CN114854488B - 野菊精油及其制备方法和应用 - Google Patents
野菊精油及其制备方法和应用 Download PDFInfo
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Abstract
本发明属于精油技术领域，具体涉及一种野菊精油及其制备方法和应用。所述野菊精油包括野菊的叶子的成分。所述制备方法，包括以下步骤：将野菊的叶子经水蒸气蒸馏，收集馏出液，即得野菊精油。研究发现，所述野菊精油能够有效抑制和/或灭杀黄单胞菌属的菌株的活性，从而防治黑腐病。
Description
技术领域
本发明属于精油技术领域，具体涉及一种野菊精油及其制备方法和应用。
背景技术
野菊(Chrysanthemum indicum)是菊科菊属植物。野菊为多年生草本，高0.25～1米，有地下长或短匍匐茎，花期6-11月，生于山坡草地、灌丛、河边水湿地、滨海盐渍地、田边及路旁。野菊花头状花序的外形与菊花相似，野生于山坡草地、田边、路旁等野生地带。野菊花性微寒，具有疏散风热、消肿解毒、消炎、保护肝脏、保护心血管等功效。
黑腐病，一种真菌，即鞭毛菌亚门棕榈疫霉菌和恶疫霉菌。黑腐病叶片发病，叶缘多处产生黄色斑，后变”V”字形向内发展，叶脉变黑呈网状，逐渐整叶变黄干枯。症状多表现为腐烂，黑斑，叶片发黄等，严重影响主要蔬菜作物、油料作物及景观植物的产量和品质，带来严重的损失。这类致病菌中部分菌株应用生产中常用的杀菌剂效果十分有限，难以抑制其对园艺作物的侵染。
野油菜黄单胞菌(Xanthomonas campestris)是一种能在全球范围内引起所有十字花科植物(如大白菜、甘蓝、花椰菜等)黑腐病的重要病原细菌，是研究植物与病原微生物之间相互关系的模式菌，其代表是野油菜黄单胞菌野油菜致病变种(Xanthomonascampestris pv.campestris,简称Xcc)。
发明内容
为了解决上述技术问题，本发明的发明人基于对本领域的研究，并经过大量的实验摸索，提供了一种野菊精油及其制备方法和应用，其能够抑制和/或灭杀黄单胞菌属的菌株的活性，从而有效防治黑腐病。
具体而言，本发明首先提供了一种野菊精油，包括野菊的叶子的成分。
本发明研究发现，将野菊的叶子进行蒸馏，所提取到的精油成分，能够有效抑制和/或灭杀黄单胞菌属的菌株的活性，从而防治黑腐病。
作为优选，所述野菊精油包括：莰烯、β-月桂烯、桉叶油醇、2-茨酮、2-茨醇、(-)-4-萜品醇、石竹烯、(Z)-β-金合欢烯、毕澄茄烯、1-乙烯基-1-甲基-2-(1-甲基乙烯基)-4-(1-甲基亚乙基)-环己烷、1,7,7-三甲基-双环[2.2.1]庚-2-基酯-3-甲基-2-丁烯酸。
作为优选，以重量份计，其包括：莰烯1～3份、β-月桂烯2～5份、桉叶油醇6～10份、2-茨酮25～30份、2-茨醇2～4份、(-)-4-萜品醇2～5份、石竹烯3～6份、(Z)-β-金合欢烯2～4份、毕澄茄烯6～10份、1-乙烯基-1-甲基-2-(1-甲基乙烯基)-4-(1-甲基亚乙基)-环己烷1～3份、1,7,7-三甲基-双环[2.2.1]庚-2-基酯-3-甲基-2-丁烯酸2～4份。
上述物质复配，共同赋予了野菊精油对黄单胞菌属的菌株的优异杀菌活性。
本发明还提供一种上述野菊精油的制备方法，包括以下步骤：
将野菊的叶子经水蒸气蒸馏，收集馏出液，即得野菊精油。本发明通过水蒸气蒸馏法对野菊的叶子进行提取，工艺简单，无污染。
作为优选，所述蒸馏的时间为1～4h。在上述蒸馏时间下，可以保证目标产物被基本上完全蒸馏出来。
作为优选，所述水蒸气蒸馏的水与野菊的叶子的质量比为1.5～2.5:1。
本发明还提供上述的野菊精油在抑制和/或灭杀黄单胞菌属的菌株的活性中的应用。
本发明还提供上述的野菊精油在防治黄单胞菌属的菌株引起的黑腐病中的应用。
作为优选，所述黄单胞菌属的菌株为野油菜黄单胞菌。
本发明还提供一种具有杀菌功效的复方精油，包括如下体积份数的组分：
上述的野菊精油4～6份，牛至2～4份，茶树1～3份，百里香0.5～1.5份。
本发明还提供上述的复方精油在抑制和/或灭杀黄单胞菌属的菌株的活性中的应用，述黄单胞菌属的菌株优选为野油菜黄单胞菌。
本发明的发明人将上述野菊精油与牛至、茶树、百里香进行合理的伍配，起到了协同增效的作用，相比使用单一野菊精油，所得复方精油针对黄单胞菌属的菌株，尤其是野油菜黄单胞菌具有更高效的抑菌/杀菌功效。
附图说明
图1为本发明实施例中野菊精油抑菌效果验证图，其中，A为野菊精油处理组，B为CK对照组。
图2为本发明实施例中野菊精油杀菌效果验证图，其中，A1～A3为野菊精油处理组，A4为CK对照组。
具体实施方式
以下结合具体实施例对本发明作进一步详细说明，但不用来限制本发明的范围。
以下实施例中所使用的实验方法如无特殊说明，均为常规方法。以下实施例中所用的实验原料和相关设备等，如无特殊说明，均可从商业途径得到。
实施例1
实施例1提供一种野菊精油，其制备方法如下：
将4100g野菊的叶子与8200mL水加入精油纯露提取设备中，采用电磁炉进行加热。先用2000瓦加热至沸腾，然后调至1500瓦，打开冷凝管中的冷凝水，蒸馏至精油没有析出为止，收集馏出液，获得野菊精油；经计算精油提取率为1.19‰。
所得野菊精油的成分分析：
1.1色谱质谱采集条件
GC-MS分析色谱条件：色谱柱为DB-5MS石英毛细管柱(30m×0.25mm×0.25μm)；升温程序：进样口的为温度250℃，柱初始温度为50℃，保持2min，8℃/min升温至220℃，保持6.7min，总时间持续30min；载气：He(99.999％)，总流量17.1mL/min，柱流量1.01mL/min，进样模式：分流，分流的比率为20:1。
GC-MS质谱条件：EI源；接口的温度为250℃；离子源的温度为：200℃；检测器电压：1KV；质量扫描范围30-500m/z，全扫描方式。
1.2成分分析结果
所得野菊精油的主要成分及其相对含量见表1。
表1
野菊精油功能验证实验：
选用黄单胞野油菜致病变种(XCC)，取浓度为3×108CFU/ml的菌液100μL均匀涂布于无抗生素的LB平板上，再加入灭菌后的6mm滤纸片放置于平板的中央，最后取10μL实施例1的野菊精油原液加至滤纸片，以不添加精油的滤纸片培养皿作为对照，28℃培养24h。野菊精油处理组和对照组各设置三组。
1、精油抑菌效果
在28℃下培养24h后，菌落生长状况如下图1。野菊精油处理组出现明显抑菌带，而CK对照组无抑菌带，说明野菊精油对野油菜黄单胞菌野油菜致病变种的生长有一定的抑制作用。
野菊精油处理组的抑菌带宽度分别为：12.65mm、11.35mm、10.38mm，平均11.46±0.93mm。
2、精油杀菌效果
分别划取野菊精油处理组抑菌带区域和非抑菌带区域的培养基(CK)，在新培养基上进行划线，然后在28℃的恒温培养箱进行培养24h。
结果见图2，从抑菌带区域选取的培养基在新培养基培养后，没有菌落长出(参见图2中A1～A3)；而从非抑菌带区域选取的培养基，在新培养基上划线培养后，细菌能正常生长并形成菌落(参见图2中A4)。说明在实施例1的野菊精油发挥作用的区域中，细菌被杀死或生长活性受到影响。
虽然，上文中已经用一般性说明、具体实施方式及试验，对本发明作了详尽的描述，但在本发明基础上，可以对其作一些修改或改进，这对本领域技术人员而言是显而易见的。因此，在不偏离本发明精神的基础上所做的这些修改或改进，均属于本发明要求保护的范围。
Claims (8)
1.野菊精油在防治黄单胞菌属的菌株引起的黑腐病中的应用，其特征在于，所述野菊精油包括野菊的叶子的成分；
所述黄单胞菌属的菌株为野油菜黄单胞菌；
所述野菊精油的制备方法包括以下步骤：
将野菊的叶子经水蒸气蒸馏，收集馏出液，即得野菊精油。
2.根据权利要求1所述的应用，其特征在于，所述野菊精油包括：莰烯、β-月桂烯、桉叶油醇、2-茨酮、2-茨醇、(-)-4-萜品醇、石竹烯、(Z)-β-金合欢烯、毕澄茄烯、1-乙烯基-1-甲基-2-(1-甲基乙烯基)-4-(1-甲基亚乙基)-环己烷、1,7,7-三甲基-双环[2.2.1]庚-2-基酯-3-甲基-2-丁烯酸。
3.根据权利要求1所述的应用，其特征在于，以重量份计，所述野菊精油包括：莰烯1~3份、β-月桂烯2~5份、桉叶油醇6~10份、2-茨酮25~30份、2-茨醇2~4份、(-)-4-萜品醇2~5份、石竹烯3~6份、(Z)-β-金合欢烯2~4份、毕澄茄烯6~10份、1-乙烯基-1-甲基-2-(1-甲基乙烯基)-4-(1-甲基亚乙基)-环己烷1~3份、1,7,7-三甲基-双环[2.2.1]庚-2-基酯-3-甲基-2-丁烯酸2~4份。
4.根据权利要求1所述的应用，其特征在于，所述蒸馏的时间为1~4 h；
和/或，所述水蒸气蒸馏的水与野菊的叶子的质量比为1.5~2.5:1。
5.一种具有杀菌功效的复方精油在抑制和/或灭杀黄单胞菌属的菌株的活性中的应用，其特征在于，包括如下体积份数的组分：
野菊精油5~7份，牛至2~4份，茶树1~3份，百里香0.5~1.5份；
所述野菊精油包括野菊的叶子的成分；
所述黄单胞菌属的菌株为野油菜黄单胞菌；
所述野菊精油的制备方法包括以下步骤：
将野菊的叶子经水蒸气蒸馏，收集馏出液，即得野菊精油。
6.根据权利要求5所述的应用，其特征在于，所述野菊精油包括：莰烯、β-月桂烯、桉叶油醇、2-茨酮、2-茨醇、(-)-4-萜品醇、石竹烯、(Z)-β-金合欢烯、毕澄茄烯、1-乙烯基-1-甲基-2-(1-甲基乙烯基)-4-(1-甲基亚乙基)-环己烷、1,7,7-三甲基-双环[2.2.1]庚-2-基酯-3-甲基-2-丁烯酸。
7.根据权利要求5所述的应用，其特征在于，以重量份计，所述野菊精油包括：莰烯1~3份、β-月桂烯2~5份、桉叶油醇6~10份、2-茨酮25~30份、2-茨醇2~4份、(-)-4-萜品醇2~5份、石竹烯3~6份、(Z)-β-金合欢烯2~4份、毕澄茄烯6~10份、1-乙烯基-1-甲基-2-(1-甲基乙烯基)-4-(1-甲基亚乙基)-环己烷1~3份、1,7,7-三甲基-双环[2.2.1]庚-2-基酯-3-甲基-2-丁烯酸2~4份。
8.根据权利要求5所述的应用，其特征在于，所述蒸馏的时间为1~4 h；
和/或，所述水蒸气蒸馏的水与野菊的叶子的质量比为1.5~2.5:1。
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