CN114600916A - 水蓼精油在抑制Neopestalotiopsis rosae中的应用 - Google Patents
水蓼精油在抑制Neopestalotiopsis rosae中的应用 Download PDFInfo
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- A—HUMAN NECESSITIES
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Abstract
本发明涉及Neopestalotiopsis rosae防治技术领域，具体涉及水蓼精油在抑制Neopestalotiopsis rosae中的应用。本发明将水蓼提取物按照不同的体积比添加至PDA培养基中，研究其对Neopestalotiopsis rosae的抑菌效果。结果表明水蓼精油在浓度（0.25‑2）μl/ml时，对于Neopestalotiopsis rosae有较好的抑菌效果，在浓度2μl/ml时，对于Neopestalotiopsis rosae的抑制率达到60%以上。本发明的水蓼精油原料易得，且提取方法简单，对于Neopestalotiopsis rosae有较好的抑制效果，同时精油具有很强的挥发性，而且具有安全、低毒等特点，而其具有的挥发性也有利于大棚种植草莓的过程中对病虫害的防治。
Description
技术领域
本发明涉及Neopestalotiopsis rosae防治技术领域，特别涉及水蓼精油在抑制Neopestalotiopsis rosae中的应用。
背景技术
植物源药剂具有易分解、安全和抗性低等特点，且其分解产物对人、畜和生态系统不能造成危害，由于其来源较广泛，能从植物的花、果、叶、茎、根中能提取得到大量的植物源提取物。前期的研究发现植物源提取物对细菌、真菌和昆虫具有很强的抑制活性，其能通过对菌细胞膜结构的破坏，导致细胞的渗透作用失调，从而抑制病原菌的生长、繁育。
草莓(Fragaria ananassa Duch.)属蔷薇科草莓属植物具有产量高、经济效益好等特点，广受人民群众的喜爱。草莓病害对草莓种植的产销量和产品质量都有影响，对于草莓病害的防治一直是广大学者研究的课题。由Neopestalotiopsis rosae起所引起的草莓根腐病在我国台湾省和其它部分省份的一些草莓产区均有发生，Neopestalotiopsisrosae能引起草莓的根腐病、冠腐病和叶斑点，其症状为棕色坏死病变，叶上肩瓣黑色，匍匐茎上轻微凹陷的深棕色坏死，果实上凹陷的红褐色坏死。病变的冠组织呈大理石红褐色坏死，边缘为深褐色，冠严重腐烂的植物通常在叶子上出现红褐色变色。台湾省报道称该疾病在2019年至2020年期间变得更加普遍和严重，并在移植后导致高达30％的植物损失。同时，在其他国家均有报道Neopestalotiopsis rosae对草莓的危害。
在大量的防治技术中，生物防治危害小，且清洁无污染，具有广阔的市场前景。尽管目前对草莓各种病害的生物防治研究有很多报道，但由于生物防治还是缺乏经验和技术，在实际生产操作中很难推广应用，目前的草莓病害研究与防治仍然主要采用化学防治。因此，研究新的防治方法是目前草莓病害防治的热点。
国内外有关植物精油作为绿色环保抑菌剂应用于植物病害病原菌的防治研究，取得了较大的进展。植物精油具有很强的挥发性，而且具有安全、低毒等特点，同时，由于其具有的挥发性也有利于大棚种植草莓的过程中对病虫害的防治。
发明内容
针对现有技术中存在的不足，本发明的目的在于提供了一种水蓼精油在抑制Neopestalotiopsis rosae中的应用。
进一步的，所述水蓼精油在应用时，将水蓼精油直接与水混合后喷洒于植株即可，所述水蓼精油与水的体积比为(0.25-2)μL：1mL，更优选为2μL：1mL。
进一步的，所述水蓼精油的主要成分包括：α-蒎烯、β-榄香烯、反式石竹烯、Α-香柠檬烯、α-律草烯、丙位依兰油烯、α-姜黄烯、甲位依兰油烯、β-红没药烯、1,2,4a,5,6,8a-六氢-1-异丙基-4,7-二甲基萘、橙花叔醇、4-(1-甲基乙基)环己醇、3,4-二甲基-3-环己烯甲醛、T-杜松醇、α-毕橙茄醇、a-红没药醇以及反式甲位佛手甘油烯。
进一步的，所述水蓼精油是由原料水蓼经过水蒸气蒸馏方式提取得到。
更进一步的，所述水蓼精油的提取方法如下：将干燥的水蓼全株粉碎后与水混合(优选的，所述干燥的水蓼全株与水的质量比为1:10)，利用微波萃取仪进行循环蒸馏(优选在800W、100℃下蒸馏2h)，然后经过油水分离得到精油。
为了确认水蓼精油对Neopestalotiopsis rosae的防治效果，本发明将水蓼提取物按照不同的体积比添加至PDA培养基中，研究其对Neopestalotiopsis rosae的抑菌效果。结果表明水蓼精油在浓度(0.25-2)μl/ml时，对于Neopestalotiopsis rosae有较好的抑菌效果，在浓度2μl/ml时，对于Neopestalotiopsis rosae的抑制率达到60％以上。本发明的水蓼精油原料易得，且提取方法简单，对于Neopestalotiopsis rosae有较好的抑制效果，同时精油具有很强的挥发性，而且具有安全、低毒等特点，而其具有的挥发性也有利于大棚种植草莓的过程中对病虫害的防治。
附图说明
图1为实施例3水蓼提取物对草莓植株的安全性评价中，实验组和对照组草莓株的外观图。
具体实施方式
下面申请人将结合具体实施例对本发明水蓼精油的新用途做进一步的详细说明。
以下实施例中，原材料水蓼(Polygonum hydropiper L.)为2021年5-6月采集于湖北省大悟县宣化镇，采用微波萃取仪，提取水蓼精油用于实验。
Neopestalotiopsis rosae来源于湖北工程学院特色果蔬质量安全控制湖北省重点实验室。植物源农药主要是利用植物中的活性成分包括植物本身和按活性结构合成的化合物及衍生物，使其免受或减轻病害等有害生物危害的植物源制剂，其优点在于：专一性强，不伤害天敌，保护环境等。
本发明将水蓼精油分别按照0.25、0.5、1和2μl/1ml(均为精油和培养基的体积比)的配比添加到PDA培养基(培养基组成为：马铃薯(去皮)200g，葡萄糖20g，琼脂 15g，蒸馏水1000ml，自然pH)中，通过计算其抑菌率，确定水蓼精油对Neopestalotiopsis rosae病菌的作用效果。
实施例1：水蓼精油的提取及成分含量鉴定
利用微波萃取仪，在功率800W的条件下，取15g干燥的水蓼全株，粉碎，向其中加入10倍重量的水，然后在100℃条件下循环蒸馏2小时，循环蒸馏产生混合体蒸气，其蒸气导入冷凝器中得到水与精油的液体混合物，在冷凝管处收集浮于水面上的水蓼提取物，即水蓼精油。
本实施例所得水蓼精油的成分含量鉴定：
通过气相色谱－质谱联用仪(7890B-5977A，安捷伦科技有限公司，美国)(GC-MS)对提取出的水蓼精油组成成分进行分析，检测条件：色谱柱为HP-5S毛细管柱 (30m×0.25mm×0.25μm)，载气为氦气，进样量为1μl，恒流不分流模式，流速为1ml/min，柱箱升温程序：起始温度50℃，保持1min，以10℃/min升温至110℃，再以5℃/min 升温至180℃，再以10℃/min升温至280℃结束，EI离子源，电子能量70eV，质子扫描范围50～550amu，四级杆温度为150℃，离子源温度为230℃，检测后得到气相色谱图和质谱图，通过与标准谱库(NIST11)的谱图进行比较鉴定化合物，解析图谱后找出与标准谱库图谱匹配度大于90％的物质作为鉴定标准，鉴定出水蓼精油中的17 种主要化学成分，并采用面积归一化法测得各组分的相对百分含量，分析鉴定结果见表 1。
表1
实施例2：水蓼精油对Neopestalotiopsis rosae的抑菌活性
为了检测实施例1得到的水蓼精油对Neopestalotiopsis rosae的抑菌效果，本实施例采取以下实验进行验证：
利用不同浓度的水蓼精油对Neopestalotiopsis rosae进行抑制实验。本实验分别将水蓼精油按照0.25、0.5、1和2μl/1ml(精油与培养基的体积比)的配比添加到PDA培养基中，通过计算其抑菌率，确定提取物对病菌的作用效果。
分别将不同量的水蓼精油加入到约50℃灭菌后的PDA固体培养基中，摇匀后倒入不同培养皿(直径为9cm)中，水蓼精油与培养基的体积比分别为：0.25μl/1ml、0.5 μl/1ml、1μl/1ml和2μl/1ml，待培养基冷却后，将在28℃培养箱黑暗培养3天后的Neopestalotiopsis rosae菌饼(直径为0.6cm)接入上述倒有含不同浓度水蓼精油的PDA培养基的培养皿中间，以PDA培养基(不含水蓼精油)作为对照，每皿三个重复，放置于生化培养箱中培养，培养条件28℃，待对照长满后，用十字交叉法测量菌落直径，计算其抑菌率，结果如表2所示。
计算方法为：抑菌率＝(对照菌落直径-处理菌落直径)/(对照菌落直径-菌饼直径) *100％。
结果表明：水蓼精油在浓度为2μl/ml时对Neopestalotiopsis rosae的抑菌率最好，达到60％以上。
表2不同浓度水蓼精油对Neopestalotiopsis rosae的抑菌率
实施例3：水蓼精油对草莓植株的安全性评价
为了检测实施例1得到的水蓼精油对草莓植株的安全性，本实施例采取以下实验进行验证：
将水蓼精油和水按照2μl/1ml比例(水蓼精油和水的体积比)均匀混合后，喷洒在草莓株上，以清水为对照，实验组和对照组的喷洒时机、用量以及光照等条件均相同， 7天后观察其表面是否有异常，确定水蓼精油对草莓植株的作用效果。
草莓植株的外观如图1，其中，A为喷洒含有水蓼精油组；B为喷洒清水的对照组，结果表明水蓼精油和水按照2μl/1ml比例混合后，对草莓不存在任何影响，因此，水蓼精油在一定浓度内是安全的。
Claims (7)
1.水蓼精油在抑制Neopestalotiopsis rosae中的应用。
2.水蓼精油在制备植物用Neopestalotiopsis rosae抑制剂中的应用。
3.水蓼精油在制备草莓用Neopestalotiopsis rosae抑制剂中的应用。
4.根据权利要求1-3中任一所述的应用，其特征在于，所述水蓼精油是由水蓼经过水蒸气蒸馏方式提取、分离得到。
5.根据权利要求4所述的应用，其特征在于，所述水蓼精油由以下方法提取得到：将干燥的水蓼全株粉碎后与水混合，利用微波萃取仪进行循环蒸馏，然后经过油水分离得到水蓼精油。
6.根据权利要求5所述的应用，其特征在于，所述水蓼精油的主要成分包括α-蒎烯、β-榄香烯、反式石竹烯、Α-香柠檬烯、α-律草烯、丙位依兰油烯、α-姜黄烯、甲位依兰油烯、β-红没药烯、1,2,4a,5,6,8a-六氢-1-异丙基-4,7-二甲基萘、橙花叔醇、4-(1-甲基乙基)环己醇、3,4-二甲基-3-环己烯甲醛、T-杜松醇、α-毕橙茄醇、a-红没药醇以及反式甲位佛手甘油烯。
7.根据权利要求4所述的应用，其特征在于，所述水蓼精油在应用时，将水蓼精油直接与水混合后喷洒于植株即可，所述水蓼精油与水的体积比为（0.25-2）μL：1mL。
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| CN106434371A (zh) * | 2016-09-29 | 2017-02-22 | 贵州医科大学 | 一株拟盘多毛孢属真菌及其用于艳山姜叶病害的鉴定方法 |
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