CN108552176B - 提高薰衣草属植物对于植食性昆虫防御性的方法 - Google Patents
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Abstract
本发明公开一种提高薰衣草属植物对于植食性昆虫防御性的方法,其为向薰衣草属植物施用有效量的茉莉酸甲酯,提高薰衣草植物花朵中的萜类化合物的含量,防御植食性昆虫,吸引植食性昆虫天敌,所述的萜类化合物为柠檬烯、β‑水芹烯和β‑罗勒烯。本发明中,喷施MeJA溶液,不仅可以起到趋避植食性昆虫的直接防御作用,还能吸引天敌昆虫来捕食植食性昆虫,起到了间接防御作用,将人工技术和自然食物链科学统一,避免了不必要的损失。
Description
技术领域
本发明公开了一种提高薰衣草属植物对于植食性昆虫防御性的方法,属于植物害虫防控领域。
背景技术
薰衣草是一种异花授粉的植物,依靠风、昆虫进行传粉。释放萜类等挥发性成分吸引传粉昆虫,是芳香植物进化而来的一种特点。但是,这也同样吸引了某些植食性昆虫。调查了田间薰衣草上的昆虫种群和数量,发现花蕾期的薰衣草上有大量的蚜虫,由于薰衣草在花蕾期枝叶都十分幼嫩,大量植食性昆虫(蚜虫)的出现,或可伤害植株体。同时田间调查中,还发现花蕾期薰衣草上面有一定数量的异色瓢虫,它是蚜虫的天敌捕食者。
采用MeJA处理植株的方法来模拟植食性昆虫对植株体的破坏,从而阐明植物对外界信号的防御响应机制,是一种常用的研究手段(Sampedro,2010;Moreira et al.,2009;Bohlmann,2008;Zeneli et al.,2006)。原因在于,JA是防御昆虫的重要信号途径之一,能通过调控萜类化合物的生物合成进而发挥防御作用(De Geyter et al.,2012)。
植食性昆虫侵害或外源MeJA处理所引起的植物体内的防御响应机制研究,一直是重要的研究领域(Erb et al.,2012)。例如,使用MeJA处理番茄植株能增加叶片上的Ⅵ型腺毛的密度,促进萜类化合物的释放,有效防御白蝇和蓟马(Escobar-Bravo et al.,2017,2016);白菜、西兰花的植株在MeJA处理后增加α-法呢烯等萜类化合物及衍生物的释放量,起到了抑制昆虫产卵的作用(Ibrahim et al.,2005);茶树植株在MeJA处理后,增加了腺毛密度,起到了防御棉铃虫的作用(Tian et al.,2014);树棉植株在MeJA处理后增加柠檬烯释放量,降低植食性昆虫的丰度(Williams et al.,2017)。
CN107690879A公开了一种经济有效地提高栽培烟草抗虫性的方法。该方法采用浓度为3mmol/L的茉莉酸对种子进行浸泡处理。茉莉酸浸泡种子不仅增强了栽培烟草对两种严重影响其产量的害虫斜纹夜蛾及烟蚜的抗性,还不影响产量。但是,该技术对于扦插繁殖的植物不适用。由于薰衣草的繁殖方式主要为扦插繁殖,而尚未解决在植株开始展叶及现蕾时,如何提高抗性又不伤害植株。
发明内容
为了解决上述问题,本发明提供一种薰衣草属植物对于植食性昆虫防御性的方法。
一种提高薰衣草属植物对于植食性昆虫防御性的方法,其特征在于,向薰衣草属植物施用有效量的茉莉酸甲酯,提高薰衣草植物花朵中的萜类化合物的含量,防御植食性昆虫,吸引植食性昆虫天敌,所述的萜类化合物为柠檬烯、β-水芹烯和β-罗勒烯。
在本发明一个实施方案中,薰衣草属植物的施用部位为植食性昆虫为害的部位,可全株施用,也可以局部施用,要防御花序的植食性昆虫,施用部位为薰衣草属植物的花序。
在本发明一个实施方案中,向薰衣草属植物的花序施用时,所述薰衣草属植物处于花蕾期。
其中,茉莉酸甲酯浓度为2-5mmol/L,优选为4mmol/L。
其中,所述的植食性昆虫包括但不限于蚜虫总科(Aphidoidea),优选为麦长管蚜(Sitobion avenae)、禾谷缢管蚜(Rhopalosiphum padi)、麦二叉蚜(Schizaphisgraminum)等。
其中,所述的植食性昆虫天敌包括但不限于瓢虫科(Coccinellidae),优选为瓢虫亚科(Coccinellinae)、红瓢虫亚科(Coccidulinae)、小艳瓢虫亚科(Sticholotidinae)、盔唇瓢虫亚科(Chilocorinae)、小毛瓢虫亚科(Scymninae)、刻眼瓢虫亚科(Ortaliinae)、隐胫瓢虫亚科(Aspidimerinae),更优选为七星瓢虫(Coccinella septempunctata)、异色瓢虫(Harmonia axyridis)、六条瓢虫(Menochilus sexmaculatus)、龟纹瓢虫(Propylaeajaponica)、澳洲瓢虫(Rodolia cardinalis)等。
其中,所述的薰衣草属植物优选为狭叶薰衣草(Lavandula angustifolia)、宽叶薰衣草(Lavandula latifolia)、杂薰衣草(Lavandula×intermedia)等。
本发明还一种提高薰衣草属植物萜类化合物含量的方法,其为向薰衣草属植物施用有效量的茉莉酸甲酯,所述的萜类化合物为柠檬烯、β-水芹烯和β-罗勒烯。
在本发明一个实施方案中,薰衣草属植物的施用部位为需要提高萜类化合物含量的部位,可全株施用,也可以局部施用,要提高花朵中的萜类化合物含量,施用部位为薰衣草属植物的花序。
在本发明一个实施方案中,向薰衣草属植物的花序施用时,所述薰衣草属植物处于花蕾期。
其中,茉莉酸甲酯浓度为2-5mmol/L,优选为4mmol/L。
本发明采用MeJA处理植株的方法来模拟植食性昆虫对植株体的破坏,使用4mmol/L的茉莉酸甲酯(MeJA)溶液处理花蕾期的狭叶薰衣草(Lavandula angustifolia)花,使用固相微萃取(SPME)收集薰衣草花萜类化合物并用气相色谱与质谱(GC-MS)手段分析化学组成,发现柠檬烯、β-水芹烯、β-罗勒烯相对百分含量显著增加,同时使用光学显微镜等仪器,发现薰衣草花萼腺毛密度显著变大,表明MeJA处理后的薰衣草花释放了更多的柠檬烯、β-水芹烯、β-罗勒烯,使用薰衣草在花蕾期的主要植食性昆虫禾谷缢管蚜(Rhopalosiphumpadi),与其天敌昆虫异色瓢虫(Harmonia axyridis)进行气味选择实验,发现了柠檬烯、β-水芹烯、β-罗勒烯具有趋避蚜虫、吸引瓢虫的显著效果。由此说明使用MeJA溶液处理薰衣草花,能够提高某些萜类的释放量,起到防御植食性昆虫,并吸引天敌昆虫,综合保护植物体的作用。
本发明中,喷施MeJA溶液,不仅可以起到趋避植食性昆虫的直接防御作用,还能吸引天敌昆虫来捕食植食性昆虫,起到了间接防御作用,将人工技术和自然食物链科学统一,避免了不必要的损失。
附图说明
图1所示为光学显微镜下薰衣草花萼腺毛数量。
图2所示为薰衣草花中萜类化合物化学组分变化。
图3所示为蚜虫气味选择。
图4所示为瓢虫气味选择。
具体实施方法
以下实施例用于说明本发明,但不用来限制本发明的范围。
实施例1
使用狭叶薰衣草(Lavandula angustifolia)扦插苗,选择两年生且长势相同的盆栽,在昼/夜温度为30℃/25℃、每天12小时自然光照的温室中培养。2017年7月中旬,从温室选择生长一致的狭叶薰衣草盆栽,每盆均有花蕾期小花花枝,分为两组,每组三盆植株。一组茉莉酸甲酯溶液喷施处理(MeJA组),早上八点左右喷施一次,均匀喷施于薰衣草花萼表面至药液欲滴为度,每株喷施量约为100mL,用硫酸纸袋子套上植株并封口防止药液挥发;另一组对照溶液喷施处理(对照组)。
MeJA组溶液的配制:茉莉酸甲酯用体积分数为0.8%的无水乙醇溶解后,用蒸馏水定容,加入1%的吐温-20(非离子型表面活性剂,作为展开剂),配制成浓度为4mmol/L的MeJA溶液。对照组溶液的配制:0.8%的乙醇和1%的吐温-20。
MeJA喷施后24小时,取新鲜小花样品2g于2mL管中,放于冰上,花冠去掉,将花萼沿沟壑方向剪成两半铺平,使用荧光显微镜、光学显微镜进行观察和计量,保证每个视野下有2~3道沟壑,随机选择5个视野,记录腺毛数量,计算平均值。本部分的取样设计是,从MeJA组、对照组盆栽中分别选择5朵花蕾期小花。
如图1所示,光学显微镜下发现MeJA喷施24小时后,单位面积下,花蕾期的薰衣草花萼上腺毛数量显著增加(MeJA组:75.6个;对照组:57.6个),约为对照组的1.5倍。
取新鲜小花样品2g于2mL管中,放于冰上,立即采用固相微萃取(SPME)的方法提取萜类化合物,使用气相色谱-质谱联用仪完成GC-MS分析,采集数据,并使用Mass Hunter(Agilent公司)软件进行成分的定性定量分析。本部分的取样设计是,从MeJA组和对照组盆栽中分别选取25朵小花,8朵为一个重复。
如图2所示,发现MeJA处理后24小时,花蕾期的萜类化合物相对百分含量显著增加的有β-水芹烯(MeJA组:8.9869%,对照组:1.9818%)、柠檬烯(MeJA组:4.8905%,对照组:1.6667%)、β-罗勒烯(MeJA组:18.5630%,对照组:11.1864%),相对百分含量显著下降的有芳樟醇(MeJA组:0.5722%,对照组:3.5514%)、乙酸薰衣草酯(MeJA组:1.6903%,对照组:11.5411%)、薰衣草醇(MeJA组:0.4837%,对照组:2.3235%)、乙酸芳樟酯(MeJA组:20.7617%,对照组:28.5245%)。此外,蒎烯的相对百分含量也有增加,但不显著(MeJA组:8.2408%,对照组:7.2387%)。
结果表明,MeJA处理后的薰衣草花释放了更多的柠檬烯、β-水芹烯、β-罗勒烯。
使用薰衣草在花蕾期的主要植食性昆虫禾谷缢管蚜(Rhopalosiphum padi),与其天敌昆虫异色瓢虫(Harmonia axyridis)进行气味选择实验。
禾谷缢管蚜(Rhopalosiphum padi)接种在生长3周的麦苗上,套上网筐,放于温暖干燥的环境中饲养,取无翅成虫用于实验。异色瓢虫(Harmonia axyridis)接种在番茄的幼嫩茎叶上,套上网筐,放于温暖干燥的环境中饲养,取成虫用于实验。
单萜的浓度配比,根据MeJA喷施后24小时,柠檬烯、β-水芹烯、β-罗勒烯的相对百分含量,单萜标准品浓度配制如表1所示。
表1单萜标准品浓度
单萜标准品 | 对照组 | MeJA组 |
β-水芹烯 | 2% | 10% |
柠檬烯 | 2% | 5% |
β-罗勒烯 | 12% | 18% |
实验仪器为气味选择***:Y型管(一条中管,两条臂管,长20厘米,直径3厘米,两臂管夹角为75°)、QC-1(S)大气采样仪、干燥塔、水塔、气味源瓶和硅胶管。
实验方法:在室温24±1℃、相对湿度80±10%的条件下,气体总流速300mL/min,使用配好浓度的标准品溶液2μL,点在气味源瓶内的滤纸上,外界空气经由干燥塔内的活性炭吸附净化后,在大气采样仪的动力推动下,通过Y型管的臂管进入中管,最后由硅胶管排出室外。
测定前,对昆虫进行饥饿处理3~4小时,测定时,昆虫从基管放虫口释放,活跃后开始记时间,10分钟内,待其过其中一臂的1/3或者停留超过1分钟时,记为选择;超过10分钟,昆虫未进行任何一臂选择时,则记录为不选择。每种气味源测试昆虫30头,单头测定,每测定完10头更新一次气味源,每测定5头调换气味源位置,以减小位置误差。测定完一种气味源后用无水乙醇清洗Y型嗅觉仪,并用蒸馏水冲洗干净直至无任何气味,并于烘箱中烘干备用于下一组测定。
如图3所示,蚜虫气味选择实验中,相比β-水芹烯、柠檬烯、β-罗勒烯标准品一侧的气味源,蚜虫更倾向选择作为对照的白油一侧的气味源,β-水芹烯、柠檬烯、β-罗勒烯均能趋避蚜虫,并且β-水芹烯10%(MeJA组)、柠檬烯2%(对照组)、柠檬烯5%(MeJA组)、β-罗勒烯18%(MeJA组)具有显著的趋避效果,喷施MeJA后,平均趋避效果增强了约15.3%,并且,成分平均百分含量每提高1%,平均趋避效果增强3.1%,其中β-水芹烯(3.7%)、柠檬烯(0.68%)、β-罗勒烯(4.9%),说明JA信号使β-水芹烯、柠檬烯、β-罗勒烯成分百分含量增加,对蚜虫趋避性能增,β-罗勒烯趋避效果提高幅度最大。
如图4所示,瓢虫气味选择实验中,β-水芹烯、柠檬烯、β-罗勒烯标准品一侧的气味源,对瓢虫均具有吸引的作用,但效果各不相同。柠檬烯5%(MeJA组)、β-水芹烯10%(MeJA组)、β-罗勒烯18%(MeJA组)、具有显著的吸引作用;而柠檬烯2%(对照组)、β-水芹烯2%(对照组)、β-罗勒烯12%(对照组)对瓢虫几乎无选择性影响或微弱的吸引作用,但不显著。喷施MeJA后,平均吸引效果增强55.1%,并且,成分平均百分含量每提高1%,平均吸引效果增强13.8%,其中β-水芹烯(31.0%)、柠檬烯(5.36%)、β-罗勒烯(4.9%),说明JA信号使β-水芹烯、柠檬烯、β-罗勒烯成分百分含量增加,对瓢虫吸引效果增强,β-水芹烯吸引效果提高幅度最大。
由此说明,喷施MeJA溶液,薰衣草花中释放的β-水芹烯、柠檬烯、β-罗勒烯百分含量提高,趋避蚜虫的效果增强,吸引瓢虫效果也增强,薰衣草植株提高了综合防御性能,更有利于保护自身。
以上所述仅是本发明的优选实施方式,应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明技术原理的前提下,还可以做出若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。
Claims (7)
1.一种提高薰衣草属植物对于植食性昆虫防御性的方法,其特征在于,向薰衣草属植物施用有效量的茉莉酸甲酯,提高薰衣草植物花朵中的萜类化合物的含量,防御植食性昆虫,吸引植食性昆虫天敌,所述的萜类化合物为柠檬烯、β-水芹烯和β-罗勒烯,其中,薰衣草属植物的施用部位为薰衣草属植物的花序,所述薰衣草属植物处于花蕾期。
2.如权利要求1所述的方法,其特征在于,茉莉酸甲酯浓度为2-5mmol/L。
3.如权利要求1所述的方法,其特征在于,所述的植食性昆虫为同翅目蚜虫总科(Aphidoidea)。
4.如权利要求1所述的方法,其特征在于,所述的植食性昆虫天敌为瓢虫科(Coccinellidae)。
5.如权利要求1所述的方法,其特征在于,所述的植物为薰衣草属植物(Lavandula)。
6.一种提高薰衣草属植物萜类化合物含量的方法,其特征在于,向薰衣草属植物施用有效量的茉莉酸甲酯,所述的萜类化合物为柠檬烯、β-水芹烯和β-罗勒烯,其中,薰衣草属植物的施用部位为处于花蕾期的薰衣草属植物的花序。
7.如权利要求6所述的方法,其特征在于,茉莉酸甲酯浓度为2-5mmol/L。
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