Inhibition of vitellogenin synthesis in Apis mellifera workers by a juvenile hormone analogue,pyriproxyfen

Insect juvenile hormone (JH) has been related to modulation of vitellogenin (Vg) synthesis, a protein produced by fat body cells, secreted in haemolymph and sequestered by developing oocytes. A stimulatory JH action has been described for the majority of species studied thus far. In some insects, however, Vg synthesis has been inhibited or unaffected by JH. The aim of this study was to re-examine the action of JH on Vg synthesis in Apis mellifera workers, since contrasting effects of this hormone were described. Newly emerged worker bees were treated with different doses of pyriproxyfen (PPN), a potent JH analogue. Vg and total protein were quantified in haemolymph samples of newly emerged up to 6-day-old worker bees. Protein synthesis activity of fat body cultured in vitro and ultrastructure of fat body cells were also examined. High doses (1.25, 2.5, 5 and 10 &mgr;g) of PPN inhibited the onset and accumulation of Vg in the haemolymph of young worker bees in a dose-dependent fashion. This inhibition was not a result of fat body cell degeneration or death, as illustrated by fat body cells ultrastructure analysis, but by impairing Vg synthesis, as demonstrated by in vitro culture of fat body cells. Low doses (0.001, 0.01 and 0.1 &mgr;g) neither affected the normal synthesis and secretion of Vg into the haemolymph nor caused an early onset of Vg in treated bees (which could be interpreted as a JH-activating effect), as shown by Vg quantification at 24-h intervals. The results suggest that a low JH titre in honey bee workers permits the onset and accumulation of Vg in haemolymph, whereas high JH levels turn off Vg synthesis.     

Vitellogenin synthesis in the lady beetle Coccinella septempunctata

To elucidate the hormonal mechanisms which regulate reproduction in a beneficial insect, we have begun to investigate the process of vitellogenesis in Coccinella septempunctata, the seven-spotted lady beetle. Vitellin (Vn) constitutes 60–70% of the total protein in C. septempunctata eggs and is composed of four polypeptides with molecular weights determined by electrophoresis in denaturing gels of 133,000, 130,000, 46,000 and 43,000. In the egg these polypeptides occur in a ratio of approx. 1:1:1:2. The two larger Vn polypeptides yielded similar peptide fragments upon partial proteolytic digestion which suggests that they are structurally related. The two smaller Vn polypeptides appear structurally distinct because they yielded unique proteolytic fragments. The Vn precursor, vitellogenin (Vg), was observed in the haemolymph of mature females, but was not detected in the haemolymph of immature females or males. The electrophoretic mobilities, antigenicity, and proteolytic fragmentation patterns of the Vg polypeptides were indistinguishable from those of their Vn counterparts. Thus the major processing of the Vn polypeptides appears to occur prior to their secretion into the haemolymph.The synthesis of Vg was examined in whole animals and in organ cultures. Vg synthesis was observed in the fat body and to a smaller extent in the ovaries of mature females. The newly synthesized Vg was rapidly secreted. Vg synthesis was not detectable in brain or thoracic muscle of mature females or in the fat body of males or immature females. Very little vitellogenin synthesis occurred in female insects reared on artificial diets. The topical application of a juvenile hormone analogue induced Vg synthesis in non-fecund females but not in males.     

Control of vitellogenin synthesis in the Monarch butterfly by juvenile hormone.

Yolk formation in the Monarch butterfly, as in many other insects, entails juvenile hormone-induced synthesis in the fat body, transport in the hemolymph, and uptake into the oocytes of specific sex-limited protein, the vitellogenin. In the Monarch, vitellogenin first appears in the hemolymph 1 day after emergence of the adult butterfly and then rises rapidly in concentration for at least 3 days. The synthesis of vitellogenin in adult females was shown by the incorporation of [³H]leucine. Active labeling was observed at the same time as the presence of vitellogenin in the hemolymph was first detectable, and the radioactivity in vitellogenin expressed as a percentage of radioactivity in total blood proteins reached 50–60% 3 days after adult emergence. The appearance of vitellogenin was prevented by ligature of freshly emerged butterflies at the neck and restored by injection of juvenile hormone. A low yet significant stimulation of vitellogenin synthesis could be detected as early as 10 hr after administration of the hormone into neck-ligated butterflies, and after 30 to 50 hr about 40% of the total blood protein label was found in the specific protein. With C₁₈-juvenile hormone (JH-I), 0.1 μg per animal was effective in inducing a low level of vitellogenin synthesis, and between 0.01 and 1 μg, a linear relationship between synthesis and log dose was observed. The synthetic analog ZR-512 was also active. In a preliminary experiment, actinomycin D effectively blocked the induction of vitellogenin synthesis.     

七星瓢虫成虫对烟蚜的捕食作用

对七星瓢虫CoccinellaseptempunctataL .成虫对烟蚜Myzuspersicae的捕食作用进行了定量研究。七星瓢虫成虫对烟蚜的功能反应属HollingⅡ型反应 ,拟合的圆盘方程为Na =1 .1 5 76Nt ( 1 +0 .0 0 3 48Nt) ,χ2 检验表明圆盘方程理论值与实测值相符。捕食选择试验表明在烟蚜、烟青虫Helicoverpaassaut(Guenee)卵和 1龄幼虫 3种猎物中 ,七星瓢虫成虫最喜好烟蚜。七星瓢虫成虫对自身密度的功能反应用Watt模型拟合为A =2 90 .0 3P-0 .7584,其捕食作用率与个体间相互干扰作用的关系用Hassell模型拟合为E =0 .8783 7P-0 .1 0 0 94。文中进一步就烟田中七星瓢虫的保护利用进行了讨论。     

七星瓢虫卵的低温贮藏条件研究

为了探究七星瓢虫卵的最适低温贮藏条件,本试验研究了七星瓢虫卵在5、7、9和11℃条件下,分别贮藏10、 20和30 d的孵化情况,结果表明七星瓢虫卵在11℃贮藏10 d条件下孵化率最高,可以达到50.67%,进一步分析最适的低温贮藏条件11℃贮藏10 d对七星瓢虫种群适合度的影响。结果显示11℃贮藏10 d对七星瓢虫化蛹率无显著影响。以豌豆修尾蚜为猎物,对其幼虫和成虫的日捕食量进行检测,结果表明七星瓢虫的卵经11℃贮藏10 d后,各个龄期幼虫和成虫的日捕食量与对照组相比无显著差异。但经11℃贮藏10 d后七星瓢虫的发育历期显著缩短,羽化率、产卵量以及子代孵化率显著降低。上述研究结果对利用低温环境提高七星瓢虫的贮藏效率提供一定的理论基础。     

七星瓢虫、大草蛉对桃粉蚜捕食功能研究

经测定,七星瓢虫成虫、4龄幼虫和大草蛉3龄幼虫对桃粉蚜的捕食功能反应属于Holling—Ⅱ型圆盘方程式,两种天敌对桃粉蚜具有较强的捕食能力。     

七星瓢虫对不同初始密度棉蚜种群的调控作用

为明确七星瓢虫成虫对棉蚜的自然控制能力,本文通过密度-天敌双因素控制试验分析了不同棉蚜初始密度(200头/株、400头/株、800头/株)和不同七星瓢虫数量(0头、1头、2头、4头)组合处理下棉蚜的种群动态;使用棉蚜种群增长率、益害比、控制效果等指标评估了天敌的控制能力,并建立简单的模型来明确天敌有效控制棉蚜的益害比....     

The nature of the changes in the pattern of RNA synthesis by the juvenile hormone analogue altosid

The effect of various concentrations of Altosid and actinomycin D under defined conditions on housefly metamorphosis was investigated with three strains of houseflies. The morphogenetic response varied with the strains and the length of time which the larvae were exposed to the juvenile hormone analogue. De novo RNA synthesis was studied with (2-¹⁴C)-glycine. Methods were developed for the isolation of nuclear, soluble, and ribosomal RNA. The procedure presented provides a DEAE-cellulose chromatographic method for the removal of high molecular weight RNA from DEAE at a neutral pH. Labelling of the RNAs was increased in the presence of the juvenile hormone analogue indicating an increase in the rate of RNA synthesis. The higher incorporation of the labelled precursor into nuclear RNA demonstrates that cytoplasmic RNA is derived from the nuclei.     

Avoidance of a generalist entomopathogenic fungus by the ladybird, Coccinella septempunctata

Fungal entomopathogens are ubiquitous within the environment and susceptible insects are predicted to be under strong selection pressure to detect and avoid virulent isolates. Beauveria bassiana is an entomopathogenic fungus with a wide host range including coccinellids. Seven-spot ladybirds, Coccinella septempunctata, overwinter predominantly in leaf litter and B. bassiana is one of their major mortality factors during winter in temperate regions. Behavioural assays were conducted to assess the ability of adult C. septempunctata to avoid lethal densities of B. bassiana conidia in soil or on leaves. Further assays considered avoidance by C. septempunctata of mycosed (B. bassiana) C. septempunctata cadavers compared with uninfected C. septempunctata cadavers or in vitro B. bassiana. Treatments in any bioassays entirely avoided by C. septempunctata were regarded as censored data, to overcome the difficulties associated with zeros in log-ratio analyses. Male and female C. septempunctata avoided contact with leaf surfaces and soil inoculated with B. bassiana and mycosed cadavers. The ability of C. septempunctata to detect and avoid B. bassiana conidia is an adaptation that undoubtedly increases survival and ultimately fitness. We predict that such behavioural responses are widespread and driven by the high cost of fungal infection to a host.     

保幼激素合成的研究进展

保幼激素 (juvenile hormone,JH) 是通过甲羟戊酸途径合成的一类倍半萜化合物。以昆虫中普遍存在的JH Ⅲ为例,从分子水平上概述了JH合成途径中的各种酶,并对其中的两个关键酶：羟甲基戊二酰辅酶A还原酶和保幼激素酸甲基转移酶作了详细介绍。还从家蚕基因组数据库(http://silkworm.genomics.org.cn)中推测出了JH合成途径中大部分酶的编码基因,初探了JH合成的调节机制,讨论了JH合成的研究趋势。     

Flight potential and oxygen uptake during early dormancy in Coccinella septempunctata

Two flight parameters (take-off and duration) and respiration level were measured, in two years in summer and early autumn, in dormant Coccinella septempunctata L. (Coleoptera: Coccinellidae) collected while hidden in grass tussocks in hibernation sites (HID) and in beetles collected on plants (PLA). The duration of tethered flight of HID beetles measured in the laboratory in late August and September 1995 (range of geometric means 190–440 s) was slightly longer than the flight of PLA beetles (80–310 s), both being much longer than trivial flight recorded in beetles foraging for prey during the breeding season (35 s). In general, the flight performance had a tendency to increase in September and to decrease in October.The oxygen consumption in HID beetles increased throughout September 1994 from 430 to 780 l g^–1 h^–1 and throughout October 1995 from 710 to 1060 l g^–1 h^–1. This increase is ascribed to a concomitant decrease in diapause intensity. A similar increase was observed also in PLA beetles in 1994 and oxygen consumption was always higher than in HID beetles, most probably due to feeding and digestion in PLA beetles.Laboratory feeding of HID beetles on aphids induced maturation of ovaries and increased oxygen uptake (from 680 to 1110 l g^–1 h^–1). Feeding on honey and pollen left their oxygen uptake unchanged. Effect of feeding on the flight parameters was mostly not significant. In agreement with its less suitable body shape and usually less distant dormancy sites, C. septempunctata was found a less apt flier than long-distance migrating coccinellid species.     

七星瓢虫成虫对绣线菊蚜捕食作用的室内测试

在实验室内测定了七星瓢虫成虫对苹果绣线菊蚜的捕食作用,结果表明,七星瓢虫成虫对绣线菊蚜密度的功能反应可用Holing-Ⅱ型圆盘方程较好地拟合,捕食作用在一定范围内随猎物密度的增加而加大;七星瓢虫成虫对绣线菊蚜的捕食作用率随着自身密度增加而减小,相互之间存在一定的干扰,自身密度的功能反应能用Hassell数学模型较好地反映。     

Antigenic and electrophoretic variants of vitellogenin in Oncopeltus blood and their control by juvenile hormone

Antigenic analysis of adult female-specific blood and yolk proteins in Oncopeltus demonstrated an incomplete vitellogenin (A), which appears in the blood prior to yolk deposition and is later modified or joined by an antigenically complete molecule (AB). Vitellogenin AB is antigenically indistinguishable from the major yolk protein of mature eggs, though the electrophoretic mobilities of the two differ in 6% acrylamide gels. Vitellogenin A alone appears in the blood of adult females in which the corpora allata are known to be inactive, i.e., during starvation or photoperiodically induced diapause. Stimulation of these females with a juvenile hormone analog restores yolk deposition, and also induces the appearance of AB in the blood. While juvenile hormone is needed for the termination of diapause and the maturation of vitellogenin in this species, diapause begins with the vitellogenin-producing mechanism already partially assembled.     

Macromolecular synthesis in wing discs of Spodoptera mauritia Boisd: effects of a juvenile hormone analogue

Last instar larvae of S. mauritia treated topically on day 0, day 1, day 2 and day 3 with a daily (dose of 25 microg juvenile hormone analogue (JHA) moulted into supernumerary larvae. The imaginal discs of the supernumerary larvae especially those of mouthparts and thoracic appendages showed pupal characteristics. However the wing discs, which showed only partial differentiation, were uneverted and highly tanned. In an effort to provide an explanation to this anomaly the RNA, DNA and protein profile in the wing discs of supernumerary larvae were studied. Quantitative analysis of DNA, RNA and protein showed a considerable increase in the amount of DNA and protein and a decline in RNA level. SDS-PAGE analysis of wing disc proteins of JHA treated larvae showed a reduction in the expression of many major proteins that were predominant in the wing discs of control larvae. The results suggest that JHA induced inactivation of genes involved in the synthesis of proteins needed for evagination process may be responsible for the formation of uneverted, partially differentiated pupal wing discs in supernumerary larvae.     

蜜蜂越冬期卵黄原蛋白和保幼激素的表达特性研究

为了探究蜜蜂越冬期体内卵黄原蛋白（Vitellogenin,Vg）和保幼激素（Juvenile hormone,JH）的表达特性。本试验分别采集夏繁期（7月）和不同越冬时期（11月、12月、1月、2月）的珲春黑蜂工蜂,通过酶联免疫吸附试验方法检测Vg、JH Ⅲ及超氧化物歧化酶（Superoxide Dismutase, SOD）的浓度变化,采用qRT-PCR方法检测Vg、JH代谢相关基因的表达量。结果表明：珲春黑蜂越冬期Vg浓度高于夏繁期,在不同越冬期Vg浓度总体呈现先上升后下降的趋势;Vg基因的变化趋势与Vg浓度相一致,而VgR基因表达量与SOD浓度变化趋势相一致,在越冬期水平高于夏繁期,且在整个越冬期呈现下降的趋势。珲春黑蜂越冬期JH Ⅲ浓度低于夏繁期,且随着越冬持续进行总体呈现持续上升的变化趋势;MFE基因的变化趋势与JH Ⅲ浓度相近;JHAMT基因表达量越冬期显著高于夏繁期,在整个越冬期呈现先上升后下降的趋势。本研究结果表明,蜜蜂越冬期体内存在Vg和JH Ⅲ间负相关的调节模型,其通过Vg、JH Ⅲ代谢相关基因的表达来提高越冬蜂抗氧化能力和抗寒性,为西方蜜蜂抗寒分子机制的深入研究提供了新的研究思路。     

Dynamics of vitellogenin synthesis in juvenile giant freshwater prawn Macrobrachium rosenbergii

The dynamics of vitellogenin (Vg) mRNA expression and patterns of Vg and vitellin distribution in the hepatopancreas and ovary of juvenile Macrobrachium rosenbergii were examined using real-time RT-PCR and immunohistochemical methods. Eyestalk ablation was seen to induce rapid development of the gonads and Vg synthesis in females. In the female hepatopancreas, Vg mRNA expression was observed several days following ablation, after which levels increased gradually with increasing gonadosomatic index (GSI). Vitellin accumulation in the oocytes also increased with increasing Vg mRNA synthesis; expression was however negligible in the ovary. Hemolymph Vg levels in females ranged from 0.04 to 2.2 mg/ml. SDS PAGE/Western blotting analysis of hemolymph samples revealed that juvenile Vg was composed of 199 and 90 kDa subunits; the 102 kDa subunit present in adult female Vg (Okuno et al., 2002. J Exp Zool 292:417-429) could not be detected at any stage of vitellogenesis in juveniles. Vg was not detectable in non-ablated juveniles. The results of this study confirmed that the mode of involvement of eyestalk factors in regulating vitellogenesis is intrinsic to both juveniles and adults, and that a basic pattern of Vg synthesis and processing is conserved. However, the fact that juveniles are not able to produce the same Vg levels observed in adult females, and do not reach high GSI levels culminating in spawning suggests that other factors and physiological conditions specific to adult females are necessary to demonstrate full reproductive ability.     

6种复配杀虫剂对七星瓢虫的毒力测定及风险评估

采用药膜法分别测定了10％甲维盐·茚虫威悬浮剂、12％甲维盐·虫螨腈悬浮剂、12％虫螨腈·虱螨脲悬浮剂、14％呋虫胺·螺虫乙酯悬浮剂、22％吡虫啉·螺虫乙酯悬浮剂和6％吡虫啉·高效氯氟氰菊酯悬浮剂6种复配杀虫剂对七星瓢虫Coccinella septempunctata Linnaeus幼虫急性毒性,并进行了初级风险评...     

Predatory effect of Coccinella septempunctata on Thrips tabaci and Trialeurodes vaporariorum

Abstract:  The predatory effect of adult ladybird Coccinella septempunctata L. on adults of thrips, Thrips tabaci Lindeman, and whiteflies, Trialeurodes vaporariorum (Westwood), was examined in controlled environment chambers, on tomato leaves, in transparent small plastic cages at proportions of 1/(10 + 10), 1/(20 + 20), 1/(30 + 30) and 1/(40 + 40) predator/number of thrips and whiteflies. We conclude that C. septempunctata could be used with success for the biological control of thrips and whiteflies in greenhouse crops, with almost the same effectiveness as for aphids, at predator/prey proportions near 1 : 30. Additionally, it was found a prey preference for T. tabaci in comparison with T. vaporariorum . According to the model used, effective predation is correlated with predator/prey ratio rather than to prey preference.