Parasitoid-host endocrine relations: self-reliance or co-optation?

High titers of juvenile hormone (JH) maintain developmental arrest in Manduca sexta larvae parasitized by Cotesia congregata. Parasitized hosts exhibit up to 9.5 times greater amounts of total hemolymph JH (from 0.6±0.09 to 2.51±0.43 ng/ml) compared to non-parasitized controls. Elevated titers are observed throughout the fifth instar, even beyond egression of the parasitoids on day 5. GC–MS analysis revealed that in hemolymph of unparasitized control larvae, JH I is the major homolog and levels of JH III are negligible; in parasitized individuals the amounts of JH I, II, and III rise, and JH III predominates. Neck ligation ensured separation of M. sexta’s corpora allata from the posterior section, which contained most of the parasitoids in the infected insects. When the posterior region was sampled, JHs were not detected in the non-parasitzed larvae, but in those parasitized, JH III was found (1.98±0.29 ng/ml, 24 h post-ligation). JH III was the only homolog produced and secreted by the parasitoid in in vitro culture. This is the first report stating that a parasitoid secretes JH III and may contribute, at least in part, to the circulating titer in the host hemocoel, concurrently promoting host production of JH I and II.     

Precocious induction of vitellogenin with JH III in the twospotted stink bug, Perillus bioculatus (Heteroptera: Pentatomidae)

The effect of juvenile hormone (JH) III on the hemolymph composition of vitellogenin was examined in Perillus bioculatus. Adult females were treated topically with JH III, and the premature presence of vitellogenin in the hemolymph was then detected using electrophoresis and Western blot analyses. JH III treatment resulted in a dose-dependent early production of vitellogenin that was detectable 48 h before vitellogenin was present in non-treated insects. Vitellogenin was not observed in the hemolymph of JH III-treated adult males. The techniques reported here may be useful for the detection, isolation and characterization of compounds with JH-like activity in P. bioculatus and other species of Heteroptera (which are thought to have JH-like substances other than the JHs with known chemical identity). These same techniques may also provide a method for researchers to investigate the interactions of JH-like compounds and other substances, such as ecdysteroid, in the regulation of vitellogenesis in Heteroptera.     

Earthworms (Clitellata, Acanthodrilidae) of the mountains of Eastern Jamaica

Fourteen species new to science are described from material collected at several sites in the Blue Mountains and the John Crow Mountains of eastern Jamaica, doubling the known endemic Jamaican earthworm fauna. New data on Dichogaster montecyanensis (Sims) are provided. All species are placed in the genus Dichogaster Beddard, which is here treated sensu lato, i.e. including Eutrigaster Cognetti. Eight of the new species have lost the posterior pair of prostates and the seminal grooves of the male field. These are D. bromeliocola, D. crossleyi, D. davidi, D. garciai, D. harperi, D. haruvi, D. hendrixi, and D. johnsoni. D. sydneyi n. sp. has independently lost the posterior prostates but not the seminal grooves. The new species D. altissima and D. manleyi have the conventional dichogastrine prostatic battery and male field characteristics. Three species described here, D. farri, D. garrawayi, and D. marleyi, all have a third pair of prostates in the 20th segment, no seminal grooves, dorsal paired intestinal caeca in segment lxv, and lack penial setae.     

叉头转录因子1基因FOXO1的特征及其在葱蝇夏滞育前期蛹体内糖脂代谢中的作用(英文)

【目的】克隆和鉴定葱蝇Delia antiqua叉头转录因子1基因,探究其在葱蝇夏滞育前期蛹体内糖代谢和脂代谢中的作用。【方法】本研究基于葱蝇转录组数据利用3′RACE法克隆叉头转录因子1基因的全长开放阅读框;利用生物信息学法分析了该基因编码蛋白的序列特征、保守结构域和二级结构,并采用最大似然法对其与其他13种昆虫来源的同源序列进行了聚类分析。通过RNA干扰技术沉默目的基因后,采用实时定量PCR法分析葱蝇夏滞育前期蛹体内目的基因下游脂肪酶brummer基因(DaBmm)和磷酸烯醇式丙酮酸羧化酶基因(DaDepck)的表达规律,并对甘油三脂(TAG)、海藻糖和葡萄糖含量及总脂肪酶的活性变化进行分析。【结果】克隆得到了葱蝇叉头转录因子1基因DaFOXO1 (GenBank登录号: MG813258),其编码蛋白含有619个氨基酸,具有典型的叉头DNA结合域,核定位信号,2个14-3-3结合区和1个富含谷氨酰胺区;DaFOXO1与铜绿蝇Lucilia cuprina FOXO1蛋白的氨基酸序列有87%的一致性,并与其聚为一支。干扰葱蝇夏滞育前期蛹DaFOXO1后,8-20 h间可显著抑制DaBmm基因的表达,12-24 h间可显著影响TAG含量和总脂肪酶的活性,8-20 h(16 h除外)间可显著降低DaPepck的表达,但对葡萄糖和海藻糖的含量没有显著影响。【结论】本研究结果表明海藻糖和葡萄糖的积累在化蛹前已完成;DaFOXO1对DaBmm和DaDepck基因的表达调控可能有利于葱蝇夏滞育前期蛹体内的脂肪积累。同时也表明靶向Bmm依赖性脂类水解过程及其下游因子可为打破昆虫滞育提供一种有效策略。     

Genetic analysis of mortality in murine angiostrongyliasis costaricensis using SMXA recombinant inbred mouse strains

The SMXA recombinant inbred mouse strain set was produced by systematic inbreeding from the F2 generation of a cross between two progenitor inbred strains, A/J and SM/J, which differed markedly with respect to the patterns of infection with Angiostrongylus costaricensis. We have applied this set to genetic analysis of mouse susceptibility to this nematode infection. The mortality was variable among substrains of the SMXA RI strains, indicating the involvement of multiple genes. Linkage analysis showed several chromosomal regions closely linked to mortality; chromosome 6 (D6Rik86, 87; P0.001), 10 (D10Rik66–D10Mit12; P=0.0058), 13 (D13Rik79, 80; P=0.0096) and 17 (D17Mit28–D17Rik76; P=0.0088).     

温度对新菠萝灰粉蚧生长发育和繁殖的影响

新菠萝灰粉蚧是近年在我国新发现的一种重要外来入侵害虫,温度是决定新菠萝灰粉蚧能否建立稳定种群的最基本因素.本研究以南瓜作为寄主,探索恒温条件对新菠萝灰粉蚧生长发育和繁殖的影响.在17、20、23、26、29和32 ℃,光周期14L∶10D,相对湿度(75±5)%的实验室条件下,测定了新菠萝灰粉蚧各虫态的发育历期、发育速率、存活率和繁殖力,组建了新菠萝灰粉蚧的实验种群生命表.结果表明: 在20～29 ℃,各虫态的发育历期均随温度的升高而缩短,在20 ℃下雌、雄若虫期的发育历期最长,分别为46.95和50.26 d;29 ℃时雌虫若虫期的发育历期最短,为20.28 d;而雄虫若虫期在32 ℃时最短,为20.70 d.各虫态的温度与发育速率的关系均符合二次回归关系.此外,温度显著影响新菠萝灰粉蚧的存活率,在29 ℃时雌、雄若虫期的存活率均最高,分别为70.3%和69.3%;雌虫世代的发育起点温度为13.80 ℃,有效积温为491.50日·度;雄虫世代的发育起点温度为11.61 ℃,有效积温为388.85日·度.新菠萝灰粉蚧的产卵前期和成虫寿命随着温度的升高而缩短;成虫产卵量在29 ℃时最高,达每雌442.2粒;最小为20 ℃,仅为每雌111.8粒;29和20 ℃下种群趋势指数分别为168.2和19.1,且在17和32 ℃时,新菠萝灰粉蚧1龄若虫和3龄若虫均表现为生长停滞.说明温度过高或过低均不利于其生长发育.温度对新菠萝灰粉蚧的生长发育、存活、繁殖及种群增长有显著的影响,23～29 ℃是最适宜新菠萝灰粉蚧生长发育和繁殖的温度范围.     

Geographic Origin and Taxonomic History of Delphastus spp. (Coleoptera: Coccinellidae) in Commercial Culture

Most of the published studies of Delphastus pusillus biology and behavior on Bemisia spp. actually refer to D. catalinae. Similarly, Delphastus species in commercial insectary cultures are probably D. catalinae and not D. pusillus. We discuss the historical reasons for the clouded identity of these native coccinellid beetles.     

Characteristics of apolipophorin-III of the southwestern corn borer, Diatraea grandiosella

Apolipophorin-III was isolated from the lipophorin-free fraction of larval plasma of the southwestern corn borer, Diatraea grandiosella, because significant amounts of apolipophorin-III were found to be present in the hemolymph not associated with lipophorin. Apolipophorin-III, purified using ammonium sulfate precipitation, cation exchange chromatography, and gel filtration, was shown to be a nonglycosylated polypeptide with 17 kDa mol. wt, as determined by SDS-PAGE and silver staining. The amino acid composition of apolipophorin-III showed similarities to published compositions of apolipophorin-III isolated from other insects. The N-terminal sequence of apolipophorin-III (DAPSTTPPQDXEKKAAEFQKTFTEQXNQLANK), is highly homologous to that of apolipophorin-III of Manduca sexta. Antiserum raised against purified apolipophorin-III was used to demonstrate an immunochemical identity between the isolated apolipophorin-III and that associated with lipophorin. This antiserum cross-reacted with apolipophorin-III of M. sexta, and antiserum raised against M. sexta apolipophorin-III cross-reacted with apolipophorin-III isolated from D. grandiosella, demonstrating an immunochemical relationship between the proteins, and providing confirmatory evidence for the identity of the isolated protein. These antisera did not react with the putative apolipophorin-III of the cricket, Acheta domesticus. Using immunoprecipitation by the apolipophorin-III antiserum of D. grandiosella, the synthesis and secretion of [³H]apolipophorin-III by the fat body in vitro was shown to be maximal in 13–15 day-old larvae, with a transit time of ca 23 min.     

Aggression among sea urchins on Caribbean coral reefs

The existence and nature of intra- and interspecific aggression were examined for five species of sea urchins inhabiting Caribbean coral reefs. The studies took place in the San Blas Islands of Panama and involved the following species: Echinometra lucunter (Linnaeus), E. viridis Agassiz, Diadema antillarum Phillipi, Lytechinus williamsi Chesher, and Eucidaris tribuloides (Lamarck). An intruder was placed next to an undisturbed resident and the behaviors and responses of both individuals were followed. All pairwise combinations of resident species and intruder species were tested except for combinations involving D. antillarum as intruder.

For E. viridis and E. lucunter, agonistic interactions occurred commonly (46–79% of trials) between conspecifics and congeners. Almost all of the agonistic encounters involved pushing. Additionally, biting occurred in 8–25 % of the trials. Residents were most often the aggressors and usually succeeded in retaining their location. Intruders only succeeded in forcing residents out of their positions if the intruder was equal to or larger in size than the resident. Surveys of an undisturbed population of E. viridis during the daytime indicated that 16% of the individuals were engaged in intraspecific agonistic interactions at any one time.

D. antillarum exhibited biting behavior against both species of Echinometra in 23–24% of the trials. Biting attacks against L. williamsi and E. tribuloides occurred rarely. L. williamsi only once demonstrated pushing behavior and never was observed biting another sea urchin. E. tribuloides occasionally exhibited pushing and biting behaviors, both as residents and as intruders, and was twice observed biting Echinometra.

These studies suggest that two kinds of agonistic interactions may commonly occur among Caribbean reef-dwelling sea urchins: (1) intraspecific and interspecific aggression among Echinometra, and (2) predatory/aggressive attacks against Echinometra by D. antillarum. The former may result in greater dispersion of Echinometra relative to food and shelter resources and control the spatial distribution and concentration of Echinometra grazing pressure within an area. The attacks by D. antillarum may result in the restriction to, or higher densities of, Echinometra in crevices and rugose microhabitats that provide shelter from the larger-bodied D. antillarum.     

The sex-peptide gene (Acp70A) is duplicated in Drosophila subobscura

A 3.1-kb region of Drosophila subobscura homologous to the Acp70A region of D. melanogaster, which contains the sex-peptide gene, was cloned and sequenced. This region contains an approximately 600-bp duplication that includes the sex-peptide and its 5′ and 3′ flanking regions. The preproteins are 54 and 56 amino acids long, respectively (as compared to 55 amino acids in D. melanogaster), and each includes a 19-amino-acid-long signal peptide. The C-terminal part of the mature peptide is highly conserved between D. melanogaster and the two copies of D. subobscura. In this species, both copies of the gene are transcribed and, like in D. melanogaster, only expressed in males. The duplicated region includes 300 bp upstream of the gene that would therefore seem sufficient for their expression in males. This region presents at its 5′ end a stretch 93-bp that has a high similarity with the corresponding region of D. melanogaster and could be part of a still unidentified regulatory element of these genes.     

Cloning and preliminary characterization of a novel cuticular antigen from the filarial parasite Dirofilaria immitis

We have described here the cloning and partial characterization of a cDNA encoding a cuticular antigen of Dirofilaria immitis. A 48-h third-stage larval D. immitis cDNA library was immunoscreened with sera raised in mice against third-stage larval cuticles (mouse anti-L3 cuticle antisera). A strongly immunoreactive clone (L3MC4) was isolated. Sequence analysis of L3MC4 showed that it was a partial length cDNA. The missing 5′ end of the clone was amplified by PCR from D. immitis adult female first-strand cDNA using the nematode 22-base splice leader sequence and a L3MC4-specific antisense primer. The composite cDNA sequence comprised 616 bases (nDiL3MC4) encoding a full-length protein of 146 amino acids (DiL3MC4). GenBank analysis showed that DiL3MC4 shared some homology to an unknown C. elegans gene product (31%) at the amino acid level. However, there were no related filarial expressed sequence tags in the current GenBank™ database. Antibodies to recombinant DiL3MC4 (rDiL3MC4) identified a 19-kDa native antigen in the adults and in the L3 and L4 larval stages of D. immitis. In addition, the antibodies bound to the cortical layers of the L3 cuticle, as revealed by immuno-gold electron microscopy. The native protein was not detected in larval and adult excretory–secretory products. Immunoblot analysis showed that serum from a rabbit that was repeatedly injected with a small number of D. immitis third stage larvae reacted with rDiL3MC4. Thus, DiL3MC4 is a novel cuticular antigen of a filarial parasite.     

Parasitism-induced effects of Glyptapanteles liparidis (Hym., Braconidae) on the juvenile hormone titer of its host, Lymantria dispar: the role of the parasitoid larvae

Parasitization by the gregarious larval endoparasitoid Glyptapantles liparidis induces a dramatic increase in the hemolymph juvenile hormone (JH) titer (especially JH III) of its host larva, Lymantria dispar. Here, we investigated the role of the parasitoid larvae in JH synthesis and release by in vitro and in vivo experiments. GC-MS analyses confirmed that the rising hemolymph JH titer coincided with the time at which the parasitoids molt to the second larval instar. Peak values in host hemolymph titers were observed prior to parasitoid emergence, and titers dropped to negligible levels within 24 h after parasitoid emergence. Whole body extracts from excised second instar parasitoids yielded JH III and trace amounts of JH II. The in vitro secretory activity of the corpora allata (CA) of L. dispar larvae was not enhanced by parasitization. When the host's CA were separated by neck ligation, we found elevated JH III titers, but no JH II in the hemolymph of the posterior section, which contained the parasitoids. Parasitoids that were kept in in vitro culture produced and released only JH III. The parasitoids’ ability to secrete JH and to molt independently from their host's molting cycles indicates that at least second instar parasitoids are hormonally self-reliant.     

Microwave-assisted, one-pot syntheses and fungicidal activity of polyfluorinated 2-benzylthiobenzothiazoles

Polyfluorinated 2-benzylthiobenzothiazoles 3a–l are prepared via a microwave-assisted, one-pot procedure. The advantages, such as good to excellent yields, shorter reaction time (14–21 min), readily available starting material, and simple purification procedure, distinguish the present protocol from other existing methods used for the synthesis of 2-benzylthiobenzothiazoles. Bioassay indicated that most of the compounds showed significant fungicidal activity against Rhizoctonia solani, Botrytis cinereapers, and Dothiorella gregaria at a dosage of 50 μg/mL. Interestingly, compared to the control of commercial fungicide, triadimefon, compound 3c exhibited much higher activities against R. solani, B. cinereapers, and D. gregaria, which showed that the polyfluorinated 2-benzylthiobenzothiazoles can be used as lead compound for developing novel fungicides.     

Heat-shock protein expression in a perennial grass commonly associated with active geothermal areas in western North America

Heat-shock protein (hsp) expression in response to short- (hours) versus long-term (weeks) heat was examined in Dichanthelium lanuginosum. Expression of cytoplasmic class I small hsps (shsps) and hsp 101 was elicited by 2 h above 40°C, along with an increase over basal levels of hsp 70. Elevated levels of heat-induced shsp and hsp 101 persisted for 5–7 days after plants were returned to control temperatures. Protein extracts from roots exposed to 42°C for several weeks displayed increased levels of shsp but decreased hsp 101 levels. This decrease in hsp 101 did not occur in D. lanuginosum from normothermic environments.