硅介导的黄瓜对棉铃虫的间接防御

发布时间：2017-12-27 08:45

本文关键词：硅介导的黄瓜对棉铃虫的间接防御　出处：《福建农林大学》2017年硕士论文　论文类型：学位论文

【摘要】：植物通过直接防御和间接防御来抵抗植食性昆虫的胁迫,硅在植物抵抗植食性昆虫的生物胁迫中扮演重要的角色。硅可以通过增强植株机械强度或组织耐磨性等来提高植物的直接防御,减轻害虫为害。然而,学界对硅如何影响植物的间接防御的认识并不深入。近年来,研究发现硅肥可以促进天敌对作物害虫的控制作用。关于硅增强植物间接防御机制的研究已开始得到学者们的关注。本研究以寄主植物-黄瓜Cucumis sativus、多食性植食性昆虫-棉铃虫Helicoverpa armigera和寄生性天敌-中红侧沟茧蜂Microplitismediator三级营养系统为研究对象,利用加硅和不加硅的处理,系统研究了硅的添加对黄瓜受棉铃虫取食胁迫的反应以及对天敌趋性的影响。利用"Y"型嗅觉仪测定了加硅和不加硅的黄瓜植株被棉铃虫为害不同时间(1h、3h、6h和12h)后对中红侧沟茧蜂的引诱反应。应用气质联用仪测定比较加/不加硅健康黄瓜植株、加/不加硅棉铃虫为害不同时间(1h、3h、6h和12h)后黄瓜植株的挥发性物质。选取棉铃虫为害6h后和12h后的黄瓜植株进行转录组测序分析,探索了硅调节植物对虫害防御的分子机制。主要结果如下:硅含量测定结果表明,添加硅之后,硅在黄瓜组织中的含量大小为:加硅的黄瓜叶片加硅的黄瓜茎干不加硅的黄瓜茎干,且不加硅的黄瓜叶片与加/不加硅的黄瓜茎干之间均没差异,可见黄瓜叶片和茎干中均能显著积累硅,且硅主要积累在黄瓜叶片中。加硅黄瓜叶片中的硅含量能够达到233.33±16.67 mg/kg,是不加硅黄瓜叶片中硅含量(20.00±10.00mg/kg)的11.67倍,同时是加硅黄瓜茎干中硅含量(33.33±0.88mg/kg)的 7 倍。"Y"型嗅觉仪实验结果表明,硅未处理下,与未被取食黄瓜植株相比,中红侧沟茧蜂趋向于被取食3h、6h和12h后的黄瓜植株;硅处理后,与未被取食黄瓜植株相比,中红侧沟茧蜂趋向被取食1h、3h、6h和12h后的黄瓜植株;在相同取食时间下,取食6h和12h后,中红侧沟茧蜂趋向硅处理的黄瓜植株。因此,硅能短时间内促进虫害黄瓜植株对中红侧沟茧蜂的吸引,并且能够增强棉铃虫为害6h后黄瓜植株对中红侧沟茧蜂对的吸引。GC-MS分析结果表明,黄瓜植株的挥发性物质主要包括醇类、萜烯类、酯类、酮类、醛类、酚类和烷烃等化合物。棉铃虫为害1h后,加硅黄瓜植株挥发物的总体含量高于不加硅黄瓜植株挥发物的总体含量,其中十二烷、2,6-二甲基十一烷、癸烷、3-甲基壬烷、十三烷和三十一烷等烷烃仅存在于加硅黄瓜植株挥发物中,并且十四烷、十五烷、十六烷、十七烷和2,6,10,14-四甲基十五烷等烷烃在加硅黄瓜植株挥发物中的含量显著升高。而棉铃虫为害3h后,加硅黄瓜植株挥发物的总体含量与不加硅黄瓜植株挥发物的总体含量基本相等,其中对氯苯亚磺酸仅在加硅植株的挥发物中检测到;十二烷在加硅植株挥发物中的含量显著高于在不加硅植株挥发物中的含量;长叶烯在加硅植株挥发物中的含量显著低于在不加硅植株挥发物中的含量。当棉铃虫为害6h后,加硅黄瓜植株挥发物的总体含量低于不加硅黄瓜植株挥发物的总体含量,其中乙醛、α-蒎烯、莰烯、β-蒎烯、甲基庚烯酮、右旋柠檬烯、桉油精和丁羟甲苯在加硅植株挥发物中的相对量显著降低;1,4-桉叶素、2-甲基萘在加硅植株挥发物中的含量显著提高。棉铃虫为害12h后,加硅黄瓜植株挥发物的总体含量又高于不加硅黄瓜植株挥发物的总体含量,臭樟脑、对氯苯亚磺酸、4-十二酮、醋酸丁酯和多种烷烃在加硅植株挥发物中的含量显著提高;4,7-二甲基十一烷在加硅植株挥发物中的含量显著降低。棉铃虫为害1h、3h、6h和12h后的加硅黄瓜植株挥发物与不加硅黄瓜植株挥发物相比,表现出总体含量升高、持平、降低和再升高的趋势。转录组数据分析表明,硅参与调控黄瓜重要诱导防御基因的表达。与健康加硅黄瓜相比,棉铃虫为害不同时间的加硅黄瓜中,硅能够调控茉莉酸信号途径,并且可以调控萜类挥发物合成关键基因TPS的表达。硅通过调控黄瓜诱导挥发物合成相关基因的表达,进而调节黄瓜虫害诱导挥发物的释放。
[Abstract]:Plants resist the stress of phytophagous insects by direct defense and indirect defense. Silicon plays an important role in plant resistance to biologic stress of phytophagous insects. Silicon can improve plant's direct defense by enhancing plant mechanical strength or tissue wear resistance to reduce pest damage. However, the academic community is not deeply aware of how silicon affects the indirect defense of plants. In recent years, the study found that silicon fertilizer can promote the control effect of natural enemies of crop pests. The research on the indirect defense mechanism of silicon reinforced plants has been paid attention by scholars. In this study, the host plant cucumber Cucumis sativus, polyphagous herbivore and cotton bollworm Helicoverpa armigera and parasitoid m.mediator - Microplitismediator three trophic system as the research object, by adding silicon and non silicon, silicon is added by cotton bollworm feeding stress reaction and the effect on natural enemies of cucumber were studied taxis. The attractant reaction of cucumber plants with silicon and without silicon on cotton bollworm at different time (1H, 3h, 6h and 12h) was detected by using the "Y" olfactory instrument. Application of GC-MS was used to compare with / without healthy cucumber plants, and silicon / silicon cotton bollworm in different time (1H, 3h, 6h and 12h) of volatile substances in Cucumber after. The transcriptional sequence analysis of cucumber plants after 6h and 12h was selected to explore the molecular mechanism of silicon regulating plant against insect pests. The main results are as follows: the results showed that addition of silicon content, silicon, the size of the content of silicon in Cucumber tissues was as follows: silicon and silicon in Cucumber Leaves of cucumber stem with silicon cucumber stem, leaf of cucumber and does not add silicon and silicon / no cucumber stem are not bad ISO, visible in cucumber leaves and stems significantly the accumulation of silicon, and silicon mainly accumulated in Cucumber leaves. Silicon content in the leaves of cucumber grown with silicon can reach 233.33 + 16.67 mg/kg, which is 11.67 times less than that of silicon cucumber leaves (20 + 10.00mg/kg), and is 7 times higher than that of silicon cucumber. The content of silicon is 33.33 times 0.88mg/kg. The experimental results of Y olfactometer showed that the untreated silicon, and have not been compared on cucumber plants, m.mediator cucumber plants tend to be 3h, 6h and 12h after feeding; silicon processing, and have not been compared on cucumber plants, m.mediator tends to be cucumber the plant, 3h, 6h and 1H on 12h; at the same time feeding, feeding on 6h and 12h, m.mediator trend of cucumber plant silicon processing. Therefore, silicon can promote the attraction of cucumber plants to the parasitoids in short time, and enhance the attraction of cucumber plants to 6h. The results of GC-MS analysis showed that the volatile compounds of cucumber plants mainly include alcohols, terpenes, esters, ketones, aldehydes, phenols and alkanes. The cotton bollworm damage after 1h, and silicon in cucumber plant volatiles is higher than the overall total content of volatile content without silicon in cucumber plant volatiles, including twelve alkanes, 2,6- two methyl eleven alkyl, 3-, methyl nonane, decane thirteen alkyl and thirty-one alkyl alkane exist only in silicon in cucumber plant volatiles, and fourteen alkyl fifteen, sixteen, seventeen alkyl alkyl, alkyl and 2,6,10,14- four methyl fifteen alkyl alkane in the volatile silicon content from cucumber plants increased significantly. The cotton bollworm 3h after adding silicon in cucumber plant volatiles and total content of volatile silicon in cucumber plant volatiles overall content is basically the same, the chlorobenzene sulfinic acid only in the volatile silicon plant was detected in twelve; in the content of volatile alkyl compounds and silicon plants is significantly higher than that in the absence of silicon plant volatile matter content of Longifolene content; in the volatile silicon plant volatiles were significantly lower than in silicon plant volatile matter content in. When the cotton bollworm 6h after adding silicon in cucumber plant volatiles is lower than the overall total content of volatile content without silicon in cucumber plant volatiles, including acetaldehyde, pinene, camphene, beta pinene, methyl heptenone, d-limonene, eucalyptol and butylated hydroxytoluene decreased significantly in the silicon plant the relative amount of volatile compounds; 1,4- cineole and 2- significantly increased the content of methyl naphthalene in the volatile silicon plant volatiles. The cotton bollworm damage after 12h, and silicon in cucumber plant volatiles and total content of volatile content is not higher than the overall silicon in cucumber plant volatiles, smelly camphor, chlorobenzene sulfinic acid, 4- twelve ketone, butyl acetate and alkanes increased significantly in content of volatile silicon plant volatiles; 4,7- two methyl eleven alkyl decreased significantly the content of volatiles in volatile silicon plant. The results showed that the concentrations of 1H, 3h, 6h and 12h in the cucumber plant volatiles increased with the increase of the total volatiles. The analysis of transcriptional data showed that silicon was involved in regulating the expression of important inducible defense genes in cucumber. Compared with healthy and silicon cucumber, silicon bolus can regulate jasmonic acid signaling pathway and regulate the expression of TPS, a key gene of terpenoid volatiles. Silicon can regulate the expression of related genes by regulating the volatiles induced by cucumber, and then regulate the release of the volatiles induced by cucumber insect pests.
【学位授予单位】：福建农林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S436.421

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