L-天冬氨酸、精氨酸多元体系中希土(Ⅲ)和钙(Ⅱ)化学形态

为有助于阐明体内希土化学形态，在前文工作基础上［1～4］，本文报道在模拟生理条件下用PH电位法和模拟计算法研究了希土和生命金属离子Ca(Ⅱ)在L-天冬氨酸(L-Asp)、L-精氨酸(L-Arg)多元体系中的化学形态，并比较Tb(Ⅲ)和Ca(Ⅱ)离子在…

Comparative expression analysis of transcription factor genes in the endostyle of invertebrate chordates

The endostyle of invertebrate chordates is a pharyngeal organ that is thought to be homologous with the follicular thyroid of vertebrates. Although thyroid-like features such as iodine-concentrating and peroxidase activities are located in the dorsolateral part of both ascidian and amphioxus endostyles, the structural organization and numbers of functional units are different. To estimate phylogenetic relationships of each functional zone with special reference to the evolution of the thyroid, we have investigated, in ascidian and amphioxus, the expression patterns of thyroid-related transcription factors such as TTF-2/MoxE4 and Pax2/5/8, as well as the forkhead transcription factors FoxQ1 and FoxA. Comparative gene expression analyses depicted an overall similarity between ascidians and amphioxus endostyles, while differences in expression patterns of these genes might be specifically related to the addition or elimination of a pair of glandular zones. Expressions of Ci-FoxE and BbFoxE4 suggest that the ancestral FoxE class might have been recruited for the formation of thyroid-like region in a possible common ancestor of chordates. Furthermore, coexpression of FoxE4, Pax2/5/8, and TPO in the dorsolateral part of both ascidian and amphioxus endostyles suggests that genetic basis of the thyroid function was already in place before the vertebrate lineage. (c) 2005 Wiley-Liss, Inc.

A serpin from Chinese shrimp Fenneropenaeus chinensis is responsive to bacteria and WSSV challenge

Arthropod defence responses (e.g. prophenoloxidase (proPO) activation and Toll pathway initiation) are mediated by serine proteinase cascades and regulated by serpins in haemolymph. A serpin (Fc-serpin) cDNA was cloned from the haemocytes of Fenneropenaeus chinensis by rapid amplification of cDNA ends (RACE) PCR and haemocyte cDNA library screening. The full-length cDNA consists of 1734 bp, encoding 411 amino acids with a calculated molecular mass of 46.55 kDa and a theoretical isoelectric point of 7.70. Fc-serpin contains a typical serpin-like homologue (serine proteinase inhibitors domain). The deduced protein contains a putative signal peptide of 19 amino acids and the serpin's signature sequence ((FHCNRPFLFLI389)-F-379). Fc-serpin showed some identity with Pacifastacus leniusculus serpin (42%) and Manduca sexta serpin-6 (34%). The reactive centre loop (RCL) sequences of Fc-serpin, P leniusculus serpin, M. sexta serpin-6 and Bombyx mori serpin-2 are highly similar. An Arg at the PI position of the reactive site indicates that Fc-serpin may have inhibitory activity against prophenoloxidase activating proteinase (PAP) and clotting enzyme. Transcripts of Fc-serpin mRNA were mainly detected in haemocytes and the lymphoid organ by RT-PCR. The variation of the mRNA transcription level in haemocytes followed by artificial infection with bacteria OF white spot syndrome virus (WSSV) was quantified by SYBR Green real-time PCR analysis. Expression profiles of Fc-serpin greatly fluctuated after challenge. This work represents the first report Of a serpin in penaeid shrimp. The data provide clues that Fc-serpin might play potential roles in the innate immunity of shrimp. (C) 2008 Elsevier Ltd. All rights reserved.

Molecular cloning and characterisation of a pattern recognition protein, lipopolysaccharide and beta-1,3-glucan binding protein (LGBP) from Chinese shrimp Fenneropenaeus chinensis

A pattern recognition protein (PRP), lipopolysaccharide and beta-1,3-glucan binding protein (LGBP) cDNA was cloned from the haemocyte of Chinese shrimp Fenneropenaeus chinensis by the techniques of homology cloning and RACE. Analysis of nucleotide sequence revealed that the full-length cDNA of 1,275 bp has an open reading frame of 1,098 bp encoding a protein of 366 amino acids including a 17 amino acid signal peptide. Sequence comparison of the deduced amino acid sequence of F. chinensis LGBP showed a high identity of 94%, 90%, 87%, 72% and 63% with Penaeus monodon BGBP, Litopenaeus stylirostris LGBP, Marsupenaeu japonicus BGBP, Homarus gammarus BGBP and Pacifastacus leniusculus LGBP, respectively. The calculated molecular mass of the mature protein is 39,857 Da with a deduced pI of 4.39. Two putative integrin binding motifs, RGD (Arg-Gly-Asp) and a potential recognition motif for beta-1,3-linkage of polysaccharides were observed in LGBP sequence. RT-PCR analysis showed that LGBP gene expresses in haemocyte and hepatopancreas only, but not in other tissues. Capillary electrophoresis RT-PCR method was used to quantify the variation of mRNA transcription level during artificial infection with heat-killed Vibrio anguillarum and Staphylococcus aureusin. A significant enhancement of LGBP transcription was appeared at 6 h post-injection in response to bacterial infection. These results have provided useful information to understand the function of LGBP in shrimp.

Alteration of metallothionein mRNA in bay scallop Argopecten irradians under cadmium exposure and bacteria challenge

Metallothionein (MT) is a superfamily of cysteine-rich proteins contributing to metal metabolism, detoxification of heavy metals, and immune response such as protecting against ionizing radiation and antioxidant defense. A metallothionein (designated AiMT2) gene was identified and cloned from bay scallop, Argopecten irradians. The full length cDNA of AiMT2 consisted of an open reading frame (ORF) of 333 bp encoding a protein of 110 amino acids. with nine characteristic Cys-X-Cys, five Cys-X-X-Cys, five Cys-X-X-X-Cys and two Cys-Cys motif arrangements and a conserved structural pattern Cys-x-Cys-x(3)-Cys-Tyr-x(3)Cys-x-Cys-x(3)-Cys-x-Cys-Arg at the C-terminus. The cloned ANT showed about 50% identity in the deduced amino acid sequence with previously published MT sequences of mussels and oysters. The conserved structural pattern and the close phylogenetic relationship of AiMT2 shared with MTs from other mollusc especially bivalves indicated that AiMT2 was a new member of molluscan MT family. The mRNA transcripts in hemolymph of AiMT2 under cadmium (Cd) exposure and bacteria challenge were examined by real-time RT-PCR. The mRNA expression of AiMT2 was up-regulated to 3.99-fold at 2 h after Listonella anguillarum challenge, and increased drastically to 66.12-fold and 126.96-fold at 16 and 32 h post-challenge respectively. Cadmium ion exposure could induce the expression of AiMT2, and the expression level increased 2.56-fold and 6.91-fold in hemolymph respectively after a 10-day exposure of 100 mu g L-1 and 200 mu g L-1 CdCl2. The sensitivity of AiMT2 to bacteria challenge and cadmium stress indicated it was a new Cd-dependent MT in bay scallop and also regulated by an immune challenge. The changes in the expression of AiMT2 could be used as an indicator of exposure to metals in pollution monitoring programs and oxidative stress, and bay scallop as a potential sentinel organism for the cadmium contamination in aquatic environment. (C) 2008 Elsevier Inc. All rights reserved.

Characterization of amphioxus GDF8/11 gene, an archetype of vertebrate MSTN and GDF11

MSTN, also known as growth and differentiation factor 8 (GDF8), and GDF11 are members of the transforming growth factor-beta (TGF-beta) subfamily. They have been thought to be derived from one ancestral gene. In the present study, we report the isolation and characterization of an invertebrate GDF8/11 homolog from the amphioxus (Branchiostoma belcheri tsingtauense). The amphioxus GDF8/11 gene consists of five exons flanked by four introns, which have two more exons and introns than that of other species. In intron III, a possible transposable element was identified. This suggested that this intron might be derived from transposon. The amphioxus GDF8/11 cDNA encodes a polypeptide of 419 amino acid residues. Phologenetic analysis shows that the GDF8/11 is at the base of vertebrate MSTNs and GDF11s. This result might prove that the GDF8/11 derived from one ancestral gene and the amphioxus GDF8/11 may be the common ancestral gene, and also the gene duplication event generating MSTN and GDF11 occurred before the divergence of vertebrates and after or at the divergence of amphioxus from vertebrates. Reverse transcriptase polymerase chain reaction results showed that the GDF8/11 gene was expressed in new fertilized cell, early gastrulation, and knife-shaped embryo, which was different from that in mammals. It suggested that the GDF8/11 gene might possess additional functions other than regulating muscle growth in amphioxus.

Complete mitochondrial genome of the Asian paddle crab Charybdis japonica (Crustacea: Decapoda: Portunidae): gene rearrangement of the marine brachyurans and phylogenetic considerations of the decapods

Given the commercial and ecological importance of the Asian paddle crab, Charybdis japonica, there is a clearly need for genetic and molecular research on this species. Here, we present the complete mitochondrial genome sequence of C. japonica, determined by the long-polymerase chain reaction and primer walking sequencing method. The entire genome is 15,738 bp in length, encoding a standard set of 13 protein-coding genes, two ribosomal RNA genes, and 22 transfer RNA genes, plus the putative control region, which is typical for metazoans. The total A+T content of the genome is 69.2%, lower than the other brachyuran crabs except for Callinectes sapidus. The gene order is identical to the published marine brachyurans and differs from the ancestral pancrustacean order by only the position of the tRNA (His) gene. Phylogenetic analyses using the concatenated nucleotide and amino acid sequences of 13 protein-coding genes strongly support the monophyly of Dendrobranchiata and Pleocyemata, which is consistent with the previous taxonomic classification. However, the systematic status of Charybdis within subfamily Thalamitinae of family Portunidae is not supported. C. japonica, as the first species of Charybdis with complete mitochondrial genome available, will provide important information on both genomics and molecular ecology of the group.

Chromosomal rearrangement in Pectinidae revealed by rRNA loci and implications for bivalve evolution

Karyotype and chromosomal localization of major (18-5.8-28S) and minor (5S) ribosomal RNA genes were studied in two species of Pectinidae, zhikong (Chlamys farreri) and bay (Argopecten irradians irradians) scallops. using fluorescence in situ hybridization (FISH). C. farreri had a haploid number of 19 with a karyotype of 3m + 4sm + 7sm-st + 4st + 1st-t, and A. i. irradians had a haploid number of 16 with a karyotype of 5st + 11t. In C. farreri, the major and minor rRNA genes had one locus each and were mapped to the same chromosome-Chromosome 5. In A. i. irradians, the major rRNA genes had two loci, located on Chromosomes 4 and 8, and the 5S rRNA gene was found at a third chromosome-Chromosome 10. Results of this and other studies indicate that karyotype of A. i. irradians (n = 16, 21 arms) is secondary and derived from an ancestral karyotype similar to that of C. farreri (n = 19, 38 arms) through considerable chromosomal loss and rearrangements. The ability to tolerate significant chromosomal loss suggests that the modal karyotype of Pectinidae and possibly other bivalves with a haploid number of 19 is likely tetraploid; i.e., at least one genome duplication has occurred during the evolution of Bivalvia.

Inbreeding depression for various traits in two cultured populations of the American bay scallop, Argopecten irradians irradians Lamarck (1819) introduced into China

This paper examines the effect of inbreeding level of population on the magnitude of inbreeding depression expressed by comparing them between two cultured populations (A and B) in the hermaphroditic animal of the bay scallop Argopecten irradians irradians. Population A is expected to have less genetic variations and higher inbreeding level due to longer cultured history (20 generations) and less "ancestral" individuals (26 individuals) than population B due to shorter cultured history (4 generations) and more "ancestral" individuals (406 individuals). Two groups within each population were produced, one using self-fertilization and one using mass-mating within the same population. Selfed offspring (AS and BS) from two populations both had lower fitness components than their mass-mated counterparts (AM and BM) and exhibited inbreeding depression for all examined traits, e.g. lower hatching, less viability and slower growth, indicating that inbreeding depression is a common feature in this animal. Fitness components in all traits of offspring from population A significantly differed those from population B and the magnitude of inbreeding depression for all traits in population A with higher inbreeding level was significantly smaller than that in population B with lower inbreeding level, indicating that both fitness components and magnitude of inbreeding depression were significantly affected by inbreeding level of populations and genetic load harbored in population A may be partially purged through inbreeding. Moreover, the magnitude of inbreeding depression in the two populations both varied among traits and life history stages. The present results support the partial-dominance hypothesis of inbreeding depression. (C) 2008 Elsevier B.V. All rights reserved.

Potential refugium on the Qinghai-Tibet Plateau revealed by the chloroplast DNA phylogeography of the alpine species Metagentiana striata (Gentianaceae)

Metagentiana striata is an alpine annual herbaceous plant endemic to the east of the Qinghai-Tibet (Q-T) Plateau and adjacent areas. The phylogeography of M. striata was studied by sequencing the chloroplast DNA (cpDNA) trnS-trnG intergenic spacer. Ten haplotypes were identified from an investigation of 232 individuals of M. striata from 14 populations covering the entire geographical range of this species. The level of differentiation amongst populations was very high (G(ST) = 0.746; N-ST = 0.774) and a significant phylogeographical structure was observed (P < 0.05). An analysis of molecular variance found a high variation amongst populations (76%), with F-ST = 0.762 (highly significant, P < 0.001), indicating that little gene flow occurred amongst the different regions; this was explained by the isolation of populations by high mountains along the Q-T Plateau and adjacent areas (N-m = 0.156). Only one ancestral haplotype (A) was common and widespread throughout the distributional range of M. striata. The populations of the Hengduan Mountains region of the south-eastern Q-T Plateau showed high diversity and uniqueness of haplotypes. It is suggested that this region was the potential refugium of M. striata during the Quaternary glaciation, and that interglacial and postglacial range expansion occurred from this refugium. This scenario was in good agreement with the results of nested clade analysis, which inferred that the current spatial distribution of cpDNA haplotypes and populations resulted from range expansion, together with past allopatric fragmentation events. (c) 2008 The Linnean Society of London.

Phylogeography of the Qinghai-Tibetan Plateau endemic Juniperus przewalskii (Cupressaceae) inferred from chloroplast DNA sequence variation

The vegetation of the northeast Qinghai-Tibetan Plateau is dominated by alpine meadow and desert-steppe with sparse forests scattered within it. To obtain a better understanding of the phylogeography of one constituent species of the forests in this region, we examined chloroplast trnT-trnF and trnS-trnG sequence variation within Juniperus przewalskii, a key endemic tree species. Sequence data were obtained from 392 trees in 20 populations covering the entire distribution range of the species. Six cpDNA haplotypes were identified. Significant population subdivision was detected (G(ST) = 0.772, N-ST = 0.834), suggesting low levels of recurrent gene flow among populations and significant phylogeographic structure (N-ST > G(ST), P < 0.05). Eight of the nine disjunct populations surveyed on the high-elevation northeast plateau were fixed for a single haplotype (A), while the remaining, more westerly population, contained the same haplotype at high frequency together with two low frequency haplotypes (C and F). In contrast, most populations that occurred at lower altitudes at the plateau edge were fixed or nearly fixed for one of two haplotypes, A or E. However, two plateau edge populations had haplotype compositions different from the rest. In one, four haplotypes (A, B, D and E) were present at approximately equivalent frequencies, which might reflect a larger refugium in the area of this population during the last glacial period. Phylogenetic analysis indicated that the most widely distributed haplotype A is not ancestral to other haplotypes. The contrasting phylogeographic structures of the haplotype-rich plateau edge area and the almost haplotype-uniform plateau platform region indicate that the plateau platform was recolonized by J. przewalskii during the most recent postglacial period. This is supported by the findings of a nested clade analysis, which inferred that postglacial range expansion from the plateau edge followed by recent fragmentation is largely responsible for the present-day spatial distribution of cpDNA haplotypes within the species.

A preliminary analysis of the phylogeny of the Swertiinae (Gentianaceae) based on ITS data

To better understand the floral diversity and the phylogeny of the Swertiinae (Gentianaceae), the internal transcribed spacer (ITS) region of nrDNA for 17 species and one outgroup was sequenced. Our data suggest that corolla type, gland shape, and corolla appendage are poorly correlated with the ITS phylogeny. The genus Swertia s.l. and the rotate group previously recognized based on the corolla types and gland shapes are polyphyletic. Four genera with simple protruding glands and three taxa with corolla appendages are not clustered as the monophyletic groups. Four separate clades, corresponding to the four sections, were identified in Swertia s.1. Lomatogoniopsis with the simple protruding gland type of the tubular group closely related to Lomatogonium of the rotate group. The deeply lobing corolla and concave foveae may be ancestral in the Swertiinae, while the tubular corolla and the protruding glands may have undergone convergent evolution.

空间注意系统中的动物特异性模块

Human being’s visual attentional system is the direct results of millions of years of evolutionary selection. As an adaptation to the environment, the most prominent function of attentional system is to facilitate the effective selection and subsequent processing of the most critical information and events from the environment with the aim of enhancing a given individual’s chance of passing his/her gene to the next generation. In the living environment of ancestral human beings, animals were undoubtedly one of those stimulus categories of great evolutionary significance. Since the process of animal-related information had life-or-death consequences for ancestral human beings, some researchers proposed a so-called animate monitoring hypothesis which states that there exists a category-specific module in the attentional system of human beings which specializes in the detection and frequent re-inspection of animal stimuli. Drawing on the available findings and theories regarding the inhibition-of-return effect, the present study utilized several variants of the spatial cueing paradigm to test the two main predictions of animate monitoring hypothesis：（1） animal stimuli in the environment are capable of summon attention in a reflexive way; （2） the inhibitory effect of attentional process on animal stimuli is less pronounced when compared to stimuli of other categories. The results of the present study provide supportive evidence to the existence of a category-specific module for animals in the attentional system. The present study contributes to the further understanding of the important role played by attentional mechanism in solving the critical adaptive problem faced by ancestral human beings during the course of evolution.

Cloning and structure analysis of hydrogenase gene from Chlamydomonas reinhardtii SE

The cDNA of Chlamydomonas reinhardtii SE encoding hydrogenase (HydA2) was obtained from the total RNA of C reinhardtii SE by RT-PCR. The DNA of hydrogenase was amplified by PCR from the genomic DNA of C reinhardtii SE. The cDNA and DNA of hydrogenase were sequenced, respectively. The structure of hydrogenase gene was analyzed by biology software. The open reading frame predicts that the hydrogenase is composed of 3584 bp encoding 505 amino acids in length with a predicted M.W. of 53.69 kDa. Ten exons (including 1518 bp) and nine introns (including 2066 bp) have been found in the hydrogenase, and there were two potential N-glycosylate sites, eight protein kinase C phosphorylation site, eight casein kinase H phosphorylation site and one sulphorylation in the sequence. The theory pI was 6.15. Total number of negatively charged residues (Asp + Glu) and positively charged residues (Arg + Lys) were 55 and 61, respectively. (c) 2005 Elsevier Ltd. All rights reserved.

Desempenho de clones de seringueira sob diferentes sistemas de sangria.

Este trabalho teve como objetivo avaliar o crescimento e a producao da borracha de clones seringueira [Havea brasiliensis ( Wild. Ex Adr. De Juss.) Muell. Arg.] sob diferentes sistemas de sangria, em condição de Cerrado dos Municipios de Barro Alto Goianesia no Estado de Goais. O plantio foi feito em fevereiro de 1992, no espacamento de 8,0 x 2,5 m (500 plantas/ha), em talhoes de 8 a 10 hectares para cada um dos clones RRIM 600, GT 1, PB 217, PB 235, PR 107 e PR 255 os quais receberam as mesmas praticas de manejo. Aos oito anos de idade, foram feitas as seguintes avaliacoes: estande final; circunferencia do caule a 1,20 m do solo; porcentagens de plantas aptas a sangria; producao de borracha acumulada na caneca pesada mensalmente; incidencia de seca de painel. A producao foi avaliada em nove sistemas de sangria em meia espiral (1/2 S), praticados cinco dias por semana (5d/7) e 10 meses ao ano (10m/12), variando na frequência de sangria (d/4 e d/7 = a cada 4 e 7 dias), a concentracao de Ethephon (ET 0,25%, 2,5%,3,3% e 5,0%) e sua frequencia de aplicacao durante o periodo chuvoso ( a cada 22, 28 e 35 dias), como segue: 1) 1/2S, d/7, ET 2,5% a cada 22 dias; 2)1/2S, d/7, ET 2,5% a cada 30 dias (referncia); 3) 1/2S. d/4, ET 2,5% a cada 30 dias; 4) 1/2S, d/7, ET 3,3% a cada 22 dias; 5)1/2S, d/7, ET 3,3% a cada 30 dias; 6) 1/2S. d/7, ET 5,0% a cada 22 dias 7) 1/2S, d/7, ET 5,0% a cada 30 dias; 8) 1/2S, d/7, ET 5,0% a cada 35 dias; 9) 1/2S, d/7, ET 0,25% (pulverizando 10 ml por painel) a cada 22 dias. Nos sistemas 1 a 8, o Ethephon foi pincelado ( 1mL) na canaleta de corte e ate 2 cm acima dela (Pa e La). O delineamento experimental foi de blocos ao acaso, com quatro repeticoes de 10 plantas poe parcela. Cada clone constitui um experimento separado, sendo os resultados de producao acumulada anual submetidos a analise de variancia e, nos caso de significancia, as medias dos sistemas foram comparadas pelo teste Tukey, ao nível de 5% de probabilidade. Nao foi constatada qualquer incidencia de seca de painel e os resultados possibilitaram as seguintes conclusoes para as condicoes da regiao: 1) o sistema 1/2S, d/7, ET 2,5% a cada 30 dias e o mais indicado par a sangria dos clones PR 255, PR 107, PB 235, PB 217 e GT 1; 2) o sistema 1/2S, d/7, ET 3,3% a cada 30 dias e o mais indicado para a sangria do clone RRIM 600; 3) a producao individual de borracha em kg/planta/ano e maior nos clones RRIM 600, PB 217 e PR 255, enquanto a producao total em kg/ha/ano e superior nos clones RRIM 600 e PB 235; 4)os clones PB 217 e PR 255 sao menos adaptados a regiao, apresentando menores valores de estande final, circunferencia do caule, porcentagem de plantas em sangria e de producao total de borracha por hectare.