Characteristics of Cellulases from Anoplophora glabripennis Motsch (Coleoptera: Cerambycidae)

The property of major cellulases from the guts of Anoplophora glabripennis larvae have been characterized. The optimal temperatures of both β 1,4 glucosidase (β glucosidase) and endo β 1,4 endoglucanase (endoglucanase, Cx) are 40℃. The β glucosidase was optimally active at pH 4\^8, while the optimal activity of the endoglucanase occurred at pH 4 4 5 6 The endoglucanase was active with a wide range of pH and temperature, the levels of activity from 25℃ to 50℃ were more than 80%, and the activity remained 60% between pH 3 2 and pH 7 2. The endoglucanase exhibited higher thermal stability than β glucosidase. Both enzymes lose their activities by heat treatment at 60℃. Two isozymes of endoglucanase were detected in sodium carboxymethylcellulose polyacrymide gels (CMC gel) by chemical colorization, and purified by elution from the gel slices. The molecular weights of the two isozymes were estimated as 26kD and 39kD respectively. Moreover molecular characteristics of the two isozymes are currently underway.

Phylogenetic Analysis of the 16S rDNA of a Strain Isolated from Diseased Larva of Anoplophora glabripennis (Motsch.)

[ Objective] Study on the phylogenetic analysis of the 16S rDNA and insecticidal characteristics of strain BH-1 isolated from diseased larva of Anoplophora glabripennis （Motseh.） [ Method ] The strain was identified by routine method and inoculated onto healthy Anoplophora glabripennis （Motseh.） for observing insecticidal effect, further 16S DNA was amplified by the specific primers for sequencing and homology analysis. [ Result] The mortality of second instar ofAnoplophora glabripennis（ Motseh. ） reached 72.7% 8 d after 10^10cfu/ml BH-1 was inoculated. The homology of 16S DNA sequences between BH-1 and Serratia marcescens accessed in GenBank reached 99.5%. Combined with the results of routine identification, BH-1 was identified as S. marcescens. [Conclusion] BH-1 could be used for biological control ofAnoplophora glabripennis （Motsch.）.

茉莉酸甲酯喷施和光肩星天牛(Anoplophora glabripennis(Motschulsky)咬食后五角枫释放的挥发物

茉莉酸甲酯喷施和光肩星天牛(Anoplophora glabripennis(Motschulsky)咬食后五角枫(Acer mono Maxim.)植株均诱导产生了乙酸丁酯、2-壬醇、乙酸己酯、3-甲基-2-丁醇、1-辛烯、里那醇、3-蒈烯、1-丁醇、(Z)-2-己烯醛9种正常植株中没有检测到的挥发物。两种处理均诱导或促进了萜烯类、醇类和醛类挥发物的大量释放,且乙酸-3-己烯酯、乙酸乙酯、己醇、反-2-己烯醇、乙基己醇、1-辛烯、石竹烯、法尼烯等挥发物的时序变化趋势也非常相似。因而推测光肩星天牛咬食五角枫后在植物体内诱导产生了茉莉酸信号传导途径。

Electrophysiological and Behavioral Responses of Adult Anoplophom glabripennis (Motschulsky) to Volatile Components of Host-Plant

Electrophysiological and behavioral responses of adult A. glabripennis (Motsch.) to volatiles from ashleaf maple (Acer negundo L.) were investigated to identify semiochemicals involved in host location. Measurable electroantennogram (BAG) responses were elicited to all compounds tested, the most effective antennal stimulants were trans-2-hexen-1-al,decyl aldehyde and trans-2-hexenl-ol.These profiles were similar between males and females. In Y-tube olfactometer bioassays, above three compounds with certain concentration, trans-2-hexen-l-al (1%), trans-2-hexen-l-ol (1%) and decyl aldehyde (10%), were significantly attractive to the adults in laboratory. The results show that either BAG or olfactory responding to a particular volatile compound are markedly influenced by the concentration.

Combining WV-2 images and tree physiological factors to detect damage stages of Populus gansuensis by Asian longhorned beetle (Anoplophora glabripennis) at the tree level

Background:Anoplophora glabripennis(Motschulsky),commonly known as Asian longhorned beetle(ALB),is a wood-boring insect that can cause lethal infestation to multiple borer leaf trees.In Gansu Province,northwest China,ALB has caused a large number of deaths of a local tree species Populus gansuensis.The damaged area belongs to Gobi desert where every single tree is artificially planted and is extremely difficult to cultivate.Therefore,the monitoring of the ALB infestation at the individual tree level in the landscape is necessary.Moreover,the determination of an abnormal phenotype that can be obtained directly from remote-sensing images to predict the damage degree can greatly reduce the cost of field investigation and management.Methods:Multispectral WorldView-2(WV-2)images and 5 tree physiological factors were collected as experimental materials.One-way ANOVA of the tree’s physiological factors helped in determining the phenotype to predict damage degrees.The original bands of WV-2 and derived vegetation indices were used as reference data to construct the dataset of a prediction model.Variance inflation factor and stepwise regression analyses were used to eliminate collinearity and redundancy.Finally,three machine learning algorithms,i.e.,Random Forest(RF),Support Vector Machine(SVM),Classification And Regression Tree(CART),were applied and compared to find the best classifier for predicting the damage stage of individual P.gansuensis.Results:The confusion matrix of RF achieved the highest overall classification accuracy(86.2%)and the highest Kappa index value(0.804),indicating the potential of using WV-2 imaging to accurately detect damage stages of individual trees.In addition,the canopy color was found to be positively correlated with P.gansuensis’damage stages.Conclusions:A novel method was developed by combining WV-2 and tree physiological index for semi-automatic classification of three damage stages of P.gansuensis infested with ALB.The canopy color was determined as an abnormal phenotype that could be directly assessed using remote-sensing images at the tree level to predict the damage degree.These tools are highly applicable for driving quick and effective measures to reduce damage to pure poplar forests in Gansu Province,China.

Cellulase in Anoplophora glabripennis adults emerging from different host tree species

In order to investigate different kinds of cellulase in insect pests, we selected male and female adults of Anoplophora glabripennis （Motschulsky） emerging from four different host species （Populus alba var. pyramidalis, P. nigra var. thevestina （Dode） Bean., P. simonii × P. pyramidliscr cv. Opera 8277 Hsu. and Salix matsudana f. lobato-glandulosa） as our research material. The enzyme activitives of three kinds of cellulase in the intestines of the adult insects were measured. The Cx-cellulase isozymes were detected with a CMC-incorporated polyacrylamide gel. The results show that： there are no statistically significant differences between the enzyme activities of males and females emerging from the different host species. The order of magnitude in activity is： Cx-cellulase 〉 β-glycosidase 〉 C1-cellulase. When the adults emerge from the same host species, there are no statistically significant differences between male and female enzyme activities of β-glycosidase and C1-cellulase, but the enzyme activity of Cx-cellulase of males is clearly higher than that of females. The patterns and migration of Cx-cellulase isozymes of males and females emerging from differ-ent hosts trees are clearly not different, and neither are they different when emerging from the same hosts.

Cellulase in Anoplophora glabripennis adults fed on original host tree species and non-original host trees

Cellulase activities of Anoplophora glabripennis （Motsch.） adults from two host plants （Populus simonii x p pyramidliscr cv. Opera Hsu. and Salix matsudana Koidz） fed on three different host tree species （Acer negundo Linn., S. matsudana Koidz and P simonii x P pyramidliscr cv. Opera Hsu.） were investigated. Enzyme activities of endoglucanase and β-glucosidase in the intestines of the insects were measured. The results show that there are no statistically significant differences in the enzyme activities of endoglucanase between male and female insects from the two host plants fed on three host trees, neither inβ-glucosidase. No statistically significant differences in the enzyme activities of endoglucanase and β-glucosidase were found between males and females from the two plants fed on three host tree species, except that the endoglucanase activity in males from the opera poplar fed on Hankow willow is lower than that fed on ash-leaf maple and opera poplar.

Oviposition Site Selection of Anoplophora glabripennis in Sugar Maple (Acer saccharum) and Its Gnawing Nidus Oviposition Habits

[Objective] The paper was to explore the oviposition site selection of Anoplophora glabripennis in sugar maple and its gnawing nidus oviposition habits. [Method] Through field and indoor rearing observation, the oviposition site selection of A. glabripennis in sugar maple was studied, and the gnawing nidus oviposition habits were analyzed. [Result] The height of oviposition site of A. glabripennis in sugar maple, the number observation holes （gnawing nidus oviposition points） and bark thickness in oviposition site showed linear correlation with diameter of branch at breast height. The gnawing nidus oviposition habit of A. glabripennis in sugar maple was mainly divided into 4 stages： gnawing nidus site selection, gnawing nidus, oviposition and nidus sealing, the duration from gnawing nidus to nidus sealing was（38.29±16.47）min. [Conclusion] The study provided reference for comprehensive prevention of A. glabripennis in greening tree species sugar maple, as well as further study on reproductive behavior and regulation mechanism of A. glabripennis.

光肩星天牛成虫光行为反应

对光肩星天牛(Anoplophora glabripennis)成虫光行为进行研究,探讨利用灯光诱捕监控其种群的可能性。在室内暗光周期下,以高压氙灯和不同光波的发光二极管(LED)红光灯为光源,配合滤光片和中性衰减片,测试光肩星雌雄两性对不同光强度、不同波长光源的行为反应。结果表明:当相对光强度为100%时,可以引起距离光源2.0 m雌雄两性的飞行趋光行为,其中雄性的起飞率高于雌性;随着光强度的降低,雌雄两性的趋光活性随之减弱;相对光强度为100%和50%时,雌雄两性的都表现出明显的避光行为,雄性的避光距离大于雌性;随着光强度的降低,雌雄两性的避光活性随之减弱,当相对光强度降为25%时,雌雄两性均无避光反应。雌雄两性对370 nm的紫外光和600~700 nm的红光趋向性最强,其次是400~500 nm蓝紫光,对500~600 nm黄绿光趋向性最弱;对620~630、640~650及655~665 nm波段的LED红光行为反应没有差别。高强度的光源能引起光肩星天牛远距离趋光行为,但也会引起明显的避光反应,这个行为特性表明灯光诱捕不适合用于监控该天牛种群。

杨-药复合经营林主要挥发物对光肩星天牛和美国白蛾的驱诱作用初步研究

[目的]筛选杨-知母、杨-亳菊和杨-薄荷复合经营试验林中主要挥发物成分,室内测定其对光肩星天牛和美国白蛾的驱诱作用。[方法]采用动态顶空吸附法收集3种杨-药复合经营试验林内药用植物地上、整株及空气中的挥发物,利用GC-MS分析鉴定挥发物组分并筛选主要挥发物,室内测定光肩星天牛和美国白蛾对它们的EAG和嗅觉行为反应。[结果]分别从杨-知母、杨-亳菊、杨-薄荷试验林中筛选出主要挥发物乙酸叶醇酯、β-石竹烯、薄荷醇。3种化合物在测试浓度范围内均能引起光肩星天牛和美国白蛾成虫的EAG反应,且反应值随浓度升高而增强。薄荷醇对光肩星天牛成虫、美国白蛾成虫和幼虫具有明显驱避作用,且在60μg·μL^(-1)浓度对光肩星天牛雌、雄成虫驱避率最高,分别为65%和47.5%;在500μg·μL^(-1)浓度对美国白蛾雄成虫和幼虫驱避率最高,分别为60%和70%;β-石竹烯仅对2种昆虫雄虫产生引诱作用,引诱率在40%~47.5%;不同浓度乙酸叶醇酯对光肩星天牛雌、雄虫和美国白蛾雄成虫驱诱作用产生分化,表现为低浓度引诱(引诱率为45%左右),高浓度趋避(趋避率为50%左右)。对美国白蛾雌成虫具有驱避作用(52.5%),对美国白蛾幼虫具有吸引作用(45%~65%)。[结论]3种试验林中主要挥发物成分乙酸叶醇酯、β-石竹烯、薄荷醇均能引起光肩星天牛和美国白蛾的EAG和嗅觉行为反应。薄荷醇效果最为明显,对两种昆虫驱避作用显著;乙酸叶醇酯作用效果次之,主要表现为低浓度引诱,高浓度驱避作用;β-石竹烯效果最差,仅对2种昆虫雄虫产生引诱作用。研究结果可为植物源引诱剂和植物源驱避剂的开发及基于“推-拉”防治策略的绿色环保杨药复合经营模式构建提供基础理论依据。

光肩星天牛胁迫声及种内空间运动行为

为了利用行为调制手段防治害虫,以光肩星天牛(Anoplophora glabripennis)为研究对象,在实验室内对光肩星天牛雌雄两性胁迫声进行录制和分析,分别在光周期和暗周期对光肩星天牛进行行为生测,分析胁迫声对光肩星天牛空间运动行为的影响。结果表明:光肩星天牛雄性胁迫声音组持续时间短于雌性、频率明显高于雌性,但雌雄胁迫声对空间运动行为的影响没有差异。在光周期,胁迫声对天牛向上爬行及水平运动均没有影响;对雄性起飞行为有抑制作用,但对雌性起飞行为没有影响。在暗周期,胁迫声对雌雄两性水平爬行有驱离作用,降低了天牛趋光爬行的活性;对雄性起飞行为没有影响,但对雌性起飞行为有促进作用。光肩星天牛胁迫声对种内个体具有警戒信号作用,这种作用与光周期相关并对雌雄两性的行为影响有所不同。

生态自控光肩星天牛灾害的诱杀树沙枣与抗性树种新疆杨配植模式研究

为筛选诱杀树沙枣Elaeagnus angustifolia和抗性树种新疆杨Populus alba var.pyramidalis合理配植比例,实现生态自控光肩星天牛Anoplophora glabripennis灾害,以甘肃河西走廊和新疆部分地区的已有光肩星天牛长期危害,且包括沙枣树和新疆杨等多树种混交林为研究对象,调查、对比和分析沙枣自然调控天牛种群的能力,筛选和提炼出可实现“有虫不成灾”的诱杀树沙枣与抗性树种新疆杨的合理配植模式。结果显示:混交林内沙枣树上光肩星天牛的刻槽数远高于抗性树种新疆杨,表现出显著的引诱功能;沙枣树对光肩星天牛的卵和初孵幼虫的杀灭率在99.8%以上,具有很强的诱杀能力,发挥了对新疆杨的高效保护作用;在不同沙枣配比的林带中,沙枣树对光肩星天牛种群的调控能力有较大差别,防护林带中沙枣树与新疆杨行间或株间配植比例约为2∶8,园林绿化林中沙枣树与新疆杨以块状或带状配植比例约为3∶7时,可达到“有虫不成灾”,实现光肩星天牛灾害的生态自控。在防护林建设中,应充分发挥沙枣对光肩星天牛的诱杀功能,合理配植诱杀树种沙枣与抗性树种新疆杨,使光肩星天牛种群数量稳定在成灾水平之下,为服务“三北”防护林工程区林业有害生物防治提供借鉴。

空间体积及光照对光肩星天牛4种行为活动的影响

为明确空间体积对光肩星天牛成虫交配、取食、移动和静息行为的影响,室内利用5种体积规格不同的容器(1000、4000、8000、16000、32000 cm^(3))进行连续5 d的试验观察。研究结果显示:(1)光肩星天牛雌、雄虫交尾行为、取食行为、移动行为发生的频次均随着体积的增大而减小;在不同空间体积中除雌、雄成虫静息活动不存在差异外,交配、取食和移动活动雌、雄成虫均存在差异;(2)1000 cm^(3)空间下取食活动、4000 cm^(3)移动活动、16000 cm^(3)取食活动的雌、雄成虫之间存在显著差异;(3)在有光照时,1000 cm^(3)空间下的取食活动、静息活动和32000 cm^(3)的静息活动以及无光照时,4000 cm^(3)的移动、静息活动、8000 cm^(3)的移动活动和32000 cm^(3)静息活动的雌、雄成虫之间存在显著差异;(4)空间体积对光肩星天牛不同行为活动的日节律存在影响。综上所述,光肩星天牛的行为活动会受到空间体积大小及光照的影响。

Is there hybridization between 2 species of the same genus in sympatry?——The genetic relationships between Anoplophora glabripennis,Anoplophora chinensis,and putative hybrids

Anoplophora glabripennis(Asian longhorn beetle,ALB)and Anoplophora chinensis(Citrus longhorn beetle,CLB)are native forest pests in China;they have become important international quarantine pests.They are found using the same Salix aureopendula host tree of Cixi,Zhejiang province,China.On this host tree,we collected additional beetles that appeared to be morphologically intermediate between ALB and CLB.By using a stereoscope,we observed that there were several bumps on the base of the elytra,which was inconsistent with ALB,which typically has a smooth elytral base,but was more like CLB,which has numerous short tubercles on the elytral base.Given their sympatry and intermediate morphology,we hypothesized that these may represent ALB×CLB hybrids.We studied the genomic profiles for 46 samples(ALB,CLB,and putative hybrids)using genotyping-by-sequencing(GBS)providing a reduced representation of the entire genome.Employing principal component analyses on the 163 GBS-derived single nucleotide polymorphism data,we found putative hybrids tightly clustered with ALB,but genetically distinct from the CLB individuals.Therefore,our initial hybrid hypothesis was not supported by genomic data.Further,while mating experiments between adult ALB and CLB were successful in 4 separate years(2017,2018,2020,and 2021),and oviposition behavior was observed,no progeny was produced.Having employed population genomic analysis and biological hybridization experiments,we conclude that the putative hybrids represent newly discovered morphological variants within ALB.Our approach further confirmed the advantage of genome-wide information for Anoplophora species assignment in certain ambiguous classification cases.

不同表面活性剂对药液性状及光肩星天牛防治效果的研究

为了明确不同表面活性剂对吡虫啉药液穿透树体及对光肩星天牛的防治效果,该研究在25%吡虫啉可湿性粉剂30倍液中分别添加2.5%杰效利、2.5%壬基酚聚氧乙烯醚(NP-10)、2.5%锐透3种表面活性剂,测定分析其表面张力、润湿性、铺展性等物理性状,并通过田间应用,测定分析叶片吡虫啉含量和光肩星天牛防治效果。结果显示:吡虫啉药液中添加2.5%锐透其各项物理性状最优,叶片药液含量最佳为6.52 mg/kg,防治效果最佳,矫正死亡率高达97.97%。因此,添加锐透作为表面活性剂效果最优。

辐照对光肩星天牛幼虫发育的影响

用高能X射线辐照处理3、4、5龄光肩星天牛幼虫，结果表明：辐照引起幼虫的生活期延长，发育迟缓；辐照抗性随虫龄的增加而增强，5龄幼虫最耐辐照，阻止其化蛹的剂量为55～60Gy。γ射线辐照杨树木段中光肩星天牛幼虫的结果也表明，60Gy能够100％阻止老熟幼虫化蛹。因此，可以推荐60Gy作为光肩星天牛幼虫检疫辐照处理的有效剂量，用于原木和木质包装的检疫除害处理。

西藏南部光肩星天牛发生情况调查报告

20 0 2年 8月 ,在对西藏南部进行林木考察时 ,首次发现了光肩星天牛 (AnoplophoraglabripennisMotsch .)在林芝、拉萨、日喀则等地危害多种林木 ,并在当地见到了该虫的活虫道、羽化孔和产卵的被害状。当前光肩星天牛还处于潜伏期 ,应引起足够重视 ,防患于未然。

自流式树干注药防治杨树光肩星天牛幼虫研究

利用自流式树干注药技术防治杨树光肩星天牛幼虫。结果表明 :在同种剂量下 ,同种药剂以一次性注药的防治效果明显高于两次性注药的防治效果。对供试的 4种药剂进行防治效果比较 ,以天牛敌对光肩星天牛幼虫的防效最高。并对天牛敌以不同的注药剂量对杨树光肩星天牛幼虫的防效进行分析 ,得出其防效与单位胸断面积注药量呈对数关系。回归方程为 :P=1 .0 4 65+0 .3 952 lnx。施药剂量 0 .5ml/cm2～ 0 .7ml/cm2 为佳。

光肩星天牛取食后复叶槭挥发物的释放机制

阔叶植物叶片通常释放少量的挥发物 ,但是当受到害虫取食后植物会释放大量的物质 ,并且其挥发物的指纹图谱与未被损伤的植株相比大不相同 .这些挥发物在植物 -害虫 -害虫天敌之间的关系中起着非常重要的作用 .为了进一步了解昆虫取食植物后挥发物的释放机制 ,作者选用复叶槭 (Acernegundo)和光肩星天牛 (Anoplophoraglabripennis (Motsch .) )为试验材料 ,利用气相色谱和质谱联用 (GC MS)检测挥发物 .在天牛取食后大多数物质在 9h的释放率达到第一个峰值 (β 蒎烯、六氢法尼烯基丙酮、乙酸苯乙酯、苯甲酸甲酯、水杨酸甲酯、syn 2 甲基丁醛肟除外 ) .在经过短时间下降后又逐渐上升 ,至天牛取食后 48h达到最大值 .只有 β 蒎烯、3 蒈烯、桉树脑、D 柠檬烯、trans 香叶基丙酮、苯甲酸甲酯、苯乙酮、苯并噻唑等物质在 72h达到最大 ,而syn 丁醛肟、1 ,3 ,3 三甲基 2 甲烯二氢吲哚在咬食 1h后就达到峰值 .天牛咬食诱导出的复叶槭挥发物 (即在正常植株中未被检测到的物质 )种类随时间的变化逐渐增加 ,在 48h达到 1 7种 .

罹病光肩星天牛幼虫分离菌株的16S rDNA系统发育分析

[目的]研究从罹病光肩星天牛[Anoplophora glabripennis(Motsch.)]幼虫刻槽中分离的菌株BH-1的16S rDNA系统发育及该菌株杀虫特性。[方法]常规细菌鉴定,将BH-1接种健康天牛后观察杀虫效果并通过设计特异引物扩增其16S rDNA序列,进行测序及同源性分析。[结果]用1010cfu/ml BH-1菌液接种2龄天牛幼虫8 d后致死率达72.7%。BH-1与GenBank收录的Serratia marcescens的16S rDNA序列的同源性达99.5%,同时结合细菌的部分常规鉴定结果,鉴定出BH-1菌株为粘质沙雷氏菌(S.marcescens)。[结论]BH-1的发现为天牛的生物防治提供了参考。