Ecdysteroid-responsive genes: E75 and Retinoid X Receptor (RXR), essential molt-regulating genes in the shrimp, Fenneropenaeus chinensis

本论文主要研究两种重要的调节蜕皮过程的基因—蜕皮激素效应基因E75和RXR在中国明对虾蜕皮中的作用。利用RT-PCR和RACE技术获得了编码FcE75和FcRXR的全长cDNA序列。FcRXR包含7个内含子，在对虾中存在不同的异形体，命名为RXR-1和RXR-2。应用荧光实时定量PCR分析表明FcE75和FcRXR基因在中国明对虾蜕皮前期（D3）其转录表达量明显上调。另外，FcE75和FcRXR基因在不同组织中的转录表达存在明显的差异。利用FcE75和FcRXR基因的双链RNA注射对虾能有效降低FcE75和FcRXR的表达水平。FcE75和FcRXR的体内沉默完全抑制了对虾的蜕皮过程，并且引起对虾的死亡。对不能正常蜕皮个体进行观察的结果表明，FcE75沉默的对虾，其上皮的收缩、新的刚毛及新表皮的形成均收到限制。在FcE75双链RNA沉默后的对虾中，我们检测了与蜕皮相关的一些效应因子，如chitinase等的转录，发现这些效应因子的转录明显受到抑制，说明FcE75和FcRXR在蜕皮过程中起到非常重要的作用。本论文首次阐明了这些基因在十足目甲壳动物蜕皮过程中的功能。

Effects of different light sources and illumination methods on growth and body color of shrimp Litopenaeus vannamei

Shrimps Litopenaeus vannamei with initial body weight of 2.108 +/- 0.036 g were sampled for specific growth rates (SGR) and body color measurements for 50 days under different light sources (incandescent lamp, IL; cool-white fluorescent lamp, FL; metal halide lamp, MHL; and control without lamp) and different illumination methods (illumination only in day, IOD, and illumination day and night, IDN). Body color of L. vannamei was measured according to the free astaxanthin concentration (FAC) of shrimp. The SGR, food intake (FI), feed conversion efficiency (FCE) and FAC of shrimps showed significant differences among the experimental treatment groups (P < 0.05). Maximum and minimum SGR occurred under IOD by MHL and IDN by FL, respectively (difference 56.34%). The FI of shrimp for the control group did not rank lowest among treatments, confirming that shrimp primarily use scent, not vision, to search for food. FI and FCE of shrimps were both the lowest among treatment groups under IDN by FL and growth was slow, thus FL is not a preferred light source for shrimp culture. Under IOD by MHL, shrimps had the highest FCE and the third highest FI among treatment groups ensuring rapid growth. FAC of shrimp were about 3.31 +/- 0.20 mg/kg. When under IOD by MHL and IDN by FL, FAC was significantly higher than the other treatments (P < 0.05). To summarize, when illuminated by MHL, L. vannamei had not only vivid body color due to high astaxanthin concentration but also rapid growth. Therefore, MHL is an appropriate indoor light source for shrimp super-intensive culture. SGR of shrimp was in significantly negative correlation to FAC of shrimp (P < 0.05). Thus, when FAC increased, SGR did not always follow, suggesting that the purpose of astaxanthin accumulation was not for growth promotion but for protection against intense light. (c) 2005 Elsevier B.V. All rights reserved.

Purification and characterisation of a natural lectin from the serum of the shrimp Litopenaeus vannamei

A natural lectin from the serum of the shrimp Litopenaeus vannamei was purified to homogeneity by a single-step affinity chromatography using fetuin-coupled agarose. The purified serum lectin (named LVL) showed a strong affinity for human A/B/O erythrocytes (RBC), mouse RBC, chicken RBC and its haemagglutinating (HA) activity was specifically dependent on Ca2+ and reversibly sensitive to EDTA. LVL inactive form had a molecular mass estimate of 172 kDa and was composed of two non-identical subunits (32 and 38 kDa) cross-linked by interchain disulphide bonds. Significant LVL activity was observed between pH 7 and 11. In HA-inhibition assays performed with several carbohydrates and glycoproteins, LVL showed a distinct and unique specificity for GalNAc/GluNAc/NeuAc which had an acetyl group, while glycoproteins fetuin and bovine submaxillary mucin (BSM) had sialic acid. Moreover, this agglutinin appeared to recognise the terminal N- and O-acetyl groups in the oligosaccharide chain of glycoconjugates. The HA activity of L. vannamei lectin was also susceptible to inhibition by lipopolysaccharides from diverse Gram-negative bacteria, which might indicate a significant in vivo role of this humoral agglutinin in the host immune response against bacterial infections. (C) 2006 Elsevier Ltd. All rights reserved.

Purification and characterization of a natural lectin from the plasma of the shrimp Fenneropenaeus chinensis

A natural lectin from the plasma of the shrimp Fenneropenaeus chinensis was purified by singlestep affinity chromatography using fetuin-coupled agarose. The purified plasma lectin showed a strong affinity for human A/B/O erythrocytes (RBC), mouse RBC and chicken RBC. The hemagglutinating (HA) activity of the lectin was dependent on Ca2+ and reversibly sensitive to EDTA. This lectin was named FC-L and its inactive form had a molecular mass estimate of 168 kDa. Fifteen N-terminal amino acid sequences of this protein were determined. We performed HA-inhibition assays with several carbohydrates and glycoproteins. FC-L showed a distinct and unique specificity to N-acetylated sugars, particularly sialic acid and sialoproteins. The FC-L also has binding activity to some Gram-negative bacteria which caused disease in shrimp and fish. The activity of FC-L was inhibited at temperatures greater than 75 degrees C and at a pH less than 7 or greater than 11. These results suggest that FC-L may play a role as pattern recognition proteins in the reorganization and clearance of invaders in shrimp F. chinensis. Crown Copyright (c) 2008 Published by Elsevier Ltd. All rights reserved.

PCR-DGGE analysis of intestinal bacteria and effect of Bacillus spp. on intestinal microbial diversity in kuruma shrimp (Marsupenaeus japonicus)

In this study, the intestinal microbiota of kuruma shrimp (Marsupenaeus japonicus) was examined by molecular analysis of the 16S rDNA to identify the dominant intestinal bacteria and to investigate the effects of Bacillus spp. on intestinal microbial diversity. Samples of the intestines of kuruma shrimp fed normal feed and Bacillus spp. amended feed. PCR and denaturing gradient gel electrophoresis (DGGE) analyses were then performed on DNA extracted directly from the guts. Population fingerprints of the predominant organisms were generated by DGGE analysis of the universal V3 16S rDNA amplicons, and distinct bands in the gels were sequenced. The results suggested that the gut of kuruma shrimp was dominated by Vibrio sp. and uncultured gamma proteobacterium. Overall, the results of this study suggest that PCR-DGGE is a possible method of studying the intestinal microbial diversity of shrimp.

Genetic improvement on Chinese shrimp (Fenneropenaeus chinensis): growth and viability performance in F1 hybrids of different populations

Fenneropenaeus chinensis distributed in the Yellow Sea and Bohai Sea of China and the west coast of the Korean Peninsula. Different geographical populations represent potentially different genetic resources. To learn further the characteristics of different geographical population, crosses among two wild and three farmed populations were produced. The two wild populations were from the Yellow Sea and Bohai Sea (WYP), and the west coast of the Korean Peninsula and coast (WKN). The three farmed populations included the offspring of first generation of wild shrimp from coast in Korea (FKN), the Huang Hai (the Yellow Sea in Chinese) No.1 (HH1), and JK98. The phenotypes growth and survival rates of these populations were compared to confirm the feasibility for crossbreeding. The body length (BL), carapace length (CL), carapace width (CW), height of the second and third abdominal segment (HST), width of the second and third abdominal segment (WST), length of the first abdominal segment (LF), length of the last abdominal segment (LL), live body weight (BW), and survival rate were measured. Different combinations were statistically performed with ANOVA and Duncan's Multiple Range Test. The results show that the survival rate of JK98(a (TM) Euro)xWKN(a (TM),) was the highest, followed by WYP(a (TM) Euro)xWKN(a (TM),), FKN(a (TM) Euro)xWYP(a (TM),), FKN(a (TM) Euro)xHH1(a (TM),) and WYP(a (TM) Euro)xFKN(a (TM),); the body weight of FKN(a (TM) Euro)sxHH1(a (TM),) was the highest, followed by FKN(a (TM) Euro)xWYP(a (TM),), WYP(a (TM) Euro)xWKN(a (TM),), WYP(a (TM) Euro)xFKN(a (TM),) and JK98(a (TM) Euro)xWKN(a (TM),); the total length had the same ranking as the body weight. All growth traits in hybrids JK98(a (TM) Euro)xWKN(a (TM),) were the lowest among all combinations. F1 hybrids had significant difference (P < 0.05) in BL, CL, HST, LL, and BW; and insignificant difference (P > 0.05) in other growth traits and survival rate. The results of Duncan's Multiple Range Test are that BL and CL of JK98(a (TM) Euro)xWKN(a (TM),) were significantly different from the other combinations; HST different from the combination of FKN(a (TM) Euro)xWYP(a (TM),), FKN(a (TM) Euro)xHH1(a (TM),) and WYP(a (TM) Euro)xWKN(a (TM),); and BW different from FKN(a (TM) Euro)xWYP(a (TM),) and FKN(a (TM) Euro)xHH1(a (TM),). As a whole, the results indicate that the FKN(a (TM) Euro)xHH1(a (TM),) was the best combination in all growth traits. Therefore, hybridization can introduce the variation to base populations. The systematic selection program based on additive genetic performance may be more effective than crossbreeding.

滇东南发现新元古代岩浆岩：SHRIMP锆石U-Pb年代学和岩石地球化学证据

滇东南老君山地区出露了一套穹隆状变形-变质岩系,新近完成的1：5万都龙幅、麻栗坡县幅区域地质矿产调查报告,将其分解为晚志留纪南温河花岗岩、新元古界新寨岩组和古元古界猛硐岩群.首次对猛硐岩群的石英角闪斜长片麻岩进行了SHRIMP锆石U-Pb定年,结合阴极发光图像分析,获得两组岩浆锆石的结晶年龄分别为761±12Ma和829±10Ma,指示滇东南老君山地区存在新元古代岩浆活动,同时从同位素年代学上,表明猛硐岩群是一套前寒武纪的变质沉积-岩浆杂岩.此外,还获得一些年龄为≈1.83Ga的残留锆石,表明该区存在古元古代的结晶基底.石英角闪斜长片麻岩及共存的斜长角闪岩,它们的原岩具有大陆裂谷背景下玄武质-玄武安山质岩浆系列的地球化学特征,可能与新元古代Rodinia超大陆聚合-裂解过程中的岩浆活动有关,本区在新元古代可能位于南华裂谷与康滇裂谷的交汇部位。

滇西北中旬松诺含矿斑岩的锆石SHRIMP U-Pb年龄及地质意义

位于义敦岛弧南端的中甸岛弧中广泛发育印支期斑岩及斑岩型和矽卡岩型铜矿床。松诺（或称松诺力赞）复式岩体位于东斑岩带中部，由石英闪长玢岩、黑云石英二长斑岩、闪长玢岩和含矿石英二长斑岩组成，其南部为普朗超大型斑岩铜矿床，北部为地苏嘎铜矿点。本文对含矿石英二长斑岩进行了岩相学和锆石SHRIMPU-Pb定年研究，结果表明所有锆石颗粒自形较好且均发育规则的韵律环带，Th含量为180-854μg／g，U含量为270-709μg／g，Th／U比值为0．77-1．24，为典型岩浆成因锆石。获得了含矿石英二长斑岩的侵位年龄为220．9±3．5Ma（n=9，MSWD=1．6），这与中甸岛弧洋壳俯冲造山作用的时限（210-235Ma）相吻合。

SHRIMP锆石年代学对西藏玉龙斑岩铜矿成矿年龄的制约

锆石SHRIMP U-Pb定年结果表明,玉龙岩体侵入时限至少为2.6Ma,其中成矿前石英二长斑岩的年龄为43.6±0.8Ma,成矿期黑云母二长花岗斑岩的年龄为41.0Ma±1.0Ma,代表了两幕较大的岩浆活动.玉龙斑岩铜矿主体成矿年龄约为40Ma,长时限多期幕式岩浆侵入和成矿期物理化学条件的剧变,是形成玉龙超大型斑岩铜矿的主要原因.玉龙斑岩铜矿与金沙江-红河成矿带众多新生代斑岩铜矿一样,属于印度-亚欧大陆45Ma陆陆主体碰撞之后第一次大规模应力释放的产物.

西昆仑山阿卡孜岩体锆石SHRIMP定年及其地球化学特征

西昆仑山阿卡孜岩体是该区已知的最古老侵入体。岩体侵入于太古代赫罗斯坦群(TTG岩系)中。获得最新的单颗粒锆石SHRIMP年龄为2426±46Ma。岩石地球化学研究表明，岩石具有高碱度富K特征；∑REE为409．94×10^-6～787．76×10^-6，6Eu为0．48～0．73；微量元素显示富集大离子元素。综合分析该岩体地质地球化学特征，认为该岩体形成于晚太古代-古元古代初期碰撞后拉张背景，是TTG岩系(赫罗斯坦群)重熔的结果。这对研究西昆仑山早前寒武纪陆壳增生及构造演化提供了重要的地质资料。