白星花金龟捕食落叶松毛虫蛹

白星花金龟Potosia brevitarsis Lewis属鞘翅目金龟甲科的一种杂食性昆虫。近年来作者在对落叶松毛虫的调查中发现,白星花金龟对该害虫种群数量的调控起着不可忽视的作用。     

白星花金龟成虫肠道细菌的分离鉴定

目的自星花金龟是我国北方常见腐食性昆虫,为了利用其腐食性行为转化处理有机垃圾、畜禽粪便及农作物秸杆等农业有机废弃物资源,并探索其中肠消化的微生态机制,而对其肠道细菌进行系统研究。方法从白星花金龟成虫肠道中无菌操作分离获得5株细菌,对其菌体形态、培养性状、染色反应、生理生化反应等进行鉴定及数量测定。结果白星花金龟成虫肠道的5个细菌菌株分别属于气球菌属（Aerococcus）、短状杆菌属（Brachybacterium）、棍状杆菌属（Clavibacter）、李斯特菌属（Listeria）、皮杆菌属（Dermabacter）,以皮杆菌属和棍状杆菌属数量最多,分别为1．3×10^7和1．0×10^7;其次为短状杆菌属和气球菌属,分别为8×10^6和5×10^6;李斯特菌属数量最少,为7×10^5。结论各菌株之间细菌数量存在明显差异,以皮杆菌属细菌数量最多,分离未获得真菌和放线菌。     

白星花金龟人为条件生物学与应用潜力

本文选取人工饲养的东亚飞蝗虫砂作为白星花金龟Potosia brevitarsis Lewis幼虫饲料、成虫产卵基质进行人为条件下的生物学研究。测定了其在不同温度条件下各虫态发育历期、不同食物下成虫单雌产卵量、不同酵化周期条件下东亚飞蝗虫砂2龄、3龄幼虫的转化力;分析了幼虫虫体和虫砂的主要营养成分;实施了虫砂盆栽小白菜试验。结果显示,白星花金龟适宜的繁育温度为22~28℃,25℃全世代发育历期为126.62(±1.05)d;以11种水果为食料,成虫单雌产卵量顺序为:芒果>菠萝>葡萄>苹果>桃>梨>西瓜>甜瓜>李子>杏>西红柿,最高207粒,最低68粒;东亚飞蝗虫砂酵化周期与取食龄期的最佳对应组合分别为2龄30 d和3龄25 d,折合每转化100 g可增长2龄虫体3.04 g、产生虫砂81.90 g或3龄虫体4.37 g、虫砂74.48 g;3龄幼虫含蛋白质(53.8%)、脂肪(6.6%)、18种氨基酸(44.68%),是昆虫源高蛋白原料;虫砂含有机质(18.72%)和氮磷钾(5.95%)。盆栽小白菜试验表明,1 cm^2施用0.13 g,效果最佳。水果残余物作为白星花金龟成虫食物,东亚飞蝗虫砂作为成虫产卵基质和幼虫繁育饲料,保持适宜温光条件,可以实现白星花金龟周年繁育。白星花金龟虫体和虫砂在生态循环农业、医药等领域具有应用潜力。     

不同补水方式对白星花金龟水分平衡的影响

为研究水分散失和水分补充对新疆新害虫白星花金龟Potosia brevitarsis Lewis的影响,在30℃恒温条件下,采用重量法测定白星花金龟脱水过程以及不同补水方式下(补蒸馏水;补盐水;补糖水)体内水分含量的变化,并与步甲、拟步甲的水分代谢进行比较。结果表明,白星花金龟的脱水与拟步甲科的网目拟地甲Opatrum subaratum Fald相似,在10h内脱水率均约为5%,而斑步甲Anisodactylus signatus 10h内脱水率约为20%。不同补水处理后,白星花金龟体内含水量迅速增加,随后继续脱水;脱水10d后,补蒸馏水、补盐水和补糖水处理的白星花金龟脱水率分别为28%,27%和21%,而未补水白星花金龟的脱水率为34%;未补水处理的甲虫脱水率和补蒸馏水和补盐水处理之间无显著性差异,而和补糖水处理之间存在显著性差异。未补水、补蒸馏水、补盐水和补糖水处理的LT50分别约为9,12,13和17d,补糖水能有效延长甲虫的存活时间。糖能有效地增加白星花金龟体内含水量,对维持其体内水分平衡起重要作用。     

不同农业有机废弃物对白星花金龟生物学特性影响研究

【目的】白星花金龟Protaetia (Liocola) brevitarsis (Lewis)幼虫取食农作物秸秆、畜禽粪便等农业废弃物,在自然界中发挥着净化环境、变废为宝的作用。【方法】本文选用8种农牧业有机废弃物,包括腐熟沼渣、平菇菌糠、猪粪、牛粪、玉米秸秆、25%玉米秸秆+牛粪、50%玉米秸秆+牛粪、75%玉米秸秆+牛粪作为饲料,研究不同农业有机废弃物对白星花金龟生物学特性的影响。【结果】沼渣组和平菇菌糠组单雌产卵量较高,分别为130.17粒和117.00粒,显著高于猪粪、牛粪和玉米秸秆处理;在牛粪与玉米秸秆混配组,各处理单雌产卵量为101.50-108.67粒,不同混合比例间无显著差异。从幼虫孵化率比较,平菇菌糠组和猪粪组幼虫孵化率较高,分别为87.78%和82.22%;玉米秸秆及其与牛粪不同比例混合处理的幼虫孵化率均很低,仅为46.67%-57.78%。从幼虫化蛹率、羽化率比较,玉米秸秆添加不同比例牛粪处理组的化蛹率和羽化率均较高,分别达到90.00%-95.56%和88.89%-91.11%;沼渣、猪粪等单一物质处理的化蛹率和羽化率均较低,分别低于90%和70%。单一物质处理中,沼渣、平菇菌糠、牛粪、玉米秸秆处理组羽化后均为雌虫比例大于雄虫比例。玉米秸秆与牛粪混配组中,仅50%玉米秸秆+牛粪处理组为雌虫比例大于雄虫比例;雌、雄成虫的体长及体宽均在(17-19)mm、(10-11)mm之间,沼渣、猪粪和牛粪处理组体长略小。以不同农业有机废弃物饲喂的白星花金龟全世代发育历期中,牛粪饲喂组白星花金龟历期最短,为(315.29±0.55)d;沼渣饲喂组白星花金龟历期最长,为(330.75±1.76)d。【结论】不同农业有机废弃物均可作为白星花金龟繁育基质,并可为其幼虫转化处理农业有机废弃物提供理论依据。     

白星花金龟绿僵菌分离及毒力测定

目的 2008年在山东泰安市黄庄堆肥中发现白星花金龟幼虫被病原菌寄生死亡,病虫体上长满灰绿色霉层,对其进行分离,经纯化获得1株真菌,编号为M-08。通过对幼虫接种,证明其致病性。方法对分离纯化出的菌株M-08,根据菌体形态特征、培养性状,按常规方法鉴定。结果 M-08菌株鉴定结果为金龟子绿僵菌长孢型变种Metarhiziumnaisopliae[Metsch Sorokin Var major（Johnston）Tulloch],毒力测定结果表明,对白星花金龟幼虫致死率高达93.3%~100%。结论金龟子绿僵菌长孢型毒力测定表明具有很高的毒性,今后有良好的应用前景。     

产卵基质模式和性比对白星花金龟生殖力的影响

[目的]为探究产卵基质模式和雌雄比对白星花金龟生殖力影响,寻求提高白星花金龟繁殖力的技术参数.设计复合产卵基质模式的思想,可为其他产卵于固体基质中的昆虫提高生殖力提供借鉴.[方法]本研究设计了5种产卵基质模式(牛粪、细沙单一基质、粪沙混、粪沙分和粪液沙复合基质)和5个雌雄比(早∶♂=4∶8、5∶7、6∶6、7∶5和8∶4),在室外敞口棚和自然温光条件下,开展对白星花金龟成虫产卵量和死亡率、幼虫孵化率的影响研究.[结果]5种产卵基质下白星花金龟成虫产卵量的顺序为粪沙分＞粪沙混＞牛粪＞粪液沙＞细沙,其中,白星花金龟在粪沙分复合产卵基质40 d的单雌产卵量达129.0粒,是原生产卵基质——牛粪的1.72倍;5个雌雄比处理比较,白星花金龟群体产卵量的顺序为7∶5＞8∶4＞6∶6＞5∶7＞4∶8;单雌产卵量的顺序为4∶ 8＞6∶6＞5∶7＞7∶5＞8∶4,最高达3.60粒/d.2项试验下成虫死亡率和幼虫孵化率在各处理间无显著差异,幼虫孵化率均在90％以上.综合分析和结合群体实际雌雄比可知,雌雄比为1∶1时,对于保障白星花金龟的群体繁殖力效果最佳.[结论]牛粪和细沙两复合体系作为产卵基质和维持雌雄比1∶1,可显著提高白星花金龟的繁殖力.     

白星花金龟聚集信息素的分离鉴定及引诱效果

[目的]明确白星花金龟Protaetia brevitarsis Lewis聚集信息素的有效成分,为白星花金龟的绿色防控提供支持.[方法]通过动态顶空法收集白星花金龟雌虫、雄虫和雌雄虫混合挥发物,用气相色谱-质谱(Gas chromatography-mass spectrometry,GC-MS)联用技术对收集的挥发物进行分析鉴定,并利用触角电位(Electroantennogram,EAG)仪和Y型嗅觉仪测定白星花金龟对8种化合物的触角电位和行为选择反应.[结果]白星花金龟雌虫、雄虫和雌雄虫混合挥发物中皆鉴定出8类15种化合物,包括6种烷烃类、2种酚类、2种芳香烃类、1种烯类、1种吡啶类、1种胺类、1种醇类和1种酮类.EAG试验结果表明,白星花金龟对8种化合物都有EAG反应,且不同挥发物间、同种挥发物不同浓度间的EAG反应存在差异.白星花金龟对蒎烷胺、3,4-二甲基-6-乙基苯酚、2,3-二甲基苯乙酮和2,6,10,14-四甲基十七烷4种挥发物的EAG反应相对值在1μg/μtL时达到最大,对萘、苯并噻唑、百里酚和正十四烷4种挥发物的EAG反应相对值在10 μtg/μtL时达到最大.行为选择反应表明,白星花金龟雌成虫、雄成虫对2,3-二甲基苯乙酮有明显趋向性,选择反应率大于70％,对2,6,10,14-四甲基十七烷、萘、苯并噻唑、百里酚和正十四烷5种物质均表现出显著或极显著的驱避效果,对3,4-二甲基-6-乙基苯酚和蒎烷胺表现出了性别上的差异,雄虫对3,4-二甲基-6-乙基苯酚表现出显著的趋向性,对蒎烷胺表现出显著的驱避性,而雌虫对这2种挥发物却无明显反应.[结论]2,3-二甲基苯乙酮对白星花金龟雌成虫、雄成虫有明显的引诱效果,2,6,10,14-四甲基十七烷、萘、苯并噻唑、正十四烷和百里酚有较强的驱避作用,推测2,3-二甲基苯乙酮是白星花金龟聚集信息素的有效组分.     

白星花金龟对寄主植物挥发物的电生理反应

[目的]白星花金龟Protaetia brevitarsis是我国经济上重要的农业害虫.本研究拟鉴定苹果、葡萄和玉米3种果实在两种状态(健康与受白星花金龟成虫为害)下的挥发物中能引起白星花金龟雌成虫触角电生理反应的挥发物成分.[方法]采用动态顶空吸附法收集苹果、葡萄和玉米3种果实的挥发物,用气相色谱-触角电位联用技术(...     

中国白星花金龟地理变异的几何形态学分析及其新疆种群的入侵来源推断

【目的】定量比较中国及周边国家地区白星花金龟 Protaetia (Liocola) brevitarsis 不同地理种群的形态差异,重点探索新疆种群与其他地区种群间的形态差异,为植物保护和生产活动提供指导,丰富微进化理论。【方法】基于几何形态学的主成分分析（principal component analysis, PCA）和典型变量分析（canonical variate analysis, CVA）方法,对国内外30个地区共700头白星花金龟的鞘翅和前胸背板进行分析比较,研究白星花金龟不同地理种群的形态变异。【结果】白星花金龟前胸背板和鞘翅形态变异主成分分析中,新疆标本都明显聚在一起,但同时与新疆周边省份标本有部分重叠,与远离新疆省市有少部分重叠。新疆标本和南方地区种群标本明显分离。在各地区种群形态差异显著性检验中,基于马氏距离（Mahalanobis distance）和普氏距离（Procrustes distance）分析显示,新疆白星花金龟与远离新疆地区标本的前胸背板和鞘翅在马氏距离和普氏距离上均为显著性差异,而与其他地区标本的前胸背板和鞘翅的差异上,并非总是显著性差异。【结论】证实了前胸背板和鞘翅上的白斑可以作为白星花金龟地理种群变异的评价指标。就新疆白星花金龟而言,地理距离在一定程度上与形态距离相吻合。我们推测新疆白星花金龟是通过青海省北部以外的宁夏、内蒙古和甘肃三省入侵到新疆自治区,或者与上述三省该害虫的入侵来源一致。本研究所呈现的研究方式,在丰富微进化理论方面具有广阔的前景,且有望丰富分子生态地理学的研究内涵。     

Hepatoprotective and antineoplastic properties of Protaetia brevitarsis larvae

Protaetia brevitarsis larvae have been used as a traditional medicine to treat various liver diseases. Freeze‐dried powder and solvent partitioned fractions of Protaetia brevitarsis larvae, in this study, were examined for hepatoprotective and antitumor activities in vivo and in vitro. Oral administration of the powder reduced signs of acute and chronic liver injuries in diethylnitrosamine‐induced hepatotoxic mouse model. Various cancer cell types, but not normal primary hepatocytes, were killed by the hexane, ethyl acetate and butanol fractions, which were partitioned from larval extracts. Apoptosis and autophagy were primarily induced in cancer cells by hexane and ethyl acetate fractions, respectively, and can be the cytotoxic mechanisms of the fractions. Further analyses of the insect fractions could lead to identification of novel bioactive substances that can be exploited as therapeutics for various human liver diseases.     

白星花金龟(Protaetia brevitarsis)幼虫抗菌物质的分离纯化

以药用昆虫白星花金龟(Protaetia brevitarsis)幼虫为研究对象,通过Sephadex G-50、RP-HPLC等技术分离纯化天然抗菌活性物质.经初步鉴定,该系列抗菌活性物质对枯草芽孢杆菌(Bacillus subttilis)表现出较强的抑菌活性.用MALDI-TOF MS对样品进行分析,检测到相对分子质量分别为2254.045 9、2622.4438的两种抗菌活性物质.推测这些抗菌活性物质在白星花金龟幼虫抵御微生物的侵袭中起到重要作用.     

The effects of feed materials on the nutrient composition of Protaetia brevitarsis larvae

To find out the effect of feed materials on the nutrient composition of Protaetia brevitarsis larvae (Pbl), three different feed materials were prepared and fed to the Pbl populations. After the Pbl population reached the third instar stage, they were boiled and dried for the nutrient analysis. The results showed that the nutrient composition of Pbl varied in response to the feed materials. Therefore, it is critical to set a feed material standard to guarantee the efficacy of Pbl medicinal property.     

Male and female Popillia quadriguttata (Fabricius) and Protaetia brevitarsis (Lewis) (Coleoptera: Scarabaeidae) response to Japanese beetle floral and pheromone lures

Popillia quadriguttata (Fabricius), and Protaetia brevitarsis (Lewis) adults were captured with Japanese beetle, Popillia japonica Newman, sex attractant and floral lures at Changchun, China during July–August 2012. The floral lure (phenethyl propionate:eugenol:geraniol, 3:7:3) was attractive to male and female P. quadriguttata (AV: 1.2 ± 0.9; 1.1 ± 0.3; total: 2.3 ± 0.8), and was similar in attraction to the combination of the sex attractant (SA) [(R, Z)-5-(1-decenyl) dihydro-2(3H)-furanone] plus the floral lure for male (1.60 ± 0.2), female (1.30 ± 1.1) and total captures (2.9 ± 3.0). However, the SA alone captured only males in much higher numbers than when combined with the floral lure (10.0 ± 6.4). In a separate earlier test, the greatest number of P. quadriguttata males (12.5 ± 3.0), female (12.2 ± 1.5) and total captures (24.7 ± 2.5) was in yellow, laboratory-made, bottle traps. The floral lure also attracted female Pro. brevitarsis (10.0 ± 3.4), while the SA attracted only few male beetles (1.0 ± 0.2). The combination SA + floral lure captured similar females (11.0 ± 2.0) and total (14.2 ± 2.2) Pro. brevitarsis as the floral lure alone. Two butterflies, Colias erate poliographus (Motschulsky) and Pieris rapae (Linnaeus), were also attracted to the floral lure. These studies indicate a potential for replacing pesticides by using the Japanese beetle lures for monitoring and control of several insects in China, and that they would be useful in monitoring and eradication of two potential scarab pests, P. quadriguttata and Pro. brevitaris, in the United States and Europe.     

Purification and Properties of Ferritin from the Last Instar Larval Hemolymph of Protaetia brevitarsis (Coleoptera)

Ferritin, an iron storage protein, has been purified from the last larval hemolymph of Protaetia brevitarsis (coleoptera) by KBr density gradient ultracentrifugation and resource Q (anion exchange chromatography) using fast performance liquid chromatography (FPLC) system. The iron content of ferritin was determined by atomic emission spectroscopy and FereneS stain. Ferritin of P. brevitarsis is shown to have two different subunits presented on a SDSPAGE in normal (N) and ironinjected (I) hemolymph. SDS PAGE revealed that the ferritin consists of two major polypeptides of 27 and 28 kDa in normal hemolymph. Interestingly, however, 30 kDa subunit was substituted for 28 kDa when iron was injected into the hemolymph. Apporximate isoelectric points of 27 kDa, 28 kDa, and 30 kDa ferritin subunits were 6.7, 6.75, and 6.8, respectively. Ferritin of P. brevitarsis was detected by FereneS stain and confirmed by Western blotting using its polyclonal antibody. Other characteristics such as amino acid composition and Nterminal amino acid sequence were investigated. Amino acid composition of ferritin (N and I) was rich in alanine, glycine, glutamine or glutamic acid and serine, but poor in histidine, arginine, methionine and phenylalanine.     

Characterization of the Hemocytes in Larvae of Protaetia brevitarsis seulensis: Involvement of Granulocyte-Mediated Phagocytosis

Hemocytes are key players in the immune response against pathogens in insects. However, the hemocyte types and their functions in the white-spotted flower chafers, Protaetia brevitarsis seulensis (Kolbe), are not known. In this study, we used various microscopes, molecular probes, and flow cytometric analyses to characterize the hemocytes in P. brevitarsis seulensis. The circulating hemocytes were classified based on their size, morphology, and dye-staining properties into six types, including granulocytes, plasmatocytes, oenocytoids, spherulocytes, prohemocytes, and adipohemocytes. The percentages of circulating hemocyte types were as follows: 13% granulocytes, 20% plasmatocytes, 1% oenocytoids, 5% spherulocytes, 17% prohemocytes, and 44% adipohemocytes. Next, we identified the professional phagocytes, granulocytes, which mediate encapsulation and phagocytosis of pathogens. The granulocytes were immunologically or morphologically activated and phagocytosed potentially hazardous substances in vivo. In addition, we showed that the phagocytosis by granulocytes is associated with autophagy, and that the activation of autophagy could be an efficient way to eliminate pathogens in this system. We also observed a high accumulation of autophagic vacuoles in activated granulocytes, which altered their shape and led to autophagic cell death. Finally, the granulocytes underwent mitotic division thus maintaining their number in vivo.