昆虫神经活性物质章鱼胺受体激动剂的设计、合成及其构效关系研究

以章鱼胺能系统为靶标,设计并合成了72个取代苄基亚胺基-2-(口恶)唑烷和取代苯胺基2-(口恶)唑啉化合物。测定了目标化合物对美州大蠊神经索腺苷酸环化酶的活化作用。结果发现配体取代基的变化可引起对章鱼胺受体活性的较大幅度改变。邻氯苄基亚胺基-4-苯基-2-(口恶)唑啉显示与 OA 受体的较高亲和力(V_(max)=33.39,Km=0.33μmol·L~(-1)。应用 Hansch-Fujita 方法、RSA、MFA、药效团分析和分子模型技术等计算机辅助设计方法对它们的结构活性关系进行剖析并建立了一些可信度高的模型。     

Age distribution of the sea urchin Strongylocentrotus nudus (A. Agassiz) in relation to algal zonation in a rocky coastal area on Oshika Peninsula, northern Japan

ABSTRACT: The age distribution of the sea urchin Strongylocentrotus nudus in relation to algal zonation was studied in a sublittoral rocky area on Oshika Peninsula, northern Japan, from June 1995 through to August 1996. The algal zonation observed there extends from a shallow area of cobbles to deeper areas dominated successively by Sargassum yezoense , Eisenia bicyclis and, finally, crustose coralline red algae. One-year-old sea urchins occurred only on the cobbles and crustose corallines in contrast to the presence of 2-year-old individuals from all areas and the restricted distribution of individuals aged 3 years and over to Eisenia and crustose corallines. Calculated mean body sizes in the Eisenia area became larger after the age of 2 years compared with those in the area of crustose corallines. Spawning was ascertained histologically in both the Eisenia and crustose coralline areas, although gonad indices were higher in the former area than in the latter throughout the year. Gut contents were dominated by E. bicyclis in the Eisenia area, and by crustose coralline red algae and small algae in the crustose coralline area. Thus, the sea urchin may settle most abundantly on crustose corallines and lives there for 1–2 years. Thereafter, they expand their habitat toward the Eisenia area, where there is richer sources of food for attaining a higher growth rate and to generate heavier gonads.     

Expressed sequence tags from organ-specific cDNA libraries of tea (Camellia sinensis) and polymorphisms and transferability of EST-SSRs across Camellia species

Tea is one of the most popular beverages in the world and the tea plant, Camellia sinensis (L.) O. Kuntze, is an important crop in many countries. To increase the amount of genomic information available for C. sinensis, we constructed seven cDNA libraries from various organs and used these to generate expressed sequence tags (ESTs). A total of 17,458 ESTs were generated and assembled into 5,262 unigenes. About 50% of the unigenes were assigned annotations by Gene Ontology. Some were homologous to genes involved in important biological processes, such as nitrogen assimilation, aluminum response, and biosynthesis of caffeine and catechins. Digital northern analysis showed that 67 unigenes were expressed differentially among the seven organs. Simple sequence repeat (SSR) motif searches among the unigenes identified 1,835 unigenes (34.9%) harboring SSR motifs of more than six repeat units. A subset of 100 EST-SSR primer sets was tested for amplification and polymorphism in 16 tea accessions. Seventy-one primer sets successfully amplified EST-SSRs and 70 EST-SSR loci were polymorphic. Furthermore, these 70 EST-SSR markers were transferable to 14 other Camellia species. The ESTs and EST-SSR markers will enhance the study of important traits and the molecular genetics of tea plants and other Camellia species.     

Effects of replacing fish oil with canola oil on the growth performance,fatty acid composition and metabolic enzyme activity of juvenile yellowtail Seriola quinqueradiata (Temminck & Schlegel, 1845)

In this study, fish oil (FO) was replaced with canola oil (CO) in juvenile yellowtail diets to establish the optimal replacement levels that ensure adequate feed quality for aquaculture. Juvenile yellowtails (initial body weight: 104.7 g) were fed one of four diets for 10 weeks: 100% FO (FO group) or FO replacement with 25%, 50% or 100% CO (CO25, CO50 and CO100 groups respectively). Body weight, specific growth rate and feed efficiency were significantly affected by the replacement of FO with CO. The final body weight of the CO100 group was significantly lower than that of the FO (control) and CO50 groups. Furthermore, the CO50 group showed significantly higher feed efficiency than the FO group during the first 4 weeks (long photoperiod and high water temperature conditions). Increasing dietary CO increased serum glucose and triglyceride concentrations in a dose‐dependent manner. Regarding the proximate composition of fillet and liver, only crude protein in fillet was affected by the replacement of FO with CO. Fatty acid compositions of fillet and liver were correlated to the amount of CO in each diet. The activities of the metabolic enzymes phosphofructokinase and alanine aminotransferase were lowest in the CO25 and CO50 groups respectively. Thus, FO replacement with CO affected growth performance, serum components, fatty acid composition and the activity of metabolic enzymes. Overall, results obtained in the present study suggest that partial replacement (50%) of FO with CO is effective and might increase juvenile yellowtail growth under long photoperiod and/or high water temperature conditions.     

Genetic population evaluation of two closely related flatfish species, the rare barfin flounder and spotted halibut, along the Japanese coast

ABSTRACT:   Barfin flounder and spotted halibut have been selected as target species for stock enhancement in Japan. Understanding the genetic condition of the wild stock is a principal requirement in any stock enhancement program. The genetic variability of barfin flounder and spotted halibut, and the population structure of spotted halibut were evaluated using microsatellite DNA markers (msDNA) and the control region of the mitocondrial DNA (mtDNA). Barfin flounder and spotted halibut showed high genetic variability at the msDNA level. Barfin flounder A was 16.7 and H _e was 0.860; spotted halibut A _n ranged from 7.7 to 10.2 and H _e ranged from 0.710 to 0.774. At the mtDNA level, high haplotype ( h  = 0.922) and low nucleotide (π = 0.002) diversities were observed for barfin flounder; however, low haplotype and nucleotide diversities ( h  = 0.603–0.620 and π = 0.001–0.002), and very low haplotype and nucleotide diversities ( h  = 0.193 and π = 0.0003) were observed for spotted halibut in the north and south locations, respectively. Slight genetic differentiation among spotted halibut sampling locations was observed from the msDNA. MtDNA analyses showed genetic differentiation between north and south locations, but not within them. The designation of north-specific and south-specific management units in the future stock enhancement activities of spotted halibut is recommended.     

Evaluation of genetic diversity of eight grouper species <Emphasis Type="Italic">Epinephelus</Emphasis> spp. based on microsatellite variations

ABSTRACT:   Genetic diversity among eight species of grouper, Epinephelus bleekeri , E. coioides , E. malabaricus , E. ongus , E. akaara , E. maculatus , E. merra and E. fuscoguttatus , was studied using six microsatellite loci, Em-01 *, Em-03 *, Em-07 *, Em-08 *, Em-10 *, and CA-07 *, with the aim of exploring the feasibility of using microsatellite data for species identification. The results showed high levels of genetic differentiation among species ( F _ST  = 0.4403 and R _ST  = 0.4954). Species identification based on fixed allelic differences was possible between E. coioides , E. ongus , and E. fuscoguttatus at Em-01 * and between E. fuscoguttatus and E. ongus at Em-08 *. Private alleles were found in all species, except for E. ongus . Pairwise F _ST ranged 0.238–0.578 ( P  < 0.008 Bonferroni correction), and Nei's genetic distance ranged 0.433–2.710. Size homoplasy was observed at Em-03 * 157 allele, which was characterized by a T–C transition at the 119th nucleotide site of PCR products. The genetic assignment test unambiguously assigned each individual to the correct species. Thus, this test can be used for species identification of unknown individuals when the multilocus genotypes of the six microsatellite loci are available. The phylogenetic (neighbor-joining) tree, which was constructed based on the genetic distance matrix, separated the eight grouper species into two main groups.     

3个群体鳙鱼mtDNA D-loop区段的限制性片段长度多态性分析

鳙鱼（Aristichthys nobilis）采自江西瑞昌、湖南氐沙的天然群体以及天津宁河的人工繁殖群体共127尾，对其线粒体DNA的D-loop区段进行PCR扩增，使用12种核酸内切限制酶酶切。酶切结果显示，在12种内切酶中，8种有酶切位点，2种个体间有变异，为AfaⅠ和HinfⅠ，共得到7种单倍型。依据单倍型频率的组成情况，计算出瑞昌与长沙、宁河群体间的Rogers遗传距离分别为0．7283和0．5007，长沙和宁河群体间的Rogers遗传距离为0．8135。Afa Ⅰ的酶切类型C和HinfⅠ的酶切类型B组成的单倍型，在长沙群体中占87．5％，而在瑞昌和宁河群体中均为零。由此可以认为鳙鱼瑞昌群体和长沙群体可能是两个隔离的独立群体。     

Microsatellite multiplex panels for population genetic analysis of red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis>

In this study, multiplex PCR panels were developed to amplify 20 microsatellite markers for red sea bream, Pagrus major, with the aim of reducing labor and material costs associated with genetic analysis of this species. The usefulness of these panels was validated in 200 fish collected at five sampling locations. The occurrence of null alleles was suggested at five markers, which were dropped from further analysis. The remaining 15 microsatellite loci showed high levels of heterozygosity (H _E range 0.34–0.97, H _O range 0.32–1) and a wide range in the number of alleles per locus (A; range 5–46). Increasing the number of microsatellite loci from three to ten and 15 improved the performance of the panels for population differentiation (F _ST) and estimation of Nei’s (D _S) genetic distance. The results of this study confirm the usefulness of genotyping a large number of microsatellite DNA markers to expand our knowledge of red sea bream population genetics.     

Genetic differences in some haematological traits of communally reared clonal ayu, Plecoglossus altivelis Temminck & Schlegel, under stressed and non-stressed conditions

Four clonal lines of ayu, Plecoglossus altivelis Temminck & Schlegel, were produced through gynogenesis, mixed before hatching and reared communally. After 10 months, a randomly taken sample was subjected to a standardized shallow-water stressor. Haematocrit, haemoglobin, red blood cells count (RBC) and mean corpuscular volume (MCV) were obtained from stressed and non-stressed fish. DNA fingerprinting was used to confirm the clonal nature of the organisms and to identify the clonal line to which each fish belonged. We observed significant differences between clonal lines mostly in the haematocrit and MCV measured under no-stress conditions. Such differences are suggested to represent mainly genetic variance, on account of the common environment provided to all the experimental groups. The stress response ratio was lower than expected, mainly due to some unexpectedly high non-stress values. Heritability values (h²) were medium to high for the no-stress measurements (mean 0.238) and very low or zero for the stressed groups’traits (excepting one high value of 0.484). We conclude that the use of communally reared clonal lines represents a good tool for the characterization of the physiological traits, thus allowing for their utilization as genetic selection criteria.