阶段。随着监测预警技术、精准施药技术、生物防治、物理防治和转基因技术等多种害虫防治技术的不断发展，先后构建了以单个作物生产过程和重大致灾害虫为对象的综合防治技术体系，在保障我国粮食产量与食品安全，推动农业害虫绿色防控方面发挥了关键作用。近年来，受全球气候变化、耕作模式变化、产业结构调整和国际贸易交流带来的生物入侵日趋频繁等多种因素的影响，我国农业害虫的发生种类和为害规律也产生了明显变化，农业害虫防治面临新的挑战。
化学防治是目前应用较为广泛的一种措施，但长期使用化学农药带来的农药残留、生物抗药性等问题日益突出。近年来，随着人们对环境生态和食品安全的日益关注，以生态友好为目的的新农药开发成为必然趋势。鉴于Iturins家族脂肽抑真菌谱广、毒性低、易于生物降解、不会引起过敏反应等诸多优点，适合开发为害虫防治药物。
发明内容
本发明的目的是提供一种含有脂肽的组合物及其在害虫防治中的应用，以解决上述现有技术存在的问题，该组合物能够防治半翅目害虫、鞘翅目害虫、直翅目害虫、蜚蠊目害虫以及鳞翅目害虫，为害虫防治新药物开发奠定基础。
为实现上述目的，本发明提供了如下方案：
本发明提供一种组合物，所述组合物包括丰原素家族脂肽、伊枯草素家族脂肽和表面活性素家族脂肽。
优选的是，所述组合物中丰原素家族脂肽、伊枯草素家族脂肽和表面活性素家族脂肽的有效含量分别为0.01％-10％
优选的是，所述组合物还包括杀虫剂。
优选的是，所述杀虫剂包括化学杀虫剂、生物杀虫剂、植物杀虫剂。
优选的是，所述化学杀虫剂包括丙氨酸甲酯、苯醚甲环性、速克灵、进口甲托、苯并咪唑44号、密霉胺、福美霜、进口蓝粉、氯虫苯甲酰胺、高效氯氟氰菊酯、乙基多杀菌素、灭幼脲氯氰菊酯、吡虫啉、溴氰菊酯、赤霉酸、吲哚乙酸、吲哚丁酸、烯腺嘌呤、羟烯腺嘌呤、苄氨基嘌呤、24-表芸苔素内酯、22,23,24-表芸苔素内酯、28-表高芸苔素内酯、28-高芸苔素内酯、14-羟基芸苔素甾醇、三十烷醇、S-诱抗素、抗坏血酸、糠氨基嘌呤、二氢卟吩铁、尿囊素、超敏蛋白、极细链格孢激活蛋白、氨基寡糖素、香菇多糖、几丁聚糖、葡聚烯糖、低聚糖素、β-羽扇豆球蛋白多肽、胆钙化醇、米尔贝霉素、二化螟性诱剂、斜纹夜蛾诱集性信息素、绿盲蝽性信息素、梨小性迷向素以及苹果蠹蛾性信息素中至少一种(以上化学杀虫剂均购自于中国农科院植保所中保绿农集团)。
优选的是，所述生物杀虫剂包括枯草芽孢杆菌、短稳杆菌、多粘类芽孢杆菌、苏云金芽孢杆菌、甲基营养型芽孢杆菌、海洋芽孢杆菌、坚强芽孢杆菌、球形芽孢杆菌、蜡质芽孢杆菌、荧光假单胞杆菌、侧孢短芽孢杆菌、短稳杆菌、地衣芽孢杆菌、解淀粉芽孢杆菌、沼泽红假单胞菌、嗜硫小红卵菌、白僵菌、球孢白僵菌、金龟子绿僵菌、哈茨木霉菌、木霉菌、淡紫拟青霉、厚孢轮枝菌、耳霉菌、盾壳霉、小盾壳霉、寡雄腐霉菌、蟑螂病毒、多角体病毒、颗粒体病毒、多杀霉素、阿维菌素以及蝗虫微孢子虫中至少一种(以上生物杀虫剂均购自于农保生物厂家)。
优选的是，植物杀虫剂包括印楝素、苦参碱、藜芦碱、烟碱、鱼藤酮、除虫菊素、苦皮藤素、桉油精、八角茴香油、狼毒素、雷公藤甲素、莪术醇、蛇床子素、丁子香酚、大黄素甲醚、香芹酚、小檗碱、甾烯醇、茶皂素、螺威、大蒜素、d-柠檬烯、萜烯醇、异硫氰酸烯丙酯、十五烯苯酚酸、十三烷苯酚酸以及苯丙烯菌酮中至少一种(以上植物杀虫剂均购自于盛源农资)。
本发明还提供所述的组合物在害虫防治中的应用，所述害虫包括半翅目害虫、鞘翅目害虫、直翅目害虫、蜚蠊目害虫和鳞翅目害虫。
优选的，所述的半翅目害虫为豆蚜、棉蚜、甘蓝蚜和麦长管蚜中的一种或多种；所述鞘翅目害虫为白星花金龟、暗黑鳃金龟、铜绿丽金龟和小云斑鳃金龟中的一种或多种；所述直翅目害虫为蝗虫、花生大蟋、蝼蛄、棉斑草螽中的一种或多种；所述蜚蠊目害虫为黑胸大蠊、美洲大蠊、澳洲大蠊、德国小蠊、日本大蠊中的一种或多种；所述鳞翅目害虫为甜纹夜蛾、斜纹夜蛾、小菜蛾、棉铃虫中的一种或多种。
本发明还包括一种保护种子及后来形成的植物器官免受害虫侵害的方法，包括利用所述的组合物处理种子的过程。优选使用包含所述组合物的包衣剂对种子包衣，所述包衣剂中还包括成膜剂和粘合剂。但不限于用所述组合物处理种子，也可以用组合物直接处理后来形成的植物或者植物器官或者土壤等。
优选的是，所述组合物的浓度为0.1μg/mL-100μg/mL。
本发明公开了以下技术效果：
本发明公开了包含三种脂肽的组合物及其新应用，即在害虫防治中的应用。经过实验证明，脂肽复配杀虫剂的组合物对半翅目昆虫或鞘翅目昆虫的杀虫活性非常明显，24小时在低浓度时杀虫效果可达90％以上，可以应用于农业害虫的防治中。
具体实施方式
现详细说明本发明的多种示例性实施方式，该详细说明不应认为是对本发明的限制，而应理解为是对本发明的某些方面、特性和实施方案的更详细的描述。
应理解本发明中所述的术语仅仅是为描述特别的实施方式，并非用于限制本发明。另外，对于本发明中的数值范围，应理解为还具体公开了该范围的上限和下限之间的每个中间值。在任何陈述值或陈述范围内的中间值以及任何其他陈述值或在所述范围内的中间值之间的每个较小的范围也包括在本发明内。这些较小范围的上限和下限可独立地包括或排除在范围内。
除非另有说明，否则本文使用的所有技术和科学术语具有本发明所述领域的常规技术人员通常理解的相同含义。虽然本发明仅描述了优选的方法和材料，但是在本发明的实施或测试中也可以使用与本文所述相似或等同的任何方法和材料。本说明书中提到的所有文献通过引用并入，用以公开和描述与所述文献相关的方法和/或材料。在与任何并入的文献冲突时，以本说明书的内容为准。
在不背离本发明的范围或精神的情况下，可对本发明说明书的具体实施方式做多种改进和变化，这对本领域技术人员而言是显而易见的。由本发明的说明书得到的其他实施方式对技术人员而言是显而易见的。本申请说明书和实施例仅是示例性的。
关于本文中所使用的“包含”、“包括”、“具有”、“含有”等等，均为开放性的用语，即意指包含但不限于。
按照授权专利“CN113115777B伊枯草素家族脂肽在防治害虫中的应用”“CN113100237B丰原素家族脂肽在害虫防治中的应用”、“CN113142213B表面活性素家族脂肽在防治害虫中的应用”说明书中公开的方法，分别制备获取伊枯草家族脂肽、丰原素家族脂肽和表面活性素家族脂肽相关化合物。
实施例1脂肽组合物对半翅目害虫的杀虫效果
将伊枯草素、丰原素、表面活性素三种脂肽依次按照1:1:1的比例进行配制，构成的组合物用水溶解，配成2000μg/mL的母液，再用水将母液稀释成5μg/mL、10μg/mL、20μg/mL、200μg/mL、500μg/mL、1000μg/mL、2000μg/mL的药液，以清水处理做对照。豆蚜(Aphiscraccivora Koch)杀虫实验使用的蚕豆幼茎、棉蚜(Aphis gossypii)杀虫实验使用的棉花幼茎、甘蓝蚜(Brevicoryne brassicae)杀虫实验使用的甘蓝嫩叶、麦长管蚜(Macrosiphumavenae)杀虫实验使用的小麦幼茎，取上述各浓度药液20mL，将植物幼茎、叶均浸泡在上述药液10min后用滤纸吸干水分，取出放在垫有滤纸的培养皿后，加入2mL去离子水保湿，再将平皿用保鲜膜覆盖，扎孔，每12h加1mL水。每个浓度下供试蚜虫各50头，3次重复，用毛笔头挑每种各50头年龄大小相近的蚜虫放在准备好的培养皿上饥饿4h，分别加入上述药液处理过的蚕豆幼茎、棉花幼茎、甘蓝嫩叶、和小麦幼茎，每12h更换一次，于25℃培养箱培养过夜，记录蚜虫的死亡头数，计算不同稀释倍数下蚜虫的死亡率。
测定昆虫的死亡率：昆虫处理过夜后，用毛笔头触碰蚜虫看出，脂肽组合物对半翅目昆虫有良好的的防治作用。若昆虫不动则视为死亡，统计并记录死亡的昆虫头数结果如表1所示。
表1组合物对半翅目害虫的防治效果
实施例2脂肽组合物的鞘翅目害虫的杀虫效果
将伊枯草素、丰原素、表面活性素三种脂肽依次按照1:1:1的比例进行配制，构成的组合物用水溶解，配成2000μg/mL的母液，再用水将母液稀释成5μg/mL、10μg/mL、20μg/mL、200μg/mL、500μg/mL、1000μg/mL、2000μg/mL的药液，以清水作为对照，根据土壤的湿润度配置药土，一般按照药液与土壤质量重量比1:1配置药土。用碎花生作为食料，将容量4L的塑料桶作容器，每桶放置混合均匀药土2L，并加入适量等量的食料拌匀。采用药土法，选取虫体大小一致的白星花金龟、暗黑鳃金龟、铜绿丽金龟、小云斑鳃金龟三龄幼虫，在相对湿度为50％～70％、温度为25℃～28℃的环境中保持24小后。每处理试虫数量各10头，重复3次。接虫后置于室内常温培养，12h后统计死亡率，如表2所示。
表2组合物对鞘翅目害虫的防治效果
实施例3脂肽组合物对直翅目害虫的杀虫效果
将伊枯草素、丰原素、表面活性素三种脂肽依次按照1:1:1的比例进行配制，构成的组合物用水溶解，配成2000μg/mL的母液，再用水将母液稀释成5μg/mL、10μg/mL、20μg/mL、200μg/mL、500μg/mL、1000μg/mL、2000μg/mL的药液，以清水处理做对照。蝗虫杀虫实验使用的小麦幼苗、花生大蟋杀虫实验使用的花生幼茎、蝼蛄和棉斑草螽的杀虫实验使用的棉花幼茎浸泡在上述药液10min后用滤纸吸干水分，取出放在垫有滤纸的培养皿后，加入2mL去离子水保湿，再将培养箱用保鲜膜覆盖，扎孔，每12h加1mL水。每个浓度下供试昆虫各10头，3次重复，用镊子挑每种各10头年龄大小相近的昆虫放在准备好的培养箱中饥饿4h，分别加入上述药液处理过的小麦幼苗、花生幼茎、棉花幼茎，每12h更换一次，置于室内常温培养过夜，记录昆虫的死亡头数，计算不同稀释倍数下昆虫的死亡率。
测定昆虫的死亡率：昆虫处理过夜后，用镊子触碰昆虫，若昆虫不动则视为死亡，统计并记录死亡的昆虫头数由表3实施数据结果可以看出，脂肽组合物对直翅目昆虫有良好的的防治作用。
表3组合物对直翅目害虫的防治效果
实施例4脂肽组合物对蜚蠊目害虫的杀虫效果
将伊枯草素、丰原素、表面活性素三种脂肽依次按照1:1:1的比例进行配制，构成的组合物用水溶解，配成2000μg/mL的母液，再用水将母液稀释成5μg/mL、10μg/mL、20μg/mL、200μg/mL、500μg/mL、1000μg/mL、2000μg/mL的药液，以清水处理做对照。用镊子分别挑每种各10头年龄大小相近的美洲大蠊、黑胸大蠊、澳洲大蠊、日本大蠊放在准备好的培养箱中，设置三个重复，分别喷洒药液至昆虫体表，接虫后置于室内常温培养，间隔4小时观察昆虫，24小时后记录昆虫的死亡头数，计算不同稀释倍数下昆虫的死亡率。
测定昆虫的死亡率：昆虫处理过夜后，用镊子触碰昆虫，若昆虫不动则视为死亡，统计并记录死亡的昆虫头数由表4实施数据结果可以看出，脂肽组合物对直翅目昆虫有良好的的防治作用。
表4组合物对蜚蠊目害虫的防治效果
实施例5脂肽组合物对鳞翅目害虫的杀虫效果
将伊枯草素、丰原素、表面活性素三种脂肽依次按照1:1:1的比例进行配制，构成的组合物用水溶解，配成2000μg/mL的母液，再用水将母液稀释成5μg/mL、10μg/mL、20μg/mL、200μg/mL、500μg/mL、1000μg/mL、2000μg/mL的药液，以清水处理做对照。用镊子分别挑每种各10头年龄大小相近的甜纹夜蛾、斜纹夜蛾、小菜蛾、棉铃虫放在准备好的培养箱中，设置三个重复，分别喷洒药液至昆虫体表，接虫后置于室内常温培养，间隔4小时观察昆虫，24小时后记录昆虫的死亡头数，计算不同稀释倍数下昆虫的死亡率，实验数据如表5所示。
表5组合物对鳞翅目害虫的防治效果
实施例6-实施例111脂肽组合物复配农药对农业害虫的防治效果
将伊枯草素、丰原素、表面活性素三种脂肽构建的组合物用水溶解，配成50μg/mL的药液，将脂肽组合物与杀虫剂按照协同重量比的500:1复配如下不同农药丙氨酸甲酯、苯醚甲环性、速克灵、进口甲托、苯并咪唑44号、密霉胺、福美霜、进口蓝粉、氯虫苯甲酰胺、高效氯氟氰菊酯、乙基多杀菌素、灭幼脲氯氰菊酯、吡虫啉、溴氰菊酯、赤霉酸、吲哚乙酸、吲哚丁酸、烯腺嘌呤、羟烯腺嘌呤、苄氨基嘌呤、24-表芸苔素内酯、22,23,24-表芸苔素内酯、28-表高芸苔素内酯、28-高芸苔素内酯、14-羟基芸苔素甾醇、三十烷醇、S-诱抗素、抗坏血酸、糠氨基嘌呤、二氢卟吩铁、尿囊素、超敏蛋白、极细链格孢激活蛋白、氨基寡糖素、香菇多糖、几丁聚糖、葡聚烯糖、低聚糖素、β-羽扇豆球蛋白多肽、胆钙化醇、米尔贝霉素、括二化螟性诱剂、斜纹夜蛾诱集性信息素、绿盲蝽性信息素、梨小性迷向素、苹果蠹蛾性信息素；枯草芽孢杆菌、短稳杆菌、多粘类芽孢杆菌、苏云金芽孢杆菌、甲基营养型芽孢杆菌、海洋芽孢杆菌、坚强芽孢杆菌、球形芽孢杆菌、蜡质芽孢杆菌、荧光假单胞杆菌、侧孢短芽孢杆菌、短稳杆菌、地衣芽孢杆菌、解淀粉芽孢杆菌、沼泽红假单胞菌、嗜硫小红卵菌、白僵菌、球孢白僵菌、金龟子绿僵菌、哈茨木霉菌、木霉菌、淡紫拟青霉、厚孢轮枝菌、耳霉菌、盾壳霉、小盾壳霉、寡雄腐霉菌、蟑螂病毒、多角体病毒、颗粒体病毒、多杀霉素、阿维菌素、蝗虫微孢子虫；印楝素、苦参碱、藜芦碱、烟碱、鱼藤酮、除虫菊素、苦皮藤素、桉油精、八角茴香油、狼毒素、雷公藤甲素、莪术醇、蛇床子素、丁子香酚、大黄素甲醚、香芹酚、小檗碱、甾烯醇、茶皂素、螺威、大蒜素、d-柠檬烯、萜烯醇、异硫氰酸烯丙酯、十五烯苯酚酸、十三烷苯酚酸、苯丙烯菌酮，分别对应表6中的实施例6-实施例111，实验方法同实施例1和2，分别测定豆蚜和白星花金龟的死亡率。
将伊枯草素、丰原素、表面活性素三种脂肽构建的组合物用水溶解，配成1g/L的药液，将脂肽组合物与杀虫剂按照协同重量比500:1复配不同农药，分别对应表6中的实施例6-实施例111，实验方法同实施例3-5分别测定美洲大蠊、蝗虫和甜菜夜蛾的死亡率，实验数据如表6所示。
表6脂肽组合物复配农药对农业害虫的防治效果
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实施例112利用组合物制作种衣剂测定其杀虫率及花生完整率
将组合物用清水溶解，配成200μg/mL的母液，再用水将母液稀释成0.1μg/mL、0.5μg/mL、1μg/mL、5μg/mL、10μg/mL、20μg/mL、50μg/mL、100μg/mL的药液浸泡花生30min，以清水浸泡作为对照。将容量4L的塑料桶作容器，每桶放置混合均匀土壤2L，并加入50粒处理花生拌匀。选取虫体大小一致的暗黑鳃金龟(Holotrichia parallela)三龄幼虫蛴螬，处理重复3次，在相对湿度为50％～70％、温度为25℃～28℃的环境中保持24小后。每处理试虫数量各为15头，重复3次。接虫后置于室内常温培养，56h后统计死亡率及花生完整率，如表7所示。
表7组合物处理种子的杀虫及保护作用
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上述实施例1-实施例112的实验结果说明，本发明伊枯草素、丰原素、表面活性素三种脂肽组合构建的组合物，对半翅目、鞘翅目、直翅目、蜚蠊目、鳞翅目的害虫具具有较好的杀虫作用，并且将该组合物与不同的杀虫或者杀菌剂进行组合后，对豆蚜、白星花金龟、美洲大蠊、蝗虫和甜纹夜蛾表现出较好的杀虫效果，尤其是对豆蚜和白星花金龟的杀虫效果更为显著。所以，本发明公开的伊枯草素、丰原素、表面活性素三种脂肽组合构建的组合物单独或者与杀虫杀菌剂结合对于防治不同目害虫具有明显的作用，为害虫防治提供了新的农药。
以上所述的实施例仅是对本发明的优选方式进行描述，并非对本发明的范围进行限定，在不脱离本发明设计精神的前提下，本领域普通技术人员对本发明的技术方案做出的各种变形和改进，均应落入本发明权利要求书确定的保护范围内。

Claims (8)

1.一种组合物在害虫防治中的应用，其特征在于，所述组合物包括丰原素家族脂肽、伊枯草素家族脂肽和表面活性素家族脂肽，所述组合物中丰原素家族脂肽、伊枯草素家族脂肽和表面活性素家族脂肽的有效含量分别为0.01％-10％，所述害虫为直翅目害虫、蜚蠊目害虫和鳞翅目害虫。
2.根据权利要求1所述的组合物在害虫防治中的应用，其特征在于，所述组合物还包括杀虫剂或者杀菌剂。
3.根据权利要求2所述的组合物在害虫防治中的应用，其特征在于，所述杀虫剂包括化学杀虫剂、生物杀虫剂、植物杀虫剂。
4.根据权利要求3所述的组合物在害虫防治中的应用，其特征在于，所述化学杀虫剂包括丙氨酸甲酯、苯醚甲环性、速克灵、进口甲托、苯并咪唑44号、密霉胺、福美霜、进口蓝粉、氯虫苯甲酰胺、高效氯氟氰菊酯、乙基多杀菌素、灭幼脲氯氰菊酯、吡虫啉、溴氰菊酯、赤霉酸、吲哚乙酸、吲哚丁酸、烯腺嘌呤、羟烯腺嘌呤、苄氨基嘌呤、24-表芸苔素内酯、22,23,24-表芸苔素内酯、28-表高芸苔素内酯、28-高芸苔素内酯、14-羟基芸苔素甾醇、三十烷醇、S-诱抗素、抗坏血酸、糠氨基嘌呤、二氢卟吩铁、尿囊素、超敏蛋白、极细链格孢激活蛋白、氨基寡糖素、香菇多糖、几丁聚糖、葡聚烯糖、低聚糖素、β-羽扇豆球蛋白多肽、胆钙化醇、米尔贝霉素、二化螟性诱剂、斜纹夜蛾诱集性信息素、绿盲蝽性信息素、梨小性迷向素以及苹果蠹蛾性信息素中至少一种。
5.根据权利要求3所述的组合物在害虫防治中的应用，其特征在于，所述生物杀虫剂包括枯草芽孢杆菌、短稳杆菌、多粘类芽孢杆菌、苏云金芽孢杆菌、甲基营养型芽孢杆菌、海洋芽孢杆菌、坚强芽孢杆菌、球形芽孢杆菌、蜡质芽孢杆菌、荧光假单胞杆菌、侧孢短芽孢杆菌、短稳杆菌、地衣芽孢杆菌、解淀粉芽孢杆菌、沼泽红假单胞菌、嗜硫小红卵菌、白僵菌、球孢白僵菌、金龟子绿僵菌、哈茨木霉菌、木霉菌、淡紫拟青霉、厚孢轮枝菌、耳霉菌、盾壳霉、小盾壳霉、寡雄腐霉菌、蟑螂病毒、多角体病毒、颗粒体病毒、多杀霉素、阿维菌素以及蝗虫微孢子虫中至少一种。
6.根据权利要求3所述的组合物在害虫防治中的应用，其特征在于，所述植物杀虫剂包括印楝素、苦参碱、藜芦碱、烟碱、鱼藤酮、除虫菊素、苦皮藤素、桉油精、八角茴香油、狼毒素、雷公藤甲素、莪术醇、蛇床子素、丁子香酚、大黄素甲醚、香芹酚、小檗碱、甾烯醇、茶皂素、螺威、大蒜素、d-柠檬烯、萜烯醇、异硫氰酸烯丙酯、十五烯苯酚酸、十三烷苯酚酸以及苯丙烯菌酮中至少一种。
7.一种保护种子及后来形成的植物器官免受害虫侵害的方法，其特征在于，包括利用权利要求1-6任一项中所述组合物处理种子的过程。
8.根据权利要求7所述的方法，其特征在于，所述组合物的浓度为0.1μg/mL-100μg/mL。
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