Enhanced carotenoid production by a mutant of the marine yeast Rhodotorula sp. hidai

After a serial of UV, EMS and NTG mutagenesis, a mutant named MM of the red marine yeast strain Rhodotorula sp. hidai was obtained. The mutant MM could produce 603.93 μg g⁻¹ of carotenoid within 5 days in the medium containing 4.0 g sucrose, 1.5 g yeast extract, 0.1 g MgSO₄, and 100 mL of sea water, with pH 6.0 and at 30 °C, while only 213.18 μg g⁻¹ of carotenoid was produced by the wild type under the same conditions.     

不同条件下波吉卵囊藻对氨氮的吸收速率

在实验室条件下模拟多种氮源共存的虾池水环境,利用稳定同位素示踪剂,研究温度、光照、盐度、pH、藻浓度对波吉卵囊藻氨氮吸收速率的影响。结果表明:温度、光照和藻浓度对波吉卵囊藻氨氮吸收速率影响极显著(P<0.01)。当温度为20℃,照度为4 500 lx,藻浓度为5.5×105/mL,波吉卵囊藻对氨氮均有最大的吸收速率,分别为1.114、1.400、1.482μg g-1 h-1。单因素方差分析得出波吉卵囊藻吸收氨氮的最优组合为:温度20℃,照度4 500lx,盐度15,pH 7.5,藻浓度5.5×105/mL。可通过对环境因子的调控,提高波吉卵囊藻对水体中氨氮的吸收速率。     

云斑尖塘鳢和线纹尖塘鳢自交与杂交子一代幼鱼生长性状及杂交优势分析

分析了由云斑尖塘鳢(Oxyeleotris marmoratus)、线纹尖塘鳢(Oxyeleotris lineolatus)、云斑尖塘鳢(♀)×线纹尖塘鳢(♂)的杂交子一代3个实验组鱼的生长性状以及杂交子代的杂交优势。结果表明,养殖35 d后,云斑尖塘鳢与线纹尖塘鳢的杂交子一代幼鱼较云斑尖塘鳢幼鱼、线纹尖塘鳢幼鱼生长速率大;杂交尖塘鳢体质量为(1.6496±0.0501)g,比云斑尖塘鳢体质量(0.9353±0.1426)g和线纹尖塘鳢体质量(0.6690±0.0687)g差异显著(p<0.05);杂交尖塘鳢的全长为(53.63±0.83)mm,比云斑尖塘鳢全长(42.99±2.21)mm差异显著(p<0.05);从生长速率上看,杂交尖塘鳢幼鱼的杂种优势十分明显,平均为105.65%,超过云斑尖塘鳢亲本的超亲杂种优势为76.37%,超过线纹尖塘鳢亲本的超亲杂种优势为146.58%。在相同的人工养殖环境下,杂交使云斑尖塘鳢和线纹尖塘鳢幼鱼的生长得到了改良。     

Genome Sequence of a Marine Carotenoid Producing Yeast Rhodotorula mucilaginosa CYJ03

Carotenoids are valuable pigments that have been widely used in food,pharmaceutical,animal breeding and cosmetics industries.Due to the increasing demand for carotenoids of natural origin,the trend for production of carotenoids by red yeast has become popular.Strain Rhodotorula mucilaginosa CYJ03 was isolated from northern Yellow Sea of China for its carotenoid producing potential.It was found that the whole genome of CYJ03 was 19.03 Mb in size and contained 6301 protein-coding genes including a gene cluster for the carotenoids biosynthesis.The genome sequence would be valuable for exploring the potential biological properties of CYJ03,as well as for facilitating the molecular genetic analysis and the manipulation of carotenoids accumulation in this strain,and for the development of it as an engineered host for carotenoid production.     

温度、光照对小环藻生长和叶绿素a含量的影响

用单因子和正交实验方法研究了温度和光照对小环藻生长和叶绿素a含量的影响。结果表明,温度对小环藻生长和叶绿素a含量有极显著影响(P<0.01),当温度在30~40℃时比增长率和叶绿素a含量分别为0.37~0.38 d-1和1 084.4~1 214.2μg.L-1,均显著高于其他温度组。光照对小环藻生长和叶绿素a含量有极显著影响(P<0.01),当光照强度从29.25~146.26μmol.m-2.s-1时比增长率最高,为0.30~0.32 d-1;而叶绿素a含量则在9.75~58.50μmol.m-2.s-1时最高,达709.7~731.8μg.L-1,极显著高于其他照度组。正交实验结果表明:在35℃、87.75μmol.m-2.s-1条件下种群增长最快,比增长率为0.4402 d-1;在30℃、29.25μmol.m-2.s-1时,叶绿素a含量最高,为1 746.6μg.L-1。     

无定形碳酸钙结合蛋白对鼠前成骨细胞MC3T3-E1碱性磷酸酶活性的影响

以 MC3T3-E1细胞系为实验材料,探讨了真核表达的重组蛋白 rACCBP 对成骨细胞的作用.结果表明:rACCBP 对 MC3T3-E1的分化具有抑制作用,用含有0.001 g/mL rACCBP 的 DMEM 培养液,培养8 d 后细胞的ALP 活性仅为7.187 U/mg,远低于参照的11.917 U/mg; rACCBP 削弱了 dexamethane 和 ascorbic acid 对 MC3T3-E1的诱导作用     

Characterization of halophilic C50 carotenoid-producing archaea isolated from solar saltworks in Bohai Bay,China

Halophilic archaea comprise the majority of microorganisms found in hypersaline environments. C50 carotenoids accumulated in archaea cells are considered potential biotechnological products and possess a number of biological functions. Ten red colordes were isolated from brine water in a saltem crystaltizer pond of the Hangu Saltworks, China. 16S rRNA gene sequence analysis showed that the colonies belonged to the extremely halophilic archaea genera Halobacterium and Halorubrum. Two representative strains, Halobacterium strain SP-2 and Halorubrum strain SP-4, were selected for further study on the phenotypic characteristics and effects of salinity and pH on accumulation and composition of pigments in their cells. The archaeal strains were isolated and grown in a culture medium prepared by dissolving yeast extract （10 g/L） and acid-hydrolyzed casein （7.5 g/L） into brine water obtained from a I.ocal salt pond. Their optimum salinity and pH for growth were 250 and 7, respectively, although pigment accumulation （OD490/ mL broth） was highest at pH 8. In addition, at 150-300 salinity, increasing salinity resulted in decreasing pigment accumulation. Analysis of the UV-Vis spectrum, TLC and HLPC chromatograms showed that C50 carotenoid bacterioruberin is the major pigment in both strains.     

盐度对威氏海链藻生长和生化指标的影响

【目的】研究威氏海链藻在不同盐度条件下的生长和生化组分变化。【方法】设置盐度梯度为5、15、25、35、45,采用血细胞计数板计数法、热乙醇提取法、尼罗红染色法、苯酚硫酸法和考马斯亮蓝法,分别测定威氏海链藻在不同盐度下的生物量、叶绿素a、类胡萝卜素、中性脂质、总糖和蛋白质含量。【结果】盐度对威氏海链藻细胞密度影响显著(P<0.05),当盐度25和盐度35时,培养8 d细胞密度最高,均为2.0×10⁶ mL^-1;盐度对威氏海链藻叶绿素a和类胡萝卜素含量有显著影响(P<0.05),当盐度15时,细胞叶绿素a和类胡萝卜素含量均最高,分别为3.4814μg/mL(4 d)、2.3549μg/mL(8 d)。盐度对威氏海链藻可溶性总糖含量有显著影响(P<0.05),当盐度15时,8 d时可溶性总糖积累含量最高,为123μg/mL。当盐度35时,6 d时可溶性蛋白积累含量最高,为142.9μg/mL,且显著高于其他实验组(P<0.05)。盐度45时,8 d时中性脂含量最高,显著高于其他实验组(P<0.05)。【结论】盐度为15~35时,威氏海链藻生长速度较快,生化组成较为稳定,盐度15时最有利于藻细胞叶绿素a和类胡萝卜素、总糖的合成;盐度25和盐度35时,藻细胞平均增长速度较快,盐度35时最有利于藻细胞可溶性蛋白积累。     

Bioaccumulation of Pb^2＋ and Its Effects on Growth, Morphology and Pigment Contents of Spirulina （Arthrospira）platensis

A laboratory experiment was conducted to assess the bioaccumulation of Pb2 and its effects on growth,morphology andpigment contents of Spirulina (Arthrospira) platensis.The specimen cultured in Zarrouk liquid medium was treated with variousinitial metal concentrations (0,5,10,30,50 and 100 μgmL-1).The growth of S.platensis was adversely affected by Pb2 at high con-centrations (30,50 and 100 μg mL-1).However,at low concentrations (5 μg mL-1),Pb2 could stimulate its growth slightly.The pig-ment contents (chlorophyll α and β carotene) were decreased in a dose-dependent manner.The highest reductions (67% and 53%respectively in chlorophyll α and β carotene) were observed in 100 μg mL-1 treatment group.The LC50 (96 h) of Pb2 was measured as75.34μg mL-1.Apart from a few cases of filament breakages at elevated concentrations (50 and 100μg mL-1),morphological abnor-malities are not specific.Metal bioaccumulation increased with Pb2 concentrations,but decreased with exposure time.The maxi-mum accumulated amount was 188 mgg-1 dry weight.The bioconcentration factor (BCF) reached to a peak at day 2,followed by agradual reduction for all the exposure concentrations.S.platensis is able to tolerate considerably high Pb2 concentrations.Conse-quently it can be used as a potential species to remove heavy metal from contaminated waters.     

Effects of Pseudoalteromonas sp. BC228 on digestive enzyme activity and immune response of juvenile sea cucumber (Apostichopus japonicus)

A marine bacterium, Pseudoalteromonas sp. BC228 was supplemented to feed in a feeding experiment aiming to determine its ability of enhancing the digestive enzyme activity and immune response of juvenile Apostichopus japonicus. Sea cucumber individuals were fed with the diets containing 0(control), 105, 107 and 109 CFU g-1 diet of BC228 for 45 days. Results showed that intestinal trypsin and lipase activities were significantly enhanced by 107 and 109 CFU g-1 diet of BC228 in comparison with control(P 0.01). The phagocytic activity in the coelomocytes of sea cucumber fed the diet supplemented with 107 CFU g-1 diet of BC228 was significantly higher than that of those fed control diet(P 0.05). In addition, 105 and 107 CFU g-1 diet of BC228 significantly enhanced lysozyme and phenoloxidase activities in the coelomic fluid of sea cucumber, respectively, in comparison with other diets(P 0.01). Sea cucumbers, 10 each diet, were challenged with Vibrio splendidus NB13 after 45 days of feeding. It was found that the cumulative incidence and mortality of sea cucumber fed with BC228 containing diets were lower than those of animals fed control diet. Our findings evidenced that BC228 supplemented in diets improved the digestive enzyme activity of juvenile sea cucumber, stimulated its immune response and enhanced its resistance to the infection of V. splendidus.     

TraceeElements in Leccinum scabrum mushrooms and topsoils from Kłodzka Dale in Sudety Mountains, Poland

In the current study, we determined concentrations and transfer rates of Ag, Al, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, Pb, P, Rb, Sr and Zn to Brown Birch Scaber Stalks (Leccinum scabrum) mushrooms emerged in the area of Sudety Mountains (Sudetes) in Poland. Fruiting bodies and topsoil samples beneath L. scabrum were collected form the K odzka Dale. The trace elements were determined using validated method and inductively coupled plasma - atomic emission spectroscopy (ICP-AES) for final measurement. Mushrooms contained Ag, Cr, Hg, Co, Ni and Sr at < 1.0 μg/g dry weight; Ba and Pb at ~1.0 μg/g dw; Cd at < 5 μg/g dw; Cu and Mn at > 10 μg/g dw; Al and Ca at ~100 μg/g dw; Fe, Na, Rb and Zn at 100 to 500 μg/g dw, Mg at ~1,000 μg/g dw; P at ~5,000 μg/g dw and K at ~30,000 μg/g dw. Ca, Mn and Ni were nearly equally distributed between stipes and caps; stipes compared to caps were enriched in Ba, Na and Sr, while caps were enriched in Ag, Al, Cd, Co, Cr, Cu, Fe, K, P, Pb, Rb and Zn. The values of bioconcentration factor (BCF) varied highly depending on chemical element and were >1 for Ag, Cd, Cu, K, Mg, Na, P, Rb and Zn, while <1 for Al, Ba, Ca, Co, Cr, Fe, Mn, Ni, Pb and Sr. Topsoil showed elevated content of lead and mean concentration was 99 ± 32 μg/g dw, while cadmium was at 0.41 ± 0.15 and those two highly toxic to human elements occurred in edible caps of L. scabrum at 4.5 ± 2.2 and 2.9 ± 2.0 μg/g dw, respectively.     

环境胁迫对栉孔扇贝(Chlamys farreri)血浆蛋白及其抗菌活性的影响

采集采自山东某养殖场的栉孔扇贝(Chlamys farreri)健康贝和病贝的血淋巴,测定其血浆蛋白浓度、血浆蛋白图谱和相对分子质量,以及血浆抗菌活性、凝集素活性等几种重要的体液免疫因素,研究环境胁迫对血浆蛋白及其抗菌活性的影响。结果表明:栉孔扇贝(C.farreri)血浆蛋白质量浓度约1 260~1 650μg/mL;血浆蛋白SDS-PAGE图谱显示3条蛋白带,相对分子质量分别为34 000、25 000和14 000;健康贝血浆抗菌活性指数BI为0.34,病贝BI为0.21;凝集素活力水平为1.602~2.204。氨氮胁迫下血浆蛋白浓度下降约780μg/mL,凝集素活力下降为1.301;高温胁迫下血浆蛋白质量浓度下降约为900μg/mL,凝集素活力为1.903;高盐胁迫下血浆蛋白浓度下降约为990μg/mL,凝集素活力为2.204。     

Speciation analysis of arsenic compounds in seafood by ion chromatography-atomic fluorescence spectrometry

Ion chromatography-ultra violet-hydride generation-Atomic Florescence Spectrometry was applied to detect 5 arsenic species in seafoods. The arsenic species studied include arsenobetaine(As B), arsenite(As(III)), dimethylarsinic acid(DMA), monomethylarsonic acid(MMA), and arsenate(As(V)), which were extracted from samples using 2% formic acid. Gradient elution using 33 mmol L~(-1) CH_3COONH_4 and 15 mmol L~(-1) Na_2CO_3 with 10 mL CH_3CH_2OH at pH 8.4 allowed the chromatographic separation of all the species on a Hamilton PRP-X100 anion-exchange column in less than 8 min. In this study, an ultrasound extraction method was used to extract arsenic species from seafood. The extraction efficiency was good and the recoveries from spiked samples were in the range of 72.6%–109%; the precision between sample replicates was higher than 3.6% for all determinations. The detection limits were 3.543 μg L~(-1) for As B, 0.4261 μg L~(-1) for As(III), 0.216 μg L~(-1) for DMA, 0.211 μg L~(-1) for MMA, and 0.709 μg L~(-1) for As(V), and the linear coefficients were greater than 0.999. We also developed an application of this method for the determination of arsenic species in bonito, Euphausia superba, and Enteromorpha with satisfactory results. Therefore, it was confirmed that this method was appropriate for the detection of arsenic species in seafood.     

湛江海域文昌鱼(Branchiostoma)线粒体Cyt b基因的全序列分析

测定了邻近湛江的茂名海区和硇洲岛海域文昌鱼各2个样本线粒体Cytb基因全序列,以尾索动物亚门的2种海鞘(Ciona spp.)和脊椎动物亚门的七鳃鳗(Petromyzon marinus)作为外群,利用分子系统学方法与GenBank收录的14个文昌鱼(Branchiostoma)的Cytb基因全序列进行了比较,UPGMA法构建了系统发生树。全序列分析表明:主产于大西洋(批针文昌鱼B.lanceolatum和佛罗里达文昌鱼B.floridae)、厦门文昌鱼(B.belcheri belcheri Gray)和湛江文昌鱼线粒体Cytb基因全序列长度为1143bp,终止密码子同为UAG;主产于青岛文昌鱼(B.belcheri tsingtaunese)和日本文昌鱼为1141bp,以T结尾,转录过程中构成UAA终止密码子。系统分析表明:16个文昌鱼样品体现出太平洋和大西洋、太平洋南北明显分化。确认采自茂名和硇洲岛的文昌鱼为厦门文昌鱼(B.belcheribelcheriGray);支持将白氏文昌鱼青岛亚种(B.belcheri tsingtaunese)提升为种,命名为日本文昌鱼(B.japonicus)。     

饥饿对褐菖鮋代谢水平的影响

在水温18℃±1℃和盐度27.0±0.5条件下,研究了饥饿对褐菖鮋耗氧率和排氨率的影响。结果表明:对照组的平均耗氧率为1.082 mg.g-1.h-1;饥饿组耗氧率随着饥饿时间的延长而下降,饥饿期间,褐菖鲉的平均耗氧率为0.893 mg.g-1.h-1,总体上低于对照组。排氨率随着饥饿时间的延长下降趋势呈阶段性变化:饥饿1 d、3 d后的排氨率略低于对照组但差异不显著(P>0.05);饥饿3～7 d,排氨率下降幅度变大,饥饿7d较饥饿3 d的排氨率下降了185.075%;饥饿14 d、21 d后排氨率分别为0.053 mg.g-1.h-1和0.028 mg.g-1.h-1,呈继续下降趋势,但下降幅度减缓。褐菖鮋的nO∶nN比值始小幅度上升,7 d时开始迅速上升,21 d后升高至27.754,差异显著(P<0.05)。     

Strain and culture medium optimization for production enhancement of prodiginines from marine-derived Streptomyces sp. GQQ-10

A mutant (GQQ-M6) of a Sponge-Derived streptomyces sp. GQQ-10 obtained by UV-induced mutation was used for producing prodiginines (PGs). Single factor experiments and orthogonal array design (OAD) methods were employed for medium optimization. In the single factor method, the effects of soluble starch, glucose, soybean flour, yeast extract and sodium acetate on PGs production were investigated individually. In the subsequent OAD experiments, the concentrations of these 5 key nutritional components combined with salinity were further adjusted. The mutant strain GQQ-M6 gave a 2.2-fold higher PGs production than that of the parent strain; OAD experiments offered a PGs yield of 61mg L-1, which was 10 times higher than that of the initial GQQ-10 strain under the original cultivation mode.     

An Investigation on Tolerance and Accumulation of a Facultative Marine Fungus Aspergillus flavus to Pentavalent Arsenic

Tolerance of a facultative marine fungus Aspergillus flavus towards As (V) was tested.Luxuriant growth of the test isolate was observed in culture media with As (V) concentrations of 25 mg L-1 and 50 mg L-1,indicating its tolerance to the metal.Accumulation rate of arsenic was always higher when exposed to As (V) at 50 mg L-1 than at 25 mg L-1.The study reveals Aspergillus flavus as a promising candidate for environmental bioremediation.Arsenic contents (mg g-1) in the fungus when exposed to 50 mg L-1 As (V) were measured as 11.1773,4.0983,and 8.0000 mg g-1 on day 3,6 and 9,respectively.The highest content was observed initially,i.e.on day 3,followed by a decline and a rise again.These results provide baseline information for further explorations regarding the exploitation of the fungus for arsenic removal.     

TCDD对斑马鱼脂质过氧化作用的初步研究

将150条斑马鱼随机等分为5组:2,3,7,8-四氯代二苯并二噁英(TCDD)4个染毒组(染毒剂量分别为0.1μg/L、0.2μg/L、0.4μg/L和0.8μg/L)和空白对照组,水质接触染毒5d后,采用分光光度法测定其肝组织中丙二醛(MDA)含量和超氧化物歧化酶(SOD)、谷胱甘肽-S-转移酶(GST)的活力,研究TCDD对斑马鱼的脂质过氧化作用。结果发现,TCDD各剂量染毒组MDA含量均有所增加,其中0.2μg/L组差异显著(P<0.05),0.4μg/L和0.8μg/L剂量组差异极显著(P<0.01);各染毒组SOD活性均有所下降,其中0.2μg/L、0.4μg/L和0.8μg/L剂量组差异极显著(P<0.01);各染毒组GST活力均降低且差异极显著(P<0.01)。一定剂量的TCDD能引起MDA含量增加,使SOD和GST活力降低,对斑马鱼具有脂质过氧化作用。     

Effects of exposure to four endocrine disrupting-chemicals on fertilization and embryonic development of Barbel chub (Squaliobarbus curriculus)

The toxicities of 4 common endocrine-disrupting chemicals (EDCs), 17β-estradiol (E2), p,p′-dichlorodiphenyldichloro-ethylene (DDE), 4-nonylphenol (NP) and tributyltin (TBT), to sperm motility, fertilization rate, hatching rate and embryonic development of Barbel chub (Squaliobarbus curriculus) were investigated in this study. The duration of sperm motility was significantly shortened by exposure to the EDCs at the threshold concentrations of 10 ng L^?1 for E2 and TBT, 1 μg L^?1 for NP and 100 μg L^?1 for DDE, respectively. The fertilization rate was substantially reduced by the EDCs at the lowest observable effect concentrations (LOECs) of 10 ng L^?1 for E2 and TBT and 10 μg L^?1 for DDE and NP, respectively. Of the tested properties of S. curriculus, larval deformity rate was most sensitive to EDC exposure and was significantly increased by DDE at the lowest experimental level of 0.1 μg L^?1. Other EDCs increased the larval deformity rate at the LOECs of 1 ng L^?1 for E2, 10 ng L^?1 for TBT and 1 μg L^?1 for NP, respectively. Despite their decreases with the increasing EDC concentrations, the hatching rate and larval survival rate of S. curriculus were not significantly affected by the exposure to EDCs. The results indicated that all the 4 EDCs affected significantly and negatively the early life stages of the freshwater fish S. curriculus. Overall, E2 and TBT were more toxic than NP and DDE, while DDE might be more toxic to larval deformity rate than to other measured parameters. Thus, the 4 EDCs showed potential negative influences on natural population dynamics of S. curriculus. Our findings provided valuable basic data for the ecological risk assessment of E2, DDE, NP and TBT.     

Grazing of Salpa thompsoni on phytoplankton in summer in the Prydz Bay region, Antarctica

During the CHINARE-Ⅺ cruise in austral summer of 1998/1999, the abundance and feeding activity of Salps thompsoni were investigated in the Prydz Bav region. Salps samples were collected by vertical tows from the depth of 200 m to the surface with a conical net of 330 μm mesh-size. At Ⅵ-3 station, the grazing rates were studied by the gut fluorescence method and culture experiments. S. Thompsoni was mainly distributed in the northern part of the survey area, and its maximum densities reached to 2795 ind. · 1000 m-3. A dramatic decrease in salp stock was observed at the Marginal Ice Zone. The results of the feeding experiments at the Ⅵ-3 show that the gut pigment contents of S. thompsoni ranged from (0. 14 - 1.27) μig ind. -1(average 0. 98 μg ind.-1). The individual ingestion rate is 7. 9 μg ind. -1 · day-1, and the filtration rate is 28 L ind. -1 · day-1 Through the daily grazing rate of S. thompsoni, which takes account of less than 1% of the phytoplankton standing stock, it shows arelatively higher grazing impress on the primary production (72. 2 % ).