CN115191449B - 一种吉龙草提取物在防治南方根结线虫中的应用 - Google Patents
一种吉龙草提取物在防治南方根结线虫中的应用 Download PDFInfo
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Abstract
本发明公开了一种吉龙草提取物在防治南方根结线虫中的应用。所述吉龙草提取物是用乙醇对吉龙草中有益物质进行提取浓缩得到的混合物。该提取物对南方根结线虫具有较高的杀灭效果，与润湿剂、分散剂、杀虫类农药混合施用可增强其作用效果，减缓南方根结线虫抗药性的产生。本发明创造性地将吉龙草提取物应用于南方根结线虫的杀灭，制备方法简单，原料成本低，制剂天然环保，抗虫效果显著，为吉龙草的高价值开发利用提供了方向，为南方根结线虫的防治提供了一种植物源环保型制剂，具有良好的开发应用前景与意义。
Description
技术领域
本发明属于农业病虫害绿色环保防治技术领域，具体涉及一种吉龙草提取物在防治南方根结线虫中的应用。
背景技术
线虫是一类低等的无脊椎动物，种类繁多，其中寄生在植物体内的称之为植物寄生线虫。植物寄生线虫是植物侵染性病害的重要病原物之一，在全世界普遍发生，为害严重。根结线虫是一类严重危害农业生产的植物寄生线虫，主要寄生在蔬菜、粮食作物、经济作物、果树、观赏植物及杂草等2000多种寄主上，热带、亚热带和温带的植物受害尤其严重，该病害发生后一般减产10％左右，严重的高达75％以上。尤其是南方根结线虫发病面积广，能侵染几乎所有栽培植物。南方根结线虫J2幼虫具有侵染植株的能力，它在土壤中感受到寄主分泌物的信号分子，并向寄主根尖部分趋向性迁移。迁移至根尖伸长区进行侵染。线虫钻入寄主根部后，利用口针将其食管腺细胞内的分泌物注射到寄主细胞内，诱导形成增大的、多核的、新陈代谢比较旺盛的并且为线虫持续提供营养物质的巨细胞。此外其细胞壁内突生长并形成典型的传递细胞，这些细胞可以增加对维管束内营养物质的吸收。根系受害后会形成大小不一性状不同的瘤状根结，开始为白色，质地柔软，后来加深成淡褐色，表面有裂纹。根系发病后其吸收和运输养分和水分的能力会下降，导致植株病弱产量降低。重病株地上部分表现更为明显，植株严重营养不良，瘦弱萎蔫，生长缓慢，叶片发黄变小，果实畸形变小或花果脱落，严重的会全株枯死。
目前，化学防治在根结线虫的防治过程中占有重要地位，在实际生产中能达到立竿见影的效果，是深受果农、菜农青睐的防治方法。但目前使用的化学杀线剂存在毒性大，污染环境，威胁人畜健康等弊端。随着公众环保意识的增强和线虫抗药性的增加，许多有效的杀线剂如神农丹、呋喃丹等已被禁用，人们更加迫切地需要安全环保的防治措施。生物防治是从生态学角度出发寻找的防治方法，以其环境相容性、绿色环保等优势成为无公害防治根结线虫的研究热点。采用生物防治的方法控制线虫，不仅能达到除害增产的效果，还可以减少化学防治带来的灾害，具有广阔的应用前景。
吉龙草，唇形科香薷属植物，也称暹罗香菜、吉笼草，一年生草本。主要生长于我国云南南部、西南部，多为人工种植，鲜见野生。《新华本草纲要》记载，吉龙草有清热、解毒、解表之功效，主要用于感冒，头痛，发热，消化不良等。由于该植物具有特殊香味，当地居民还将吉龙草当作蔬菜或佐料食用，是一种天然的植物香料。
目前对于吉龙草开发应用的相关报道较少，且多集中于提取精油的增香、抗菌方向。本发明首次公布了吉龙草提取物在杀灭南方根结线虫中的应用，为吉龙草的高价值开发利用提供了借鉴，为南方根结线虫的防治提供了一种天然植物源制剂。
发明内容
本发明的目的在于提供一种吉龙草提取物在防治南方根结线虫中的应用。其特征在于制备方法简单，原料成本低，制剂天然环保，抗虫效果显著，为吉龙草的高价值开发利用提供了方向，为南方根结线虫的防治提供了一种植物源环保型制剂。
为实现上述发明目的，本发明以阴干吉龙草为原料，经粉碎、过筛，乙醇浸提，减压浓缩等步骤，得到对南方根结线虫具有良好杀灭作用的植物提取物。具体技术方案步骤如下：
1)将阴干吉龙草进行粉碎、过筛，吉龙草粉末与70％～85％乙醇溶液以1:4～10(v/w)料液比进行混匀，置于40～50℃下，避光超声振荡40～90min，经0.22μm滤膜抽滤，滤渣重复浸提2次，合并3次抽滤滤液。
2)乙醇浸提滤液经旋转蒸发仪65℃、-0.1MPa下浓缩至原液体积的1/4，得吉龙草提取物。
取上述吉龙草提取物，以乙醇-吐温20溶液(9:1，v/v)为溶剂进行稀释，吉龙草提取物终浓度分别为62.5、125、250、500、1000μg/mL，取96孔板，向各孔中加入90μL南方根结线虫溶液(含线虫40～50条)，以16孔为一组分别加入5个浓度的吉龙草提取物溶液各10μL，以乙醇-吐温20溶液(9:1，v/v)为阴性对照，置于25℃培养箱中避光培养，24h、48h、72h时分别观察计数线虫死亡情况，计算各组致死率，公式如下：
结果表明上述吉龙草提取物在500μg/mL浓度下72h对南方根结线虫的致死率达到93.74％。
上述吉龙草提取物经GC-MS联用分析(色谱柱选用HP-5MS，以氦气为载气，流量1.0mL/min)，测定其主要组分为柠檬醛、香芹酮、β-蒎烯等。
本发明的有益效果是克服了现有技术的不足，提供一种具有杀灭南方根结线虫的以天然植物提取物及其应用。该提取物制备原料成本低，制备方法简单，所制备提取物安全环保，抗虫效果显著，其中的杀灭南方根结线虫活性成分的确定，可为揭秘防治南方根结线虫内在机理提供研究途径，为吉龙草的高价值开发利用提供了方向，为南方根结线虫的防治提供了一种植物源环保型制剂。
具体实施方式
以下结合实例对本发明的原理和特征进行描述，所举实例只用于解释本发明，并非用于限定本发明的范围。
实施例1
本发明提供一种吉龙草提取物，其制备方法如下：
1)将阴干吉龙草进行粉碎、过筛，吉龙草粉末与75％乙醇溶液以1:9(v/w)料液比进行混匀，置于45℃下，避光超声振荡90min，经0.22μm滤膜抽滤，滤渣重复浸提2次，合并3次抽滤滤液。
2)乙醇浸提滤液经旋转蒸发仪65℃、-0.1MPa下浓缩至原液体积的1/4，得吉龙草提取物。
实施例2
本发明提供一种吉龙草提取物，对其杀灭南方根结线虫进行浓度、时间梯度试验
取吉龙草提取物，以乙醇-吐温20溶液(9:1，v/v)为溶剂进行稀释，吉龙草提取物终浓度分别为62.5、125、250、500、1000μg/mL，取96孔板，向各孔中加入90μL南方根结线虫溶液(含线虫40～50条)，以16孔为一组分别加入5个浓度的吉龙草提取物溶液各10μL，以乙醇-吐温20溶液(9:1，v/v)为阴性对照，置于25℃培养箱中避光培养，24h、48h、72h时分别观察计数线虫死亡情况，计算各组致死率，公式如下：
结果表明上述吉龙草提取物在500μg/mL浓度下72h对南方根结线虫的致死率达到93.74％。
实施例3
本发明提供一种吉龙草提取物，对其杀灭不同地域、不同作物上南方根结线虫进行效果试验
2021年7月，将采集自潍坊章丘、烟台牟平高陵、烟台栖霞蛇窝泊、烟台海阳留格、烟台蓬莱村里集等地的患线虫病的西红柿、黄瓜、土豆、南瓜、茄子根系组织进行处理，通过清洗、浸泡、过筛、离心等步骤，分离得到5种作物上寄生的线虫虫体，经鉴定5种作物寄生线虫均为南方根结线虫。
取本发明吉龙草提取物，以乙醇-吐温20溶液(9:1，v/v)为溶剂进行稀释，吉龙草提取物终浓度范围在10～500μg/mL，取96孔板5个，分别向各板中加入5种作物分离到的线虫溶液90μL，再向各孔中按梯度加入各浓度吉龙草提取物稀释液，以乙醇-吐温20溶液(9:1，v/v)为阴性对照，置于25℃培养箱中避光培养，24h、48h时分别观察计数线虫死亡情况，计算各组致死率，公式如下：
将统计计算的到的24h及48h不同浓度下致死率数据进行统计分析(软件SPSS22.0)，结果显示，本发明吉龙草提取物对5种作物提取的南方根结线虫的半致死浓度(IC50)在24h时，范围在121～148μg/mL，在48h时，范围在66～101μg/mL，证明本发明吉龙草提取物对南方根结线虫具有较好的防治作用。
上述对所公开的实例的说明，仅为了使本领域专业技术人员能够实现或使用本发明。对这些实例的多种修改对本领域的专业技术人员来说将是显而易见的，且本文中所定义的一般原理可以在不脱离本发明的精神或范围的情况下，在其它实施例中实现。因此凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
Claims (4)
1.一种吉龙草提取物在防治南方根结线虫中的应用，其特征在于，所述吉龙草提取物制备工艺如下：吉龙草经阴干、粉碎得到吉龙草干粉，与质量分数70％～85％的乙醇溶液按料液比g:ml为1:4～10的比例充分混匀，在40～50℃下经超声仪超声提取40～90min，经0.22μm孔径的滤膜抽滤，抽滤后的滤渣重复施用乙醇溶液提取2～3次，将几次抽滤得到的所有滤液进行合并，合并后的滤液经旋转蒸发仪在65℃、-0.1MPa下浓缩至原液体积的1/4，得吉龙草提取物。
2.根据权利要求1所述一种吉龙草提取物在防治南方根结线虫中的应用，其特征在于，吉龙草提取物在与润湿剂、分散剂混配施用，其中润湿剂添加质量为吉龙草提取物质量的0.2‰～2‰，分散剂添加质量为吉龙草提取物质量的0.1‰～15‰。
3.根据权利要求1所述一种吉龙草提取物在防治南方根结线虫中的应用，其特征在于，应用范围为吉龙草提取物对黄瓜、西红柿、土豆、姜、葡萄、大豆、茄子、桃中南方根结线虫的防治。
4.根据权利要求1～3所述一种吉龙草提取物在防治南方根结线虫中的应用，其特征在于，施用方法如下：
1)制剂加水稀释为杀虫液，其中吉龙草提取物质量分数为1.25‰～3‰；
2)根据南方根结线虫虫害发生阶段选择不同施用量，杀虫液施用量范围是8～30kg；
3)根据不同防治需求选择施用方式为喷施或冲施；
4)根据施用土壤干湿情况追施0～2倍杀虫液质量的清水；
5)根据虫害发生程度及防治需求追施0～2遍杀虫液，相邻两遍喷施操作间隔2～5天进行。
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