Complete nuclear ribosomal DNA sequence amplification and molecular analyses of <Emphasis Type="Italic">Bangia</Emphasis> (Bangiales,Rhodophyta) from China

Filamentous Bangia, which are distributed extensively throughout the world, have simple and similar morphological characteristics. Scientists can classify these organisms using molecular markers in combination with morphology. We successfully sequenced the complete nuclear ribosomal DNA, approximately 13 kb in length, from a marine Bangia population. We further analyzed the small subunit ribosomal DNA gene (nrSSU) and the internal transcribed spacer (ITS) sequence regions along with nine other marine, and two freshwater Bangia samples from China. Pairwise distances of the nrSSU and 5.8S ribosomal DNA gene sequences show the marine samples grouping together with low divergences (00.003; 0–0.006, respectively) from each other, but high divergences (0.123–0.126; 0.198, respectively) from freshwater samples. An exception is the marine sample collected from Weihai, which shows high divergence from both other marine samples (0.063–0.065; 0.129, respectively) and the freshwater samples (0.097; 0.120, respectively). A maximum likelihood phylogenetic tree based on a combined SSU-ITS dataset with maximum likelihood method shows the samples divided into three clades, with the two marine sample clades containing Bangia spp. from North America, Europe, Asia, and Australia; and one freshwater clade, containing Bangia atropurpurea from North America and China.     

Heavy metals in sediments and their bioaccumulation in <Emphasis Type="Italic">Phragmites australis</Emphasis> in the Anzali wetland of Iran

Accumulation of metals in both sediments and Phragmites australis organs was studied. Samples were collected from seven stations located in Anzali wetland, Iran. The samples were analyzed by inductively coupled plasma mass spectrometry (ICP-MS). The results showed that concentration of the studied metals (except As and Cd) were higher in sediments than in P. australis organs. Metal accumulation was found to be significantly (P <0.05) higher in roots than in above-ground organs of P. australis. The bioaccumulation factor (BAF) and the transfer factor (TF) also verified the highest rate of metal accumulation in roots and their reduced mobility from roots to the above-ground organs. Pearson correlation coefficient showed significant relationships between metal concentrations in sediments and those in plant organs. It should be pointed out that sediment and plant samples exhibited higher metal concentrations in eastern and central parts than in western and southern parts of the wetland. The mean concentrations of all studied elements (except for Fe, V and Al) were higher in these sediment samples than in the Earth’s crust and shale. High accumulation of metals in P. australis organs (roots and shoots) is indicative of their high bioavailability in sediments of the wetland. The correlation between metal concentrations in sediments and in P. australis indicates that plant organs are good bioindicators of metal pollution in sediments of Anzali wetland.     

Antialgal and antilarval activities of bioactive compounds extracted from the marine dinoflagellate <Emphasis Type="Italic">Amphidinium carterae</Emphasis>

With the global ban on the application of organotin-based marine coatings by the International Maritime Organization, the development of environmentally friendly, low-toxic and nontoxic antifouling compounds for marine industries has become an urgent need. Marine microorganisms have been considered as a potential source of natural antifoulants. In this study, the antifouling potential of marine dinoflagellate Amphidinium carterae, the toxic and red-tide microalgae, was investigated. We performed a series of operations to extract the bioactive substances from Amphidinium carterae and tested their antialgal and antilarval activities. The crude extract of Amphidinium carterae showed significant antialgal activity and the EC₅₀ value against Skeletonema costatum was 55.4 μg mL^?1. After purification, the isolated bioactive substances (the organic extract C) exhibited much higher antialgal and antilarval activities with EC₅₀ of 12.9 μg mL^?1 against Skeletonema costatum and LC₅₀ of 15.1 μg mL^?1 against Amphibalanus amphitrite larvae. Subsequently, IR, Q-TOFMS, and GC-MS were utilized for the structural elucidation of the bioactive compounds, and a series of unsaturated and saturated 16- to 22-carbon fatty acids were detected. The data suggested the bioactive compounds isolated from Amphidinium carterae exhibited a significant inhibiting effect against the diatom Skeletonema costatum and Amphibalanus amphitrite larvae, and could be substitutes for persistent, toxic antifouling compounds.     

Antifouling effect of bioactive compounds from selected marine organisms in the Obhur Creek,Red Sea

Three species of sponges and a tunicate were collected from Obhur creek of Jeddah coast for this bioactivity study. In order to assess the antifouling efficacy of selected marine organisms, methanolic extracts of these organisms were tested against different fouling bacterial forms and II-instar stage of the barnacle, Balanus amphitrite. Antibiosis, bioactivity and followed by multivariate analyses were carried out to check the efficacy of antifouling effect of the selected marine organisms. Principal component analysis revealed the exemplary antifouling efficacy of the sponge extracts of Stylissa sp. observed followed by Hyrtios sp. against bacterial forms in the laboratory study. De-trended correspondence analysis confirmed that the contribution of antifouling efficacy of the selected sponge extracts was observed to be more towards Bacillus sp., Vibrio sp. and Alteromonas sp. Moreover, the efficacy of Hyrtios sp. extract(20.430 μg m L~(-1)) followed by Stylissa sp.(30.945 μg m L~(-1)) showed higher against barnacle instar compared with other extracts in the bioactivity assay. Bray-Curtis cluster analysis under paired linkage categorized all the sponge extracts into one major cluster with 75% similarity, and one outlier tunicate. More than 80% similarity observed between Hyrtios sp. and Stylissa sp. Fourier transform infrared spectroscopy(FTIR) showed that the contribution of major peaks found in the marine organisms were towards sulfones, sulfoxides, cyanates and ketones.     

Spatio-temporal Variation of Soil Respiration and Its Driving Factors in Semi-arid Regions of North China

Soil respiration (SR) is the second-largest flux in ecosystem carbon cycling. Due to the large spatio-temporal variability of environmental factors, SR varied among different vegetation types, thereby impeding accurate estimation of CO₂ emissions via SR. However, studies on spatio-temporal variation of SR are still scarce for semi-arid regions of North China. In this study, we conducted 12-month SR measurements in six land-use types, including two secondary forests (Populus tomentosa (PT) and Robinia pseudoacacia (RP)), three artificial plantations (Armeniaca sibirica (AS), Punica granatum (PG) and Ziziphus jujuba (ZJ)) and one natural grassland (GR), to quantify spatio-temporal variation of SR and distinguish its controlling factors. Results indicated that SR exhibited distinct seasonal patterns for the six sites. Soil respiration peaked in August 2012 and bottomed in April 2013. The temporal coefficient of variation (CV) of SR for the six sites ranged from 76.98% to 94.08%, while the spatial CV of SR ranged from 20.28% to 72.97% across the 12-month measurement. Soil temperature and soil moisture were the major controlling factors of temporal variation of SR in the six sites, while spatial variation in SR was mainly caused by the differences in soil total nitrogen (STN), soil organic carbon (SOC), net photosynthesis rate, and fine root biomass. Our results show that the annual average SR and Q₁₀ (temperature sensitivity of soil respiration) values tended to decrease from secondary forests and grassland to plantations, indicating that the conversion of natural ecosystems to man-made ecosystems may reduce CO₂ emissions and SR temperature sensitivity. Due to the high spatio-temporal variation of SR in our study area, care should be taken when converting secondary forests and grassland to plantations from the point view of accurately quantifying CO₂ emissions via SR at regional scales.     

Inbreeding depression on growth and survival of full-sib family of Manila clam (<Emphasis Type="Italic">Ruditapes philippinarum</Emphasis>)

In present study, the inbreeding depression (ID) of growth and survival of Manila clam (Ruditapes philippinarum) was investigated at larval and juvenile stages. Nine inbred families (A ₂, B ₂, C ₂, D ₂, E ₂, F ₂, G ₂, H ₂ and I ₂) were established by mating within nine full-sib families with expected inbreeding coefficient of 0.25. Inbred families showed significant differences in shell length and hatching rate of D-larvae (straight-hinged larvae). The larvae of the nine inbred families grew slower than those of control group (CG), and their ID value ranged from 0.81% ± 6.09% to 16.10% ± 1.49%. The ID value of larval survival rate varied between 27.47% ± 9.36% and 70.50% ± 13.66%. The ID was also detected for juvenile growth in A ₂, B ₂, C ₂, and D ₂, which ranged from 4.60 ± 2.21 to 17.71 ± 7.73. The A ₂ family maintained the highest juvenile survival rate, whereas the other inbred families exhibited ID values varying between 62.79% ± 4.54% and 96.14% ± 0.87%. The linear relationship of estimated ID between growth and survival was negatively correlated (R = ?0.434, P < 0.05). The results of this study suggested that the ID of growth was common at the larval stage but was less prevalent at juvenile stage. In contrast, the ID of survival increased from larval to juvenile stage. A better understanding of the effect of inbreeding may aid to selective breeding of Manila clam.     

Unusual hydrocarbon waxes detected in <Emphasis Type="Italic">Salix oritrepha</Emphasis> leaf from Nianbaoyeze Mountains,eastern Qinghai-Tibetan Plateau

Plant biomarkers, such as hydrocarbon waxes, are frequently found in various sediments and could be adopted as paleovegetation and paleoclimate indicators. Nevertheless, scarce researches have focused on leaf waxes in higher plants of alpine region. Herein, hydrocarbon leaf wax components of Salix oritrepha, which flourish in Nianbaoyeze Mountains in eastern Tibetan Plateau were fully discussed. The n-alkane distribution in leaves ranges from n-C₂₁ to n-C₂₉ with maxima at n-C₂₅, which were entirely different with Salix taxa displayed in previous surveys in non-alpine regions. The unusual even carbon nalkenes from n-C_22:1 to n-C_30:1, which were thought to appear only in aquatic organisms, were firstly reported in an alpine plant. Additionally, iso-(2-methyl) alkanes, ranging from i-C₂₃ to i-C₂₉ with maxima at i-C₂₅, which have been commonly reported in microorganisms, were also identified in an alpine plant for the first time. Unusual hydrocarbon distribution detected in Salix oritrepha leaf from Nianbaoyeze Mountains is most likely due to the extreme environment in such alpine region.     

Global water vapor content decreases from 2003 to 2012: An analysis based on MODIS data

Water vapor in the earth′s upper atmosphere plays a crucial role in the radiative balance, hydrological process, and climate change. Based on the latest moderate-resolution imaging spectroradiometer(MODIS) data, this study probes the spatio-temporal variations of global water vapor content in the past decade. It is found that overall the global water vapor content declined from 2003 to 2012(slope b = –0.0149, R = 0.893, P = 0.0005). The decreasing trend over the ocean surface(b = –0.0170, R = 0.908, P = 0.0003) is more explicit than that over terrestrial surface(b = –0.0100, R = 0.782, P = 0.0070), more significant over the Northern Hemisphere(b = –0.0175, R = 0.923, P = 0.0001) than that over the Southern Hemisphere(b = –0.0123, R = 0.826, P = 0.0030). In addition, the analytical results indicate that water vapor content are decreasing obviously between latitude of 36°N and 36°S(b = 0.0224, R = 0.892, P = 0.0005), especially between latitude of 0°N and 36°N(b = 0.0263, R = 0.931, P = 0.0001), while the water vapor concentrations are increasing slightly in the Arctic regions(b = 0.0028, R = 0.612, P = 0.0590). The decreasing and spatial variation of water vapor content regulates the effects of carbon dioxide which is the main reason of the trend in global surface temperatures becoming nearly flat since the late 1990 s. The spatio-temporal variations of water vapor content also affect the growth and spatial distribution of global vegetation which also regulates the global surface temperature change, and the climate change is mainly caused by the earth's orbit position in the solar and galaxy system. A big data model based on gravitational-magmatic change with the solar or the galactic system is proposed to be built for analyzing how the earth's orbit position in the solar and galaxy system affects spatio-temporal variations of global water vapor content, vegetation and temperature at large spatio-temporal scale. This comprehensive examination of water vapor changes promises a holistic understanding of the global climate change and potential underlying mechanisms.     

Genetic transformation of <Emphasis Type="Italic">Nannochloropsis oculata</Emphasis> with a bacterial phleomycin resistance gene as dominant selective marker

The gene ble from Streptoalloteichus hindustanus is widely used as a selective antibiotic marker. It can control the phleomycin resistance, and significantly increase the tolerance of hosts to zeocin. The unicellular marine microalga Nannochloropsis oculata is extremely sensitive to zeocin. We selected ble as the selective marker for the genetic transformation of N. oculata. After the algal cells at a density of 2×10⁷ cells mL^?1 was digested with 4% hemicellulase and 2% driselase for 1 h, the protoplasts accounted for 90% of the total. The ble was placed at the downstream of promoter HSP70A-RUBS2 isolated from Chlamydomonas reinhardtii, yielding a recombinant expression construct pMS188. The construct was transferred into the protoplasts through electroporation (1 kV, 15 μS). The transformed protoplasts were cultured in fresh f/2 liquid medium, and selected on solid f/2 medium supplemented with 500 ng mL^?1 zeocin. The PCR result proved that ble existed in the transformants. Three transformants had been cultured for at least 5 generations without losing ble. Southern blotting analysis showed that the ble has been integrated into the genome of N. oculata. The ble will serve as a new dominant selective marker in genetic engineering N. oculata.     

Effect of benzo[a]pyrene on detoxification and the activity of antioxidant enzymes of marine microalgae

The objective of this study was to examine the effect of benzo[a]pyrene(Ba P) on the detoxification and antioxidant systems of two microalgae,Isochrysis zhanjiangensis and Platymonas subcordiformis.In our study,these two algae were exposed to Ba P for 4 days at three different concentrations including 0.5 μg~(L-1)(low),3 μg~(L-1)(mid) and 18 μg~(L-1)(high).The activity of detoxification enzymes,ethoxyresorufin O-deethylase(EROD) and glutathione S-transferase(GST) increased in P.subcordiformis in all Ba P-treated groups.In I.zhanjiangensis,the activity of these two enzymes increased at the beginning of exposure,and then decreased in the groups treated with mid-and high Ba P.The activity of antioxidant enzyme superoxide dismutase(SOD) increased in I.zhanjiangensis in all Ba P-treated groups,and then decreased in high Ba P-treated group,while no significant change was observed in P.subcordiformis.The activity of antioxidant enzyme catalase(CAT) increased in I.zhanjiangensis and P.subcordiformis in all Ba Ptreated groups.The content of malondialdehyde(MDA) in Isochrysis zhanjiangensis increased first,and then decreased in high Ba P-treated group,while no change occurred in P.subcordiformis.These results demonstrated that Ba P significantly influenced the activity of detoxifying and antioxidant enzymes in microalgae.The metabolic related enzymes(EROD,GST and CAT) may serve as sensitive biomarkers of measuring the contamination level of Ba P in marine water.     

Distribution of diatoms and silicoflagellates in surface sediments of the Yellow Sea and offshore from the Changjiang River,China

The spatial distribution of siliceous microfossils (diatoms and silicoflagellates) in the surface sediments was mapped at 113 sites in the Yellow Sea and sea areas adjacent to the Changjiang (Yangtze) River, China. In total, 267 diatom taxa and two silicoflagellate species were identified from the sediments. The spatial variations in abundance and diversity were classified into three distinct geographic patterns using Q mode clustering: a south-north geographic pattern, a coastal-offshore pattern and a unique pattern in the Changjiang River mouth. The south-north geographic pattern was related to the spatial variations in sea temperature. Coscinodiscus oculatus, a warm-water species, indicated these variations by a gradual decrease in abundance from the south to the north. The coastal-offshore pattern was in response to the spatial variations in salinity. Cyclotella stylorum, Actinocyclus ehrenbergii and Dictyocha messanensis, the dominant brackish species in coastal waters, significantly decreased at the isobaths of approximately 30 m, where the salinity was higher than 31. Paralia sulcata and Podosira stelliger indicated the impact of the Yellow Sea Warm Current in the central Yellow Sea. The unique pattern in the Changjiang River mouth showed the highest species diversity but lower abundance, apparently because: freshwater input can significantly increase the proportion of brackish species; nutrients can supply the growth of phytoplankton; and high sedimentation rates can dilute the microfossil abundance in the sediments. Our results show that an integration of environmental factors (e.g., nutrient levels, sedimentation rate, sea temperature, salinity and water depth) determined the spatial characteristics of the siliceous microfossils in the surface sediments.     

Taxonomy,Morphology and Phylogeny of a New Oligotrich Ciliate–<Emphasis Type="Italic">Omegastrombidium hongkongense</Emphasis> n. sp. (Protozoa: Ciliophora) from Clear Water Bay,Hong Kong

One new marine oligotrich ciliate, Omegastrombidium hongkongense n. sp., was isolated from a bloom of Noctiluca scuntillans near Port Shelter, Hong Kong. The morphology and infraciliature of this new species were studied on both living and protargol-stained specimens. Its phylogenetic position was discussed based on the sequence of the small subunit rRNA gene. O. hongkongense is different from its congeners with special characters. The cells are usually heart-shaped, and the cell size usually is (20–35) × (20–30) μm in vivo. Its deep buccal cavity extends obliquely to about 1/2 of cell length. It shows prominent apical protrusion. The adoral zone of membranelles is divided into 17–19 collar membranelles and four buccal membranelles. It has one ball-like macronucleus. The girdle kinety forms a closed loop which obliquely surrounds the body. The ventral kinety and thigmotactic membranelles are not observed. The SSU rRNA sequence of O. hongkongense was close to those of Strombidium paracalkinsi and Varistrombidium kelum with approximately 99% similarity. In the phylogenetic trees, O. hongkongense can be grouped with O. elegans and V. kielum species with very low support (16% ML).     

Optimization of protectant,salinity and freezing condition for freeze-drying preservation of <Emphasis Type="Italic">Edwardsiella tarda</Emphasis>

Novel preservation condition without ultra-low temperature is needed for the study of pathogen in marine fishes. Freeze-drying is such a method usually used for preservation of terrigenous bacteria. However, studies using freeze-drying method to preserving marine microorganisms remain very limited. In this study, we optimized the composition of protectants during the freeze-drying of Edwardsiella tarda, a fish pathogen that causes systemic infection in marine fishes. We found that the optimal composition of protectant mixture contained trehalose (8.0%), skim milk (12.0%), sodium citrate (2.0%), serum (12.0%) and PVP (2.0%). Orthogonal and interaction analyses demonstrated the interaction between serum and skim milk or sodium citrate. The highest survival rate of E. tarda was observed when the concentration of NaCl was 10.0, 30.0 and between 5.0 and 10.0 g L?1 for preparing TSB medium, E. tarda suspension and protectant mixture, respectively. When E. tarda was frozen at ?80°C or ?40°C for 6 h, its survival rate was higher than that under other tested conditions. Under the optimized conditions, when the protectant mixture was used during freeze-drying process, the survival rate (79.63%–82.30%) of E. tarda was significantly higher than that obtained using single protectant. Scanning electron microscopy (SEM) image indicated that E. tarda was embedded in thick matrix with detectable aggregation. In sum, the protectant mixture may be used as a novel cryoprotective additive for E. tarda.     

Interspecific competition and allelopathic interaction between <Emphasis Type="Italic">Karenia mikimotoi</Emphasis> and <Emphasis Type="Italic">Dunaliella salina</Emphasis> in laboratory culture

Algal allelopathy is a manifold ecological/physiological phenomenon that is focused on chemical interactions and autotoxicity. We investigated the allelopathic interactions between Karenia mikimotoi and Dunaliella salina in laboratory cultures based on diff erent temperature (15°C, 20°C, and 25°C) and lighting (40, 80, and 160 μmol/(m²·s)) conditions. The growth of D. salina in bi-algae culture (1:1 size/density) was significantly restrained. The results of cell-free filtrate culture indicate that direct cell-tocell contact was not necessary in interspecific competition. Further experimental results demonstrated that allelochemicals released from K. mikimotoi were markedly influenced by both temperature (P =0.013) and irradiance (P =0.003), resulting in diff erent growth characteristics of D. salina in filtrate mediums. Compared with the plateau period, K. mikimotoi exudates in the exponential phase had a stronger short-term inhibition effect on D. salina in normal conditions. A clear concentration-dependent relationship was observed in the effect of allelochemicals released from K. mikimotoi with low-promoting and high-repressing effects on D. Salina in a short time-scale. In addition, allelopathic substances remain stable and effective under high temperature and pressure stress. Many flocculent sediments adhering with D. salina cells were observed in all filtrate mediums, while the quantity and color depended on the original culture conditions.     

Identification and bioinformatics analysis of microRNAs from the sporophyte and gametophyte of <Emphasis Type="Italic">Pyropia haitanensis</Emphasis>

Pyropia haitanensis (T. J. Chang et B. F. Zheng) N. Kikuchi et M. Miyata (Porphyra haitanensis) is an economically important genus that is cultured widely in China. P. haitanensis is cultured on a larger scale than Pyropia yezoensis, making up an important part of the total production of cultivated Pyropia in China. However, the majority of molecular mechanisms underlying the physiological processes of P. haitanensis remain unknown. P. haitanensis could utilize inorganic carbon and the sporophytes of P. haitanensis might possess a PCK-type C₄-like carbon-fixation pathway. To identify microRNAs and their probable roles in sporophyte and gametophyte development, we constructed and sequenced small RNA libraries from sporophytes and gametophytes of P. haitanensis. Five microRNAs were identified that shared no sequence homology with known microRNAs. Our results indicated that P. haitanensis might posses a complex sRNA processing system in which the novel microRNAs act as important regulators of the development of different generations of P. haitanensis.     

Phylogenetic diversity of culturable bacteria in surface seawater from the Drake Passage,Antarctica

The Drake Passage is located between the Antarctic Peninsula and Tierra del Fuego in the south of South America. Surface seawater samples were collected at seven sites in the Drake Passage during the austral summer of 2012. The 16S rRNA sequences were analyzed from 187 isolated bacterial strains. Three phyla, 29 genera and 56 species were identified. The three phyla were Actinobacteria, Firmicutes and Proteobacteria; the Proteobacteria included α-Proteobacteria, β-Proteobacteria and γ-Proteobacteria. γ-Proteobacteria, Actinobacteria and Firmicutes were the dominant class or phyla in terms of quantity and species. Gram-positive bacteria (Actinobacteria and Firmicutes) accounted for 57.8% of all types identified. There were nine dominant genera, including Curtobacterium, Staphylococcus, and Halomonas, and 14 dominant species including Curtobacterium flaccumfaciens, Curtobacterium pusillum, and Staphylococcus sciuri. Of the strains identified, 87.2% were catalase positive or weakly positive.     

Biodegradation of <Emphasis Type="Italic">Enteromorpha</Emphasis> polysaccharides by intestinal micro-community from <Emphasis Type="Italic">Siganus oramin</Emphasis>

Micro-communities are supposed to have more potential functions of biodegradation of polysaccharides than single strain; however, the intestinal micro-communities involved in the biodegradation of Enteromorpha polysaccharides (EP) were seldom reported. In order to obtain the EP-degrading micro-community, the intestines of Siganus oramin was obtained to isolate the micro-communities, which were enriched by 0.3% of EP as the sole carbon source. A stable micro-community with EP degradative capability was achieved after seven generations of subculture, named H1. Results showed that H1 was able to degrade 75% of EP within 24 hours, and the activity of EP lyases reached 500 U mL^?1 in 32 hours. With denaturing gradient gel electrophoresis (DGGE) and 16S rRNA gene clone library analysis, ten bacteria closely related to Marinomonas pontica, Microbacterium sp., Leucobacter chironomi, Cyclobacterium sp., Algoriphagus winogradskyi, Pseudoalteromonas sp. and Vibrio sp. were determined. Furthermore, compared with the DGGE bands sequence and the clone library analysis, the dominant bacteria of the EP-biodegrading micro- community were Pseudoalteromonas sp. and Vibrio sp., with the respective proportion of 38% and 46%, and they should play an important role in EP degradation together with other degrading bacteria in the micro-community H1.     

Effects of dietary genistein on GH/IGF-I axis of Nile tilapia <Emphasis Type="Italic">Oreochromis niloticus</Emphasis>

There is considerable concern that isoflavones, such as genistein in fish feed composed of soybean protein, aff ects somatic growth in fish. Our previous works demonstrated that 30 and 300 μg/g dietary genistein had no significant eff ect on growth performance in Nile tilapia (Oreochromis niloticus), but the higher level of genistein (3 000 μg/g) significantly depressed growth. This study was conducted to further examine the eff ects of dietary genistein on the endocrine disruption on growth hormone/insulin-like growth factor-I (GH/IGF-I) axis in Nile tilapia (O. niloticus). Juvenile fish were fed by hand twice daily to satiation with one of four isonitrogenous and isoenergetic diets, each containing either 0, 30, 300 or 3 000 μg/g genistein. Following an 8-week feeding period, plasma GH and IGF-I levels were investigated by radioimmunoassay and gene expression levels of gh, ghrelin, gnrhs, ghr, npy, npyrs, pacap, ghrs, igf-I, igf-Ir, and igfbp3 were examined by real-time PCR. The results show that no significant change in plasma GH and IGF-I levels in fish fed with diets containing 30 μg/g and 300 μg/g genistein. mRNA expression of genes along the GH/IGF-I axis remained unaff ected, except for igf-Ir, which was stimulated by the 300 μg/g genistein diet. While in fish fed the 3 000 μg/g genistein diet, the plasma GH and IGF-I levels decreased, and mRNA expression of gh, ghr2, npyr1, igf-I, and igf-Ir were also significantly depressed. In contrast, npy and igfbp3 mRNA expression were enhanced. This study provides convincing evidence for growth impediment by genistein by disturbing the GH/IGF-I axis in Nile tilapia O. niloticus.