Mutations in exons of the CYP17-II gene affect sex steroid concentration in male Japanese flounder (Paralichthys olivaceus)

As a specific gene of fish,cytochrome P450c17-Ⅱ(CYP17-Ⅱ) gene plays a key role in the growth,development and reproduction level of fish.In this study,the single-stranded conformational polymorphism(SSCP) technique was used to characterize polymorphisms within the coding region of CYP17-Ⅱ gene in a population of 75 male Japanese flounder(Paralichthys olivaceus).Three single nucleotide polymorphisms(SNPs) were identified in CYP17-Ⅱ gene of Japanese flounder.They were c.G594A(p.G188R),c.G939A and c.G1502A(p.G490D).SNP1(c.G594A),located in exon 4 of CYP17-Ⅱ gene,was significantly associated with gonadosomatic index(GSI).Individuals with genotype GG of SNP1 had significantly lower GSI(P < 0.05) than those with geno-type AA or AG.SNP2(c.G939A) located at the CpG island of CYP17-Ⅱ gene.The mutation changed the methylation of exon 6.Indi-viduals with genotype AA of SNP2 had significantly lower serum testosterone(T) level and hepatosomatic index(HSI) compared to those with genotype GG.The results suggested that SNP2 could influence the reproductive endocrine of male Japanese flounder.How-ever,the SNP3(c.G1502A) located in exon 9 did not affect the four measured reproductive traits.This study showed that CYP17-Ⅱ gene could be a potentially useful candidate gene for the research of genetic breeding and physiological aspects of Japanese flounder.     

Effects of BPA and E2 on expression profiles of genes related to hypothalamic-pituitary-gonadal axis of half-smooth tongue sole Cynoglossus semilaevis

Endocrine disrupting chemicals (EDCs) are increasingly viewed as persistent pollutants, similar to natural hormones in function. This paper describes the expression profiles of 7 genes (DMRT, VTG, GnRHR, FSHR, CYP17A, CYP19A, and CYP19B) involved in sex steroid synthesis and action as well as sexual development in adult male and female Cynoglossus semilaevis, after exposure to different concentrations of Bisphenol A (BPA) and 17β-estradiol (E2). Both BPA (1, 10, 50, 125, and 250 mg/kg) and E2 (0.5, 5, and 10 mg/kg) induced changes in target gene expression, although the estrogenic effects of E2 as a model estrogen were stronger. Among the 7 genes, VTG, CYP17A and CYP19 responded strongly to BPA or E2 exposure and can thus serve as reference biomarkers for estrogenic EDCs exposure in marine teleosts. These data will provide a window to establish a hypothalamic-pituitary-gonadal model in C. semilaevis to better understand the effect pathways of EDCs.     

Dietary vitamin A,ascorbic acid and α-tocopherol affect the gonad development and reproductive performance of starry flounder Platichthys stellatus broodstock

The present trial was conducted with starry fl ounder Platichthys stellatus broodstock to evaluate the effects of dietary vitamin A,ascorbic acid,and α-tocopherol on the gonadal development and reproductive performance.8 000 IU/kg diet vitamin A(VA group),500 mg/kg diet ascorbic acid(Vcpp group),or 250 mg/kg diet α-tocopherol(α-TA group)was added into basal diet to create 3 vitamin experimental diets,respectively.Each diet was fed to 450 starry fl ounder broodstock for 104 days.Samples were collected weekly.The gonadosomatic index(GSI)of 3 vitamin experimental groups fi rst decreased and then increased.Maximum GSI of Vcpp group was higher than that of α-TA group but lower than that of VA group.The spawning periods of 3 vitamin experimental groups lasted 49,56,and 45 days,respectively.No mature eggs were observed in the control group during the trial.The absolute fecundity(AF)and relative fecundity(RF)of α-TA group was higher than that of Vcpp group but lower than that of VA group.The results suggest that different vitamins play different roles in the fi sh reproductive process.Vitamin A stimulated the maturation of the ovary,ascorbic acid prolonged the spawning period,and α-tocopherol affected the development of the eggs.     

双棘黄姑鱼卵巢发育的周年变化

观察了双棘黄姑鱼卵巢发育的周年变化,结果表明,双棘黄姑鱼雌鱼性腺成熟系数(GSI)一年有两个峰值,分别出现在5月份(9.38%±0.32%)和10月份(7.56%±0.43%);HSI与GSI密切相关[R=0.76];卵巢发育水温在19～29℃范围,繁殖水温为24～28℃,在广东省饶平海区,双棘黄姑鱼繁殖时间在每年的5月和10月,属于春、秋二季产卵类型的鱼类。     

Molecular characterization and expression profiles of four transformer-2 isoforms in the Chinese mitten crab Eriocheir sinensis

The transformer-2(tra-2) gene plays a key role in the regulatory hierarchy of sexual differentiation in somatic tissues and in the germline of Drosophila melanogaster.In this study,sequences and expression profiles of tra-2 in the Chinese mitten crab Eriocheir sinensis were characterized.Four tra-2 isoforms,designated as Estra-2a,Estra-2b,Estra-2c,and Estra-2d,were isolated.They all contained an RNA-recognition motif(RRM) and a linker region,which shared high similarity with other reported tra-2s.Sequence analysis revealed that Estra-2a,Estra-2b and Estra-2c are encoded by the same genomic locus and are generated by alternative splicing of the pre-mRNA.Compared with the other three isoforms,Estra-2d lacks the RS2 domain.Quantitative real-time PCR showed that all four isoforms were highly expressed in the fertilized egg,and in the 2-4 cell and blastula stages compared with larval stages(P0.01),suggesting their maternal origin in early embryonic developmental stages.Notably,Estra-2a was highly expressed in male somatic tissues,while Estra-2c was significantly highly expressed in the ovary.These results suggest that Estra-2c is involved in sexual differentiation of the Chinese mitten crab.Our findings provide basic information for further functional studies of the tra-2 gene/protein in this species.     

PRELIMINARY STUDY ON THE HOLOCENE ENVIRONMENT CHANGES IN XINJIANG──GEOLOGIC RECORDS AND SEQUENCE

ＰＲＥＬＩＭＩＮＡＲＹ ＳＴＵＤＹ ＯＮ ＴＨＥ ＨＯＬＯＣＥＮＥ ＥＮＶＩＲＯＮＭＥＮＴ ＣＨＡＮＧＥＳ ＩＮ ＸＩＮＪＩＡＮＧ──ＧＥＯＬＯＧＩＣ ＲＥＣＯＲＤＳ ＡＮＤ ＳＥＱＵＥＮＣＥ ＺｈｏｎｇＷｅｉ（钟巍）（ＤｅｐａｒｔｍｅｎｔｏｆＧｅｏ...     

Preliminary study on the holocene environment changes in xinjiang—geologic records and sequence

A great number of geologic records has shown that the climatic environment in Xinjiang in the Holocene was mainly characterized by warm-dry, although alternated with sub-fluctuations, several important boundaries of environmental changes can also be determined as follows: 12,000 a B.P., 9000 a B. P., 3000 a B.P., 1500 a B.P. and 400 a B.P.. Since the Holocene, Xinjiang has undergone the influence of two kinds of environmental development patterns (the westerly type and the monsoon type), so the hydro-thermal disposal here was much more complicated. Generally speaking, the neoglacials of Xinjiang are characteristics of cold (cool)-humid and in the Climatic Optimal, it is dominated by warm-humid conditions in spite of being intercalated several sub-fluctuations of warm-dry intervals.     

Marine bacterium strain screening and pyrethroid insecticide-degrading efficiency analysis

A pyrethroid insecticide-degrading bacterium, strain HS-24, was isolated from an offshore seawater environment. The strain, which can degrade cypermethrin(CYP) and deltamethrin(DEL), was identified as Methylophaga sp. The optimal culture and degradation conditions for CYP and DEL by strain HS-24 is pH 7 at 28°C. Under optimum culture conditions, strain HS-24 exhibited a broad degradation concentration range of 100, 200, 400, 600, and 800 mg/L for CYP and DEL. The metabolic intermediates were analyzed by NMR, which provided strong evidence that CYP and DEL removal occurred mainly because of a biological process. The toxicity of the degradation products of strain HS-24 was studied simultaneously by measuring the light output of the luminescence bacterium. This demonstrated that the biodegradation ability of strain HS-24 significantly decreased the toxicity of CYP- and DEL-contaminated aquaculture seawater. Finally, the findings of this paper indicate that strain HS-24 is thus revealed as a biological agent for the remediation of marine aquatic environments.     

Assessment of internal controls for data normalization of gene expression after different bacterial stimulation by quantitative real-time PCR in golden pompano Trachinotus blochii

Trachinotus blochii is one of the important commercial fish species.In this study,we aim to confirm the reliability reference genes in T.blochii during different bacterial challenge through quantitative real-time PCR(qRT-PCR).The expression of the seven selected genes in four immune organs(i.e.,spleen,kidney,intestine,and gill) stimulated with Vibrio harveyi,Edwardsiella tarda,and Streptococcus agalactiae were determined by qRT-PCR.The PCR data was analyzed using the geNorm and NormFinder algorithms.The results showed the selection of the internal controls should be tissue specific when studying gene expression in response to bacterial stimulation.After 48 h of stimulation with V.harveyi,geNorm ranked EF1 A/Actin,18 S rRNA/B2M,UBCE/B2M,and 18 S rRNA/B2M,as the most stably expressed genes in spleen,kidney,intestine,and gill,respectively.After 48 h of stimulation with E.tarda,geNorm ranked 18 S rRNA/EF1 A,18 S rRNA/B2M,B2M/RPL13,and 18 S rRNA/EF1 A,as the most stably expressed genes in spleen,kidney,intestine,and gill,respectively.After 48 h of stimulation with S.agalactiae,18 S rRNA/EF1 A,18 S rRNA/B2 M,B2 M/Actin,and 18 S rRNA/B2M were ranked as the most stably expressed genes in spleen,kidney,intestine,and gill,respectively.Compared to the results analyzed by geNorm,reference genes received similar rankings when using NormFinder software.The results showed that the reference genes appeared to be not only tissue specific,but also specific to the infecting species of bacteria.If one gene is preferred when T.blochii were infected by bacteria,18 S rRNA,B2M,B2M,18 S rRNA may be used in spleen,kidney,intestine,and gill,respectively.     

A new high-resolution Late Glacial-Holocene climatic record from eastern Nanling Mountains in South China

A 350-cm-long sediment core sequence from Dahu Swamp situated in the eastern Nanling Mountains was selected for high-resolution paleoclimatic reconstruction since the Late Glacial period. The multi-proxy records of this paper reveal several evidently dry and cold events that may coincide with the Oldest Dryas, the Older Dryas, the Younger Dryas in the late deglacial period. Two relatively wetter and warmer phases occurred in ca. 15,000–14,400 cal yr B.P. and 13,500–12,800 cal yr B.P. respectively may correspond to the Bølling and Allerød warming events. The Younger Dryas event (ca. 12,800–11,500 cal yr B.P.) revealed by multi-proxies was characterized by relatively colder and drier climate. A warmer and wetter climate, occurred in ca. 10,000–6000 cal yr B.P., was consistent with the Holocene Optimum, which coincided with the maximum Northern Hemisphere insolation. The “8.2kyr cool event” and even the “8.8kyr cool event” were indicated as well from our sediment core. A dry mid-Holocene period (ca. 6000–3000 cal yr B.P.) indicated by multi-proxies does not follow the traditional concept of the wet mid-Holocene conditions observed in other regions in China.     

Expression and localization of a novel phosducin-like protein from amphioxus Branchiostoma belcheri

A full length amphioxus cDNA, encoding a novel phosducin-like protein (Amphi-PhLP),was identified for the first time from the gut cDNA library of Branchiostoma belched. It is comprised of 1 550 bp and an open reading frame (ORF) of 241 amino acids, with a predicted molecular mass of approximately 28 kDa. In situ hybridization histocbemistry revealed a tissue-specific expression pattern of Amphi-PhLP with the high levels in the ovary, and at a lower level in the hind gut and testis, hepatic caecum, gill, endostyle, and epipharyngeal groove, while it was absent in the muscle, neural tube and notochord. In the Chinese Hamster Ovary (CHO) cells transfected with the expression plasmid pEGFP-NIIAmphi-PhLP, the fusion protein was targeted in the cytoplasm of CHO cells, suggesting that Amphi-PhLP is a cytosolic protein. This work may provide a framework for further understanding of the physiological function of Amphi-PhLP in B. belcheri.     

Diurnal expression of circadian clock genes period 1 and period 3 in Pelteobagrus vachellii

Circadian clock genes are crucial for generating and sustaining most rhythmic daily functions in the animal kingdom,which entrain the rhythms of biochemical,physiological,and behavioural processes.To better understand the molecular oscillations of the circadian rhythms in darkbarbel catfish(Pelteobagrus vachellii),we isolated and characterized two circadian clock genes in P.vachellii,period 1(perl),and period3(per3).The circadian clock gene perl was found to encode a 1 428-amino acid polypeptide,including PER-ARNT-SIM(PAS) dimerisation domains,a PAS-associated C-terminal motif(PAC),a short mutable domain(S/M),and a nuclear export signal(NES).The 4 902-bp per3 cDNA includes an open reading frame encoding a 1 292-amino acid residue polypeptide with a PER-ARNT-SIM(PAS) domain,cytoplasmic localisation domain(CLD),interaction site(TIS),and a nuclear localisation signal(NLS).The perl and per3 gene was constitutively expressed in all examined tissues.Moreover,perl expression within a light/dark cycles showed rhythmic expression in the diencephalon,brain,liver and intestine,with,the acrophase at 15:15,12:52,7:51,and 12:55,respectively.Daily expression of per3 was rhythmic in the diencephalon,brain,liver and intestine,with the acrophase at 8:15,9:54,10:39,and 10:25 h,respectively.These findings expand our understanding of circadian mechanism at the molecular level in this species.     

Study on vegetation and climate changes in Beijing region since late pleistocene

１　ＩＮＴＲＯＤＵＣＴＩＯＮＳｉｎｃｅｔｈｅ１９８０ｓａｓｅｒｉｅｓｏｆｒｅｓｅａｒｃｈｐｌａｎｉｎｃｌｕｄｉｎｇＩＧＢＰ（ｔｈｅＩｎｔｅｒｎａｔｉｏｎａｌＧｅｏｓｐｈｅｒｅ．ＢｉｏｓｐｈｅｒｅＰｒｏｇｒａｍｍｅ）ｈａｖｅｂｅｅｎｓｅｔｕｐ,ｏｆｔｈｅｍ,ＰＡＧＥＳ（ｔｈｅＰａｓｔＧｌｏｂａｌｃｈａｎｇｅｓ）ｉｓａｎｉｍｐｏｒｔａｎｔｃｏｒｅｐｒｏｊｅｃｔ．Ｔｈｅｎａｔｕｒｅｒｅｃｏｒｄｓｏｆｏｃｅａｎｄｅｐｏｓｉｔ,ｔｅｒｒｅｓｔｒｉａｌ（ｌｏｅｓｓ,ｌａｋｅｓａｎｄｓｗａｍｐ）ｄｅｐｏｓｉ…     

Antioxidant enzyme activities in different genders and tissues of amphioxus Branchiostoma belcheri tsingtauense

Information regarding antioxidant enzymes in amphioxus remains lacking, and this study was carried out to examine the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) in different genders and tissues of amphioxus Branchiostoma belcheri tsingtauense. Results show that (1) CuZn-SOD, CAT and GPX activities in the whole amphioxus B. belcheri tsing- tauense were basically at the same levels in male and female amphioxus, whereas both T-SOD and Mn-SOD activities in male amphioxus were significantly higher than that in the female (P<0.05); (2) The testis had significantly higher T-SOD and CuZn-SOD activities than the ovary (P<0.05); (3) CuZn-SOD activity was undetectable in the guts of male and female amphioxus; (4) For both male and female am- phioxus, the activities of CAT and GPX in the gonads including testis and ovary were the lowest (P<0.05) among the tissues examined; (5) The gut and gill had the same level GPX activities while the gut had a higher CAT activity; (6) There was no clear difference in CAT and GPX activities in the corresponding tissues between male and female amphioxus. The study on SOD, CAT and GPX activities in different genders and tissues of the protochordate provides data for future comparison of amphioxus antioxidant enzymes with those of invertebrates and vertebrates.     

Sporo-pollen Assemblage and Paleoclimate Events in Shelf Area of the Southern Yellow Sea Since 1.5 ka B.P.

Based on the authors‘ 1986 to 1994 sporo-pollen assemblage analysis in the southern Yellow Sea area, data from 3 main cores were studied in combination with ^14C, palaeomagnetic and thermoluminescence data. The evolution of the paleoclimate environments in the southern Yellow Sea since 15ka B.P. was revealed that, in deglaciation of the last glacial period, the climate of late glaciation transformed into that of postglaciation, accompanied by a series of violent climate fluctuations. These evolution events happened in a global climate background and related to the geographic changes in eastern China. We distinguished three short-term cooling events and two warming events. Among them, the sporo-pollen assemblage of subzone A1 showed some cold climate features indicating that a cooling event occurred at about 15 - 14ka. B .P. in early deglaciation. This subzone corresponds to the Oldest Dryas. In subzone A3 , many drought-enduring herbal pollens and some few pollens of cold-resistant Picea, Abies, etc. were found, which indicated that a cooling event, with cold and arid climate, occurred at about 12- 11ka. B.P. in late deglaciation. This subzone corresponds to the Younger Dryas. The sporo-pollen assemblage of zone B showed warm and arid climate features in postglaciation. Although the assemblage of subzone B2 indicated a cold and arid climate environment, the development of flora in subzone B2 climate was less cold than that in A3 . Subzone B2 indicated a cooling event which occurred at about 9ka B.P. in early Holocene. Subzone A2, with some distinct differences from subzone A1 and A3 , indicated a warming event which occurred at 14 - 13ka. B.P. and should correspond to a warming fluctuation. The sporo-pollen assemblage of zone C showed features of warm-moist flora and climate, and indicated a warming event which universally occurred along the coast of eastern China at 8 - 3ka B.P. in middle Holocene, and its duration was longer than that of any climate events mentioned above. This period was climatic optimum and belonged to an altithermal period in postglaciation.     

脂多糖、苯酚、硫酸铜对红笛鲷非特异性细胞毒性细胞受体基因表达的影响

应用Real-time PCR技术,研究脂多糖(lipopolysaccharide,LPS)、苯酚、硫酸铜刺激红笛鲷(Lutjanussanguineus)后非特异性细胞毒性细胞受体(NCCRP-1)基因在不同组织里的表达差异。结果发现,LPS刺激红笛鲷24 h后NCCRP-1在红笛鲷头肾、脾脏、胸腺、肝脏、心脏、脑、肌肉和肠组织中均有表达,其中头肾表达量最高,脾脏次之,然后依次是肝脏、脑、肌肉、胸腺和肠,心脏表达量最少。LPS、苯酚和CuSO4刺激红笛鲷后,随着刺激时间的增长,NCCRP-1表达量在各组织达到峰值的时间不同。以头肾为模式组织,RT-PCR的结果显示,红笛鲷NCCRP-1在LPS、苯酚和CuSO4的刺激下的表达模式相似,随着时间的增加NCCRP-1表达量逐渐增加,分别在24、9、12 h处达到最高,达到对照组的52、30、24倍左右,之后表达量开始下降。免疫组织化学表明,NCCRP-1只在头肾、脾脏和胸腺的特定细胞中表达。     

Characterization,expression and function analysis of DAX1 gene of scallop (Chlamys farreri jones and preston 1904) during its gametogenesis

DAX1, a member of nuclear receptor superfamily, has a function in the sex determination and gonadal differentiation of several vertebrate species. However, little information about DAX1 of invertebrates is available. Here we cloned a homolog of scallop (Chlamys farreri Jones and Preston 1904) dax1, Cf-dax1, and determined its expression characteristics at mRNA and protein levels. The cDNA sequence of Cf-dax1 was 2093 bp in length, including 1404 bp open reading frame (ORF) encoding 467 amino acids. Unlike those of vertebrates, no conserved LXXLL-related motif was found in the putative DNA binding region of Cf-DAX1. Fluorescence in situ hybridization showed that Cf-dax1 located on the short arm of a pair of subtelocentric chromosomes. Tissue distribution analysis using semi-quantitative RT-PCR revealed that Cf-dax1 expressed widely in adult scallop tissues, with the highest expression level found in adductor muscle, moderate level in mantle, gill and testis, and low level in kidney, ovary and hepatopancreas. The result of quantitative real-time PCR indicated that the expression of Cf-dax1 was significantly higher (P<0.05) in testis than in ovary at the same stage, showing a sex-dimorphic expression pattern. Furthermore, immunohistochemical detection found that Cf-DAX1 mainly located in spermatogonia and spermatocytes of testis and in oogonia and oocytes of ovary, implying that DAX1 may involve in gametogenesis of bivalves.