棉花—棉铃虫—寄生蜂三级营养链间中红侧沟茧蜂的实时趋性反应
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摘要
本文以寄主植物-棉花Gossypium hirsutum、植食性昆虫-棉铃虫Helicoverpa armigera和寄生性天敌-中红侧沟茧蜂Microplitis mediator三营养级系统为研究对象,研究了中红侧沟茧蜂对不同龄期棉铃虫幼虫及其危害棉花植株的趋性行为反应以及棉铃虫幼虫取食诱导棉株挥发物的成分。另外,首次应用电子鼻zNoseTM,构建棉花植株常见挥发性化合物特征谱库,建立棉铃虫幼虫危害棉花植株的指纹图谱,实时监测了棉铃虫幼虫危害诱导棉花挥发物中β-蒎烯和β-月桂烯的释放节律。主要结果如下:
1、采用“Y”形昆虫嗅觉仪测定中红侧沟茧蜂对不同龄期棉铃虫幼虫及其危害棉株的趋性行为反应。结果表明,中红侧沟茧蜂对3、4、5龄棉铃虫幼虫没有明显的趋向行为反应,1龄和2龄棉铃虫幼虫对中红侧沟茧蜂有显著的引诱作用,但中红侧沟茧蜂对1龄和2龄棉铃虫幼虫的趋性差异不显著。在中红侧沟茧蜂对不同龄期棉铃虫幼虫危害棉株的趋性行为反应测试中,设置两种类型处理,分别为棉铃虫幼虫为害棉株后去除棉铃虫幼虫(简称去虫)和棉铃虫幼虫为害棉株+棉铃虫幼虫复合处理(简称未去虫)。结果显示,1-5龄棉铃虫幼虫危害后去除棉铃虫幼虫及其虫粪处理的棉株与健康植株均对中红侧沟茧蜂有吸引作用,但二者对中红侧沟茧蜂的引诱力差异不显著。在未去虫的处理中,虫害诱导的植物挥发物比健康植株释放的挥发物更能吸引中红侧沟茧蜂,与健康植株存在显著性差异,而且不同龄期棉铃虫幼虫危害处理间差异较大,对中红侧沟茧蜂引诱作用的相对顺序为:5龄危害植株>2龄、4龄危害植株>1龄和3龄危害植株(2、4龄危害处理间差异不显著;1、3龄危害处理间差异不显著)。
2、利用气质联用系统(GC-MS)对不同龄期棉铃虫幼虫危害棉株挥发物成分进行定性及定量分析,结果表明,1-5龄棉铃虫幼虫处理棉株与对照棉株相比挥发性信息物质在种类和含量上均有差异,本研究中,在健康植株、虫害诱导的棉株挥发物中均检测鉴定出近30种挥发性信息物质;健康棉株检测不到β-月桂烯和3,7-二甲基-1-辛醇的释放,而1-5龄棉铃虫幼虫取食处理均可检测到;去虫与未去虫两处理间挥发物的种类基本相同,只是在挥发物含量上存在差异。虫害诱导的植物挥发物含量明显高于健康植株释放的挥发物含量,说明棉铃虫幼虫取食对棉花挥发物有显著诱导作用;棉铃虫幼虫为害棉株+棉铃虫幼虫复合处理中的α-蒎烯、β-蒎烯、β-月桂烯、(E)-乙酸-3-己烯酯和(D)-柠檬烯等挥发物的释放量明显高于去除棉铃虫幼虫及其虫粪处理,且两处理间差异显著。
3、采用电子鼻对棉花植株常见挥发物进行了定性分析,确定了每类化合物的相对保留指数,构建了棉花植株常见挥发性化合物特征谱库。同时利用电子鼻构建棉铃虫幼虫危害棉株的指纹图谱库,比对健康植株与棉铃虫幼虫危害棉株的色谱图及指纹图谱发现:棉铃虫幼虫危害棉株挥发物的种类和含量均比对照棉株多,而且未去除棉铃虫幼虫的处理与去除棉铃虫幼虫的处理相比,被诱导挥发物的含量显著升高,这与GC-MS的测定结果基本一致。
4、通过电子鼻实时监测不同处理棉株挥发性化合物β-蒎烯、β-月桂烯的释放节律。结果发现,当棉铃虫幼虫危害棉花植株2 h后,β-蒎烯的释放量明显增加,与健康植株相比差异显著;β-月桂烯在棉铃虫幼虫危害2 h的释放量与对照相比没有显著差异,在棉铃虫幼虫危害2-4 h其释放量显著增加,在危害6-8 hβ-蒎烯和β-月桂烯分别达到各自最大释放量,随后缓慢回落,但仍然保持较高释放水平。
This study was based on a tritrophic system including cotton Gossypium hirsutum, cotton bollworm Helicoverpa armigera, and parasitoid Microplitis mediator, the behavioral responses of M. mediator to different instars cotton bollworm (CBW) larvae and cotton plant damaged by different instars CBW larvae were studied, cotton plant volatiles induced by CBW larvae were collected and analyzed in the laboratory. Furthermore, by using electronic nose zNoseTM for the first time, the cotton plant common volatiles library was constructed. We also got the fingerprint database of cotton plant infested by CBW larvae, and the real-time release rhythm ofβ-pinene andβ-myrcene from cotton plant damaged by CBW larvae treatment were monitored. The major results are listed as follows:
1. The taxis responses of M. mediator to different instars CBW larvae and cotton plant damaged by different instars CBW larvae were measured in a Y-shaped olfactometer. The result showed that 3rd, 4th and 5th instar CBW larvae had non-significant attraction for M. mediator, while 1st and 2nd instar CBW larvae had significant attraction for M. mediator, but there was no significant differences between them. Among the behavioral responses of M. mediator to cotton plants damaged by different instars CBW larvae, there are two kinds of treatment, wiping out CBW larvae from cotton plant damaged by CBW larvae (dislodge larvae treatment) and damaged cotton plants +larval CBW (retain larvae treatment). The results revealed that both cotton plant damaged by CBW larvae and healthy cotton plant were attractive to M. mediator, yet there was no significant differences between them. As for damaged cotton plants + larval CBW, the volatiles from cotton plant infested by CBW larvae were more attractive to M. mediator than those of healthy plant, and there existed significant differences between them. There was notable differences among different instars, The relative selective preferences order was plant infested by 5th instar larvae of CBW> plant infested by 2nd and 4th instar larvae of CBW> Plant infested by 1st and 3rd instar larvae of CBW.
2. The qualitative and quantitative analyses of the cotton plant volatiles induced by different instars CBW larvae were conducted by using GC-MS, the results showed that the kinds and contents of volatiles from cotton plant infested by CBW larvae were more than those of healthy plant. About 30 kinds of volatiles were qualitatively identified, healthy plant did not detect the release ofβ-myrcene and 1-octanol, 3,7-dimethyl-, but cotton plant damaged by CBW larvae could be detected. The type of volatiles from cotton plant damaged by CBW larvae (dislodge larvae treatment) were the same as those of holding CBW larvae treatment, there was difference in the aspect of content. The content of volatiles from cotton plants infested by CBW larvae were markedly higher than healthy plants, this stated that CBW larvae had striking induction to cotton plant volatiles. Moreover, compared with cotton plant damaged by CBW larvae (dislodge larvae treatment), the contents ofα-pinene,β-pinene,β-myrcene, 3-hexen-1-ol, acetate, (E)-and (D)-limonene in retaining CBW larvae treatment were extraordinary higher, and there was significant differences between them.
3. By using electronic nose for the first time, cotton plant common volatile compounds were qualitative analyzed in laboratory. We determined relative retention index for each type of compounds, then constructed the cotton plant common volatiles library. based on the library, the volatiles from cotton plant infested by CBW larvae were collected, and we also got the fingerprint database of cotton plant damaged by CBW larvae. Comparing the healthy plant with cotton plant infested by CBW larvae, it was found that both the kinds and contents of volatiles from cotton plant damaged by CBW larvae treatment were notably higher than those of control (no treatment). Furthermore, the contents of volatiles from cotton plant damaged by CBW larvae (retaining larvae treatment) were significantly higher than those of cotton plant damaged by CBW larvae (dislodge larvae treatment), which was basically the same as the results obtained from GC-MS.
4. Moreover, by using electronic nose, the real-time release rhythm ofβ-pinene andβ-myrcene from cotton plant infested by CBW larvae treatment were monitored. The results showed that when cotton plant damaged by CBW larvae for two hours,β-pinene released significantly increased compared with the control. However, there was no difference in the contents ofβ-myrcene between the cotton plant damaged by CBW for two hours and healthy plant. After invested for 2-4 hours, theβ-myrcene contents markedly increased. And after 6-8 hours, the contents ofβ-pinene andβ-myrcene reached their maximum release respectively, and then slow down, but still maintained a high level of release.
1、采用“Y”形昆虫嗅觉仪测定中红侧沟茧蜂对不同龄期棉铃虫幼虫及其危害棉株的趋性行为反应。结果表明,中红侧沟茧蜂对3、4、5龄棉铃虫幼虫没有明显的趋向行为反应,1龄和2龄棉铃虫幼虫对中红侧沟茧蜂有显著的引诱作用,但中红侧沟茧蜂对1龄和2龄棉铃虫幼虫的趋性差异不显著。在中红侧沟茧蜂对不同龄期棉铃虫幼虫危害棉株的趋性行为反应测试中,设置两种类型处理,分别为棉铃虫幼虫为害棉株后去除棉铃虫幼虫(简称去虫)和棉铃虫幼虫为害棉株+棉铃虫幼虫复合处理(简称未去虫)。结果显示,1-5龄棉铃虫幼虫危害后去除棉铃虫幼虫及其虫粪处理的棉株与健康植株均对中红侧沟茧蜂有吸引作用,但二者对中红侧沟茧蜂的引诱力差异不显著。在未去虫的处理中,虫害诱导的植物挥发物比健康植株释放的挥发物更能吸引中红侧沟茧蜂,与健康植株存在显著性差异,而且不同龄期棉铃虫幼虫危害处理间差异较大,对中红侧沟茧蜂引诱作用的相对顺序为:5龄危害植株>2龄、4龄危害植株>1龄和3龄危害植株(2、4龄危害处理间差异不显著;1、3龄危害处理间差异不显著)。
2、利用气质联用系统(GC-MS)对不同龄期棉铃虫幼虫危害棉株挥发物成分进行定性及定量分析,结果表明,1-5龄棉铃虫幼虫处理棉株与对照棉株相比挥发性信息物质在种类和含量上均有差异,本研究中,在健康植株、虫害诱导的棉株挥发物中均检测鉴定出近30种挥发性信息物质;健康棉株检测不到β-月桂烯和3,7-二甲基-1-辛醇的释放,而1-5龄棉铃虫幼虫取食处理均可检测到;去虫与未去虫两处理间挥发物的种类基本相同,只是在挥发物含量上存在差异。虫害诱导的植物挥发物含量明显高于健康植株释放的挥发物含量,说明棉铃虫幼虫取食对棉花挥发物有显著诱导作用;棉铃虫幼虫为害棉株+棉铃虫幼虫复合处理中的α-蒎烯、β-蒎烯、β-月桂烯、(E)-乙酸-3-己烯酯和(D)-柠檬烯等挥发物的释放量明显高于去除棉铃虫幼虫及其虫粪处理,且两处理间差异显著。
3、采用电子鼻对棉花植株常见挥发物进行了定性分析,确定了每类化合物的相对保留指数,构建了棉花植株常见挥发性化合物特征谱库。同时利用电子鼻构建棉铃虫幼虫危害棉株的指纹图谱库,比对健康植株与棉铃虫幼虫危害棉株的色谱图及指纹图谱发现:棉铃虫幼虫危害棉株挥发物的种类和含量均比对照棉株多,而且未去除棉铃虫幼虫的处理与去除棉铃虫幼虫的处理相比,被诱导挥发物的含量显著升高,这与GC-MS的测定结果基本一致。
4、通过电子鼻实时监测不同处理棉株挥发性化合物β-蒎烯、β-月桂烯的释放节律。结果发现,当棉铃虫幼虫危害棉花植株2 h后,β-蒎烯的释放量明显增加,与健康植株相比差异显著;β-月桂烯在棉铃虫幼虫危害2 h的释放量与对照相比没有显著差异,在棉铃虫幼虫危害2-4 h其释放量显著增加,在危害6-8 hβ-蒎烯和β-月桂烯分别达到各自最大释放量,随后缓慢回落,但仍然保持较高释放水平。
This study was based on a tritrophic system including cotton Gossypium hirsutum, cotton bollworm Helicoverpa armigera, and parasitoid Microplitis mediator, the behavioral responses of M. mediator to different instars cotton bollworm (CBW) larvae and cotton plant damaged by different instars CBW larvae were studied, cotton plant volatiles induced by CBW larvae were collected and analyzed in the laboratory. Furthermore, by using electronic nose zNoseTM for the first time, the cotton plant common volatiles library was constructed. We also got the fingerprint database of cotton plant infested by CBW larvae, and the real-time release rhythm ofβ-pinene andβ-myrcene from cotton plant damaged by CBW larvae treatment were monitored. The major results are listed as follows:
1. The taxis responses of M. mediator to different instars CBW larvae and cotton plant damaged by different instars CBW larvae were measured in a Y-shaped olfactometer. The result showed that 3rd, 4th and 5th instar CBW larvae had non-significant attraction for M. mediator, while 1st and 2nd instar CBW larvae had significant attraction for M. mediator, but there was no significant differences between them. Among the behavioral responses of M. mediator to cotton plants damaged by different instars CBW larvae, there are two kinds of treatment, wiping out CBW larvae from cotton plant damaged by CBW larvae (dislodge larvae treatment) and damaged cotton plants +larval CBW (retain larvae treatment). The results revealed that both cotton plant damaged by CBW larvae and healthy cotton plant were attractive to M. mediator, yet there was no significant differences between them. As for damaged cotton plants + larval CBW, the volatiles from cotton plant infested by CBW larvae were more attractive to M. mediator than those of healthy plant, and there existed significant differences between them. There was notable differences among different instars, The relative selective preferences order was plant infested by 5th instar larvae of CBW> plant infested by 2nd and 4th instar larvae of CBW> Plant infested by 1st and 3rd instar larvae of CBW.
2. The qualitative and quantitative analyses of the cotton plant volatiles induced by different instars CBW larvae were conducted by using GC-MS, the results showed that the kinds and contents of volatiles from cotton plant infested by CBW larvae were more than those of healthy plant. About 30 kinds of volatiles were qualitatively identified, healthy plant did not detect the release ofβ-myrcene and 1-octanol, 3,7-dimethyl-, but cotton plant damaged by CBW larvae could be detected. The type of volatiles from cotton plant damaged by CBW larvae (dislodge larvae treatment) were the same as those of holding CBW larvae treatment, there was difference in the aspect of content. The content of volatiles from cotton plants infested by CBW larvae were markedly higher than healthy plants, this stated that CBW larvae had striking induction to cotton plant volatiles. Moreover, compared with cotton plant damaged by CBW larvae (dislodge larvae treatment), the contents ofα-pinene,β-pinene,β-myrcene, 3-hexen-1-ol, acetate, (E)-and (D)-limonene in retaining CBW larvae treatment were extraordinary higher, and there was significant differences between them.
3. By using electronic nose for the first time, cotton plant common volatile compounds were qualitative analyzed in laboratory. We determined relative retention index for each type of compounds, then constructed the cotton plant common volatiles library. based on the library, the volatiles from cotton plant infested by CBW larvae were collected, and we also got the fingerprint database of cotton plant damaged by CBW larvae. Comparing the healthy plant with cotton plant infested by CBW larvae, it was found that both the kinds and contents of volatiles from cotton plant damaged by CBW larvae treatment were notably higher than those of control (no treatment). Furthermore, the contents of volatiles from cotton plant damaged by CBW larvae (retaining larvae treatment) were significantly higher than those of cotton plant damaged by CBW larvae (dislodge larvae treatment), which was basically the same as the results obtained from GC-MS.
4. Moreover, by using electronic nose, the real-time release rhythm ofβ-pinene andβ-myrcene from cotton plant infested by CBW larvae treatment were monitored. The results showed that when cotton plant damaged by CBW larvae for two hours,β-pinene released significantly increased compared with the control. However, there was no difference in the contents ofβ-myrcene between the cotton plant damaged by CBW for two hours and healthy plant. After invested for 2-4 hours, theβ-myrcene contents markedly increased. And after 6-8 hours, the contents ofβ-pinene andβ-myrcene reached their maximum release respectively, and then slow down, but still maintained a high level of release.
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