The potential impacts of the arrival of the harlequin ladybird, Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae), in Britain

Abstract.  1. The harlequin ladybird, Harmonia axyridis , has recently arrived in Britain.
2. This species has been introduced from Asia into many parts of the world for biological control purposes.
3. In many parts of North America it has become the predominant aphidophagous coccinellid in less than 20 years, and in north-western Europe it is spreading and increasing in number rapidly.
4. Since establishment in North America and continental Europe, reports of its effectiveness as a biological control agent of aphids and coccids have been accompanied by accounts of negative effects on other aphidophagous species and humans.
5. Here the potential impacts of the arrival of the harlequin ladybird in Britain are assessed.     

异色瓢虫生物生态学研究进展

原产于亚洲的异色瓢虫[Harmonia axyridis Pallas(Coleoptera:Coccinellidae)]是最重要的捕食性瓢虫种类之一,被广泛应用于农林业害虫的生物防治.本文综合分析了国内外近几十年对异色瓢虫的研究成果,对其重要的生物学及生态学特性进行了归纳,主要涉及生活史、繁殖行为及策略、捕食行为和自残行为等,分析了相关研究对其应用前景的影响.基于异色瓢虫的人工饲养、农药交互作用以及入侵生态影响等的总结,提出了预防异色瓢虫造成生态不平衡的具体措施.     

Weak photoperiodic response facilitates the biological invasion of the harlequin ladybird Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae)

Photoperiodic regulation of reproductive diapause in two invasive and two native populations of Harmonia axyridis and in one native population of Harmonia yedoensis was investigated in laboratory at 20°C, five photoperiods (day length of 10, 12, 14, 16 and 18 h) and two diets: (i) eggs of the Angoumois grain moth Sitotroga cerealella and (ii) the green peach aphid, Myzus persicae. Laboratory strains originated from native populations of H. axyridis from Irkutsk province of Siberia and H. yedoensis from South Korea showed a strong photoperiodic response: under short photoperiods (10–14 h and 10–12 h for H. axyridis and H. yedoensis, correspondingly), all females which fed on eggs and most of those fed on aphids did not start to lay eggs during 40 days after emergence, while under long photoperiods, all females fed on aphids and most of those fed on eggs oviposited. The photoperiodic response of H. axyridis from South Korea was less strong: on the both diets, the range of the photoperiodic response (the difference in the proportion of ovipositing females between the treatments with long and short days) was ca 40%. In the European (Czech Republic) and in the Caucasian (Sochi region, Russia) invasive populations of H. axyridis, the photoperiodic response was very weak: the proportion of females that started oviposition (when fed on aphids) or at least reproductive maturation (when fed on eggs) during 40 days after emergence was close to 100%, independently of the photoperiodic conditions. Obviously, instead of a rapid micro‐evolutionary adaptation of the critical day length to a new climate, the invasive populations of the harlequin ladybird decrease their dependence on photoperiod and thus the weak photoperiodic response of SE Asian population of H. axyridis can be considered as a pre‐adaptation further developed during the invasion.     

Role of photoperiod in the development and reproduction of Harmonia axyridis (Coleoptera,Coccinellidae)

To improve the laboratory mass rearing and the winter greenhouse use of the multicoloured Asian lady beetle Harmonia axyridis (Pallas) (Coleoptera, Coccinellidae), the effects of photoperiod on the development and fecundity of H. axyridis were investigated at 20°C and photoperiods ranging from 0L:24D to 24L:0D. We foun'd photoperiod to significantly affect the pre-imaginal development and reproduction of H. axyridis. The development of the pre-imaginal stage was significantly faster at shorter photoperiods (0–14?h) than at longer ones (16–24?h). The proportion of females laying eggs within the first 30 days after eclosion, the mean number of eggs per ovipositing female, and the long axial length of the first oocyte were all lower at shorter photoperiods (6–12?h) than at longer ones (14–24?h). These results suggest that adult H. axyridis may encounter problems in preying and reproducing in the winter conditions of unheated and unilluminated greenhouses in areas with temperatures lower than 20°C and day lengths shorter than 12?h.     

异色瓢虫幼虫对卵的种内自残和种间捕食

自残现象的发生在异色瓢虫Harmonia axyridis中很是常见,为明确自残行为对其存活和生长发育的影响,在非选择性条件下我们对异色瓢虫幼虫对非姊妹卵的种内自残和对七星瓢虫Coccinella septempunctata卵的种间捕食进行了研究。结果表明,异色瓢虫取食同种卵和异种卵均可完成生长发育,与取食蚜虫相比：①其从卵到成虫及蛹期的发育历期明显缩短（P<0.05）,其中1龄幼虫发育历期缩短非常明显（P<0.01）;②自然条件下高死亡率的1龄幼虫的存活率明显提高（P<0.05）;③2-4龄幼虫、蛹和成虫的体重变轻（P<0.05）,且体长在4龄幼虫间也表现出显著差异（P<0.05）。这表明异色瓢虫种内自残和种间捕食对其在食物恶劣条件下延续种群至关重要,但对种群的繁衍却不是最理想的途径。     

多巴脱羧酶对异色瓢虫生殖力的调控机制

[目的]为了探究多巴脱羧酶(DOPA decarboxylase,DDC)对异色瓢虫Harmonia axyridis生殖力的影响及其调控机制.[方法]利用RNA干扰技术(RNA interference,RNAi),将异色瓢虫4龄幼虫的DDC基因(HaDDC)抑制表达,成虫羽化后第8天开始统计20 d内累计产卵量及产...     

异色瓢虫越冬聚集行为对其能量代谢的影响

异色瓢虫以成虫滞育越冬,越冬期间聚集行为尤为显著。为探讨异色瓢虫越冬聚集行为的适应意义,本研究利用便携式红外线CO2分析仪测定了6 种聚集度（分别用1头、2头、5头、10 头、20头和50头表示）下异色瓢虫越冬成虫的呼吸量,并通过呼吸量计算得到各聚集度下的呼吸速率（Respiration rate, Rr）和能量代谢速率（Metabolism rate, Rm）。结果表明: 不同聚集度间异色瓢虫越冬成虫的呼吸速率和能量代谢速率均存在显著性差异,相关性分析表明Rr和Rm值与聚集度之间呈显著负相关。即随着聚集度的增加,越冬成虫的呼吸速率和能量代谢速率呈逐渐降低的趋势。此外,利用“Y”形嗅觉仪测定了异色瓢虫实验种群和越冬种群成虫对不同味源的选择趋性,表明异色瓢虫均对各自的群体表现出趋性,而其越冬聚集行为的发生需要一种特殊的气味物质。本研究表明聚集行为可以有效降低异色瓢虫的新陈代谢速率,减少能量消耗,有利于其种群成功越冬,这对其越冬来说具有非常重要的意义。     

异色瓢虫对蚜虫性外激素的行为反应研究

蚜虫性外激素假荆芥内酯(Z,E-Nepetalactone,NEP)能够吸引多种捕食性和寄生性天敌昆虫,它专一性强,生物活性突出,对非靶标生物安全。异色瓢虫Harmonia axyridis(Pallas)作为一种重要的捕食性天敌,被广泛应用于农业生产中。为明确异色瓢虫对蚜虫性外激素的行为反应,本研究在室内用"Y"型嗅觉仪测定了异色瓢虫成虫对不同剂量NEP的嗅觉行为反应;并在田间,每7 d释放100μL浓度为1 mg/m L的NEP,测定了NEP对异色瓢虫种群数量变动的影响。室内嗅觉测定试验结果表明异色瓢虫雌虫对10μL剂量NEP有显著的选择性,而雄虫对1μL剂量NEP有显著的选择性;田间试验发现,释放NEP的处理区异色瓢虫数量显著高于对照区。本研究为蚜虫性外激素在害虫绿色防控中的应用提供了依据。     

异色瓢虫卵黄蛋白单克隆抗体的制备及鉴定

【目的】为了能准确地追踪异色瓢虫Harmonia axyridis (Pallas)卵黄原蛋白（vitellogenin, Vg）的合成、转运途径和吸收方式,以及卵黄蛋白（vitellin, Vn）在卵母细胞内的积累及分布情况,本研究对异色瓢虫的Vn进行了单克隆抗体（monoclonal antibody, McAb）的制备。【方法】以异色瓢虫Vn免疫BLAB/C小鼠,应用杂交瘤技术,经过3次亚克隆筛选,制备能稳定分泌抗Vn的单克隆抗体。【结果】实验获得4株能够稳定分泌抗异色瓢虫Vn的单克隆抗体,即5E2, 5E11, 1E9和5H8。其中1E9, 5E11和5E2亚型均为IgG1,5H8亚型为IgM。Western blot免疫印迹分析显示,4株单克隆抗体可以特异性地识别Vn,而与雄虫血淋巴无反应。其中,5E2和1E9可以与异色瓢虫抗原的4个亚基发生较强的免疫反应,结合腹水制备前上清效价检测结果最终选取5E2制备单克隆抗体。5E2单克隆抗体的效价为1∶81 000,SDS-PAGE分析显示5E2重链和轻链的分子量分别为50和27 kD。【结论】本实验成功制备出一株能够稳定分泌抗异色瓢虫Vn的单克隆抗体,为建立酶联免疫吸附试验（ELISA）方法测定其动态变化奠定了基础。     

Ecology and biological control application of multicoloured Asian ladybird, Harmonia axyridis: A review

The ecology of and biological control by multicoloured Asian ladybird beetle, Harmonia axyridis (Pallas) are reviewed. Our emphasis is on assembling and interpreting information on general characteristics, invasion and establishment, sexual activity, foraging and predation, development, survival and reproduction, predator-predator interactions, natural enemies, biocontrol, non-target effects and status of H. axyridis as a pest of fruits. Colonization of H. axyridis for aphid biocontrol in the USA have been successful in terms of its establishment, but its abundance is turning out to be a nuisance to humans. Its harmful non-target impact on beneficial organisms, humans and native species is becoming a debatable issue. The question on its present position, whether it is a biocontrol agent or pest, is a critical issue and discussed. Inferences from the empirical data are made and new avenues for future research are suggested.     

异色瓢虫不同色斑型成虫的耐寒性研究

【目的】研究异色瓢虫Harmonia axyridi不同色斑型成虫的耐寒能力,以明确其色斑型比例所呈现的季节性变化的适应意义。【方法】本文测定了异色瓢虫实验种群和自然种群不同色斑型成虫的过冷却点(Supercooling point,SCP)及低温存活率。【结果】同一发育温度下异色瓢虫实验种群不同色斑型成虫的SCP和低温存活率均没有显著性差异(P>0.05),但是低温暴露中饲养在25℃下成虫的存活率下降更为剧烈。异色瓢虫自然种群黑底型与黄底型成虫的SCP呈现出明显的季节性变化,其SCP在7月份最高,分别为﹣8.2℃和﹣7.5℃;在1月份达到最低,分别为﹣16.8℃和﹣18.2℃。越冬开始(11月)黑底型成虫的SCP低于黄底型成虫的,整个越冬期间黄底型成虫的SCP一直低于黑底型成虫的,但是异色瓢虫黑底型与黄底型越冬成虫的低温存活率却没有显著性差异(P>0.05)。【结论】越冬期间异色瓢虫黄底型成虫的SCP低于黑底型的,这样可以通过体液过冷却的方式来避免结冰造成的伤害,以增强其越冬种群的耐寒能力,这也可能是越冬期间异色瓢虫黄底型成虫数量显著上升的一个原因。     

不同温度条件下异色瓢虫四龄幼虫亲缘关系对自残行为的影响

在14、18、22、24℃及30℃下对异色瓢虫Harmonia axyridis(Pallas)的不同同血缘的四龄幼虫自残行为及其对生长发育的影响进行研究。结果表明,异色瓢虫四龄幼虫自残发生概率受环境温度条件影响显著。在22-24℃处理范围内同血缘个体间自残发生概率均显著小于非同血缘个体。自残滞后时间同样受环境温度条件影响显著,且在同一温度条件下同血缘个体间均显著小于非同血缘处理。个体间相互攻击率不受环境温度变化影响,而在高于22℃下受血缘关系影响显著。此外,总受攻击次数在不同温度处理间差异显著。同血缘个体间自残后死亡率随温度增加而减少,均在30%以上。而非同血缘个体间自残后死亡率不受温度变化影响,均在70%以上。自残后残存个体的化蛹率及成虫羽化率均不受环境温度变化及个体间血缘关系的影响,化蛹率除30℃处理外均在90%以上,而在各温度下两血缘处理的羽化率均大于70%。     

Honeydew volatile emission acts as a kairomonal message for the Asian lady beetle Harmonia axyridis (Coleoptera: Coccinellidae)

Abstract The Asian lady beetle Harmonia axyridis Pallas is considered as an invasive species in most territories where it has been introduced. Because aphid honeydew acts as an attractant for many aphid predators and parasitoids, the objectives of this work were to collect and identify the volatile compounds released from the aphid excretory product to evaluate how these semiochemicals could affect the H. axyridis foraging behavior. Twelve volatile chemicals were identified from the Megoura viciae Buckton honeydew including four alcohols, three ketones, three aldehydes, a pyrazine and a monoterpene. The volatiles 3‐methyl‐1‐butanol and 3‐methyl‐butanal were highlighted as the two most abundant semiochemicals released from the M. viciae honeydew. Vicia faba L. plants treated with crude honeydew attracted more than 80% of the tested individuals with 40% of attracted beetles located on the plant. Four volatile compounds (3‐hydroxy‐2‐butanone, 3‐methyl‐butanal, 3‐methyl‐1‐butanol and limonene) were also highlighted to attract more than 75% of the coccinellids toward the odor source and to locate about 35% of them on the plants. Limonene was the most efficient attractant since 89% of the H. axyridis responded to this odor. The use of the identified semiochemicals as well as the composition of an artificial honeydew could certainly be helpful to control the dispersal of the Asian lady beetle H. axyridis.     

环境颜色对异色瓢虫生长发育及繁殖能力的影响

在室内利用不同颜色的饲养容器,对异色瓢虫Harmonia axyridis (Pallas) 生长发育、体长体重变化及繁殖能力进行了比较研究。结果显示：不同环境颜色条件对异色瓢虫幼虫各龄历期均有显著影响; 尽管蛹期受不同颜色影响显著,但其总胚后发育期在各处理间差异不显著。异色瓢虫1龄和2龄幼虫体长增量在不同颜色处理间无显著差异,但3龄幼虫在红色条件下体长增量显著小于其余各处理。不同环境颜色条件下异色瓢虫各虫期体重增量均有显著差异,而4龄幼虫体重增量在各处理中均显著大于其余各虫期,并占总胚后发育期体重增量的50%以上。异色瓢虫成虫的交配持续时间在各处理间无显著差异,但其产卵前期在黄色及绿色条件下显著小于其余各处理。其首堆产卵量在各处理间无显著差异,但在红色及自然光下的48 h累计产卵量均显著小于其余处理。     

冷驯化对异色瓢虫后代生长发育及适合度的影响

为明确冷驯化处理对异色瓢虫Harmonia axyridis后代生长发育及适合度的影响, 本研究通过两性生命表的制作, 在室内条件下调查了冷处理后其后代发育历期、 成虫产卵前期、 寿命和生殖力以及后代生命表参数。结果表明: 冷驯化使异色瓢虫后代卵的发育历期延长, 幼虫（1-4龄）和蛹的发育历期则缩短; 随着冷驯化时间的延长, 后代体长和体重增量均减小。且完成发育的后代成虫产卵前期延长, 寿命缩短, 生殖力下降。后代生命表参数内禀增长率（r）、 周限增长率（λ）、 净生殖率（R₀）和年龄特征存活率（l_x）均降低, 但是后代雌虫所占比例却升高。此外, 冷驯化类型对异色瓢虫后代生长发育的影响也不相同。经相同时间（5 d）的低温诱导, 变温诱导的后代成虫寿命比恒定低温诱导的长, 但是生殖力却低; 变温诱导的后代生命表参数（r, λ和R₀）均小于恒定低温诱导的, 但是l_x却高于恒定低温诱导的。结果说明, 异色瓢虫亲代经历冷驯化, 这种对低温的可塑性反应会延伸到下一代, 而且还能够影响后代的适合度, 这对其在低温下的存活和繁殖具有重要的意义。     

Nutritional plasticity of the predatory ladybeetle Harmonia axyridis (Coleoptera: Coccinellidae): comparison between natural and substitution prey

The predatory coccinellid Harmonia axyridis is a polyphagous species, efficient at controlling certain aphid species and already commercialized in Europe for that purpose. The complete development of this predator can be accomplished using the aphid Acyrthosiphon pisum or Ephestia kuehniella eggs as substitution prey. Biochemical analyses were conducted on the proteins, lipids, and carbohydrates of these two different prey species. E. kuehniella eggs were 2 times richer in amino acids than A. pisum adults (12% of the fresh weight vs. 6%). E. kuehniella eggs were 3 times richer in lipids than the aphids but, on the contrary, the aphids were 1.5 times richer in glycogen. The impact of these two kinds of food on the body composition of the coccinellid was evaluated to appreciate the degree of nutritional plasticity of the coccinellid. The composition of the coccinellids feeding either on E. kuehniella eggs or on aphids was compared for amino acid, fatty acid and glycogen contents, revealing a good capability of H. axyridis to develop on foods that are very different in their biochemical composition. Nevertheless, when fed on aphids, the crude protein content of the predator was reduced and the lipid content decreased by a factor of two, with a change in amino and fatty acid patterns. Some biological parameters, such as larval mortality, adult weight, and fecundity, were modified according to the food eaten.     

异色瓢虫海藻糖合成酶基因的克隆及低温诱导表达分析

海藻糖是昆虫的血糖, 在昆虫体内主要通过海藻糖合成酶（trehalose-6-phosphate synthase, TPS）催化合成。本研究通过同源克隆和cDNA末端快速扩增（rapid-amplification of cDNA ends, RACE） 技术, 从异色瓢虫Harmonia axyridis中克隆得到了TPS基因的cDNA全长序列, 命名为HaTPS（GenBank登录号： FJ501960）, 全长2 949 bp, 包含3′非翻译区为505 bp, 5′非翻译区为26 bp, 开放阅读框长2 418 bp, 共编码805个氨基酸。软件分析显示该基因编码蛋白的分子量为90.58 kD, 等电点为7.01, 包含两个糖基化位点, 无信号肽和跨膜结构。同源比对分析发现, 昆虫中TPS基因高度保守, 包含两个保守的结构域。同时, 采用实时荧光定量PCR技术对异色瓢虫HaTPS在不同发育阶段、 低温诱导条件下的表达量进行了研究。结果表明： HaTPS在预蛹期的表达量最高; 在短时低温诱导条件下, HaTPS的表达量随着温度的降低而显著升高, 在降温和升温处理条件下, HaTPS的表达量呈现先升高后下降的表达趋势。结果表明, TPS基因在昆虫抗逆中起到了重要的调节作用。昆虫经过低温诱导, 其TPS基因的调控能力得到提升。     

异色瓢虫不同地理种群的ISSR种群多样性分析

异色瓢虫Harmonia axyridis(Pallas)在亚洲地区是一种重要的捕食性天敌昆虫,但是当引入到欧美后,对当地的物种产生了威胁.本文用ISSR技术对两个中国本地种群和一个引入美国的种群进行了种群多样性以及种群分化的分析.8条ISSR引物共扩增出105条带,其中多态性条带比率为96.19%.Nei's多样性指数、Shannon信息指数、多态性条带比率,在中国种群与美国种群中均非常接近,种群分化不明显(G_(st)=0.042).AMOVA分析显示种群多样性主要来源于种群内部,种群问的基因流非常强(N_m=5.537).基于Nei's多样性构建的聚类结果显示ISSR是一种快速有效的研究异色瓢虫种群多样性关系的技术.     

异色瓢虫与隐斑瓢虫的区别及其色斑型和横脊的频率

异色瓢虫Harmonia axyridis(Pallas)和隐斑瓢虫H.yedoensis(Takizawa)是亲缘关系很近的姐妹种,在中国(北京到南岭,西至甘肃南部和西藏东部)、日本(北海道以南)和南韩等地同域分布,常常在松树等植物上共存。长期以来,隐斑瓢虫被认为是异色瓢虫的一个异名。有时从外形上很难区分这2种瓢虫,因此对于依据野外数据,分析异色瓢虫色斑型和鞘翅横脊的发生频率及小进化会产生一些错误。本文从没有隐斑瓢虫分布的东北地区的材料及其他数据,报道异色瓢虫的色斑型及鞘翅横脊发生频率。异色瓢虫的花斑型在我国东部地区(从东北至广东北部,西至甘肃和云南)发生率很低,而在新疆西北部发生率较高。鞘翅横脊的发生率从东北佳木斯的98.85%降低到云南大理的78.26%。本文列出了区分这2种瓢虫的形态特征及自然分布。如果鞘翅具横脊,则属于异色瓢虫,但如果标本来自2种瓢虫的共存区,鞘翅没有横脊,鞘翅的斑纹呈花斑型、四窗型或二窗型,则很难从外部形态上对它们进行鉴定。但这2种瓢虫的幼虫很容易区分。本文还提供了2种瓢虫的雄性外生殖器形态图、幼虫和成虫。