Construction of a new gene integration platform system for theSynechococcus sp. PCC7942

According to the known sequence of iron stress-induced gene (isiAB operon), we cloned its 1.5 kb fragment by PCR, and used this fragment as integration homologous fragment. After several steps of subcloning donor DNA into theisiAB fragment, a donor plasmid pZL which could be integrated into the chromosomal DNA ofSynechococcus sp. PCC7942 was constructed. In order to express the heterologous gene at a high level through the integration platform system, we constructed the donor DNA by the following steps. We cloned the strong promoter (240 bp) of heat shock genegroESL operon fromSynechococcus sp. PCC7942 by PCR. Then subcloned the multiple cloning sites (MCS),rbcS polyA into the downstream of thegroESL promoter. The kanamycin resistance gene, as the marker gene, was also subcloned into the donor DNA. Thus, in the donor plasmid pZL, the integration homologous fragment and several expression elements, such asgroESL promoter, MCS,rbcS polyA terminator and kanamycin resistance gene, were all included.

After naturally transformed and introduced the donor plasmid pZL intoSynechococcus sp. PCC7942, as in the pZL, the donor DNA sequence is flanked by two DNA fragments (0.4 kb and 0.7 kb) homologous to theisiAB fragment ofSynechococcus sp. PCC7942, the homologous DNA can recombine with the chromosomal DNA. After screening by kanamycin, the transformants which integrated the heterologous DNA were selected. The efficiency of transformation is about 1×10⁻⁶. By southern blot analysis, it was confirmed that the donor DNA had been integrated into the chromosomal DNA ofSynechococcus sp. PCC7942, located on the site of theisiAB gene, and can be replicated with the chromosomal DNA.

     

Extracellular release of β-lactamase by osmotic shock inSynechococcus transformant

A unicellular cyanobacteriumSynechococcus sp. strain PCC 7002 was transformed with plasmid pQL1, on which β-lactamase gene (bla) and β-galactosidase gene (lacZ) were encoded. The transformant cells released β-lactamase into medium by an abrupt drop of osmotic pressure. This result indicates that this cyanobacterium recognizes and processes the signal sequence of β-lactamase, and accumulates the enzyme in periplasm. Repeated release of β-lactamase was possible by repeated osmotic shocks wihout, impairing cell viability. On the other hand, most of the β-galactosidase remained in cytoplasm under the osmotic shock.

     

Production of γ-linolenic acid and stearidonic acid by <Emphasis Type="Italic">Synechococcus</Emphasis> sp. PCC7002 containing cyanobacterial fatty acid desaturase genes

Genetic modification is useful for improving the nutritional qualities of cyanobacteria. To increase the total unsaturated fatty acid content, along with the ratio of ω-3/ω-6 fatty acids, genetic engineering can be used to modify fatty acid metabolism. Synechococcus sp. PCC7002, a fast-growing cyanobacterium, does not contain a Δ6 desaturase gene and is therefore unable to synthesize γ-linolenic acid (GLA) and stearidonic acid (SDA), which are important in human health. In this work, we constructed recombinant vectors Syd6D, Syd15D and Syd6Dd15D to express the Δ15 desaturase and Δ6 desaturase genes from Synechocystis PCC6803 in Synechococcus sp. PCC7002, with the aim of expressing polyunsaturated fatty acids. Overexpression of the Δ15 desaturase gene in Synechococcus resulted in 5.4 times greater accumulation of α-linolenic acid compared with the wild-type while Δ6 desaturase gene expression produced both GLA and SDA. Co-expression of the two genes resulted in low-level accumulation of GLA but much larger amounts of SDA, accounting for as much to 11.64% of the total fatty acid content.     

Vitreoscilla hemoglobin gene (vgb) improves lutein production in Chlorella vulgaris

Vitreoscilla hemoglobin is an oxygen-binding protein that promotes oxygen delivery and reduces oxygen consumption under low oxygen conditions to increase the efficiency of cell respiration and metabolism. In this study, we introduced a Vitreoscilla hemoglobin gene （vgb） into Chlorella vulgaris by Agrobacterium tumefaciens-mediated transformation （ATMT）. PCR analysis confirmed that the vgb gene was successfully integrated into the Chlorella vulgaris genome. Analysis of biomass obtained in shake flasks revealed transformant biomass concentrations as high as 3.28 g/L, which was 38.81% higher than that of the wild-type strain. Lutein content of transformants also increased slightly. Further experiments recovered a maximum lutein yield of 2.91 mg/L from the transformants, which was 36.77% higher than that of the wild-type strain. The above results suggest that integrated expression of the vgb gene may improve cell growth and lutein yield in Chlorella vulgaris, with applications to lutein production from Chlorella during fermentation.     

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.     

A method of batch-purifying microalgae with multiple antibiotics at extremely high concentrations

Axenic microalgal strains are highly valued in diverse microalgal studies and applications. Antibiotics, alone or in combination, are often used to avoid bacterial contamination during microalgal isolation and culture. In our preliminary trials, we found that many microalgae ceased growing in antibiotics at extremely high concentrations but could resume growth quickly when returned to an antibiotics-free liquid medium and formed colonies when spread on a solid medium. We developed a simple and highly efficient method of obtaining axenic microalgal cultures based on this observation. First, microalgal strains of different species or strains were treated with a mixture of ampicillin, gentamycin sulfate, kanamycin, neomycin and streptomycin (each at a concentration of 600 mg/L) for 3 days; they were then transferred to antibiotics-free medium for 5 days; and finally they were spread on solid f/2 media to allow algal colonies to form. With this method, five strains of Nannochloropsis sp. (Eustigmatophyceae), two strains of Cylindrotheca sp. (Bacillariophyceae), two strains of Tetraselmis sp. (Chlorodendrophyceae) and one strain of Amphikrikos sp. (Trebouxiophyceae) were purified successfully. The method shows promise for batch-purifying microalgal cultures.     

Picoplankton distribution in different water masses of the East China Sea in autumn and winter

Picoplankton distribution was investigated in different water masses of the East China Sea in November,2006 and February,2007.The autumn and winter cruises crossed three major water masses:the coastal water mass(CWM),the mixed water mass(MWM),which forms on the continental shelf,and the Kuroshio water mass(KWM).Picoplankton composition was resolved into four main groups by flow cytometry,namely Synechococcus,Prochlorococcus,picoeukaryotes,and heterotrophic bacteria.The average abundances of Synechococcus,picoeukaryotes,and heterotrophic bacteria were(0.63±10.88)×103,(1.61±1.16)×103,(3.39±1.27)×105 cells/mL in autumn and(6.45±8.60)×103,(3.23±2.63)×103,(3.76±1.37)×105 cells/mL in winter,respectively.Prochlorococcus was not found in the CWM and seldom observed in surface samples in either season.However,Prochlorococcus was observed in the MWM and KWM(approximately 10 3 cells/mL) in both autumn and winter.Synechococcus distribution varied considerably among water masses,with the highest levels in KWM and lowest levels in CWM.The depth-averaged integrated abundance of Synechococcus was approximately 5-fold higher in KWM than in CWM,which may be due primarily to water temperature.In the MWM,Synechococcus was resolved as two subgroups;the presence of both subgroups was more common in autumn.Picoeukaryote abundance varied less among water masses than Synechococcus,and heterotrophic bacteria depth-averaged integrated abundance exhibited the smallest seasonal variations with respect to water mass.Correlation analysis showed that relationships between picoplankton abundances and environmental factors(temperature,nutrients,and chlorophyll a) differed among the three water masses,suggesting that the three water masses have different effects on picoplankton distribution(particularly Synechococcus).     

Camptotheca acuminata decne residue after camptothecin extract as a substrate to produce mushroom spawn

Currently in China,no technically and economically viable methods exist to handle large quantities of Camptotheca acuminata Decne residue(CA residue) after camptothecin extract while there is a great demand for low cost alternatives to replace the cottonseed hull-based materials used in commercial mushroom culture.Hence,it is of importance for camptothecin extract factories and mushroom producers to explore the utilization of CA residue in mushroom industry.We conducted a research to study how partially or completely substituting traditional mushroom substrate by CA residue would influence the mycelial growth in mushroom spawn production.5 mushroom strains from 4 species were used in the test,i.e.,oyster mushroom(Pleurotus ostreatu) strains,Zayou No.1 and Xide 33,needle mushroom(Flammulina velutipes) strain Chuanjin No.3,hairy wood ear(Auricularia polytricha) strain Huang Er No.10,and shiitake(Lentinula edodes) strain Wuxiang.The nutrient element composition and heavy metal contents of CA residue were determined to ensure its safety and to determine its appropriate component in the substrate formulation for mushroom spawn production.The four substrate formulations(one control-CK,and three treatments,named,T1,T2,T3,) contained 0%,40%,79%,and 100% CA residue,respectively,to allow comparison of the fungal mycelial growth.The control(CK) was the popularly used formulation in Chinese commercial mushroom production,comprising of 73% cottonseed hulls,10% sawdust,15% wheat bran,1% lime,1% white sugar(percentage by weight).All mushroom spawns of the five strains in the four treatments were incubated under the same conditions.The results showed that mycelia of the five mushroom strains grew significantly faster on the substrates containing CA residue than on the substrate with no CA residue(CK).There were no significant differences in the mycelial growth rate among treatments containing CA residue for the two oyster mushrooms and the needle mushroom,but mycelial growth rate in treatments T2 and T3 was significantly higher than in treatment T1 for hairy wood ear and shiitake.The results suggest that CA residue can be used to culture oyster mushroom,needle mushroom,hairy wood ear,and shiitake spawn,and the medium containing CA residue can stimulate their mycelial growth.The commercial production of mushroom spawn using CA residue not only brings better economical benefits including lower cost to mushroom producers,but also reduces environmental pollution by providing a means to reduce dumping and piling of CA residue.     

Genetic transformation of Platymonas (Tetraselmis) subcordiformis (Prasinophyceae, Chlorophyta) using particle bombardment and glass-bead agitation

Platymonas (Tetraselmis) subcordiformis is a unicellular marine green alga. It was found to be very sensitive to the herbicide Basta through a sensitivity test indicating it could be employed as a selective agent. The bar gene is a practicable and selectable marker gene. The vector containing the expression cassette of the bar gene was transferred to P. subcordiformis by both particle bombardment and glass-bead agitation and transformants were then selected using Basta. Finally, Southern blotting analysis indicated that the bar gene had been successfully integrated into the nuclear genome of P. subcordiformis using both of the transgenic techniques, with the transformation efficiency of the glass-bead method being slightly higher than that of particle bombardment. This is the first report on stable transformation of P. subcordiformis, and will improve fundamental research and enlarge application of this alga.     

Picophytoplankton abundance and community structure in the Philippine Sea, western Pacific

Flow cytometric determinations of the abundance distribution and community structure of picophytoplankton (i.e., Prochlorococcus spp., orange fluorescence Synechococcus spp. and picoeukaryotes) were used for samples taken from the Philippine Sea in the western tropical Pacific Ocean from September to October of 2004. A fluorescence probe was employed to detect Chlorophyll a (Chl a). Abundances of Prochlorococcus spp., orange fluorescence Synechococcus spp. and picoeukaryotes ranged from 0.1 to 58×10³ cells ml^?1, 0.38 to 17×10² cells ml^?1 and 0.42 to 26×10² cells ml^?1, respectively. Synechococcus spp. and picoeukaryotes co-occurred in relatively shallow water with the maximum abundance observed at 50 to 70 m depth, while Prochlorococcus spp. only occurred in the 70 to 200 m layer. Prochlorococcus spp. was the dominant picophytoplankton population in terms of abundance and biomass. The cell size and carbon biomass content were estimated for the three picophytoplankton groups. In addition, among the three groups of picophytoplankton, the relative contribution of red fluorescence to the total red fluorescence varied with depth. The fluorescence and light scatter properties of individual cells indicated that in the upper 100 m layer, picoeukaryotes were a major contributor to total red fluorescence, while at the depth below 100 m, Prochlorococcus spp. and Synechococcus spp. made an important contribution to the total red fluorescence.     

Physiological aspects of a high-co2 requiring mutant and the high-co2 growing cells of the cyanobacteriumSynechococcus pcc7942

A new chemically mutagenic mutant ofSynechococcus PCC7942 named high-CO₂ requiring mutant, which could grow at 4% CO₂ but could not grow at air levels of CO₂, was isolated. Comparative studies on some physiological aspects of the mutant and high-CO₂ growing cells (growing at 4% CO₂) were conducted. The result showed that the mutant had lower growing rate, about 1/40th photosynthetic affinity to inorganic carbon, 25% lower carbonic anhydrase (CA) activity, lower quenching rate of chlorophyll fluorescence, and about 1/2 alkalinization rate of the medium. The CA activity responses of the two types of cells to different concentration of CO₂ were determined. Upon the addition, of inorganic carbon (Ci), the rate of active Ci uptake described by the rate of chlorophyll fluorescence quenching of the mutant was obviously lower compared with that of the high-CO₂ growing cells; the size of the internal inorganic carbon pool size detemined by the extent of fluorescence quenching of the mutant was also smaller.

     

RAPD study on some common species ofPorphyra in China

Electroporation, PEC, PEG plus electroporation and Biolistics methods were tested in gene transformation ofP. yezoensis. The exogenousgus was from plasmid of pBI121 and pCAMBIA1301, both contain the CaMV35S promoter. The receptors included the protoplasts, tissues and free-living conchocelis filaments ofP. yezoensis. Several factors, for example, the voltage, capacitance and bivalent cations, etc., were studied. Results show that these four methods are all efficient for gene transformation inP. yezoensis; and that PEG is the best one, with transformation efficiency of up to 4×10⁻⁵. GUS activity was detected 26 days after transformation by using PEG method.

     

Expression of organophosphorus-degradation gene (opd) in aggregating and non-aggregating filamentous nitrogen-fixing cyanobacteria

Genetic engineering in filamentous N₂-fixing cyanobacteria usually involves Anabaena sp. PCC 7120 and several other non-aggregating species. Mass culture and harvest of such species are more energy consuming relative to aggregating species. To establish a gene transfer system for aggregating species, we tested many species of Anabaena and Nostoc, and identified Nostoc muscorum FACHB244 as a species that can be genetically manipulated using the conjugative gene transfer system. To promote biodegradation of organophosphorus pollutants in aquatic environments, we introduced a plasmid containing the organophosphorus-degradation gene (opd) into Anabaena sp. PCC 7120 and Nostoc muscorum FACHB244 by conjugation. The opd gene was driven by a strong promoter, P _psbA . From both species, we obtained transgenic strains having organophosphorus-degradation activities. At 25°C, the whole-cell activities of the transgenic Anabaena and Nostoc strains were 0.163±0.001 and 0.289±0.042 unit/μg Chl a, respectively. However, most colonies resulting from the gene transfer showed no activity. PCR and DNA sequencing revealed deletions or rearrangements in the plasmid in some of the colonies. Expression of the green fluorescent protein gene from the same promoter in Anabaena sp. PCC 7120 showed similar results. These results suggest that there is the potential to promote the degradation of organophosphorus pollutants with transgenic cyanobacteria and that selection of high-expression transgenic colonies is important for genetic engineering of Anabaena and Nostoc species. For the first time, we established a gene transfer and expression system in an aggregating filamentous N₂-fixing cyanobacterium. The genetic manipulation system of Nostoc muscorum FACHB244 could be utilized in the elimination of pollutants and large-scale production of valuable proteins or metabolites.     

Temporal and spatial variations of abundance of phycocyanin- and phycoerythrin-rich <Emphasis Type="Italic">Synechococcus</Emphasis> in Pearl River Estuary and adjacent coastal area

Three surveys were carried out in Pearl River Estuary and adjacent coastal area in May, August, and November, 2013, to investigate the temporal and spatial variations of abundance of phycoerythrin-rich Synechococcus (PE-rich SYN) and phycocyanin-rich Synechococcus (PC-rich SYN). The effects of environmental factors on the alternation of the different Synechococcus groups were also elucidated. PE-rich SYN was detected in three surveys, whereas PC-rich SYN was detected in May and August, but not in November. The highest abundances of PE-rich SYN and PC-rich SYN were recorded in August and May, with mean values of 74.17×10³ and 189.92×10³ cells mL^?1, respectively. From May to November, the relative abundance of PE-rich SYN increased, whereas that of PC-rich SYN declined. PE-rich and PC-rich SYN presented similar horizontal distributions with high abundance in the southern estuary in May, and in the western estuary in August. The abundances of PE-rich and PC-rich SYN were high at 27–32°C and salinity of 10–20. PC-rich SYN was not detected at < 24°C, and PC:PE-rich SYN decreased in abundance with salinity increase. When less than 20 mg L^?1, suspended particulate matter (SPM) was helpful for Synechococcus growth. PE-rich SYN decreased in abundance when the concentration of dissolved inorganic nitrogen increased in May and November, and the concentration of phosphate increased in November. However, PC-rich SYN abundance and nutrients showed no correlation. Principal component analysis and regression analysis indicated that PE-rich SYN significantly correlated with the principal components that were affected by environmental factors.     

A new record of a flathead fish (Teleostei: Platycephalidae) from China based on morphological characters and DNA barcoding

A new record of Platycephalus sp.1 (sensu Nakabo, 2002) was documented based on morphological characters and DNA barcoding. We collected 174 specimens of the genus Platycephalus from Chinese coastal waters of Dongying, Qingdao, Zhoushan, and Beihai. Samples were identified as Platycephalus sp.1 morphologically. The coloration, meristic traits, and morphometric measurements are consistent with previously published records. In brief, it is an orange-brown flathead fish with dark brown spots scattered on head and body, lateral line scales 83 to 99 with one or two spine-bearing anteriormost pored scale, no yellow blotch on the caudal fin. Cytochrome oxidase I subunit (COI) gene fragments were sequenced for phylogenetic analysis. The mean evolutionary distance within the species Platycephalus sp.1 was 0.1%. Net evolutionary distances between Platycephalus sp.1 and other species of Platycephalus ranged from 10.8% to 19.7%, which is much greater than the threshold for species delimitation. The COI sequence analysis strongly supports the validity of Platyceohalus sp.1 at genetic level.     

Establishment and characterization of a testicular Sertoli cell line from olive flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

The culture of Sertoli cells has become an indispensable resource in studying spermatogenesis. A new Sertoli cell line (POSC) that consisted predominantly of fibroblast-like cells was derived from the testis of the olive flounder Paralichthys olivaceus and sub-cultured for 48 passages. Analysis of the mtDNA COI gene partial sequence confirmed that the cell line was from P. olivaceus. Cells were optimally maintained at 25°C in DMEM/F12 medium supplemented with fetal bovine serum, basic fibroblast growth factor, and epidermal growth factor. The growth curve of POSC showed a typical “S” shape. Chromosome analysis revealed that the cell line possessed the normal P. olivaceus diploid karyotype of 2n=48t. POSC expressed dmrt1 but not vasa, which was detected using RT-PCR and sequencing. Immunocytochemistry revealed that the cells exhibited the testicular Sertoli cell marker FasL. Therefore, POSC appeared to consist of testicular Sertoli cells. Bright fluorescent signals were observed after the cells were transfected with pEGFP-N3 plasmid, with the transfection efficiency reaching 10%. This research not only offers an ideal model for further gene expression and regulation studies on P. olivaceus, but also serves as valuable material in studying fish spermatogenesis, Sertoli cell-germ cell interactions, and the mechanism of growth and development of testis.     

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.     

Simultaneous estimates ofsynechococcus spp. Growth and grazing mortality rates in the English Channel

The marine chroococooid phycoerythrin-containingSynechococcus spp. cyanobacterium has been implicated as a substantial component of the photosynthetic picoplankton in the ocean. Although its importance as food source for heterotrophic nanoplankton is now recognized, information about the cycling ofSynechococcus biomass and its diel pattern is limited and study methodology varies among authors. The selective metabolic inhibitor method was used to simultaneously estimate growth and grazing disappearance rates ofSynechococcus in the English Channel where growth rates ranged from 0.25 to 0.72/d (mean ±SD=0.51±0.17/d) and grazing mortality rates ranged from 0.19 to 0.64/d (mean ±SD=0.48±0.17/d). Size-fractionated experiments demonstrated that up to 70% ofSynechococcus disappearance could be attributed to grazers going through a 2 μm Nuclepore filter.Synechococcus grazing mortality rates (mean=0.74 ±0.25/d) during the day were always higher than that (mean=0.2±0.20/d) during the night, while growth rates showed no clear diel pattern. A positive correlation was observed between growth rates andin situ temperature ranging from 9 to 17°C, while in contrast grazing was independent of temperature. The close similatiry between average growth and grazing rates suggests a rapid recycling ofSynechococcus biomass in English Channel coastal waters.     

Inhibitory activity of an extract from a marine bacterium Halomonas sp. HSB07 against the red-tide microalga Gymnodinium sp. （Pyrrophyta）

In recent years, red tides occurred frequently in coastal areas worldwide. Various methods based on the use of clay, copper sulfate, and bacteria have been successful in controlling red tides to some extent. As a new defensive agent, marine microorganisms are important sources of compounds with potent inhibitory bioactivities against red-tide microalgae, such as Gymnodinium sp. （Pyrrophyta）. In this study, we isolated a marine bacterium, HSB07, from seawater collected from Hongsha Bay, Sanya, South China Sea. Based on its 16S rRNA gene sequence and biochemical characteristics, the isolated strain HSB07 was identified as a member of the genus Halomonas. A crude ethyl acetate extract of strain HSB07 showed moderate inhibition activity against Gymnodinium sp. in a bioactive prescreening experiment. The extract was further separated into fractions A, B, and C by silica gel column chromatography. Fractions B and C showed strong inhibition activities against Gymnodinium. This is the first report of inhibitory activity of secondary metabolites of a Halomonas bacterium against a red-tide-causing microalga.     

Single nucleotide polymorphisms in intron 1 and intron 2 of Larimichthys crocea growth hormone gene are correlated with growth traits

The growth hormone gene (GH) affects animal growth and is a potential target for genetic studies of variation related to growth traits. In this study, we analyzed single nucleotide polymorphisms (SNPs) in GH intron regions and their associations with growth traits in large yellow croaker, Larimichthys crocea, from Zhejiang and Fujian stocks. The results of PCR-single strand conformation polymorphism showed two haplotypes of intron 1, named AA and AB genotypes, in Zhejiang stock. AB exhibited an SNP at position 196 (G A) that was negatively correlated with body height and positively correlated with standard length/body height (P 0.05). Two different genotypes, CC and CD, were identified in intron 2 in Fujian stock, with CD showing an SNP at position 692 (T C). The CD genotype had a significantly positive correlation with both weight and total length (P 0.01). These basic data highlight the potential for using GH as a genetic marker of fish growth in marker assisted selection.