CN112825867B - 垫状棱子芹在杀螨杀虫方面的用途及其生物防治药剂和制备方法 - Google Patents
垫状棱子芹在杀螨杀虫方面的用途及其生物防治药剂和制备方法 Download PDFInfo
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- CN112825867B CN112825867B CN202110005975.3A CN202110005975A CN112825867B CN 112825867 B CN112825867 B CN 112825867B CN 202110005975 A CN202110005975 A CN 202110005975A CN 112825867 B CN112825867 B CN 112825867B
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- A01N49/00—Biocides, pest repellants or attractants, or plant growth regulators, containing compounds containing the group, wherein m+n>=1, both X together may also mean —Y— or a direct carbon-to-carbon bond, and the carbon atoms marked with an asterisk are not part of any ring system other than that which may be formed by the atoms X, the carbon atoms in square brackets being part of any acyclic or cyclic structure, or the group, wherein A means a carbon atom or Y, n>=0, and not more than one of these carbon atoms being a member of the same ring system, e.g. juvenile insect hormones or mimics thereof
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Abstract
本发明“垫状棱子芹在杀螨杀虫方面的用途及其生物防治药剂和制备方法”属于生物防治领域。本发明涉及垫状棱子芹在杀螨杀虫方面的用途,以及基于垫状棱子芹精油的生物防治药剂。本发明的生物防治药剂可高效杀灭害螨及害虫,同时无毒无害、安全可靠、成本低廉。
Description
技术领域
本发明属于生物防治领域,具体涉及垫状棱子芹在杀螨杀虫方面的用途及其生物防治药剂和制备方法。
背景技术
二斑叶螨(Tetranychus urticae)是一种世界性害螨,属于蛛形纲。二斑叶螨寄主植物十分广泛,达50余科800多种,常见的有草莓、茄子、黄瓜、番茄、辣椒、大豆、豇豆、芸豆、高粱、西瓜、黄瓜、桃、梨、苹果、杏、李、樱桃、天竺葵、一品红和棉花等多种作物和近百种杂草。二斑叶螨成、若螨均可产生为害,发生初期害螨多聚集在叶背主脉两侧,受害叶片正面叶脉两侧表现失绿,随着螨量的增加和为害程度的加剧,叶片很快失绿,渐变为褐色,叶片逐渐变硬变脆,最后枯黄脱落。防治二斑叶螨的主要依靠施用化学合成的杀螨剂,由于长期使用,目前二斑叶螨对常用杀螨剂,如阿维菌素、哒螨灵、唑螨酯、噻螨特等多种药剂均产生较高抗性,推荐剂量基本无效。同时化学药剂的使用,对非靶标生物也具有杀伤作用,破坏生态平衡,引起害螨的再猖獗。迫切需要发现高效环境友好的杀螨剂。
西花蓟马(Frankliniella occidentalis)属缨翅目蓟马科昆虫,是一种世界性的危险害虫,其寄主范围广,食性杂,寄主植物包括马铃薯、茄子、辣椒、黄瓜、花卉、香蕉、芒果等500多种重要经济作物和粮食作物,随着西花蓟马的不断扩散蔓延,其寄主种类仍然在持续增加。西花蓟马以锉吸式口器取食植物的茎、叶、花、果,导致花瓣退色、叶片皱缩,茎和果则形成伤疤,最终可能使植株枯萎,同时还传播番茄斑萎病毒在内的多种植物病毒,对农业生产造成严重威胁。由于西花蓟马虫体小,危害活动隐秘,能进行孤雌生殖,繁殖能力强,种群增长迅速,化学防治仍然是主要的防治手段,但由于化学农药长期不合理的使用,造成了西花蓟马对有机磷、氨基甲酸酯、拟除虫菊酯类及多杀菌素等杀虫剂产生了不同程度的抗药性,这不仅增加了防治难度,同时也加大了对环境的压力。生产中迫切需要发现对西花蓟马有效的绿色无公害杀虫剂。
嗜卷书虱(Liposcelis bostrychophilus)是一种重要的储藏物害虫,属昆虫纲,啮虫目。嗜卷书虱可以生活在谷物间、植物间、树皮下、地衣及旧木上,为孤雌生殖,个体微小,发育历期短,成虫寿命长,食性杂,抗饥饿能力强,繁殖速度极快,大量发生时可引起严重的经济损失,对粮食、食品、饲料及药材等储藏物危害极大,且由于其身体上粘附的排泄物能携带传播病菌和引起过敏,加之活动性极强,因此会对人类的健康带来威胁。在实际生产中,仓储类害虫多使用化学熏蒸剂防治,然而长期单一使用磷化氢熏蒸剂导致嗜卷书虱对磷化氢的抗药性增强,且易造成害虫再猖獗,甚至会对人类健康造成危害,寻找新型安全环保的杀虫剂替代物是亟需解决的问题。
垫状棱子芹(Pleurospermum hedinii Diels)为伞形科棱子芹属植物,是我国的特有植物。多年生莲座状草本,高4-5厘米,直径10-15厘米。垫状棱子芹的根粗壮,圆锥状,直伸。茎粗短,肉质,直径1-1.5厘米,基部被栗褐色残鞘。叶近肉质,基生叶窄长椭圆形,长2-5厘米,二回羽裂;一回羽片5-7对,近无柄,小裂片倒卵形或匙形,长1-2.5毫米;叶柄长2-4厘米,扁平,基部宽达4毫米;叶柄长2-4厘米,扁平,基部宽达4毫米。顶生复伞形花序径5-10厘米,总苞片多数,叶状;伞辐多数,小总苞片8-12,倒卵形或倒披针形,长4-8毫米,不裂或羽裂。萼齿近三角形;花瓣淡红或白色,近圆形。果卵形或宽卵形,长4-5毫米,有密集小水泡状突起,果棱有宽翅,微波状褶皱,每棱槽1油管,合生面2油管。花期7-8月,果期8-9月。产地西藏东部,青海西部果洛、治多等县。生长于海拔5000米左右的山坡草地。
本领域尚未报道过垫状棱子芹在杀螨杀虫方面的应用。
发明内容
本发明基于本领域的上述空白,发现垫状棱子芹在杀灭二斑叶螨、农业害虫西花蓟马以及仓储害虫嗜卷书虱方面的作用,并提供其在生防领域杀螨杀虫方面的用途。
本发明的技术方案如下:
垫状棱子芹在杀螨杀虫方面的用途。
所述虫选自农业害虫或仓储害虫;所述螨为害螨;
优选地,所述害螨为二斑叶螨;
优选地,所述农业害虫为西花蓟马,所述仓储害虫为嗜卷书虱。
所述杀螨杀虫的方式选自触杀、或,熏杀。
采用垫状棱子芹精油杀螨杀虫;
优选地,所述垫状棱子芹精油提取自垫状棱子芹全草。所述全草指的是垫状棱子芹包括根部及地上部分的完整植株。
垫状棱子芹在制备生防药剂方面的用途。
一种生物防治药剂,其特征在于,其活性成分为垫状棱子芹精油。
所述生物防治药剂选自杀螨剂、和/或,杀虫剂;
优选地,所述垫状棱子芹精油的杀螨杀虫活性成分包括:α-蒎烯、β-蒎烯、β-月桂烯、芳樟醇、α-萜品醇、γ-榄香烯。
一种生物防治药剂的制备方法,其特征在于,将垫状棱子芹进行水煮或蒸馏、冷凝、萃取而得。
所述水煮指用水煮沸处理8-12h,优选10h;
优选地,所述冷凝指所述水煮或蒸馏后冷凝得到油水混合物;
优选地,所述蒸馏指水蒸气蒸馏法蒸馏8-12h,优选10h;
优选地,所述萃取指将油水混合物用正己烷萃取;
优选地,所述萃取的水浴温度为38-45℃,优选40℃;
优选地,萃取所得的混合物经干燥后蒸发去除正己烷;
优选地,所述干燥指采用无水硫酸钠干燥;
优选地,所述蒸发指采用旋转蒸发仪蒸发。
本发明发现垫状棱子芹的杀虫杀螨作用,并以此验证了其在生物防治领域的突出效果,因此本发明请求保护垫状棱子芹在杀螨杀虫中的用途、垫状棱子芹在制备杀螨产品中的用途、垫状棱子芹精油在防治农业害虫中的用途、垫状棱子芹精油在防治仓储害虫中的用途、垫状棱子芹精油制备以中药材垫状棱子芹为原料,垫状棱子芹精油采用水煮/蒸馏、冷凝、萃取、脱水、蒸发溶剂步骤制备而得,垫状棱子芹精油在杀螨杀虫药剂中的用途,特征在于所述药剂的药效成分为垫状棱子芹精油。同时本发明还提供一种基于垫状棱子芹精油的生物防治药剂,该药剂可高效杀灭害螨及害虫,同时无毒无害、又不产生污染,是安全可靠、成本低廉、环保高效的新型杀虫剂/杀螨剂。
具体实施方式
下面结合具体实施例对本发明的内容做进一步详细说明,但并不以此限制本发明的保护范围。
生物材料的来源
试虫来源:西花蓟马、二斑叶螨、嗜卷书虱系人工饲养。
垫状棱子芹全草,采自西藏日喀则市南木林县土布加乡。
本发明实验例所使用的试剂,未特别说明的试验试剂均为本领域常规试剂或采用本领域常规方法获得,可通过商业途径购买,均为分析纯试剂。如无特殊说明,实验例进行的实验操作均为本领域常规操作。
第1组实施例、垫状棱子芹的生物防治新用途
本组实施例提供垫状棱子芹在杀螨杀虫方面的用途。
在具体的实施例中,所述虫选自害虫,所述螨为害螨
在更具体的实施例中,所述害虫选自农业害虫或仓储害虫;所述害螨为二斑叶螨;
在优选的实施例中,所述农业害虫选自西花蓟马;
在优选的实施例中,所述仓储害虫选自嗜卷书虱。
在一些实施例中,所述防治指杀灭;
在另一些实施例中,所述对二斑叶螨和西花蓟马的作用方式为毒杀,对嗜卷书虱的作用方式为熏蒸。
在进一步的实施例中,采用垫状棱子芹精油杀螨杀虫;
优选地,所述垫状棱子芹精油以垫状棱子芹为原料,采用蒸馏法制备而得。
第2组实施例、垫状棱子芹的生物防治制药用途
本组实施例提供垫状棱子芹在制备生物防治药剂方面的用途。
在具体的实施例中,所述生物防治药剂包括但不限于:杀虫剂,杀螨剂。
更具体地,所述生物防治药剂可制成本领域的常规剂型,包括但不限于:粉剂、可湿性粉剂、滴剂、胶悬剂、乳油、喷射剂、气雾剂、毒饵、蚊香、粘捕剂、烟剂、杀虫涂料等等。第3组实施例、本发明的生物防治药剂
本组实施例提供一种生物防治药剂。本组所有的实施例都具备如下共同特征:所述生物防治药剂的活性成分为垫状棱子芹精油。
在具体的实施例中,所述生物防治药剂的剂型选自杀螨剂或杀虫剂;
优选地,所述垫状棱子芹精油的活性成分包括:α-蒎烯、β-蒎烯、β-月桂烯、芳樟醇、α-萜品醇和γ-榄香烯。
在进一步的实施例中,所述生物防治药剂还包括本领域常见的杀虫剂领域可接受的辅料,包括但不限于:乳化剂、有机溶剂、填料、湿润剂、稀释剂等。
第4组实施例、本发明生物防治药剂的制备方法
本组实施例提供一种生物防治药剂的制备方法。本组所有的实施例都具备如下共同特征:所述制备方法包括:将垫状棱子芹进行水煮或蒸馏、冷凝、萃取而得。
在具体的实施例中,所述蒸馏指水蒸气蒸馏法蒸馏8-12h,优选10h,充分提取植物材料中的可挥发成分。
所述水煮指用水煮沸处理8-12h,优选10h,这样能把精油成分尽可能完全地从植物中煮出来。
优选地,所述萃取指用正己烷萃取;
更具体地,使用2L的分液漏斗,正己烷:蒸出物=1:1,分层后保留已溶解精油的正己烷层。
优选地,所述萃取的水浴温度为38-45℃,优选40℃,正己烷在此温度下,能快速蒸发、冷凝回收。
优选地,萃取所得的混合物经干燥后蒸发去除正己烷;
优选地,所述干燥指采用无水硫酸钠干燥;
优选地,所述蒸发指采用旋转蒸发仪蒸发。
优选地,所述精油比重为0.835g/mL。
实验例1、垫状棱子芹精油的制备
3.5kg垫状棱子芹采自西藏日喀则市南木林县土布加乡,用水煮法或水蒸气蒸馏法处理8h-12h,得到的油水混合物冷凝回收后用正己烷萃取,加无水硫酸钠干燥后,使用旋转蒸发仪除去正己烷,所得成分为垫状棱子芹精油。其中,水浴温度为40℃。
垫状棱子芹精油得率0.10%(w/w),即100g垫状棱子芹可提取出0.1g垫状棱子芹精油。
实验例2、垫状棱子芹精油成分鉴定
采用GC-MS检测方法对垫状棱子芹精油成分进行鉴定:
检测仪器:气相色谱仪(Agilent6890N)和质谱检测仪(Agilent5973N)。。
检测条件:HP-5MS 30mm×0.25mm×0.25μm石英毛细管柱,初始柱温60℃,保持1min,以12℃/min的速率,程序升温至180℃保持0.5min,再以25℃/min的速率程序升温至280℃保持18min,汽化器温度280℃,载气为氦气,柱流量1.0mL/min,分流比1:10。
质谱电离方式EI,电子轰击能量70eV,扫描质量范围20-550m/z,扫描时间2s。
取垫状棱子芹精油1μL(100倍丙酮稀释液)进样,经气相色谱-质谱联用仪分析鉴定,通过HP-MSD化学工作站检索Nist05标准质谱图库,并结合有关文献,检测并鉴定到26个峰。
成分鉴定结果如下表1所示:
表1 GC-MS测定垫状棱子芹精油化学成分
上表1中鉴定出来的全部成分的总百分比未到100%,是因为垫状棱子芹精油中仍存在未知成分,它们可能是新的化合物,无法利用现有的GC-MS检测方法鉴定出来。
实验例3、本发明生物防治药剂的杀螨效果
二斑叶螨-毒杀:LD50=20.36mg/mL(19.03,21.78)
采用叶碟法对二斑叶螨进行本发明垫状棱子芹精油的杀灭效果验证,具体操作如下:12孔细胞培养板,在培养板内壁涂聚四氟乙烯,用电吹风吹干;水和琼脂以100:1.5的比例在微波炉中加热混匀,倒入12孔细胞培养板中(每孔约2-3ml),冷却至凝固;取新鲜未施用农药的芸豆叶片,用直径19mm的打孔器将叶片打孔,制成圆形叶碟;取一定量的垫状棱子芹精油加无水乙醇(量不超过药液体系的2%)溶解,加二甲基亚砜(不超过药液体系的1%)助溶,配制成母液,后用蒸馏水稀释至所需浓度并得到所述药液体系;将叶碟浸入药剂10s后取出,晾干,将沾过药液的芸豆叶片背面朝上放在倒有琼脂的培养板上,每孔接入10只二斑叶螨成螨;用宣纸将培养板有孔的一面粘贴牢固并放进培养箱中,温度26-28℃,相对湿度为65%-75%,光周期L:D=16:8;24h后在体视镜下观察记录死亡数量,用毛笔接触动者为活,不动者为死,计算LD50。
死亡率(%)=死亡数/试虫数*100
校正死亡率(%)=(处理组死亡率-对照组死亡率)/(1-对照组死亡率)*100
半致死剂量(LD50):指能引起50%供试个体死亡的所需剂量。利用Probit软件计算而得。
结果如下表2所示:
表2叶碟法测定垫状棱子芹精油杀螨活性
LD50=20.36mg/mL,95%的置信区间为(19.03,21.78),y=-8.45+6.46x。该半致死剂量的含义为:每毫升药液体系(包括蒸馏水、溶剂-酒精和助溶剂-二甲基亚砜)中含有20.36mg精油,以这样的浓度处理,半数供试个体死亡。
实验例4、本发明生物防治药剂的农业害虫杀虫效果
西花蓟马-毒杀:LD50=8.57mg/mL(7.84,9.38)
西花蓟马-毒杀:取一定量垫状棱子芹精油加无水乙醇(量不超过药液体系的1%)溶解,加吐温80(不超过药液体系的0.5%)助溶,配制成母液,后用蒸馏水稀释至所需浓度并得到所述药液体系备用;将1mL药液放入容积为22mL玻璃闪烁瓶中,均匀滚动数圈,形成药膜,将药液倒出,闪烁瓶晾干备用。取新鲜未施用农药的扁豆,将扁豆制成1cm见方长段浸入药剂10s后取出,晾干,放置于闪烁瓶中;每个处理设置5个重复,每个重复接入10头西花蓟马的二龄若虫,150目防虫网封瓶口;放置于室温下,24h后观察结果,以毛笔拨动虫体,虫体不动记为死亡。
死亡率(%)=死亡数/试虫数*100
校正死亡率(%)=(处理组死亡率-对照组死亡率)/(1-对照组死亡率)*100
半致死剂量(LD50):指能引起50%供试个体死亡的所需剂量。利用Probit软件计算而得。
结果如表3所示:
表3叶碟法测定垫状棱子芹精油杀虫活性
LD50=8.57mg/mL,95%的置信区间为(7.84,9.38),y=-4.47+4.79x。该半致死剂量的含义指:每毫升药液体系(包括蒸馏水、溶剂-酒精和助溶剂-吐温)中含有8.57mg精油,以这样的浓度处理,半数供试个体死亡。
实验例5、本发明生物防治药剂的仓储害虫杀虫效果
嗜卷书虱-熏蒸:LC50=0.80mg/L(0.73,0.87)
垫状棱子芹精油对嗜卷书虱的熏蒸活性测定:用毛笔挑取10头试虫放入直径为0.5cm,高1.6cm的小玻璃瓶中(易于观察试虫的死亡情况),瓶口内壁涂聚四氟乙烯以防止试虫逃逸,小玻璃瓶置于高12cm直径4cm的大磨口玻璃瓶(290mL),丙酮作为溶剂稀释药剂成等比系列,用移液枪吸取10微升的药剂稀释液(6个浓度)滴于滤纸条上(长7.5cm,宽1.5cm),将滤纸片用固体胶粘在磨口瓶盖内,迅速拧紧磨口瓶盖,并用封口膜缠绕一周密封。24小时后,观察和记录试虫的死亡情况。每个浓度重复5次,每个重复10头试虫,丙酮不加药剂作为空白对照。24小时后观察,将小玻璃瓶内的试虫倒在一张白纸上,用毛笔拨动虫体,虫体不动即为死亡。
死亡率(%)=100×死虫数/试虫数。
校正死亡率(%)=100×(处理组死亡率-对照组死亡率)/(1-对照组死亡率)。
半致死剂量(LC50)即引起50%嗜卷书虱死亡率的剂量,使用Probit软件包计算得到半致死剂量。
结果如表4所示:
表4密闭空间法测定垫状棱子芹精油熏蒸活性
LC50=0.80mg/L,95%的置信区间为(0.73,0.87),y=0.50+5.01x。该半致死剂量的含义为:每升密闭空间内含有0.80mg精油,以这样的浓度处理,半数供试个体死亡。
Claims (10)
1.垫状棱子芹在杀螨杀虫方面的用途,采用提取自垫状棱子芹全草的垫状棱子芹精油杀螨杀虫;所述螨为二斑叶螨;所述虫选自西花蓟马或嗜卷书虱;所述垫状棱子芹精油的杀螨杀虫活性成分包括:α-蒎烯、β-蒎烯、β-月桂烯、芳樟醇、α-萜品醇和γ-榄香烯。
2.根据权利要求1所述的垫状棱子芹在杀螨杀虫方面的用途,其特征在于,所述杀螨杀虫的方式选自触杀或熏杀。
3.垫状棱子芹在制备生防药剂方面的用途, 所述生防药剂选自杀螨剂或杀虫剂;所述螨为二斑叶螨;所述虫选自西花蓟马或嗜卷书虱;采用提取自垫状棱子芹全草的垫状棱子芹精油杀螨杀虫;所述垫状棱子芹精油的杀螨杀虫活性成分包括:α-蒎烯、β-蒎烯、β-月桂烯、芳樟醇、α-萜品醇和γ-榄香烯。
4.一种生物防治药剂,其特征在于,其活性成分为提取自垫状棱子芹全草的垫状棱子芹精油;所述生物防治药剂选自杀螨剂或杀虫剂;所述螨为二斑叶螨;所述虫选自西花蓟马或嗜卷书虱;所述垫状棱子芹精油的杀螨杀虫活性成分包括:α-蒎烯、β-蒎烯、β-月桂烯、芳樟醇、α-萜品醇和γ-榄香烯。
5.一种生物防治药剂的制备方法,其特征在于,将垫状棱子芹进行水煮或蒸馏、冷凝、萃取、溶剂蒸发而得;所述生物防治药剂选自杀螨剂或杀虫剂;所述螨为二斑叶螨;所述虫选自西花蓟马或嗜卷书虱;所述水煮指用水煮沸处理8-2h;所述蒸馏指水蒸气蒸馏法蒸馏8-12h;所述冷凝指所述水煮或蒸馏后冷凝得到油水混合物;所述萃取指将油水混合物用正己烷萃取。
6.根据权利要求5所述的一种生防药剂的制备方法,其特征在于,所述水煮指用水煮沸处理10h。
7.根据权利要求5所述的一种生防药剂的制备方法,其特征在于,所述蒸馏指水蒸气蒸馏法蒸馏10h。
8.根据权利要求5所述的一种生防药剂的制备方法,其特征在于,萃取所得的混合物经干燥后蒸发去除正己烷。
9.根据权利要求8所述的一种生防药剂的制备方法,其特征在于,所述干燥指采用无水硫酸钠干燥。
10.根据权利要求8所述的一种生防药剂的制备方法,其特征在于,所述蒸发指采用旋转蒸发仪蒸发。
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