Loss of preferred habitat and pollen limitation threatens reproduction in a rare mountain Paeonia delavayi

Paeonia delavayi is a wild tree peony species endemic to high-altitude regions in southwestern China.Recent agricultural land expansion,however,not only causes its dwindling population size,but also poses a severe threat to its long-term persistence.Since our knowledge of the reproductive biology of P.delavayi is very limited,and some management misconceptions have further exacerbated the already worrisome situation,the aim of the present study is to provide some scientific evidence regarding habitat preference and breeding system of P.delavayi,and to help correct some management misconceptions.Specifically,morphological traits of P.delavayi populations growing in two different habitats(i.e.,the open area versus forest understory),including basal stem diameter,plant height and flower numbers per plant,were measured and compared with each other.A range of floral manipulation experiments was further conducted to study the extent of self-compatibility and pollen limitation.The results showed that P.delavayi preferred the open habitat over forest understory as it generally grew bigger with thickerstems,and produced a higher number of flowers in the former habitat.Therefore,the open habitat should receive immediate priority for conservation action instead of being converted to agricultural land;also,P.delavayi is pollen limited,as evidenced by the fact that experimental flowers receiving supplemental non-self pollen generally displayed elevated seed production than flowers subjected to other treatments(e.g.,non-supplemental self pollen,non-supplemental non-self pollen and supplemental self pollen),which is consistent with reported observations that alpine plants are more likely to be pollen-limited than lowland plants.We suggest that human intervention might be necessary to guarantee the long-term persistence of P.delavayi as harsh alpine environment,intense competition for pollinators and different anthropogenic perturbations co-limit its reproductive success.     

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.     

Collecting <Emphasis Type="Italic">Ophiocordyceps sinensis</Emphasis>: an emerging livelihood strategy in the Garhwal,Indian Himalaya

In the Garhwal of Uttarakhand, India, the Bhotiya, an ethnically and culturally distinct tribal group, were historically engaged in seasonal migration (i.e. transhumance) to take advantage of scarce mountain resources and trade relations with Tibet. This livelihood practice has all but disappeared. Households are adapting to these changing circumstances by engaging in the collection and sale of the valuable alpine medicinal fungus Ophiocordyceps sinensis, widely known as Cordyceps. The collection of this fungus has exploded, emerging as a lucrative yet high-risk livelihood strategy for many Bhotiya communities. The Bhotiyas’ historic herding and trade-based interactions and knowledge of these alpine environments where Cordyceps are found uniquely positions them to access this valuable biological resource. Elsewhere in the Himalayan region, some households are earning as much as two-thirds of their income from the collection of Cordyceps; in China Cordyceps is now listed as an endangered species due to intense over-exploitation in the Tibetan Plateau. This paper seeks to fill the void in the scientific literature on the social, ecological and economic aspects of the emerging Cordyceps trade in the Garhwal. Our study investigates the socio-spatial dimensions of Cordyceps collection in the high alpine meadows. We document how a fusion of local knowledge and practice with alpine mountain systems has served to reinvigorate the economic integrity of mountain communities at a time of rapid socio-economic change and to reimagine a new relationship between alpine resources and community well-being. The article offers suggestions to address the sustainability of both Cordyceps collection and livelihood activities which hinge on this fungus population. We find a need for (1) community-based conservation measures that are rooted in (2) secure resource access rights for local communities to continue sustainable collection and sale of Cordyceps and (3) participatory-and science-based processes for determining appropriate local collection numbers.     

The mycobiota of discoloured needles of the Swiss stone pine (<Emphasis Type="Italic">Pinus cembra</Emphasis> L.) in the Tatra Mountains (Poland)

The Swiss stone Pine (Pinus cembra L.) is an alpine species, fairly commonly occurring in the Alps and the Carpathians, close to the timberline. Natural sites of the Swiss stone pine in Poland are found exclusively in the Tatra Mountains, within the area of the Tatra National Park (TNP). In 2017, the health status of the needles of P. cembra was determined and their mycological analysis was carried out. As a result, 11 species of fungi were isolated from the needles showing disease symptoms, manifested by various shades of discoloration. The most frequently observed species was Fusarium oxysporum. It probably does not bring on disease symptoms, but may affect the development of other frequently isolated species (such as Sydowia polyspora, Lophodermium sp. and Lophodermium conigenum) that are pathogenic to Pinus spp.     

Effects of different phosphorus concentrations and N/P ratios on the growth and photosynthetic characteristics of <Emphasis Type="Italic">Skeletonema costatum</Emphasis> and <Emphasis Type="Italic">Prorocentrum donghaiense</Emphasis>

The effects of different phosphorus (P) concentrations (0.36, 3.6, and 36 μmol/L corresponding to low-, middle-, and high-P concentration groups, respectively) and nitrogen (N)/P ratios on the growth and photosynthetic characteristics of Skeletonema costatum and Prorocentrum donghaiense were studied. For both species, the high-P (HP) concentration group showed the greatest algal density and highest specific growth rate. Changes in the maximum efficiency of photosystem II (F _v /F _m ) were monitored under the various P and N/P conditions. The largest decrease in F _v /F _m was in the low-P (LP) group in S. costatum and in the HP group in P. donghaiense. There were high rapid light curves and photochemical quantum yields (Φ _PSII) for S. costatum in the HP group, while the actual photosynthetic capacity was higher in P. donghaiense than in S. costatum in the MP group. Under eutrophic but relatively P-restricted conditions, P. donghaiense had higher photosynthetic activity and potential, which could cause this dinoflagellate to increasingly dominate the phytoplankton community in these conditions. Under the same P concentration and N/P ratio, P. donghaiense had a larger relative maximum rate of electron transport and higher Φ _{PS II} values than those of S. costatum. These differences between P. donghaiense and S. costatum may explain the interaction and succession patterns of these two species in the Changjiang (Yangtze) River estuary from a photosynthesis perspective.     

Changes of Biogenic Elements in <Emphasis Type="Italic">Phragmites australis</Emphasis> and <Emphasis Type="Italic">Suaeda salsa</Emphasis> from Salt Marshes in Yellow River Delta,China

Little information is available on biogenic elements (carbon, nitrogen, phosphorus and sulfur) and the ecological stoichiometric characteristics of plants in coastal wetlands. To investigate the contents of carbon, nitrogen, phosphorus and sulfur of plants, and their ecological stoichiometric characteristics in the Yellow (Huanghe) River Delta, plant samples were collected from two typical salt marshes (Suaeda salsa and Phragmites australis wetlands) during the period of from August to October in 2007, and the ratios of C/N, C/P, N/P, C/N/P and C/N/P/S were calculated. Results showed that during the studying period, plant C, N and P were lower than the global average values, and plant N and P were lower than the China’s average values. Leaf C and S in Suaeda salsa were significantly lower than those in Phragmites australis (P < 0.05), and leaf N and P in Suaeda salsa and Phragmites australis showed no significant differences (P > 0.05). Average C/N ratios were 23.75 in leaf, 73.36 in stem, 65.67 in root of Suaeda salsa, and 33.77 in leaf, 121.68 in stem, 97.13 in root of Phragmites australis. Average C/N ratios of Suaeda salsa and Phragmites australis were all great than 25, indicating the salt marsh in the Yellow River Delta is an N limitation system. Average C/P ratios were 276.78 in leaf, 709.28 in stem and 1031.32 in root of Suaeda salsa, and 536.94 in leaf, 768.13 in stem and 875.22 in root of Phragmites australis. The average N/P ratios of Suaeda salsa were 12.92 in leaf, 10.77 in stem and 10.91 in root, and the average N/P ratios of Phragmites australis were 16.40 in leaf, 7.40 in stem and 6.92 in root, indicating the Suaeda salsa wetlands were N limited and Phragmites australis wetlands were N limited in August and P limited in October in 2007. The average C/N, C/P and C/N/P ratios in Suaeda salsa and Pragmites australis were higher than the global average values, indicating the lower quality of organic matter provided by wetland plants in the Yellow River delta.     

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.     

Screening and isolation of the algicidal compounds from marine green alga <Emphasis Type="Italic">Ulva intestinalis</Emphasis>

Twenty species of seaweed were collected from the coast of Zhejiang, China, extracted with ethanol, and screened for algicidal activity against red tide microalgae Heterosigma akashiwo and Prorocentrum micans. Inhibitory effects of fresh and dried tißsues of green alga Ulva intestinalis were assessed and the main algicidal compounds were isolated, purified, and identified. Five seaweed species, U. intestinalis, U. fasciata, Grateloupia romosissima, Chondria crassicaulis, and Gracilariopsis lemaneiformis, were investigated for their algicidal activities. Fresh tissues of 8.0 and 16.0 mg/mL of U. intestinalis dissolved in media significantly inhibited growth of H. akashiwo and P. micans, respectively. Dried tissue and ethyl acetate (EtOAc) extracts of U. intestinalis at greater than 1.2 and 0.04 mg/mL, respectively, were fatal to H. akashiwo, while its water and EtOAc extracts in excess of 0.96 and 0.32 mg/mL, respectively, were lethal to P. micans. Three algicidal compounds in the EtOAc extracts were identified as 15-ethoxy-(6z,9z,12z)-hexadecatrienoic acid (I), (6E,9E,12E)-(2-acetoxy-β-D-glucose)-octadecatrienoic acid ester (II) and hexadecanoic acid (III). Of these, compound II displayed the most potent algicidal activity with IC₅₀ values of 4.9 and 14.1 µg/mL for H. akashiwo and P. micans, respectively. Compound I showed moderate algicidal activity with IC₅₀ values of 13.4 and 24.7 µg/mL for H. akashiwo and P. micans, respectively. These findings suggested that certain macroalgae or products therefrom could be used as effective biological control agents against red tide algae.     

Biodeposition,respiration, and excretion rates of an introduced clam <Emphasis Type="Italic">Mercenaria mercenaria</Emphasis> in ponds with implications for potential competition with the native clam <Emphasis Type="Italic">Meretrix meretrix</Emphasis> in Shuangtaizi estuary,China

This study aimed to evaluate the potential impacts of an introduced clam Mercenaria mercenaria on estuarine ecosystem, and implications for the niche competition with a native clam Meretrix meretrix. The biodeposition, respiration, and excretion rates of M. mercenaria were determined seasonally using a sediment trap and a closed respirator in field. The biodeposition rates of M. mercenaria were 0.06–0.37 g/ (ind.·d), and the respiration rates were 0.31–14.66 mg/(ind.·d). The ammonia and phosphate excretion rates were 0.18–36.70 and 1.44–14.87 μg/(ind.·d), respectively. The hard clam M. mercenaria may discharge dry deposits up to 2.1×10⁵ t, contribute 18.3 t ammonia and 9.0 t phosphate to culture ponds, and consume 7.9×10³ t O₂ from ponds annually. It suggested that the hard clam M. mercenaria might play an important role in pelagic-benthic coupling in pond ecosystem through biodeposition and excretion. A comparison of the key physiological parameters of the introduced clam M. mercenaria and the native clam Meretrix meretrix suggested that M. mercenaria had a niche similar to that of Meretrix meretrix in Shuangtaizi estuary and might have a potential competition with Meretrix meretrix for habitat and food if M. mercenaria species escaped from the culture pond or artificially released in estuarine ecosystem.     

Alterations to biological soil crusts with alpine meadow retrogressive succession affect seeds germination of three plant species

Biological soil crusts (BSCs) are the important components of alpine meadow ecosystems. The extent and morphology of BSCs vary greatly with alpine meadow retrogressive succession due to grazing pressure. There is significant interest in impacts of crust composition on plant seed germination, especially in (semi-) arid environments. However, little is known about the influences of BSCs, and their associations with alpine meadow succession, on germination of typical alpine meadow vascular plant species. In a full factorial common-garden experiment, we studied effects of: (1) crust type, (2) seed position, and (3) surface texture on seed germination. We chose three typical alpine meadow plant species (i.e. Poa pratensis, Tibetia himalaica and Potentillen nivea), which belonged to different functional groups (graminoids, legumes, and forbs) and play important roles in all alpine meadow succession stages. Crust type and seed position influenced seed germination, and the inhibitory effects of BSCs depended on the crust type and seed species tested. The major factors influencing seed germination were BSC type, seed position, soil texture, and the interactions between BSC type and seed position; species and seed position; species and surface texture; and species, crust type, and surface texture. Cyanobacteria crust significantly inhibited germination of all seeds. Seed position also had a significant effect on seed germination (p < 0.001). Fewer seedlings germinated on the surface than below the surface, this was especially true for P. nivea. seeds within cyanobacteria and lichen crusts. Only germination rates of T. himalaica on the soil surface were significantly correlated with plant occurrence frequency within the alpine meadow community. The poor correlation for the other two species is possibly that they are perennials. Our results clearly demonstrated that BSCs can be biological filters during seed germination, depending on the BSC succession stage. Through their influences on seed germination, BSCs can strongly influence community assemblages throughout alpine meadow retrogressive succession.     

Leaf stable carbon isotope composition in <Emphasis Type="Italic">Picea schrenkiana</Emphasis> var. <Emphasis Type="Italic">tianschanica</Emphasis> in relation to leaf physiological and morphological characteristics along an altitudinal gradient

To understand the effects of leaf physiological and morphological characteristics on δ ¹³C of alpine trees, we examined leaf δ ¹³C value, LA, SD, LNC, LPC, LKC, Chla+b, LDMC, LMA and N_area in one-year-old needles of Picea schrenkiana var. tianschanica at ten points along an altitudinal gradient from 1420 m to 2300 m a.s.l. on the northern slopes of the Tianshan Mountains in northwest China. Our results indicated that all the leaf traits differed significantly among sampling sites along the altitudinal gradient (P<0.001). LA, SD, LPC, LKC increased linearly with increasing elevation, whereas leaf δ ¹³C, LNC, Chla+b, LDMC, LMA and N_area varied non-linearly with changes in altitude. Stepwise multiple regression analyses showed that four controlled physiological and morphological characteristics influenced the variation of δ ¹³C. Among these four controlled factors, LKC was the most profound physiological factor that affected δ ¹³C values, LA was the secondary morphological factor, SD was the third morphological factor, LNC was the last physiological factor. This suggested that leaf δ ¹³C was directly controlled by physiological and morphological adjustments with changing environmental conditions due to the elevation.     

Significance of different microalgal species for growth of moon jellyfish ephyrae, <Emphasis Type="Italic">Aurelia</Emphasis> sp.1

The scyphozoan Aurelia aurita (Linnaeus) sp. l., is a cosmopolitan species-complex which blooms seasonally in a variety of coastal and shelf sea environments around the world. The effects of different microalgal species on the growth of newly-released Aurelia sp.1 ephyrae were studied under laboratory conditions. We fed ephyrae with four different microalgal species (diatom, autotrophic dinoflagellate, heterotrophic dinoflagellate, and chlorophyta) plus Artemia nauplii for 12-24 d at 18°C. Results showed that the growth rate diverged significantly for Artemia nauplii compared to other food types. In addition, there was no significant variation between the growth rates for Skeletonema costatum and Prorocentrum donghaiense, and no significant variation was found in the growth rates for N. scintillans and P. subcordiformis. Artemia nauplii could support the energy requirement for the newly-released ephyrae to develop to meduase, and the ephyrae with Artemia nauplii showed a significant average growth rate of 25.85% d^?1. Newly-released ephyrae could grow slightly with some species of microalgae in the earliest development stage. Chain diatom Skeletonema costatum and autotrophic dinoflagellate Prorocentrum donghaiense, could not support the growth of the ephyrae, while heterotrophic dinoflagellate Noctiluca scintillans and chlorophyta Platymonas subcordiformis could support the growth of the ephyrae. However, none of the ephyrae fed with the tested phytoplankton could mature to medusae.     

Trophic Interaction in a <Emphasis Type="Italic">Portunus rituberculatus</Emphasis> Polyculture Ecosystem Based on Carbon and Nitrogen Stable Isotope Analysis

Trophic interaction among various biomass groups in a swimming crab Portunus trituberculatus polyculture pond was investigated using carbon and nitrogen stable isotope analysis. The polycultured animal species also included white shrimp Litopenaeus vannamei, short-necked clam Ruditapes philippinarum, and redlip mullet Liza haematochila. The mean δ¹³C value for all the biomass groups in polyculture ecosystem ranged from ?25.61‰ to ?16.60‰, and the mean δ¹⁵N value ranged from 6.80‰ to 13.09‰. Significant difference in the δ¹³C value was found between particulate organic matter (POM) and sediment organic matter (SOM) (P < 0.05), indicating that these two organic matter pools have different material sources. Assuming that a ¹³C-enrichment factor of 1.00‰ and a ¹⁵N-enrichment factor of 2.70‰ existed between consumer and prey, diets of the four cultured animals were estimated using a stable isotope mixing model. The estimated model results indicated that P. trituberculatus mainly feed on Aloidis laevis; L. vannamei mainly feed on shrimp feed; while A. laevis, R. philippinarum and L. haematochelia mainly feed on POM. Shrimp feed was also an important food source of R. philippinarum and L. haematochelia. The diets of P. trituberculatus, L. vannamei, R. philippinarum, and L. haematochila showed complementary effects in this polyculture ecosystem. Our finding indicated that the polyculture of these four organisms with suitable farming density could make an effective use of most of the food sources, which can make a highly efficient polyculture ecosystem.     

Effects of nitrogen and phosphorus on the growth of <Emphasis Type="Italic">Levanderina fissa</Emphasis>: How it blooms in Pearl River Estuary

Effects of nitrogen (N) and phosphorus (P) from different sources and at different concentrations on the growth of Levanderina fissa (= Gyrodinium instriatum) were studied in laboratory conditions. The findings might explain the recurrent blooms of this species in Pearl River Estuary, China. Results showed that nutrient limitation significantly inhibited the growth of L. fissa. The values of specific growth rate (μ _max) and half-saturation nutrient concentration (K _S) were 0.37 divisions/d and 8.49 μmol L^?1 for N, and 0.39 divisions/d and 1.99 μmol L^?1 for P, respectively. Based on K _S values, dissolved inorganic N level in PRE was sufficient to support the high proliferation of L. fissa, while dissolved inorganic P concentration was far lower than the minimum requirement for its effective growth. L. fissa was not able to utilize dissolved organic N (DON) compounds such as urea, amino acids, and uric acid. However, it grew well by using a wide variety of dissolved organic P (DOP) sources like nucleotides, glycerophosphate, and 4-nitrophenylphosphate. The results from this study suggested that the ability in DOP utilization of L. fissa offers this species a competitive advantage in phytoplankton communities. The high level and continuous supply of DIN, enrichment of DOP, together with warm climate and low salinity in the Pearl River Estuary provided a suitable nutrient niche for the growth of L. fissa, and resulted in the recurrent blooms in the estuary.     

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.     

Immunogenicity and protective role of antigenic regions from five outer membrane proteins of <Emphasis Type="Italic">Flavobacterium columnare</Emphasis> in grass carp <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis>

Flavobacterium columnare causes columnaris disease in freshwater fish. In the present study, the antigenic regions of five outer membrane proteins (OMPs), including zinc metalloprotease, prolyl oligopeptidase, thermolysin, collagenase and chondroitin AC lyase, were bioinformatically analyzed, fused together, and then expressed as a recombinant fusion protein in Escherichia coli. The expressed protein of 95.6 kDa, as estimated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was consistent with the molecular weight deduced from the amino acid sequence. The purified recombinant protein was used to vaccinate the grass carp, Ctenopharyngodon idella. Following vaccination of the fish their IgM antibody levels were examined, as was the expression of IgM, IgD and IgZ immunoglobulin genes and other genes such as MHC Iα and MHC IIβ, which are also involved in adaptive immunity. Interleukin genes (IL), including IL-1β, IL-8 and IL-10, and type I and type II interferon (IFN) genes were also examined. At 3 and 4 weeks post-vaccination (wpv), significant increases in IgM antibody levels were observed in the fish vaccinated with the recombinant fusion protein, and an increase in the expression levels of IgM, IgD and IgZ genes was also detected following the vaccinations, thus indicating that an adaptive immune response was induced by the vaccinations. Early increases in the expression levels of IL and IFN genes were also observed in the vaccinated fish. At four wpv, the fish were challenged with F. columnare, and the vaccinated fish showed a good level of protection against this pathogen, with 39% relative percent survival (RPS) compared with the control group. It can be concluded, therefore, that the five OMPs, in the form of a recombinant fusion protein vaccine, induced an immune response in fish and protection against F. columnare.     

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.