CN114431256A - 一种含有除虫菊素与球孢白僵菌的复配组合物及其应用 - Google Patents
一种含有除虫菊素与球孢白僵菌的复配组合物及其应用 Download PDFInfo
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- CN114431256A CN114431256A CN202111551377.2A CN202111551377A CN114431256A CN 114431256 A CN114431256 A CN 114431256A CN 202111551377 A CN202111551377 A CN 202111551377A CN 114431256 A CN114431256 A CN 114431256A
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Abstract
本发明涉及一种含有除虫菊素与球孢白僵菌的复配组合物及其应用,属于农药技术领域。该复配组合物的有效成分包括除虫菊素和球孢白僵菌,所述除虫菊素与所述球孢白僵菌复配的质量比为(0.1‑1.0)∶(0.1~4.9)。通过活性成分除虫菊素与球孢白僵菌在合理的复配比例范围内,对蚜虫、烟粉虱或黄曲条跳甲的防治具备协同增效作用。
Description
技术领域
本发明涉及一种含有除虫菊素与球孢白僵菌的复配组合物及其应用,属于生物农药技术领域。
背景技术
杀虫剂在害虫防治中发挥重要作用,随着环保和健康意识的强化,低毒、天然的杀虫剂正逐渐替代化学杀虫剂,植物源和微生物杀虫剂深受市场欢迎。除虫菊素是天然的植物源杀虫剂,是除虫菊花Pyrethrum cinerariifolium Trev.的提取物,含有除虫菊素I与II(pyrethrin I,II)、瓜叶菊素I与II(cinerin I,II)、茉酮菊素I与II(jasmolin I,II)等杀虫活性成份。天然除虫菊素一般加工成水乳剂用于防治农业害虫,具有高效、快速而广谱的杀虫特性,并且环境友好。然而,天然除虫菊素价格昂贵,且对害虫的持久有效性不足,限制了其在农业领域的应用。
球孢白僵菌Beauveria bassiana是一种重要的昆虫病原真菌,有广谱杀虫性,能寄生于1000余种昆虫上。在分类学上,球孢白僵菌属于真菌界Fungi,子囊菌门Ascomycota,粪壳菌纲Sordariomycetes,肉座菌目Hypocreales,虫草菌科Cordycipitaceae,白僵菌属Beauveria。它的分生孢子常被加工成粉剂、可湿性粉剂、颗粒剂、油悬浮剂及可分散油悬浮剂,用于防治农林业害虫,如松毛虫、玉米螟等。白僵菌杀虫剂具有天然、无毒、无抗药性、易于生产、价格较便宜,以及环境友好的优点,但速效性较差、药效不稳定等缺点制约了其大规模应用。
如能发挥除虫菊素与球孢白僵菌两者的优势,弥补各自的不足,研发出具有快速、持久杀虫效果,且价格较便宜的复配剂,必将拥有广阔的市场前景,产生显著的经济效益。
关于除虫菊素混配增效研究不多,有研究论文报告了除虫菊素与植物源杀虫剂(如苦参碱、苦皮藤素)混配对蚜虫或叶螨有增效作用(林桂雁等,除虫菊素+苦参碱对苹果红蜘蛛防效试验,烟台果树,2005,(04):27-28;刘雨晴等,天然苦皮藤素和天然除虫菊素混配对三种蚜虫的毒力及田间防效,植物保护,2014,40(02):175-178;胡贵磊等,除虫菊素和苦参碱复配对麦二叉蚜的联合生物活性及田间药效,农药,2021,60(11):836-839);除虫菊素与农用抗生素阿维菌素或多杀菌素联合,则对蚜虫有增效作用(苏栩等,阿维菌素与4种生物杀虫剂复配对麦二叉蚜的联合毒力,麦类作物学报,2020,40(05):630-637;胡贵磊等,除虫菊素与多杀菌素复配对麦二叉蚜的联合毒力及田间药效,河南科技学院学报(自然科学版),2021,49(01):41-45)。
有一些中国发明专利(如申请号:CN202110087491.8,CN202110485980.9,CN201910628344.X,CN201910075192.5,CN201910628344.X,CN201810221506.3,CN201811476401.9,CN201710832259.6,CN201510631211.X,CN201310113097.2等)公开了含有除虫菊素的杀虫杀菌组合物制备技术,包括将除虫菊素与植物源物质(如苦皮藤素、异硫氰酸烯丙酯、印楝素、藜芦碱、蛇床子素、小檗碱、桉叶油、百里香酚、薄荷、茶树油、丁子香酚、茵陈油、桉叶油、迷迭香精油、薰衣草、沉香粉、大蒜、雷公藤、山辣椒、半夏等)或其他天然物质(如壳寡糖、氨基寡糖素、赤霉素、木质素磺酸钠、阿维菌素等)及化学农药(如噻虫胺)混配的技术。而另一些专利(如CN202110714348.7,CN202010832727.1,CN201911148141.7,CN201811251887.6等)则公开含有除虫菊有效成份的生物有机肥料及其制备方法。
然而,关于除虫菊素与昆虫病原真菌混配的研究则罕有报导。王定锋等研究了除虫菊素与球孢白菌的生物相容性,发现除虫菊素不影响球孢白僵菌XJBb3005菌株的菌丝生长,且促进产孢,但对孢子萌发却有抑制作用(王定锋等,5种植物源农药与球孢白僵菌相容性研究,茶叶学报,2015,56(04):244-248)。中国发明专利(除虫菊酯与生物真菌互配灭蚊剂及其制造方法,申请号:CN201811284644.2)公开了一种除虫菊酯与贵阳腐霉真菌混配的杀蚊组合物。
可见,经过综合目前研究进展,天然除虫菊素与球孢白僵菌混配制剂研发尚属空白。本发明旨在研发一种天然除虫菊素与球孢白僵菌混配杀虫剂,以促进生物农药发展,为提高环境质量和食物安全做出贡献。
发明内容
本发明的目的在于克服现有技术的不足,提供一种活性成分包括除虫菊素与球孢白僵菌的复配组合物。
本发明的另一目的是提供所述含有除虫菊素与球孢白僵菌的复配组合物在防治农业害虫中的应用。
为实现上述目的,本发明采取的技术方案为:一种含有除虫菊素与球孢白僵菌的复配组合物,所述复配组合物的有效成分包括除虫菊素和球孢白僵菌。所述复配组合物中的球孢白僵菌为高纯度的球孢白僵菌分生孢子粉(高孢粉)。
作为本发明所述含有除虫菊素与球孢白僵菌的复配组合物的优选方案,所述除虫菊素与所述球孢白僵菌高孢粉复配的质量比为(0.1-1.0)∶(0.1~4.9);即,所述除虫菊素取0.1-1.0克与所述球孢白僵菌分生孢子1.77-86.73亿个/克复配。
作为本发明所述含有除虫菊素与球孢白僵菌的复配组合物的优选方案,所述除虫菊素与所述球孢白僵菌复配的质量比为(0.2-0.8)∶(0.3-1.8);即,所述除虫菊素取02-0.8克与所述球孢白僵菌分生孢子5.31-31.86亿个/克复配。
作为本发明所述含有除虫菊素与球孢白僵菌的复配组合物的优选方案,所述除虫菊素是指天然除虫菊花的提取物,有效成份为除虫菊素I、除虫菊素II、瓜叶菊素I、瓜叶菊素II、茉酮菊素I和茉酮菊素II的混合物。
作为本发明所述含有除虫菊素与球孢白僵菌的复配组合物的优选方案,所述复配组合物的剂型为可分散油悬浮剂、油悬浮剂或超低容量剂。
另外,本发明还提供本发明所述含有除虫菊素与球孢白僵菌的复配组合物在防治农业害虫中的应用。
作为本发明所述应用的优选方案,所述农业害虫包括蚜虫、烟粉虱或黄曲条跳甲。
与现有技术相比,本发明的有益效果为:
(1)本发明提供含有除虫菊素与球孢白僵菌的复配组合物,为一种对环境友好的农药组合物,其活性成分除虫菊素与球孢白僵菌在合理的复配比例范围内,对蚜虫、烟粉虱或黄曲条跳甲的防治具备协同增效作用;
(2)通过活性成分除虫菊素与球孢白僵菌的复配,有效的解决了单一成分的天然除虫菊素价格昂贵、且对害虫的效性不持久以及球孢白僵菌速效性较差、药效不稳定等缺点;本发明提供含有除虫菊素与球孢白僵菌的复配组合物可实现大规模工业化生产。
附图说明
图1为1.5%天然除虫菊素水乳剂和1.5%除虫菊素·球孢白僵菌可分散油悬浮剂对瓜蚜的处理结果图;其中图1A为空白对照;图1B为1.5%天然除虫菊素水乳剂对瓜蚜的处理;图1C为1.5%除虫菊素·球孢白僵菌可分散油悬浮剂对瓜蚜的处理。
图2为1.5%除虫菊素·球孢白僵菌可分散油悬浮剂和0.2%除虫菊素·球孢白僵菌超低容量油悬浮剂对黄曲条跳甲的处理结果图;其中,图2A为1.5%除虫菊素·球孢白僵菌可分散油悬浮剂对黄曲条跳甲的处理;图2B为0.2%除虫菊素·球孢白僵菌超低容量油悬浮剂对黄曲条跳甲的处理;图2C为空白对。
图3为1.5%天然除虫菊素水乳剂和1.5%除虫菊素·球孢白僵菌可分散油悬浮剂对烟粉虱的处理结果图;图3A为空白对照;图3B为1.5%天然除虫菊素水乳剂对烟粉虱的处理;图3C为1.5%除虫菊素·球孢白僵菌可分散油悬浮剂对烟粉虱的处理。
具体实施方式
为更好地说明本发明的目的、技术方案和优点,下面将结合具体附图和实施例对本发明作进一步说明。
实施例1:球孢白僵菌高孢粉制备与质量检测
取球孢白僵菌BbPs01菌株(广东省微生物菌种保藏中心保藏号GDMCC NO:61494),利用大米为基质进行发酵,收获分生孢子后,通过冷冻干燥,制得高纯度分生孢子粉(高孢粉)。采用国家标准《GB/T 25864—2010,球孢白僵菌粉剂》标准方法,测得纯孢粉的含孢量为(1.77±0.31)×1011/g,分生孢子活力(萌发率)为(93.75±1.92)%,干燥减量为(2.53±0.43)%,杂菌率1.05%。
实施例2:天然除虫菊素对球孢白僵菌BbPs01菌株的生物相容性
用天然除虫菊素原油(云南南宝生物科技有限责任公司生产,总有效成份含量为71.78%)配置含除虫菊素总有效成分不同浓度梯度的植物调和油备用,浓度分别为1000mg/L、2000mg/L、5000mg/L、10000mg/L和15000mg/L,再称取30mg纯孢粉于20mL含除虫菊素的植物调和油中,以不含除虫菊素的植物调和油作对照,常温下放置,封口膜密封,每个处理重复3次。放置1d、15d和30d后测定孢子萌发率,结果如表1所示。
由表1的结果表明,天然除虫菊素原油在不大于10000mg/L剂量时,不会显著降低白僵菌的孢子萌发率,就算在高达15000mg/L剂量处理下,也只比对照降低3%左右的孢子萌发率。由此说明,天然除虫菊素原油对球孢白僵菌是安全的。
表1 天然除虫菊素原油对球孢白僵菌BbPs01菌株孢子萌发的影响
注:表中数据为平均数±标准差,同列数据后不相同字母表示在P<0.05水平差异显著。
实施例3:天然除虫菊素与球孢白僵菌BbPs01菌株的联合毒力
供试害虫为瓜蚜(棉蚜,Aphisgossypii Glover),采集自华南农业大学实验农场,饲养于在健康的黄瓜上,稳定繁殖多代后用于实验,保持适宜的光照、湿度和温度,选择生长期一致的成虫供生物活性测定。参照农业行业标准《NY/T1154.9-2008农药室内生物测定试验准则杀虫剂第9部分:喷雾法》,以及《NY/T 1154.7-2006农药室内生物测定试验准则杀虫剂第7部分:混配的联合作用测定》执行。采用离体叶片喷雾法处理,剪取大小相似且健康的黄瓜叶片放入培养皿中,每皿1片叶,皿底铺滤纸保湿,挑取大小一致的蚜虫成虫20只于叶片上,按每片叶子3mL药液用量,使用Potter喷雾塔(英国BURKARD)处理,对照叶片喷0.02%Tween-80溶液,置于光照培养箱(珠江LRH-400A-GSI-L3)中培养,培养设定条件为光照14:10(L:D)、温度25±1℃,相对湿度70±10%。每隔24h观察蚜虫死亡情况,以蚜虫四肢不动确定为死亡。统计死亡虫数,每处理重复4次。根据对数几率法求毒力方程和半致死剂量。参照Mansour等(1966)方法,用协同毒力指数(CF)评价混合剂的联合作用,其中CF≥20表示增效,CF≤-20表示拮抗,20≥CF≥-20表示相加作用。CF值根据下式计算,式中死亡率均为校正值,MM表示混合剂的病死率,PM表示除虫菊素处理的病死率,BM表示白僵菌处理的死亡率,结果如表2所示,CF的计算公式如下:
由表2的结果表明,当除虫菊素0.1-0.9mg/L与球孢白僵菌高孢粉(球孢白僵菌高纯度分生孢子粉)0.1-4.9mg/L(或0.18-3.19×108活孢子/mL)混配时,对瓜蚜联合毒力表现各异,即低剂量处理(总有效成份为1mg/L)表现出增效作用(处理号M0208、M0307、M0406、M0505、M0604、M0703),个别1mg/L剂量处理(M0218)也表现出增效作用;较高剂量处理(总有效成份为2-5mg/L)则表现出相加作用(处理号M0416、M0614、M0812、M0901、M0149、M0337、M0535、M0723、M0921),只有个别组合在处理后48h表现出拮抗作用(处理M0614、M0812和M0921)。经毒力回归,估计了供试药剂对瓜蚜的LC50,在处理后72h时,天然除虫菊素原油为2.48mg/L(95%置信区间1.96-2.89),而球孢白僵菌的LC50为0.35(0.19-0.53)mg/L,相当于0.62×108(0.33×108-0.94×108)孢子/mL。
由此可知,除虫菊素与球孢白僵菌分生孢子的复配组合物对害虫瓜蚜的防治具有协同增效作用,天然除虫菊素的质量分数为0.1%-1.0%,球孢白僵菌高孢粉0.1-4.9%(或天然除虫菊素有效成份0.1-1.0克,球孢白僵菌活孢子1.77-86.73亿个/克)。优选地,除虫菊素0.2%-0.8%,球孢白僵菌高孢粉0.3-1.8%(或天然除虫菊素有效成份0.2-0.8克,球孢白僵菌活孢子5.31-31.86亿个/克)。
表2 白僵菌孢子与除虫菊素原油混配比例对瓜蚜活性的影响
实施例4:1.5%除虫菊素·球孢白僵菌可分散油悬浮剂的配制
按表3用量称取各组分,先将天然除虫菊素原油和增效胺溶入少量油基中,再缓慢加入球孢白僵菌高孢粉,振荡混匀,补充油基,再加入各种增稠剂,混匀,再加入各种其他成份并补足油基,在匀浆机下8000rpm搅拌30min,充分混合均匀。分装,标签,低温保存或背光干燥保存,备用。
表3 1.5%除虫菊素·球孢白僵菌可分散油悬浮剂各组分用量
实施例5:0.15%除虫菊素·球孢白僵菌超低容量油悬浮剂的配制
按表4用量称取各组分,先将天然除虫菊素原油和增效胺溶入少量油基中,再缓慢加入球孢白僵菌高孢粉,振荡混匀,补充油基,再加入各种增稠剂,混匀,再加入各种其他成份并补足油基,在匀浆机下8000rpm搅拌30min,充分混合均匀。分装,标签,低温保存或背光干燥保存,备用。
表4 0.2%除虫菊素·球孢白僵菌超低容量油悬浮剂各组分用量
实施例6:含有除虫菊素与球孢白僵菌的复配组合物的制剂对瓜蚜的活性
先用0.02%的吐温80溶液,稀释供试药剂到设计浓度,然后参照前述方法,测定是实施例4和实施例5的制剂对瓜蚜的生物活性,试验结果如表5、表6和图1所示。
试验结果表明,1.5%和0.2%除虫菊素·球孢白僵菌制剂对瓜蚜的活性较高,与单一成分的1.5%除虫菊素水乳剂活性相当(表5)。
表6毒力回归分析表明,1.5%除虫菊素·球孢白僵菌可分散油悬浮剂和0.15%除虫菊素·球孢白僵菌超低容量油悬浮剂在48小时与72小时的LC50值介于4-16mg/L之间,低于单一的1.5%除虫菊素水乳剂;可见,1.5%除虫菊素·球孢白僵菌可分散油悬浮剂和0.2%除虫菊素·球孢白僵菌超低容量油悬浮剂对瓜蚜的活性高于1.5%天然除虫菊素水乳剂。
由图1可知,图1A为空白对照;图1B为1.5%天然除虫菊素水乳剂处理;图1C为1.5%除虫菊素·球孢白僵菌可分散油悬浮剂处理;结果显示,用1.5%除虫菊水乳剂处理后的瓜蚜体色由绿色转为棕红色,1.5%除虫菊素·球孢白僵菌可分散油悬浮剂处理后的瓜蚜体色不仅变为棕红,体表并长出菌丝(图1)。
表5 不同药剂对瓜蚜的致死率
注:表中数据为平均数±标准差,同列数据后不相同字母表示在P<0.05水平差异显著。
表6 不同药剂对瓜蚜的毒力
实施例7:含有除虫菊素与球孢白僵菌的复配组合物的制剂对黄曲条跳甲的活性
黄曲条跳甲Phyllotreta striolata在养虫笼内用新鲜菜心饲养,保持适宜的光照、湿度和温度,3~5d后选择生长期一致的成虫供生物活性测定。将大小基本一致,健康状态的黄曲条跳甲用CO2麻醉,然后转移到离心管中,加入药液(实施例4、实施例5的制剂和1.5%天然除虫菊素水乳剂),迅速盖好并上下颠倒50次,转移到铺有滤纸的一次性塑料碗中,置于(26±1)℃,光周期14:10(L:D),相对湿度(95±5)%的人工气候箱中饲养。浸泡处理后的黄曲条跳甲用新鲜萝卜饲养,每两天更换一次,同时记录试虫的死亡数,并将死虫转移至铺有滤纸的培养皿中,根据虫体特征确定试虫是否为真菌侵染致死。用0.02%吐温-80溶液处理作为对照,每处理10头虫,重复三次,试验结果如表7、表8和图2所示。
试验结果表明,三种供试制剂(实施例4、实施例5的制剂和1.5%天然除虫菊素水乳剂)对黄曲条跳甲的活性如表7所示,与表6的数据相比,三种试剂对黄曲条跳甲的活性低于对瓜蚜的活性。
三种供试制剂(实施例4、实施例5的制剂和1.5%天然除虫菊素水乳剂)处理黄曲条跳甲6d后,1.5%除虫菊素·球孢白僵菌可分散油悬浮剂的LC50值13.4mg/L,0.2%除虫菊素·球孢白僵菌超低容量油悬浮剂的LC50值18.4mg/L,均低于1.5%天然除虫菊素水乳剂的26.7mg/L,可见,1.5%除虫菊素·球孢白僵菌可分散油悬浮剂和0.2%除虫菊素·球孢白僵菌超低容量油悬浮剂对黄曲条跳甲的活性高于1.5%天然除虫菊素水乳剂(表8)。
由图2可知,图2A为1.5%除虫菊素·球孢白僵菌可分散油悬浮剂处理;图2B为0.2%除虫菊素·球孢白僵菌超低容量油悬浮剂处理;图2C为对照;制剂处理后的跳甲体表渐生菌丝,最终被菌丝包被(图2)。
表7 不同药剂对黄曲条跳甲的活性
注:表中数据为平均数±标准差,同列数据后不相同字母表示在P<0.05水平差异显著。
表8 不同药剂对黄曲条跳甲的毒力
实施例8:含有除虫菊素与球孢白僵菌的复配组合物的制剂对烟粉虱的活性
用吸虫器从实验室饲料传代的B型烟粉虱(Bemisia tabacii B-biotype)种群捕捉成虫,释放到内置盆栽健壮无病虫害的黄瓜苗的养虫笼中,按每片叶子50对释放虫量。在光照14:10(L:D)、温度25±1℃、相对湿度70±10%条件下,让烟粉虱成虫在笼中产卵1d,而后驱走全部烟粉虱成虫。继续培养,观察烟粉虱卵孵化情况,待发育到二龄若虫期时(约7天)进行试验,试验结果如表9、表10和图3所示。
用剪刀将被二龄烟粉虱幼虫侵染的黄瓜叶片剪下,保留其叶柄长约1cm,参照前述Potter喷雾法处理叶片,取出后用含有植物营养液的棉花包被叶柄,放入培养皿中,每皿1片叶,皿底铺滤纸保湿,对照叶片处理0.02%Tween-80溶液,置于光照培养箱中培养,培养设定条件为光照14:10(L:D)、温度25±1℃,相对湿度70±10%。从第2d开始,每48h在解剖镜下检查若虫死亡情况,虫体干瘪枯黄确定为死亡。统计死亡虫数,每处理重复3次,测定药剂对二龄烟粉虱的毒力。
试验结果表明,三种供试制剂(实施例4、实施例5的制剂和1.5%天然除虫菊素水乳剂)对烟粉虱的活性如表9所示,与表6的数据相比,三种试剂对烟粉虱的活性稍低于对瓜蚜的活性。
1.5%除虫菊素·球孢白僵菌可分散油悬浮剂和0.2%除虫菊素·球孢白僵菌超低容量油悬浮剂的LC50值(处理后6天)分别为17.7mg/L和16.0mg/L,低于1.5%天然除虫菊素水乳剂的24.8mg/L;可见,1.5%除虫菊素·球孢白僵菌可分散油悬浮剂和0.2%除虫菊素·球孢白僵菌超低容量油悬浮剂对烟粉虱的活性高于1.5%天然除虫菊素水乳剂(表10)。
由图3可知,图3A为空白对照;图3B为1.5%天然除虫菊素水乳剂对瓜蚜;图3C为1.5%除虫菊素·球孢白僵菌可分散油悬浮剂对烟粉虱的处理,菊素白僵制剂处理后的烟粉虱体表最终被菌丝包被(图3)。
表9 不同药剂对烟粉虱的致死率
注:表中数据为平均数±标准差,同列数据后不相同字母表示在P<0.05水平差异显著。
表10 不同药剂对烟粉虱的毒力
综上,实验结果表明,本发明提供含有除虫菊素与球孢白僵菌的复配组合物,其活性成分除虫菊素与球孢白僵菌在合理的复配比例范围内,对蚜虫、烟粉虱或黄曲条跳甲的防治具备协同增效作用。
最后所应当说明的是,以上实施例仅用以说明本发明的技术方案而非对本发明保护范围的限制,尽管参照较佳实施例对本发明作了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的实质和范围。
Claims (7)
1.一种含有除虫菊素与球孢白僵菌的复配组合物,其特征在于,所述复配组合物的有效成分包括除虫菊素和球孢白僵菌。
2.如权利要求1所述的复配组合物,其特征在于,所述除虫菊素与所述球孢白僵菌复配的质量比为(0.1-1.0)∶(0.1~4.9)。
3.如权利要求2所述的复配组合物,其特征在于,所述除虫菊素与所述球孢白僵菌复配的质量比为(0.2-0.8)∶(0.3-1.8)。
4.如权利要求1-3任一所述的复配组合物,其特征在于,所述除虫菊素是指天然除虫菊花的提取物,有效成份为除虫菊素I、除虫菊素II、瓜叶菊素I、瓜叶菊素II、茉酮菊素I和茉酮菊素II的混合物。
5.如权利要求1-3任一所述的复配组合物,其特征在于,所述复配组合物的剂型为可分散油悬浮剂、油悬浮剂或超低容量剂。
6.如权利要求1-5任一所述的复配组合物在防治农业害虫中的应用。
7.如权利要求6所述的应用,其特征在于,所述农业害虫包括蚜虫、烟粉虱或黄曲条跳甲。
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CN115316404B (zh) * | 2022-09-06 | 2023-09-29 | 浙江宜葆生物科技有限公司 | 一种含有除虫菊素与球孢白僵菌的可分散油悬浮剂及其应用 |
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