异色瓢虫成虫体型及体内脂肪含量对其耐寒能力的影响

为明确异色瓢虫Harmonia axyridis (Pallas)成虫体型及体内脂肪含量对其耐寒能力的影响, 本研究在2种发育温度(18℃和25℃)下通过表型可塑性诱导获得了不同体型的异色瓢虫成虫, 测定了成虫体型大小、体内脂肪含量、过冷却点(supercooling point, SCP)及其在恒定低温(constant low temperature, CLT)和变温(fluctuating thermal regime, FTR)下的存活率。结果表明: 较低发育温度(18℃)下的成虫体型比25℃下的明显大(P<0.01), 体内脂肪含量显著高于25℃下的(P<0.01); 且两2种发育温度下成虫体内脂肪含量与虫体干重呈较好的正相关关系。由SCP频次分布图可知, 饲养在18℃下的成虫SCP主要集中在-8 ~ -6℃范围内, 饲养在25℃下的成虫SCP主要集中在-10 ~ -9℃范围内; 且成虫体型大小与过冷却能力呈现负相关关系。经过相同时间低温暴露后成虫的存活率按下列顺序降低: FTR18℃＞FTR25℃＞CLT18℃＞CLT25℃。结果表明低温暴露过程中脂肪贮存对异色瓢虫成虫存活是非常重要的, 但是冷伤害是影响存活的基本因素。     

海藻糖酶基因TRE2-like和TRE2在异色瓢虫羽化过程中的表达与功能

【目的】异色瓢虫Harmonia axyridis是一种重要的捕食性天敌昆虫,海藻糖在异色瓢虫的变态发育、羽化等整个生命过程都起着重要的作用。本研究以前期获得的类似膜结合型海藻糖酶(TRE2-like)与膜结合型海藻糖酶(TRE2)基因为基础,探讨在异色瓢虫羽化阶段这两个海藻糖酶的潜在功能,为阐明异色瓢虫从蛹发育到成虫时海藻糖代谢机制提供参考。【方法】根据TRE2-like和TRE2基因序列设计双链RNA(dsRNA)区域片段并合成对应的dsRNA,通过RNAi将其注射到异色瓢虫2日龄蛹中。采用实时荧光定量PCR(RT-qPCR)检测RNAi处理后羽化第1天的异色瓢虫成虫糖代谢相关基因的表达;同时采用蒽酮比色法、酶标法等分别测定RNAi处理后羽化第1天的异色瓢虫成虫主要糖类物质含量及TRE活性变化,并观察异色瓢虫羽化后的表型变化。【结果】结果表明,与对照组(dsGFP注射组)相比,异色瓢虫2日龄蛹被注射TRE2-like或TRE2 dsRNA后,其新羽化成虫体内TRE2-like和TRE2表达量均极显著下调,且少数个体出现了蜕皮与翅形成困难等畸形表型。可溶性海藻糖酶活性在注射dsTRE2-like后显著降低,膜结合型海藻糖酶活性在注射dsTRE2后显著降低;注射dsTRE2后糖原含量显著下降,注射dsTRE2-like后糖原和海藻糖含量显著下降,注射dsTRE2-like+dsTRE2后糖原和葡萄糖含量显著下降,且海藻糖含量极显著下降。注射dsTRE2-like, dsTRE2和dsTRE2-like+dsTRE2后可溶性海藻糖酶基因TRE1-1和TRE1-2表达下降或显著下降,而TRE1-5表达上升或显著上升,海藻糖合成酶(trealose-6-phosphate synthase, TPS)、糖原磷酸化酶(glycogen phosphorylase, GP)、糖原合成酶(glycogen synthase, GS)基因的表达均显著下调。【结论】TRE2-like和TRE2基因表达被抑制后,异色瓢虫海藻糖等代谢受到影响。研究结果为探究异色瓢虫体内膜结合型海藻糖酶的潜在功能和调控机制奠定了基础。     

Trehalase in Harmonia axyridis (Coleoptera: Coccinellidae): effects on beetle locomotory activity and the correlation with trehalose metabolism under starvation conditions

Trehalose is important in the energy metabolism of insects and protects them from extreme environmental conditions. To study the synthesis and degradation of trehalose, which is hydrolyzed to glucose by trehalase, two soluble trehalase cDNAs of the beetle Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) (HaTreh1-1 and HaTreh1-2) were cloned. The HaTreh mRNA expression levels were higher during the larval than adult stages. The locomotory behavior of the beetle and its mRNA expression of HaTreh1-1 and HaTreh1-2 under starvation were then investigated. Sprint speed and maximum moving distance were increased, and the pause frequency decreased in H. axyridis adults starved for 8 h compared with control (0 h) adults. In contrast, the mRNA expression of both HaTrehs increased quickly in adults starved from 8 to 18 h, with HaTreh1-1 in particular being expressed 289-fold in adults starved for 18 h versus control. These results show that stored food reserves can provide energy to sustain vital activities for 8 h in H. axyridis adults. However, the beetles require trehalose and other energy sources to provide energy to find food and for vital activities when starved for more than 8 h.     

Dynamics of fat content during induction and termination of “trophic diapause” in Harmonia sedecimnotata Fabr. females (Coleoptera, Coccinellidae)

Experimental studies have demonstrated that onset, termination, and resumption of oviposition, same as corresponding changes in germarium size, are closely correlated with the dynamics of fat reserves. Even short-term (10 days long) termination of oviposition caused by the absence of aphid preys with the availability of carbohydrate food is accompanied by significant increase in absolute (weight) and relative (the percentage of total dry weight) fat content. This confirms the correctness of considering the studied phenomenon as a “real” reproductive diapause but not as a “short-term interruption of oviposition.” In combination with earlier published data, the results of the study once again suggest that division of insect adaptations to environmental changes into “short-term” and “long-term” adaptations based, correspondingly, on “neural” and “neurohormonal” mechanisms is rather conventional. In reality, a gradient or a spectrum of reactions is observed.     

Efficacy and nontarget effects of reduced-risk insecticides on Aphis glycines (Hemiptera: Aphididae) and its biological control agent Harmonia axyridis (Coleoptera: Coccinellidae)

The efficacy of three reduced-risk insecticides (pyrethrins, insecticidal soap, and narrow-range mineral oil) was determined for nymphs and adults of the soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), an exotic pest of North American soybean, Glycine max (L.) Merr. These insecticides also were evaluated for nontarget effects on one of the aphid's key biological control agents, multicolored Asian lady beetle, Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae), including first and third instars, pupae, and adults. A Potter Spray Tower was used to conduct direct spray laboratory bioassays. Results indicated that although pyrethrins and narrow-range mineral oil caused 100% mortality to A. glycines nymphs and adults at 72 h posttreatment, insecticidal soap caused equivalent mortality to only the nymphs during the same time period. However, A. glycines adult mortality due to the insecticidal soap (83.3%) was significantly greater than the control. Pyrethrins were highly toxic to first instars of H. axyridis (98% mortality), but they had no effect on third instars, pupae, or adults. Mineral oil and insecticidal soap were moderately lethal to first (48.9 and 40% mortality, respectively) and third (31.9 and 38.8% mortality, respectively) instars of H. axyridis, but they had no effect on pupae and adults. Our results suggest that pyrethrins, insecticidal soap, and narrow-range mineral oil may prove useful for soybean aphid management in organic soybean due to efficacy against the aphid with differential nontarget effects on select stages of H. axyridis. Additional studies will be necessary to elucidate the efficacy of these insecticides under field conditions.     

Dropping behaviour of larvae of aphidophagous ladybirds and its effects on incidence of intraguild predation: interactions between the intraguild prey, Adalia bipunctata (L.) and Coccinella septempunctata (L.), and the intraguild predator, Harmonia axyridis Pallas

Abstract.  1. Two experiments were performed in the laboratory to assess the behaviour of dropping from a host plant as a defence against intraguild predation in aphidophagous ladybird larvae.
2. In the first experiment, encounters were observed on bean plants between fourth instars of the intraguild predator species, Harmonia axyridis , and first instars of two other ladybird species, Adalia bipunctata (L.) and Coccinella septempunctata (L.). The percentages of first instars of the latter two species that dropped from the plant in response to attack differed dramatically, with 47.5% of C. septempunctata first instars dropping vs. 0% of A. bipunctata .
3. In the second experiment, first instars of A. bipunctata or C. septempunctata and a fourth instar of H. axyridis were allowed to forage together on bean plants for 3 h. During this time, 44.3% of C. septempunctata larvae dropped from the plant, but less than 2% of A. bipunctata larvae did so. In contrast, 95.0% of A. bipunctata larvae fell victim to intraguild predation by H. axyridis vs. only 54.5% of C. septempunctata larvae.
4. The significance of dropping behaviour of ladybird larvae as a defence against intraguild predation, and the relationship of dropping behaviour to species-specific habitat affinity of ladybirds, is discussed.     

Influence of feeding a fish oil-containing diet to young,lean, adult dogs: effects on lipid metabolites,postprandial glycaemia and body weight

The aim of this study was to determine the effect of feeding a fish oil (FO)-containing diet on lipid and protein metabolism, postprandial glycaemia and body weight in young, lean, adult dogs. Eight female Beagles were randomly assigned to one of two isonitrogenous and isoenergetic diets, Control or FO, in a crossover design. At the beginning of the experiment and at 30 and 60 d, a baseline blood sample was collected and the dogs then were fed their daily ration. Nitrogen balance began at 07:00 h on day 63 of each experimental period and ended at 07:00 h on day 69. On day 66 of each period, a single dose (7.5 mg/kg) of ¹⁵N-glycine was administered orally to each dog via gelatin capsule. Postprandial glycaemia did not differ between treatments or among sampling days within treatment. Cholesterol concentration was increased (p < 0.05) on the Control treatment throughout the experiment when compared to values of day 0. Dogs fed the FO treatment had higher plasma triglyceride and ghrelin concentrations than those fed the Control treatment. Body weight and food intake did not differ between dietary treatments. Faecal excretion was increased (p < 0.05) in the FO treatment. Dry matter digestibility was decreased (p < 0.05) and fat digestibility tended (p < 0.10) to decrease in the FO treatment. Overall, feeding a FO-containing diet showed a protective effect against the rise of plasma cholesterol and it increased plasma ghrelin concentration. However, FO supplementation did not appear to affect protein metabolism or postprandial glycaemia in adult lean dogs.