SECRETION OF LIPASES IN THE DIGESTIVE TRACT OF THE CRICKET Gryllus bimaculatus

Little is known concerning the sites and the ratios of the lipase secretions in insects, therefore we undertook an examination of the lipase secretion of fed and unfed adult female Gryllus bimaculatus. The ratio of triacylglyceride lipase, diacylglyceride lipase, and phosphatidylcholine lipase secreted by fed females in the caecum and ventriculus is 1:1.4:0.4. These activities decrease in the caecum by 30–40% in unfed females. The total lipase activity (TLA) in the caecum is about 10 times that in the ventriculus. Minimal lipase secretion occurs before and during the final moult, and remains at this level in unfed crickets, indicating a basal secretion rate. In 2‐day‐old fed females, about 10% of the TLA in the entire gut is found in the crop, about 70% in the caecum, 20% in the ventriculus, and 3% in the ileum. Lipases in the ventriculus are recycled back to the caecum and little is lost in the feces. Oleic acid stimulated in vitro lipase secretion, but lipids did not. Feeding stimulated lipase secretion, starvation reduced lipase secretion, but this does not prove a direct prandal regulation of secretion, because feeding also induced a size and volume increase of the caecum.     

ORIGINS AND ACTIVATION OF PROPHENOLOXIDASES IN THE DIGESTIVE TRACT OF THE CRICKET,Gryllus bimaculatus

The function of Phenoloxidases (POs) in sclerotization and defense in insects is well understood, but little is known concerning their occurrence, origins, and function in the digestive tract. In Gyrllus bimaculatus gut all of the PO activity is found in the lumen of the digestive tract, and no detectible activity is found in homogenates of the gut epithelium or secretions from incubated epithelial tissues. Prophenoloxidases (PPOs) are synthesized in the hemocytes of Bombyx mori and are transported into the cuticle. It is suggested that the PPOs in the caecal lumen of G. bimaculatus likewise are synthesized in hemocytes and are transported by unknown means into the caecal lumen, where they are activated to POs by trypsin. Peristalsis transports the POs both forward into the crop and posterior within the peritrophic membrane into the hind gut. The PPOs in the hemolymph consist of a trimer (270–280 kDa) and a tetramer (340–370 kDa). The active POs in the gut lumen consist of a monomer (85–95 kDa) in addition to an activated trimer and tetramer.     

THE SECRETION OF DIGESTIVE ENZYMES AND CAECAL SIZE ARE DETERMINED BY DIETARY PROTEIN IN THE CRICKET Gryllus bimaculatus

In Gryllus bimaculatus, the size of the caecum decreases in the latter half of each instar to a stable minimal size with a steady minimal rate of digestive enzyme secretion until feeding resumes after ecdysis. The higher the percent protein in the newly ingested food, the faster and larger the caecum grows, and as a consequent the higher the secretion rate of trypsin and amylase. When hard boiled eggs (40% protein) are eaten the caecum is 2× larger, the trypsin secretion is almost 3× greater, and amylase 2.5× greater then when fed the same amount of apples (1.5% protein). Only dietary protein increases amylase secretion, whereas dietary carbohydrates have no effect on amylase secretion. The minimal caecal size and secretion rate must be supported by utilization of hemolymph amino acids, but the growth of the caecum and increasing enzymes secretions after the molt depend upon an amino acid source in the lumen. This simple regulation of digestive enzyme secretion is ideal for animals that must stop feeding in order to molt. This basic control system does not preclude additional regulation mechanisms, such as prandal, which is also indicated for G. bimaculatus, or even paramonal regulation.     

社鼠和褐家鼠的能量代谢及消化道形态的比较

比较研究了社鼠和褐家鼠的摄入能、消化率、同化率以及消化道长度和重量的差异。结果表明,在饲养条件下,社鼠每天摄入能为1069．43kJ·（kg0.75）－1,高于褐家鼠（708．55kJ·（kg0.75）-1）,社鼠对花生的消化率（95．38％）较褐家鼠（91．74％）为高,但两种动物对大米的消化率和同化率较为接近。社鼠消化道各器官（除小肠）的长度和重量皆明显高于褐家鼠。具有较大的消化道和较高的消化率是社鼠对野外较差食物条件的适应对策。     

SEX‐SPECIFIC EVOLUTIONARY POTENTIAL OF PRE‐ AND POSTCOPULATORY REPRODUCTIVE INTERACTIONS IN THE FIELD CRICKET,TELEOGRYLLUS COMMODUS

Mate choice often depends on the properties of both sexes, such as the preference and responsiveness of the female and the sexual display traits of the male. Quantitative genetic studies, however, traditionally explore the outcome of an interaction between males and females based solely on the genotype of one sex, treating the other sex as a source of environmental variance. Here, we use a half‐sib breeding design in the field cricket, Teleogryllus commodus, to estimate the additive genetic contribution of both partners to three steps of the mate choice process: the time taken to mate; the duration of spermatophore attachment; and the intensity of mate guarding. Rather than each sex contributing equally to the interactions, we found that genetic variation for latency to mate and spermatophore attachment was sex‐specific, and in the case of mate‐guarding intensity, largely absent. For a given interaction, genetic variation in one sex also appears to be largely independent of the other, and is also uncorrelated with the other traits. We discuss how pre‐ and postcopulatory interactions have the potential to evolve as an interacting phenotype, but that any coevolution between these traits, due to sexual selection or sexual conflict, may be limited.     

温度和pH对洞庭鲇鱼消化酶活性的影响

采用酶学分析方法研究了温度和pH对洞庭鲇鱼蛋白酶、脂肪酶和淀粉酶活力的影响。结果表明,在设定的温度和pH范围内,鲇鱼各消化酶的活力均随着温度和pH的升高呈现先升后降的变化趋势。其中,胃蛋白酶的最适温度为40℃,肝胰脏、前肠、中肠和后肠蛋白酶的最适温度为45℃;脂肪酶的最适温度均为35℃;胃淀粉酶的最适温度为35℃,其他部位均为30℃。胃、肝胰脏、前肠、中肠和后肠蛋白酶的适宜pH分别为2.0、8.5、7.5、8.0和8.0;脂肪酶的适宜pH均为7.5;淀粉酶肝胰脏的适宜pH为7.5,其余部位均为7.0。鲇鱼各消化酶活力存在器官特异性。在最适温度下,蛋白酶活力顺序为前肠>肝胰脏>胃>中肠>后肠,脂肪酶的活力顺序均为肝胰脏>胃>前肠>中肠>后肠,淀粉酶的活力顺序为肝胰脏>前肠>中肠>后肠>胃,各部位之间差异显著(P<0.05)。     

DIGESTION IN ADULT FEMALES OF THE LEAF‐FOOTED BUG Leptoglossus zonatus (HEMIPTERA: COREIDAE) WITH EMPHASIS ON THE GLYCOSIDE HYDROLASES α‐AMYLASE, α‐GALACTOSIDASE,AND α‐GLUCOSIDASE

The leaffooted bug, Leptoglossus zonatus (Hemiptera: Coreidae) is an emerging pest of several crops around the World and up to now very little is known of its digestive system. In this article, glycoside hydrolase (carbohydrase) activities in the adult midgut cells and in the luminal contents of L. zonatus adult females were studied. The results showed the distribution of digestive carbohydrases in adults of this heteropteran species in the different intestinal compartments. Determination of the spatial distribution of α‐glucosidase activity in L. zonatus midgut showed only one major molecular form, which was not equally distributed between soluble and membrane‐bound isoforms, being more abundant as a membrane‐bound enzyme. The majority of digestive carbohydrases were found in the soluble fractions. Activities against starch, maltose and the synthetic substrate NPαGlu were found to show the highest levels of activity, followed by enzymes active against galactosyl oligosaccharides. Based on ion‐exchange chromatography elution profiles and banding patterns in mildly denaturing electrophoresis, both midgut α‐amylases and α‐galactosidases showed at least two isoforms. The data suggested that the majority of carbohydrases involved in initial digestion were present in the midgut lumen, whereas final digestion of starch and of galactosyl oligosaccharides takes place partially within the lumen and partially at the cell surface. The complex of carbohydrases here described was qualitatively appropriate for the digestion of free oligosaccharides and oligomaltodextrins released by α‐amylases acting on maize seed starch granules.     

THE MAINTENANCE OF GENETIC VARIATION FOR OVIPOSITION RATE IN TWO‐SPOTTED SPIDER MITES: INFERENCES FROM ARTIFICIAL SELECTION

Despite the directional selection acting on life‐history traits, substantial amounts of standing variation for these traits have frequently been found. This variation may result from balancing selection (e.g., through genetic trade‐offs) or from mutation‐selection balance. These mechanisms affect allele frequencies in different ways: Under balancing selection alleles are maintained at intermediate frequencies, whereas under mutation‐selection balance variation is generated by deleterious mutations and removed by directional selection, which leads to asymmetry in the distribution of allele frequencies. To investigate the importance of these two mechanisms in maintaining heritable variation in oviposition rate of the two‐spotted spider mite, we analyzed the response to artificial selection. In three replicate experiments, we selected for higher and lower oviposition rate, compared to control lines. A response to selection only occurred in the downward direction. Selection for lower oviposition rate did not lead to an increase in any other component of fitness, but led to a decline in female juvenile survival. The results suggest standing variation for oviposition rate in this population consists largely of deleterious alleles, as in a mutation‐selection balance. Consequently, the standing variation for this trait does not appear to be indicative of its adaptive potential.     

DIFFERENCES IN THE REGULATION OF GROWTH AND BIOMINERALIZATION GENES REVEALED THROUGH LONG‐TERM COMMON‐GARDEN ACCLIMATION AND EXPERIMENTAL GENOMICS IN THE PURPLE SEA URCHIN

Across heterogeneous landscapes, populations may have adaptive differences in gene regulation that adjust their physiologies to match local environments. Such differences could have origins in acclimation or in genetically fixed variation between habitats. Here we use common‐garden experiments to evaluate differences in gene expression between populations of the purple sea urchin, Strongylocentrotus purpuratus, spanning 1700 km and average temperature differences of 5°C to 8°C. Across expression profiles from 18,883 genes after 3 years of common conditions, we find highly correlated expression patterns (Pearson's r = 0.992) among most genes. However, 66 genes were differentially expressed, including many ribosomal protein and biomineralization genes, which had higher expression in urchins originally from the southern population. Gene function analyses revealed slight but pervasive expression differences in genes related to ribosomal function, metabolism, transport, “bone” development, and response to stimuli. In accord with gene expression patterns, a post‐hoc spine regrowth experiment revealed that urchins of southern origin regrew spines at a faster rate than northern urchins. These results suggest that there may be genetically controlled, potentially adaptive differences in gene regulation across habitats and that gene expression differences may be under strong enough selection to overcome high, dispersal–mediated gene flow in this marine species.     

PHARMACOLOGICAL REGULATION OF DIGESTION IN THE ANAUTOGENOUS FLESH FLY,Sarcophaga crassipalpis,BY SIMPLE INJECTION OF 6‐HYDROXYDOPAMINE

Female anautogenous Sarcophaga flesh flies need a protein meal to start large‐scale yolk polypeptides (YPs) production and oocyte maturation. Protein meal rapidly elicits a brain‐dependent increase in midgut proteolytic activity. Trypsin and chymotrypsin together represent over 80% of protease activity in liver‐fed flies. Abdominal injection of 6‐hydroxydopamine (6‐OHDA) dose‐dependently prohibits this increase in proteolytic activity at translational level in a similar way as post liver feeding decapitation. Delayed injection of 6‐OHDA later than 6 h post liver meal has no effect. In flesh flies, chemical decapitation by 6‐OHDA, by interrupting the brain‐gut dopaminergic signaling, can be used as tool for the controlled inhibition of midgut proteolytic activity and subsequent ovarial development. Inhibition of ovarial development is probably indirect due to a deficit in circulating amino acids needed for YPs synthesis.     

SUPPRESSION OF AcMNPV REPLICATION BY ADF AND THYMOSIN PROTEIN UP‐REGULATION IN A NEW TESTIS CELL LINE,Ha‐shl‐t

Host cytoskeletons facilitate the entry, replication, and egress of viruses because cytoskeletons are essential for viral survival. One mechanism of resisting viral infections involves regulating cytoskeletal polymerization/depolymerization. However, the molecular mechanisms of regulating these changes in cytoskeleton to suppress viral replication remain unclear. We established a cell line (named Ha‐shl‐t) from the pupal testis of Helicoverpa armigera (Lepidoptera: Noctuidae). The new testis cell line suppresses Autographa californica multiple nucleocapsid nucleopolyhedrovirus (AcMNPV) replication via disassembly of cytoskeleton. Up‐regulation of thymosin (actin disassembling factor) and adf (actin depolymerizing factor) reduces F‐actin. Silencing thymosin or adf or treating cells with the F‐actin stabilizer phalloidin led to increased AcMNPV replication, while treating cells with an F‐actin assembly inhibitor cytochalasin B decreased viral replication. We infer that Ha‐shl‐t cells utilize F‐actin depolymerization to suppress AcMNPV replication by up‐regulating thymosin and adf. We propose Ha‐shl‐t as a model system for investigating cytoskeletal regulation in antiviral action and testicular biology generally.     

ROLE OF GLUTAMATE DEHYDROGENASE AND GLUTAMINE SYNTHETASE IN CHLORELLA VULGARIS DURING ASSIMILATION OF AMMONIUM WHEN JOINTLY IMMOBILIZED WITH THE MICROALGAE‐GROWTH‐PROMOTING BACTERIUM AZOSPIRILLUM BRASILENSE1

Enzymatic activities of glutamate dehydrogenase (GDH) and glutamine synthetase (GS) participating in the nitrogen metabolism and related ammonium absorption were assayed after the microalga Chlorella vulgaris Beij. was jointly immobilized with the microalgae‐growth‐promoting bacterium Azospirillum brasilense. At initial concentrations of 3, 6, and 10 mg · L^?1 NH₄⁺, joint immobilization enhances growth of C. vulgaris but does not affect ammonium absorption capacity of the microalga. However, at 8 mg · L^?1 NH₄⁺, joint immobilization enhanced ammonium absorption by the microalga without affecting the growth of the microalgal population. Correlations between absorption of ammonium per cell and per culture showed direct (negative and positive) linear correlations between these parameters and microalga populations at 3, 6, and 10 mg · L^?1 NH₄⁺, but not at 8 mg · L^?1 NH₄⁺, where the highest absorption of ammonium occurred. In all cultures, immobilized and jointly immobilized, having the four initial ammonium concentrations, enzymatic activities of Chlorella are affected by A. brasilense. Regardless of the initial concentration of ammonium, GS activity in C. vulgaris was always higher when jointly immobilized and determined on a per‐cell basis. When jointly immobilized, only at an initial concentration of 8 mg · L^?1 NH₄⁺ was GDH activity per cell higher.     

SEX LINKAGE,SEX‐SPECIFIC SELECTION,AND THE ROLE OF RECOMBINATION IN THE EVOLUTION OF SEXUALLY DIMORPHIC GENE EXPRESSION

Sex‐biased genes—genes that are differentially expressed within males and females—are nonrandomly distributed across animal genomes, with sex chromosomes and autosomes often carrying markedly different concentrations of male‐ and female‐biased genes. These linkage patterns are often gene‐ and lineage‐dependent, differing between functional genetic categories and between species. Although sex‐specific selection is often hypothesized to shape the evolution of sex‐linked and autosomal gene content, population genetics theory has yet to account for many of the gene‐ and lineage‐specific idiosyncrasies emerging from the empirical literature. With the goal of improving the connection between evolutionary theory and a rapidly growing body of genome‐wide empirical studies, we extend previous population genetics theory of sex‐specific selection by developing and analyzing a biologically informed model that incorporates sex linkage, pleiotropy, recombination, and epistasis, factors that are likely to vary between genes and between species. Our results demonstrate that sex‐specific selection and sex‐specific recombination rates can generate, and are compatible with, the gene‐ and species‐specific linkage patterns reported in the genomics literature. The theory suggests that sexual selection may strongly influence the architectures of animal genomes, as well as the chromosomal distribution of fixed substitutions underlying sexually dimorphic traits.     

DIFFERENCES IN THE PRODUCTION AND EXCRETION KINETICS OF OKADAIC ACID,DINOPHYSISTOXIN‐1, AND PECTENOTOXIN‐2 BETWEEN CULTURES OF DINOPHYSIS ACUMINATA AND DINOPHYSIS FORTII ISOLATED FROM WESTERN JAPAN1

We established clonal cultures of Dinophysis acuminata Clap. et Lachm. and D. fortii Pavill. isolated from western Japan and examined toxin production in them, focusing on intracellular production and extracellular excretion. At the end of incubations, the total amounts of pectenotoxin‐2 (PTX‐2), dinophysistoxin‐1 (DTX‐1), and okadaic acid (OA) in the D. acuminata cultures reached up to 672.7 ± 14.7 (mean ± SD), 88.1 ± 2.8, and 539.3 ± 39.7 ng · mL^?1, respectively, and the excreted extracellular amounts were equivalent to 5.1, 79.5, and 79.5% of the total amounts, respectively. Similarly, at the end of incubations, the total amounts of PTX‐2, DTX‐1, and OA in the D. fortii cultures reached up to 526.6 ± 52.6 (mean ±SD), 4.4 ± 0.4, and 135.9 ± 3.9 ng · mL^?1, respectively, and the excreted extracellular amounts were equivalent to 1.8, 80.1, and 86.6% of the total amounts, respectively. Further, we tested the availability of cell debris and dissolved organic substances that originated from the ciliate prey Myrionecta rubra for growth and toxin production in D. acuminata. Although no significant growth was observed in D. acuminata in the medium containing the cell debris and organic substances originated from M. rubra, the toxicity was significantly greater than that in the control (P < 0.05–0.001); this finding suggested the availability of organic substances for toxin production. However, toxin productivity was remarkably lower than that of Dinophysis species feeding on living M. rubra.     

LOCAL ADAPTATION IN A CHANGING WORLD: THE ROLES OF GENE‐FLOW,MUTATION, AND SEXUAL REPRODUCTION

In spatially heterogeneous environments, the processes of gene flow, mutation, and sexual reproduction generate local genetic variation and thus provide material for local adaptation. On the other hand, these processes interchange maladapted for adapted genes and so, in each case, the net influence may be to reduce local adaptation. Previous work has indicated that this is the case in stable populations, yet it is less clear how the factors play out during population growth, and in the face of temporal environmental stochasticity. We address this issue with a spatially explicit, stochastic model. We find that dispersal, mutation, and sexual reproduction can all accelerate local adaptation in growing populations, although their respective roles may depend on the genetic make‐up of the founding population. All three processes reduce local adaptation, however, in the long term, that is when population growth becomes balanced by density‐dependent competition. These relationships are qualitatively maintained, although quantitatively reduced, if the resources are locally ephemeral. Our results suggest that species with high levels of local adaptation within their ranges may not be the same species that harbor potential for rapid local adaptation during population expansion.     

DECOUPLING OF SHORT‐ AND LONG‐DISTANCE DISPERSAL PATHWAYS IN THE ENDEMIC NEW ZEALAND SEAWEED CARPOPHYLLUM MASCHALOCARPUM (PHAEOPHYCEAE,FUCALES)1

The processes that produce and maintain genetic structure in organisms operate at different timescales and on different life‐history stages. In marine macroalgae, gene flow occurs through gamete/zygote dispersal and rafting by adult thalli. Population genetic patterns arise from this contemporary gene flow interacting with historical processes. We analyzed spatial patterns of mitochondrial DNA variation to investigate contemporary and historical dispersal patterns in the New Zealand endemic fucalean brown alga Carpophyllum maschalocarpum (Turner) Grev. Populations bounded by habitat discontinuities were often strongly differentiated from adjoining populations over scales of tens of kilometers and intrapopulation diversity was generally low, except for one region of northeast New Zealand (the Bay of Plenty). There was evidence of strong connectivity between the northern and eastern regions of New Zealand’s North Island and between the North and South Islands of New Zealand and the Chatham Islands (separated by 650 km of open ocean). Moderate haplotypic diversity was found in Chatham Islands populations, while other southern populations showed low diversity consistent with Last Glacial Maximum (LGM) retreat and subsequent recolonization. We suggest that ocean current patterns and prevailing westerly winds facilitate long‐distance dispersal by floating adult thalli, decoupling genetic differentiation of Chatham Island populations from dispersal potential at the gamete/zygote stage. This study highlights the importance of encompassing the entire range of a species when inferring dispersal patterns from genetic differentiation, as realized dispersal distances can be contingent on local or regional oceanographic and historical processes.     

IDENTIFICATION AND EXPRESSION ANALYSIS OF TWO 14‐3‐3 PROTEINS IN THE MOSQUITO Aedes aegypti,AN IMPORTANT ARBOVIRUSES VECTOR

The 14‐3‐3 proteins are evolutionarily conserved acidic proteins that form a family with several isoforms in many cell types of plants and animals. In invertebrates, including dipteran and lepidopteran insects, only two isoforms have been reported. 14‐3‐3 proteins are scaffold molecules that form homo‐ or heterodimeric complexes, acting as molecular adaptors mediating phosphorylation‐dependent interactions with signaling molecules involved in immunity, cell differentiation, cell cycle, proliferation, apoptosis, and cancer. Here, we describe the presence of two isoforms of 14‐3‐3 in the mosquito Aedes aegypti, the main vector of dengue, yellow fever, chikungunya, and zika viruses. Both isoforms have the conserved characteristics of the family: two protein signatures (PS1 and PS2), an annexin domain, three serine residues, targets for phosphorylation (positions 58, 184, and 233), necessary for their function, and nine alpha helix‐forming segments. By sequence alignment and phylogenetic analysis, we found that the molecules correspond to ? and ζ isoforms (Aeae14‐3‐3ε and Aeae14‐3‐3ζ). The messengers and protein products were present in all stages of the mosquito life cycle and all the tissues analyzed, with a small predominance of Aeae14‐3‐3ζ except in the midgut and ovaries of adult females. The 14‐3‐3 proteins in female midgut epithelial cells were located in the cytoplasm. Our results may provide insights to further investigate the functions of these proteins in mosquitoes.     

INTERACTIONS AMONG PHOSPHATE UPTAKE,PHOTOSYNTHESIS, AND CHLOROPHYLL FLUORESCENCE IN NUTRIENT‐LIMITED CULTURES OF THE CHLOROPHYTE MICROALGA DUNALIELLA TERTIOLECTA1

Phosphate‐limited and phosphate‐sufficient continuous cultures of the marine chlorophyte microalga Dunaliella tertiolecta Butcher were examined for their responses to the addition of phosphate. Phosphate‐limited cultures showed a marked quenching of chl fluorescence following a pulse of phosphate. This response was absent from cells growing under phosphate‐sufficient conditions. Both the extent of fluorescence quenching (where present) and the initial rate of change in quenching were dependent on the concentration of phosphate added to cell suspensions and on the degree of limitation (growth rate in continuous culture). The addition of phosphate also brought about a transient decrease in photosynthetic oxygen evolution and a stimulation in respiration, which were relaxed as the added phosphate was depleted from the external medium. The applicability of using nutrient‐induced fluorescence transients as a tool to identify the nutrient status of phytoplankton populations is discussed.     

THALLUS DIFFERENTIATION OF PHOTOSYNTHESIS,GROWTH, REPRODUCTION,AND UV‐B SENSITIVITY IN THE GREEN ALGA ULVA PERTUSA (CHLOROPHYCEAE)1

The chlorophyte Ulva is perceived as a simple and uniform algal form, with little functional differentiation within a thallus. We compared morphology, pigmentation, photosynthesis, growth, reproduction, and UV‐B sensitivity between different thallus regions of Ulva pertusa Kjellman. Thallus thickness and cell size were significantly greater, whereas cell number was less in the basal region than in other regions. Photosynthetic pigment contents were lowest in the basal region and increased toward the marginal region. Photosynthetic capacity and photosynthetic efficiency normalized to fresh weight, area, volume, and cell number showed a progressive increase from the basal to marginal parts; however, on a chl basis those values were equal regardless of thallus part. Values of light saturation point were not statistically different between regions. Growth rates increased from marginal to basal and to middle parts of the thallus, whereas sporulation was highest in marginal (100%) followed by middle (30%) and basal parts (0%). Daily observation over 9 days showed that 56% of the basal cells divided once and did not produce spores, whereas every marginal cell went through its first division and 89% of the primary daughter cells also divided, resulting in 100% sporulation. A 7‐day treatment with PAR and PAR + UV‐A caused a significant decrease in the effective quantum yield of all thallus regions, followed by a recovery toward the initial values, whereas PAR + UV‐A + UV‐B irradiation led to greater photoinhibition and less recovery. Marked differences in the UV‐B sensitivity were observed, with marginal parts being more sensitive and basal parts most resistant.