河北地区熊蜂物种多样性与蜂群繁育特性

基于2005-2009年从河北地区（包括河北省、北京市和天津市）所采集的1893号熊蜂标本资料,结合中国科学院动物研究所馆藏记录,分析了河北地区熊蜂物种多样性和蜂群繁育特性. 结果表明：河北地区共有熊蜂属昆虫8亚属32种,其中河北省32种,北京市18种,天津市5种;西部太行山区、北部燕山山区和坝上高原地区熊蜂种类丰富度和多度较高;河北地区熊蜂的访花植物涉及到21科80种,其中,菊科、豆科和唇形科植物是大多数熊蜂种类访问的主要对象;小峰熊蜂、红光熊蜂、密林熊蜂、火红熊蜂和重黄熊蜂5种熊蜂群势强大,平均单群蜂的工蜂数量在110只以上,雄蜂数量在160只以上,子代蜂王数在30只以上;这5种熊蜂的繁育成群率均在50%以上,易于人工驯养,具有重要的传粉利用价值.     

Cryptic bumblebee species: consequences for conservation and the trade in greenhouse pollinators

Commercial greenhouse growers in both Japan and China are increasingly using reared orange-tailed bumblebees known previously as Bombus hypocrita Pérez as pollinators. Phylogenetic analysis of the DNA (COI) barcodes with Bayesian methods shows that this "species" is a long-standing confusion of two cryptic species. We find that the orange-tailed bumblebees in North China are actually part of the widespread Russian (otherwise white-tailed) B. patagiatus Nylander (as B. patagiatus ganjsuensis Skorikov, n. comb.), whereas the orange-tailed bees in Japan are true B. hypocrita. This situation has been further complicated because two other cryptic species from North China that were previously confused with the Russian B. patagiatus are now recognised as separate: B. lantschouensis Vogt n. stat. and B. minshanensis Bischoff n. stat.. As demand for pollination services by greenhouse growers inevitably increases, these bees are more likely to be transported between countries. In order to conserve genetic resources of pollinator species for their option value for future food security, we advocate preventing trade and movement of B. patagiatus from China into Japan and of B. hypocrita from Japan into China.     

甘肃麦积山风景区熊蜂物种多样性调查

熊蜂属昆虫是山区植物的重要传粉者,在保持生物多样性和维护生态系统平衡方面发挥着十分重要的作用。麦积山风景区位于甘肃东南部的西秦岭山区,地处北部温带黄土高原向南部亚热带河谷山地和西部高寒青藏高原过渡的地带,地理位置独特,生物资源丰富。为了探明该景区内熊蜂物种多样性现状,笔者于2007—2010连续4年对该景区的熊蜂属昆虫进行了系统调查。结果表明:2007—2010年,在该景区12个采样点共采集熊蜂属昆虫1 765头,隶属于7亚属20种;在12个采样点中,石门和净土寺的熊蜂种类丰富度最高;在20种熊蜂中,火红熊蜂Bombus pyrosoma、红光熊蜂B.ignitus、疏熊蜂B.remotus、小峰熊蜂B.hypocrita s.l.和重黄熊蜂B.picipes的物种多度较高,依次位居前5位;麦积山风景区熊蜂区系成分以东洋区+古北区共有种为主(55%),其中东洋区成分高于古北区成分;麦积山熊蜂区系成分和南部亚热带湿润气候区的白水江自然保护区以及西部高寒气候区的扎尕那自然保护区比较接近,和北部温带半干旱气候区的崆峒山风景区相距较远。     

小峰熊蜂蜂毒磷脂酶A2基因的克隆及表达分析

磷脂酶A2 （phospholipase A2, PLA2）是蜂毒主要成分, 也是蜂毒的主要过敏原, 在熊蜂个体和群体防御方面具有重要功能。为了探究熊蜂A2基因的生物学功能, 本研究以小峰熊蜂Bombus hypocrita为材料进行了蜂毒PLA2基因的克隆、 鉴定与表达特性分析。结果表明： 该基因全长为2 272 bp, GenBank登录号为KF214771, 由4个外显子和3个内含子组成, 编码区（CDS）长为543 bp, 共编码180个氨基酸残基。氨基酸序列相似性分析显示, 成熟的小峰熊蜂PLA2（含有136个氨基酸）与其他蜂类PLA2的氨基酸序列相似性较高, 均包含10个保守的半胱氨酸残基、 1个保守的Ca2+结合位点和1个酶活性中心。基于PLA2氨基酸序列的系统进化树分析表明, 熊蜂属Bombus与蜜蜂属Apis在不同分支上, 属单系群, 且蜜蜂属分化较早。荧光定量PCR结果表明, PLA2基因在小峰熊蜂各日龄均有表达, 且随日龄增长, 表达量呈先上升后下降的趋势, 10日龄时出现峰值, 其表达量显著高于其他日龄（P<0.05）。半定量PCR结果表明, PLA2基因在毒腺、 卵巢、 中肠中表达量较高, 在足、 触角、 食道腺中表达量较低, 在脂肪体、 肌肉、 神经、 气管、 复眼、 脑中未表达。本研究探明了小峰熊蜂PLA2的基因结构及其表达特性, 丰富了熊蜂PLA2的生物学基础, 为进一步深入研究熊蜂PLA2生物学功能和作用机制以及开发蜂毒生物制剂等鉴定了基础。     

不同蜂在温室草莓园的访花行为和传粉生态学比较

应用意大利蜜蜂、本土小峰熊蜂和引自欧洲的地熊蜂3种蜂在北京为温室草莓传粉,对其传粉行为和生态特性进行研究。结果表明:意大利蜜蜂的出巢温度、工作起点温度、平均单花停留时间和访花间隔时间均显著高于小峰熊蜂和地熊蜂(P<0.01),而这些行为在2种熊蜂之间差异不显著(P>0.05);3种蜂携带花粉回巢的比例依次为意大利蜜蜂>小峰熊蜂>地熊蜂,而其携带的花粉活性依次为意大利蜜蜂>小峰熊蜂=地熊蜂;2种熊蜂开始活动的时间比意大利蜜蜂早,日活动时间比意大利蜜蜂长;地熊蜂在一天中的活动频率最高(P<0.01);经3种蜂授粉的草莓果实营养品质差异不显著(P>0.05),与对照相比也无差异,而经过3种蜂授粉的草莓果实畸形率降低、单果质量增大;经2种熊蜂授粉的草莓果实畸形率低于意大利蜜蜂(P<0.05)。本土小峰熊蜂能够有效替代意大利蜜蜂或地熊蜂为温室草莓授粉。     

意大利蜜蜂和小峰熊蜂在温室桃园的传粉行为及其影响因素

意大利蜜蜂Apis mellifera ligustica和小峰熊蜂Bombus hypocrita是我国设施农业中的两种主要授粉昆虫, 二者的传粉方式和传粉效率因不同作物而各有不同。为了选用最理想的蜂种为温室作物授粉, 提高作物的授粉效率, 我们于2008-2010年连续3年在北京的温室桃园进行了意大利蜜蜂和小峰熊蜂的传粉行为及其影响因素的研究。结果表明, 两种蜂的日活动规律不同, 和意大利蜜蜂相比, 小峰熊蜂的活动起点温度低、 日工作时间长、 单花访问时间长, 采集蜂在温室内距离蜂巢不同距离之间扩散比较均匀。而意大利蜜蜂采集蜂的数量随授粉半径的增大而明显减少。温室内环境因子对两种蜂传粉活动的影响基本一致, 温度对两种蜂的传粉活动影响最大, 其次是湿度、 单花花蜜浓度和光照强度, 而单花花蜜体积对蜂活动无明显影响。本研究认为对于温室桃授粉应优先选用授粉性能稳定的小峰熊蜂, 并且适当调节温室内环境条件, 以提高其授粉效率。     

华北地区六种熊蜂的地理分布及生态习性

2005～2007年连续3年对华北地区小峰熊蜂Bombus hypocrita、密林熊蜂B.patagiatus、红光熊蜂B.ignitus、明亮熊蜂B.lucorum、火红熊蜂B.pyrosoma和重黄熊蜂B.picipes等6种熊蜂的地理分布和生态习性进行研究。结果表明,这6种熊蜂在华北地区的地理分布差异很大,火红熊蜂分布最广,重黄熊蜂分布最窄。6种熊蜂在垂直分布上也存在差异,红光熊蜂分布于较低海拔高度地区,明亮熊蜂分布于较高海拔高度地区,但在海拔1300～1800m范围内6种熊蜂都有分布。6种熊蜂中,火红熊蜂的年活动周期较长,为180天左右。小峰熊蜂、明亮熊蜂、密林熊蜂和重黄熊蜂的年活动周期居中。红光熊蜂的年活动周期最短,为140天左右。6种熊蜂采访植物涉及到17科63种,采访植物存在一定的偏好性,主要采访豆科、菊科、蔷薇科和唇形科等植物;而且,不同种熊蜂采集植物的种类也存在较大差异。     

Bumblebee commercialization will cause worldwide migration of parasitic mites

We investigated natural populations of three Japanese native bumblebee species to determine the status of infestation by a tracheal mite, Locustacarus buchneri, which we had earlier detected in introduced commercial colonies of the European bumblebee, Bombus terrestris. We also investigated mite infestation in commercial colonies of a Japanese native species, B. ignitus, which are mass-produced in the Netherlands and reimported into Japan. We detected the mite in both natural and commercial colonies of the Japanese species. Comparison of 555 bp sequences of the mitochondrial DNA (mtDNA) cytochrome oxidase 1 (CO1) gene from the mite showed that there were seven haplotypes, on the basis of combinations of substitutions at eight sites in the gene. The haplotypes of the mites in the Japanese native bumblebees and the haplotypes of the mites in B. terrestris did not overlap; however, mtDNA of mites detected in the commercial colonies of B. ignitus possessed the same sequence as a European haplotype. These results indicate that transportation of bumblebee colonies will cause overseas migration of parasitic mites of different origins.     

细胞色素b基因序列与11种熊蜂的系统进化

通过比较一段357b的细胞色素b的序列,分析了熊蜂属5亚属11种熊蜂系统发育关系。5亚属包括Bombus( 5种）,Thoracobombus(3种）,Mendacibombus(1种）,Fervidobombus(1种）和Pyrobombus(1种）,该序列有65个单变异态位点和71个简约信息多态位点,翻译成119个氨基酸序列后有45个氨基酸变异位点。根据P－距离构建的邻接树（NJ tree)和最大简约树（MP tree)都显示同样的结果：Mendacibombus (B.avinovielllus)分化最早;Fervidobombus(B.pensylvanicus)次之;Pyrobombus(B.impatiens)和Bombus形成姊妹群;Bombus亚属是单系群,其中B.ignitus在所研究的5个种中分化最早。     

Antifibrinolytic role of a bee venom serine protease inhibitor that acts as a plasmin inhibitor

Bee venom is a rich source of pharmacologically active substances. In this study, we identified a bumblebee (Bombus ignitus) venom Kunitz-type serine protease inhibitor (Bi-KTI) that acts as a plasmin inhibitor. Bi-KTI showed no detectable inhibitory effect on factor Xa, thrombin, or tissue plasminogen activator. In contrast, Bi-KTI strongly inhibited plasmin, indicating that it acts as an antifibrinolytic agent; however, this inhibitory ability was two-fold weaker than that of aprotinin. The fibrin(ogen)olytic activities of B. ignitus venom serine protease (Bi-VSP) and plasmin in the presence of Bi-KTI indicate that Bi-KTI targets plasmin more specifically than Bi-VSP. These findings demonstrate a novel mechanism by which bumblebee venom affects the hemostatic system through the antifibrinolytic activity of Bi-KTI and through Bi-VSP-mediated fibrin(ogen)olytic activities, raising interest in Bi-KTI and Bi-VSP as potential clinical agents.     

Morphological variation in relation to flower use in bumblebees

To understand resource partitioning in a bumblebee community, we analyzed various morphological characters. A total of 1269 individuals of six bumblebee species, Bombus ardens, B. hypocrita, B. diversus, B. ignitus, B. honshuensis and B. beaticola, were examined and principal component analysis showed that the bumblebee species were clearly differentiated. Glossa, prementa and head lengths were positively correlated with the second component, and a longer proboscis was associated with a narrower body, which may help bees to intrude into and access deep‐lying nectar sources. Bombus diversus, with a long proboscis and narrow body, preferred flowers with a long corolla tube, whereas B. hypocrita and B. ignitus, which have short proboscises and wide bodies, visited flowers with short corollas or dish‐shaped flowers. Two pairs of consubgeneric species that have similar morphological characteristics, B. ardens and B. beaticola, and B. hypocrita and B. ignitus, divided flower resources by habitat selection and seasonal partitioning. For resource partitioning among bumblebee species, not only morphology but also other factors, such as habitat and seasonal preference, flower use, foraging behavior, and interspecific interactions, are responsible.     

Distribution and forage use of exotic bumblebees in South Island, New Zealand

The rapid decline in bumblebee populations within Europe has been linked to habitat loss through agricultural intensification, and a consequential reduction in the availability of preferred forage plants. The successful introduction of four European Bombus species to the South Island of New Zealand from England (in 1885 and 1906) provides an opportunity to determine how important different forage plants (also introduced from the U.K.) are to two severely threatened European bumblebee species (Bombus ruderatus and B. subterraneus). In January 2003 we conducted a survey of bumblebee populations across 70 sites in the central and southern South Island, recording which plant species were being used as pollen and nectar sources for each Bombus species. All four bumblebee species showed a clear preference for plants of European origin. Only B. terrestris, the most polylectic species, was recorded feeding on native plant species. The longer-tongued bumblebees, B. hortorum, B. ruderatus, and B. subterraneus, foraged predominantly on just two plant species; Trifolium pratense for both nectar and pollen, and Echium vulgare for nectar. These plant species are now declining in abundance in the U.K. Our results provide support for the hypothesis that the loss of flower-rich meadows, particularly those containing populations of Fabaceae species with long corollae, is responsible for the decline of bumblebee species across Europe. Comparison with earlier bumblebee surveys suggests that long-tongued bumblebees may also be in decline in New Zealand, particularly B. subterraneus which is now very localised and scarce.     

红光熊蜂卵黄原蛋白基因的cDNA全长序列克隆和表达分析

红光熊蜂Bombus ignitus Smith是许多经济作物和野生植物的重要授粉昆虫之一。卵黄原蛋白基因(vitellogenin,Vg)在昆虫的生殖调控中和行为方面起到重要的作用,本试验对Vg基因全长cDNA的克隆和测序及在蜂王、工蜂和雄性蜂三型蜂中的表达分析得出:Vg基因的全长cDNA为5 481 bp,GenBank中的登录号为FJ913883,有一个完整的开放阅读框(ORF),编码1 772个氨基酸,N-末端的前16个氨基酸为一个信号肽。接近C-末端区域存在保守的GL/ICG基元,其后含有9个半胱氨酸,而且DGXR位于GL/ICG基元上游18个氨基酸残基处。其氨基酸序列与韩国的熊蜂B.ignitus和B.hypocrita相似性高达95%,与西方蜜蜂Apis mellifera的相似性达到51%。Vg的mRNA首先在蜂王蛹期的白眼蛹(Pw)时期出现,其表达量在蜂王整个蛹期发育过程中呈上升趋势,且在黑眼蛹(Pbd)时期达到最高,在成年蜂的脂肪体中的表达量仍在升高,甚至更高。Vg也在工蜂蛹期的白眼蛹(Pw)时期被检测到,然后在整个蛹期发育过程中呈现上升趋势,在刚羽化出房时达到高峰,Vg的mRNA水平随着成年蜂日龄的增加而增加,到15日龄时达到最高,然后呈现下降趋势。对于雄性蜂,Vg的mRNA虽然卵黄原蛋白基因的mRNA水平几乎在整个蛹期发育阶段都表达,但是表达水平非常低,只有在刚羽化出房时期表达水平较高。     

Toxicity of six commercially formulated insecticides and biopesticides to third instar larvae of mushroom sciarid, Lycoriella ingenua Dufour (Diptera: Sciaridae), in New South Wales, Australia

Abstract  Lycoriella ingenua is one of the major pests of cultivated mushrooms, Agaricus bisporus (Lange) Imbach . Insecticide resistance among mushroom sciarid populations has been reported from other countries, and there is a need to determine the toxicity of currently approved and potential pesticides to sustain control of mushroom sciarid populations in Australia. The present study investigated the toxicity of six commercial formulations of insecticides or biopesticides against third instar larvae of L .  ingenua using laboratory bioassays. Insecticide treatments were incorporated into the growing medium for sciarid larvae and the concentration of the pesticide, which killed 90% of the test population (LD₉₀) determined the efficacy of selected insecticides. Triflumuron was the most effective insecticide against L. ingenua with an LD₉₀ of 53.12 mg active ingredient (a.i.)/m² followed by cyromazine (LD₉₀, 179.68 mg a.i./m²) and diazinon (LD₉₀, 261.72 mg a.i./m²). Abamectin and Bacillus thuringiensis ssp. israelensis were ineffective against L. ingenua . Steinernema feltiae , an entomopathogenic nematode, reduced the number of third instar larvae of L. ingenua only when applied at a higher rate (LD₉₀, 732 422 nematodes/m²) than was recommended on the label.     

Molecular cloning and fibrin(ogen)olytic activity of a bumblebee (Bombus hypocrita sapporoensis) venom serine protease

Although several bee venom serine protease genes have been previously described, fibrin(ogen)olytic activity of these serine proteases has been reported for only two bumblebees to date, Bombus ignitus and B. terrestris. Here, we cloned venom serine proteases from the other bumblebee species, B. hypocrita sapporoensis and B. ardens ardens. The venom serine protease genes of B. h. sapporoensis and B. a. ardens consist of 358 amino acids and 357 amino acids, respectively. We compared the predicted mature protein sequences of these serine protease genes to those previously reported for other bees. A phylogenetic analysis shows that B. h. sapporoensis venom serine protease is further immediately close to B. ignitus and B. terrestris venom serine proteases, excluding the venom serine protease of B. a. ardens. Using B. h. sapporoensis venom serine protease (Bs-VSP), we identified that Bs-VSP acts as a fibrin(ogen)olytic enzyme. We also found that Bs-VSP activates prothrombin and directly degrades fibrinogen into fibrin degradation products. Our results further define roles for bumblebee venom serine proteases as fibrin(ogen)olytic agents.     

Induction of colony initiation by Japanese native bumble bees using cocoons of the exotic bumblebee Bombus terrestris

The colony initiation rates of Bombus hypocrita (a native Japanese bumblebee) and Bombus terrestris (a European species) foundresses were compared after 4 weeks of exposure to B. terrestris cocoons. The B. terrestris cocoons, when replaced weekly, were effective for inducing oviposition by foundresses of both species. There were no significant differences in the colony initiation rates of B. terrestris and B. hypocrita, either with the control treatment or with the cocoons. The cocoon method was also tested for five species and two subspecies of native Japanese bumblebees. The colony initiation rate was higher for foundresses of the subgenus Bombus s. str. than for foundresses of the subgenera Pyrobombus, Diversobombus, and Thoracobombus. When replaced weekly, the cocoons of B. terrestris are effective inducers of colony foundation in three Japanese native species, namely B. ignitus, B. hypocrita hypocrita, and B. h. sapporoensis.     

Bumblebee Venom Serine Protease Increases Fungal Insecticidal Virulence by Inducing Insect Melanization

Insect-killing (entomopathogenic) fungi have high potential for controlling agriculturally harmful pests. However, their pathogenicity is slow, and this is one reason for their poor acceptance as a fungal insecticide. The expression of bumblebee, Bombus ignitus, venom serine protease (VSP) by Beauveria bassiana (ERL1170) induced melanization of yellow spotted longicorn beetles (Psacothea hilaris) as an over-reactive immune response, and caused substantially earlier mortality in beet armyworm (Spodopetra exigua) larvae when compared to the wild type. No fungal outgrowth or sporulation was observed on the melanized insects, thus suggesting a self-restriction of the dispersal of the genetically modified fungus in the environment. The research is the first use of a multi-functional bumblebee VSP to significantly increase the speed of fungal pathogenicity, while minimizing the dispersal of the fungal transformant in the environment.