Oviposition preferences of <i>Anoplophora glabripennis</i> on three host plants and composition analysis of host plant volatiles

ZHU Ning; ZHANG Dongyong; SHEN Hongxia; HU Qin; FAN Jianting

doi:10.11833/j.issn.2095-0756.2017.06.013

Volume 34 Issue 6

Nov. 2017

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ZHU Ning, ZHANG Dongyong, SHEN Hongxia, HU Qin, FAN Jianting. Oviposition preferences of Anoplophora glabripennis on three host plants and composition analysis of host plant volatiles[J]. Journal of Zhejiang A&F University, 2017, 34(6): 1059-1064. doi: 10.11833/j.issn.2095-0756.2017.06.013

Citation:

ZHU Ning, ZHANG Dongyong, SHEN Hongxia, HU Qin, FAN Jianting. Oviposition preferences of Anoplophora glabripennis on three host plants and composition analysis of host plant volatiles[J]. Journal of Zhejiang A&F University, 2017, 34(6): 1059-1064. doi: 10.11833/j.issn.2095-0756.2017.06.013

Oviposition preferences of Anoplophora glabripennis on three host plants and composition analysis of host plant volatiles

doi: 10.11833/j.issn.2095-0756.2017.06.013

1.
School of Forestry and Biotechnology, Zhejiang A & F University, Hangzhou 311300, Zhejiang, China
2.
Zhejiang Changshan Oil-Tea Research Institute, Quzhou 324200, Zhejiang, China
3.
Ninghai Forest Science and Technology Promotion Station of Zhejiang, Ninghai 315600, Zhejiang, China

Received Date: 2016-11-17
Rev Recd Date: 2017-03-14
Publish Date: 2017-12-20

Abstract

Anoplophora glabripennis (Motschulsky) have a wide range of host plants and endanger both of the health host and the stressed host. In order to find the plant volatiles which affect oviposition behaviors and lay the foundation for the development of green attractant of A. glabripennis. The oviposition preference of three host plants, Salix ohsidare, Melia azedarach and Acer negundo, was tested by laboratory the selective and no-selective oviposition test. The volatile components of three host plants were analyzed by the method of dynamic headspace adsorption to combine Gas Chromatography-Mass Spectrometry (GC/MS). The results of both the selective and the no-selective test showed that S. ohsidare was the most suitable for ovipositing by A. glabripennis, followed by M. azedarach, and finally A. negundo. The success rate of oviposition of A. glabripennis on S. ohsidare was 69.4%, and the one on A. negundo was 0. The results of host volatiles analysis showed that 16 compounds were found in S. ohsidare, and 11 compounds were detected in M. azedarach and A. negundo. 3-Carene and 2-ethyl-Hexanal only tested in S. ohsidare. The contents of nonanal, styrene, and acetophenone in S. ohsidare were significantly higher than M. azedarach and A. negundo. In the above five compounds, 3-carene and nonanal were reported to show the attraction on A. glabripennis, so we speculate that 3-carene and nonanal may be the effective components for attracting oviposition of A. glabripennis females.
- forest protection,
- Anoplophora glabripennis,
- oviposition,
- host,
- volatiles,
- composition analysis

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光肩星天牛Anoplophora glabripennis(Motschulsky)属鞘翅目Coleoptera天牛科Cerambycidae沟胫天牛亚科Lamiinae星天牛属Anoplophora，是一种多食性蛀干性害虫，主要危害15个科几十种阔叶树种，包括杨属Populus，柳属Salix和槭属Acer等，广泛分布于华东和华北地区，其中在“三北”防护林和沿海防护林危害最为严重^[1-5]。光肩星天牛成虫取食寄主植物的小枝和嫩叶，幼虫蛀食寄主植物的韧皮部和木质部，破坏寄主植物的输导组织，严重时甚至导致树木衰弱，最终死亡^[6-7]。植食性昆虫通过寄主植物释放的挥发性次生物质，寻找、定位和识别寄主植物，进行选择性取食和产卵^[8-11]。天牛亚科特殊的下口式口器，形成了一个独特的产卵方式，通过在寄主植物的树皮上咬一条缝或一个洞进行刻槽产卵^[12]。光肩星天牛最喜欢在寄主植物树皮相对光滑的树干上部或主枝上产卵，但是光肩星天牛雌虫沿着树干进行产卵，甚至把卵产在因树冠枯死而暴露在外的树根上^[13-14]。此外，光肩星天牛雌虫产卵的刻槽数与寄主植物的品质和种类有很大关系^{[12, 15]}。光肩星天牛的寄主植物范围广泛，寄主挥发物成分复杂，并且光肩星天牛既危害衰弱寄主也危害健康寄主^[15-16]，对寄主的生理状态没有显著的偏好性，这跟一般的天牛习性不同^{[3, 17]}，这些特点都给光肩星天牛引诱剂的研究带来了很大的难度。目前，有关对光肩星天牛的防治主要还是物理防治、药剂防治和生物防治^[18-19]，市场上急需一种高效、绿色、无公害的光肩星天牛引诱剂。本研究期望通过研究光肩星天牛对适宜寄主的产卵行为差异，寻找影响其产卵的植物源信息素物质，为寻找一种绿色无公害的光肩星天牛引诱剂奠定基础。

1. 材料与方法

1.1. 供试虫源

试验所用的光肩星天牛采集于浙江省慈溪市沿海防护林(30°17′N，121°23′E，平均海拔为2.2 m)的青皮垂柳Salix ohsidare上，用捕虫网进行人工捕捉。防护林为青皮垂柳，夹竹桃Nerium indicum，海滨木槿Hibiscus hamabo，木麻黄Casuarina equisetifolia，苦楝Melia azedarach和白蜡Fraxinus chinensis等树种组成的混交林带，林地地势较为平坦，其中青皮垂柳受到光肩星天牛和星天牛危害较为严重，平均树高约为10 m，平均胸径约为30 cm，林下无其他植被。光肩星天牛成虫在5月中下旬、6月上旬陆续羽化出孔。

1.2. 供试树种

复叶槭Acer negundo，青皮垂柳和苦楝树干均采自于浙江农林大学生物测定中心旁的苗圃地。

1.3. 室内产卵试验

产卵试验在浙江农林大学养虫室内进行，温度设为25~28 ℃，光周期14 h:10 h(光照:黑暗)。产卵试验分为无选择性产卵试验和有选择性产卵试验。无选择性产卵试验：在木质养虫笼(50 cm × 40 cm × 40 cm)内分别放入1根复叶槭、青皮垂柳和苦楝的树干(长30 cm ×直径8 cm)和大量新鲜的1年生枝条，且树干放在装有水的培养皿内，防止树干水分流失。每个养虫笼内再放入1对性成熟的光肩星天牛成虫(1雌+1雄)，重复5次。有选择性产卵试验：将上述3种树干放入同一个养虫笼内，每个养虫笼再放入1对性成熟的光肩星天牛成虫(1雌+1雄)，重复5次。该实验2016年6月12日开始，7月12日结束，记录每根树干上光肩星天牛的刻槽数和实际产卵数。

1.4. 挥发物的收集与分析

采用动态顶空吸附法分别抽取复叶槭、青皮垂柳和苦楝树干的气体挥发物。用无色无味的聚乙烯袋(55.8 cm × 44.3 cm, Reynolds, Richmond, VA, 美国)包裹树干部位，聚乙烯袋内的空气通过含有活性炭(20~40目)的玻璃管(6 mm × 150 mm内径)纯化后，将含有Super-Q吸附剂(60~80目)的TVOC不绣钢采样管(6 mm × 150 mm)与大气采样仪(QC-1S型，中国北京劳动保护研究所)进气口相连接，含有活性炭的玻璃管与大气采样仪的出气口相连接，大气采样仪的所有部件通过硅胶管连接。流速1.5 L·min^-1，采样时间为6 h(10：00-16：00)，重复3次·处理^-1。收集的挥发物样品用含有以150.0 μg·L^-1正十二烷为内标的色谱纯正己烷溶剂洗脱3次，500.0 μL·次-1，洗脱下来的样品放在-5 ℃的冰箱保存待用。

采用安捷伦气相色谱-质谱联用仪Agilent 7890A-5975C分析复叶槭、青皮垂柳、苦楝等3种寄主植物的挥发物。色谱柱为HP-5MS[30 m (长)× 0.25 mm (内径)× 0.25 μm (膜)]，进样量1.0 μL，不分流进样。升温程序：初始温度40 ℃，恒温1 min，以6 ℃·min^-1的速率升到250 ℃，保持3 min，再以10 ℃·min^-1的速率升到270 ℃，保持5 min，共运行46 min。溶剂延迟3 min，离子源温度为230 ℃，四级杆温度为150 ℃。检测器温度为280 ℃，前进样口温度为220 ℃，电子轰击电压为70 eV，质量扫描范围为30~500。分流出口吹扫流量为60.0 mL·min^-1，隔垫吹扫流量为3.0 mL·min^-1。样品中挥发物成分的鉴定是通过与美国科学技术研究所(NIST)质谱库的出峰时间对比完成，定量分析通过内标正十二烷的峰面积计算。

1.5. 统计分析

用统计软件SPSS 16.0进行数据分析，刻槽数、产卵量和挥发物成分在不同寄主之间的差异显著性通过方差(ANOVA)分析，采用Duncan比较法。

3. 结论与讨论

光肩星天牛在青皮垂柳、苦楝和复叶槭等3种寄主植物树干上有选择性产卵和无选择性产卵试验结果都表明：光肩星天牛最喜欢在青皮垂柳上产卵，且产卵成功率较高，而最不喜欢在复叶槭上产卵，产卵成功率较低，在有选择性产卵试验中，光肩星天牛在复叶槭上的产卵成功率甚至为0。罗亚萍^[20]也报道，光肩星天牛在4种寄主上成功产卵量依次为柳树Salix babylonica＞苦楝＞木麻黄＞黄山栾树Koelreuteria bipinnata，说明柳树是光肩星天牛最偏好产卵的一种寄主，与本研究结果一致。而在每种寄主植物上的尝试刻槽数都大于实际成功产卵数，说明光肩星天牛并没有在每一个刻槽里产卵。KEENA^[21]试验也表明：光肩星天牛雌虫在寄主植物树皮上咬出产卵刻痕，但最后并没有选择去产卵。这是光肩星天牛造出刻槽后发现不适合产卵，还是有意的给天敌制造一些假想从而躲避天敌，还值得进一步研究。另外，我们的观察发现：光肩星天牛的每个卵槽里最多只有1个卵，这与LINGAFELTER等^[14]报道的光肩星天牛1个卵槽只产1个卵相符合。我们推测，天牛是一种具有自残性的昆虫，1个刻槽最多只有1粒卵是为了避免后代之间为竞争食物和生存环境而进行自残。

植食性昆虫通过寄主植物释放的挥发性气味对寄主植物进行识别和定位，从而进行取食和产卵^[22]。在寄主植物树干气体挥发物试验中，3-蒈烯是只在青皮垂柳中检测到的2种化合物之一，产卵试验也显示，光肩星天牛最喜欢在青皮垂柳上产卵，而NEHME等^[23]通过Y型嗅觉仪证明：3-蒈烯能够引起光肩星天牛雌虫的嗅觉反应，且野外试验进一步验证，3-蒈烯对光肩星天牛有吸引作用。因此，我们可以推测，3-蒈烯是一种吸引光肩星天牛产卵的植物源信息素物质。另外，壬醛只在青皮垂柳和苦楝中检测到，而且在青皮垂柳中的质量浓度明显高于苦楝。WICKHAM等^[24]通过Y型嗅觉仪和野外诱捕试验表明，光肩星天牛雌虫体表物质庚醛、壬醛和十六醛按一定比例混合后对光肩星天牛雌雄都有显著的引诱作用，其中雄虫的比例更高。这些结果肯定了壬醛是一种对光肩星天牛具有引诱作用的植物源物质。最后，2-乙基己醛只在青皮垂柳中检测到，而苯乙烯和苯乙酮在青皮垂柳中的质量浓度也显著高于苦楝和复叶槭，但是没有文献报道这3种化合物对光肩星天牛行为的研究结果，需要我们进一步的试验验证。

在植物源化合物中，还有哪些是关键的引诱物质，更重要的是在什么样的比例下，才能达到最好的引诱效果，都有待于我们进一步试验，其中青皮垂柳挥发物中几种特殊成分或者主要成分的自然比例非常值得进行下一步的野外试验效果检验。

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化合物	寄主植物树干气体挥发物质量浓度/(μg·L^-1)
化合物	青皮垂柳	苦楝	复叶槭
正己醛hexanal	0b	3.85±1.04 a	0b
3-己酮3-hexanone	0b	4.86±1.59 a	0b
苯乙烯styrene	50.09±6.35 a	8.41±1.44 b	6.70±1.43 b
α-薇烯a-pinene	70.80±2.93 a	74.83±5.95 a	76.10±8.94 a
莰烯camphene	11.25±1.42 a	9.38±1.57 a	10.35±1.44 a
2-乙基已醛2-ethyl-hexanal	12.06±3.63 a	0b	0b
正辛醛octanal	0b	9.24±1.71a	0b
3-蒈烯3-carene	4.91±1.05 a	0b	0b
丁酸丁酯butanoic acidbutyl ester	14.44±3.36 b	25.19±1.14 a	0c
2-乙基己醇2-ethyl-1-hexanol	161.14±29.28 a	153.36±17.06 a	121.42±5.02 b
苯乙酮acetophenone	19.73±0.94 a	11.69±0.47 b	9.48±2.79 b
十一烷undecane	17.96±2.75 a	13.15±2.76 a	0b
壬醛nonanal	107.02±17.04 a	45.74±3.45 b	0c
棒脑camphor	81.52±22.94 a	83.23±21.99 a	85.78±7.65 a
十三烷tridecane	105.12±16.35 a	93.95±7.77 a	100.59±8.53 a
十四烷tetradecane	78.17±0.93 a	65.35±2.8 ab	55.32±16.84 b
环癸烷cyclodecane	0b	48.18±7.17 a	0b
正十六烷hexadecane	33.02±6.70 a	0b	18.41±4.01 a
正十七烷heptadecane	33.09±15.23 a	0b	11.22±2.53 a
酞酸二乙酯diethyl phthalate	131.89±12.41a	137.13±18.98 a	121.20±7.86 a
说明:表中数据为平均值±标准误, 数据后标有不同字母表示差异显(P＜0.05, Duncan's多重比较)。

Oviposition preferences of Anoplophora glabripennis on three host plants and composition analysis of host plant volatiles

doi: 10.11833/j.issn.2095-0756.2017.06.013