Reciprocal hybrids of tench Tinca tinca (L.) × bream Abramis brama (L.), and tench × carp Cyprinus carpio L., and some characteristics of their early development

Reciprocal hybrids between tench Tinca tinca (L.) and carp Cyprinus carpio L., and tench and bream Abramis brama (L.) were produced artificially. The survival of all these hybrids during embryogenesis was quite high. The highest survival rate (over 60%) at the eyed stage was observed for tench and bream hybrids (both sex combinations). The hatching rates of these hybrids were also over 60%. The number of larvae with some abnormalities (i.e. deformed body) was low. In contrast, the hatching rates of tench and carp hybrids were very low (0.2%). From over 1000 fertilized eggs, only three specimens started swimming, and only one specimen survived to juvenile stage. Embryos of hybrids and their parental species differ in morphological features. These differences were also visible in the juveniles. Body parameters of juvenile hybrids produced from three species had intermediate values in comparison to parental fish.     

Studies on sperm of diploid and triploid tench, Tinca tinca (L.)

The tench Tinca tinca is an interesting fish from the viewpoint of polyploidy and related atypical reproduction aspects. Triploid tench were produced artificially. Studies of spermiation as well as of sperm motility and structure were performed on several triploid and diploid males simultaneously with individual experimental crosses with diploid females to define their reproductive capacities. The testes of triploids visually looked less developed in the most of cases with lower sperm production (0.05 cm³ sperm per male), GSI and weight of testes compared to diploids (0.58 cm³ sperm per male). Analysis of variance showed significant influence of ploidy level on the percentage of motile spermatozoa. Triploidy did not change percentage of live spermatozoa and velocity of spermatozoa at the first time of sperm movement. The study of sperm structure by scanning electron microscopy revealed that most sperm cells were of normal structure with some anomalies. Sperm heads of triploid and diploid males were mostly round-shaped, 1.86±0.2 and 1.6±0.18 μm in diameter. The midpiece of triploid spermatozoa was slightly narrower than that of diploid ones with typical cylindrical shape. Flow cytometry revealed sperm cells of triploids to be largely aneuploid (1.47 n) with high mosaic DNA, oscillating from haploid DNA content (1.0 n) to diploid DNA content (1.9 n). Experimental crosses between triploid males and diploid females revealed that these males were capable to stimulate effective development with relatively high level of fertilization and hatching rates from 0 to 70%. In conclusion, triploidization does not seem to guarantee sterility of tench.     

Insemination, fertilization and gamete management in tench, Tinca tinca (L.)

Various procedures for artificial insemination in tench, Tinca tinca (L.) were re-examined with evaluation of fecundity of males and females among different tench strains. The objectives of this study were to enhance fertilization and hatching rates through optimization of the activation solution, the insemination process, the activation of gametes, and the elimination of eggs stickiness. Sperm for all experiments was collected directly into immobilization solution of modified Kurokura solution containing 180 mM of NaCl and stored at 2 °C for 2.5–5 h prior to the experiment. When dechlorinated tap water was used for activation a gamete ratio of 1150 spermatozoa per egg showed the best significant fertilisation and hatching rates. Optimal ratio between eggs (weight in g) and activation solution (in cm³) was 1:1. Different concentrations of activation solutions such as NaCl from 0 to 68 mM (0–136 mOsmol kg⁻¹) without buffer statistically decreased fertilization and hatching rates. The activation solution containing 17 mM of NaCl, 10 mM Tris–HCl, pH 8 and 9 significantly increased fertilization and hatching rates compared to dechlorinated tap water of pH 7 or activation solution containing 17 mM of NaCl, 10 mM Tris–HCl, pH 6 and 7. Adhesiveness of the eggs was successfully removed by incubation in Alcalase and activity: 3.16 Anson units per cm³.     

Does overfeeding affect tench Tinca tinca (L.) juveniles?

Duplicate groups of tench (Tinca tinca) juveniles (mean weight 0.69 g) were fed at 28 °C for 70 days either a commercial formulated diet, Futura, or frozen Chironomidae larvae at four levels: below satiation, at about satiation, above satiation and well above satiation. These are represented by the feeding groups F1–F4 and C1–C4, respectively. No mortality was observed throughout the experiment. In groups F2–F4 a considerable incidence of uneaten food (>40% observations), deformities of the caudal penduncle, retarded growth, elevated condition coefficient (>1.2), reduced amounts of minerals in the tissue (<10% dry matter), high C/N ratio (>5) and high caloric value of the tissue (>28 J mg⁻¹ dry matter) (the latter two suggesting excessive fat deposits), were found. The combination of these indices was indicative of overfeeding in fish fed the Futura diet at the daily doses ≥2.7% fish biomass. First symptoms of overfeeding: a considerable incidence of uneaten food, retarded growth, and elevated C/N ratio were observed in the group fed Chironomidae larvae at the highest level, (C4, wet Chironomidae 20.7% of the fish biomass daily, that is, 3.9% of dry chironomids per fish biomass and day). Thus, restricted daily doses of formulated diet not exceeding 2.5% fish biomass are recommended for tench juveniles aged 130–200 days post hatch. Safe daily doses of frozen Chironomidae (in terms of larvae dry weight) remain below 3.5% of fish biomass.     

Induced meiotic gynogenesis in tench, Tinca tinca (L.) using irradiated heterogenic sperm

Effects of the starting time and duration of cold shock, as well as the source of heterogenic sperm on the percentage of viable gynogenic larvae (PVGL) in tench were studied. The DNA in sperm of red common carp (Cyprinus carpio var singuonensis) was inactivated by ultraviolet radiation prior to use to induce meiotic gynogenetic development in tench. In experiment 1, tench eggs were cold-shocked for 30 min starting at 1, 3, 5, 7, 10 and 15 min post activation. In experiment 2, cold shock began 5 min after activation and lasted for 10, 20, 30, 40, 50 min, respectively. Each experiment was run in triplicate using 3 tench females, and one group not treated with cold shock was included in each experiment to serve as a control group. In experiment 3, sperm of bigmouth buffalo (Ictiobus cyprinellus), red common carp or grass carp (Ctenopharyngodon idella) was used to induce gynogenesis in tench with a 20-min cold shock starting 5 min post activation. The results showed that PVGL from the control group was very low (0.11–0.47%). In experiment 1, the highest average PVGL (9.60%) was observed when cold shock treatment was applied 5 min post activation. When cold shock treatment was started 5 min post activation, duration of cold shock affected PVGL. Cold shock lasting 20 min resulted in the highest average PVGL (12. 57%) among the selected duration of cold shock studied in experiment 2. The average PVGL was 2.3, 8.6, and 9.3%, respectively, for eggs induced by sperm of bigmouth buffalo, red common carp and grass carp. Average PVGL was significantly lower for eggs induced by sperm of bigmouth buffalo, compared with that for eggs induced by sperm of the other two species. However, average PVGL were similar for eggs induced by sperm of red common carp and grass carp. In summary, the optimal conditions for gynogenesis in tench include the use of irradiated sperm of grass carp to activate the eggs and cold shock of 20 min starting 5 min post activation. Since female tench grow much faster to a larger size than male tench, gynogenesis of tench holds great potential for production enhancement.     

The effect of feeding on oxygen consumption and ammonia excretion of juvenile tench Tinca tinca (L.) reared in a water recirculating system

The effect of feeding frequency (one, three, and continuous feeding), feed ration (0.2, 0.5, 0.8% of total fish biomass), and feeding per se on the oxygen consumption (OC, mg O₂ kg⁻¹ h⁻¹) and ammonia excretion (AE, mg TAN kg⁻¹ h⁻¹) of juvenile tench (body weight 15–19 g) and variations in these parameters in daily cycles were examined. Fish metabolism was studied in a recirculating system (rearing tanks of 0.2 m³, water temperature 23 °C). It was found that oxygen consumption and ammonia excretion depended significantly on feed ration. An increase of feed ration from 0.2 to 0.8% of fish biomass caused an increase of OC and AE from 126.80 mg O₂ kg⁻¹ h⁻¹ and 1.95 mg TAN kg⁻¹ h⁻¹ to 187.35 mg O₂ kg⁻¹ h⁻¹ and 8.80 mg TAN kg⁻¹ h⁻¹ (p<0.05). There was no dependence between feeding frequency and the mean rate of oxygen consumption. However, the relationship between feeding frequency and ammonia excretion by juvenile tench was statistically significant (p<0.05). Feeding frequency significantly affected daily fluctuations of AE and OC. It was found that diurnal variations in metabolic rates were strictly related to tench feeding, and the daily variations of AE were significantly higher than OC.     

一种免疫诱导型鲤启动子的克隆与功能分析

为发掘适用于基因工程抗病育种的鱼类启动子,通过实时荧光定量PCR实验对鲤Rab GTP酶(Ras-associated binding-GTPases 1a3,Rab1a3)基因的表达模式进行了分析,证实该基因在鳃、头肾等与机体免疫防御功能密切相关的组织内转录水平较高,且免疫激活后转录显著增强,符合基因工程抗病育种所需的外源免疫基因转录模式。从鲤细菌人工染色体文库中,使用Rab1a3基因特异引物筛选获得包含该基因区域的文库克隆,测序获得该基因完整序列,以及上下游调控序列。通过生物信息学手段,预测到长度为1014 bp的鲤Rab1a3基因启动子序列,该启动子不具有典型的TATA盒或CpG岛特征,存在多个免疫相关转录因子结合位点。在草鱼肾组织细胞系内验证该启动子活性,结果显示,绿色荧光蛋白基因和萤火虫荧光素酶基因都能够在该启动子驱动下表达,证实该片段具有启动子活性,且启动子活性在受到免疫诱导后增强,双荧光素酶报告基因检测结果显示,该启动子活性在免疫刺激后增强至免疫刺激前的8.67倍。研究表明,鲤Rab1a3基因启动子有望被开发成为免疫诱导型的基因工程元件,驱动外源免疫基因在鱼体内适时表达,抵御外界病原感染,同时避免非必要条件下的过度表达形成生长负担。     

Morphological changes of digestive structures in starved tench Tinca tinca (L.) juveniles

The aim of the study was to evaluate the effect of short-term starvation on the morphology of the digestive system of juvenile tench Tinca tinca (L.). The morphological changes in digestive structures of juveniles, fed 70 days starter or Chironomidae larvae, were examined after 1, 12, 24 and 48 h of starvation at 28 °C. The morphological changes in the organisation of the digestive structures were as follows: (1) Intestine – proteolysis of the intestinal mucosa, especially of enterocytes in apical part of mucosal folds; (2) Liver – progressive reduction of lipid vacuoles and glycogen granules, and, in effect, volume reduction of hepatocytes cytoplasm; (3) Pancreas – progressive degeneration of exocrine pancreatic cells, parallel with decrease of zymogen activity. The fish fed natural food prior to starvation seem to be more resistant to starvation than those fed starter.     

Effects of different diets on juvenile tench, Tinca tinca (L.) reared under controlled conditions

In a laboratory experiment at 28 °C, four commercial starters (Carp Starter, Uni Starter, Perla Plus, Asta) were fed intensively to 11-month-old juvenile tench Tinca tinca of initially 41 mm TL and 0.70 g BW, over a period of 120 days. Frozen chironomid larvae were fed to fish below satiation as a reference diet for comparison of the fish body chemical composition. Final survival rates of 99–100% and satisfactory growth were attained with all dry diets, ranging from 73.3 (Carp Starter) to 84.8 mm TL (Asta) and from 5.39 (Carp Starter) to 8.09 g BW (Perla Plus). External body deformities of up to 96.4% specimens appeared in those fed Uni Starter, Carp Starter and Perla Plus, whereas 0% were recorded for Asta and chironomids. Fish in groups with deformities showed significantly elevated lipid content in their bodies, lower ash content and higher condition coefficient values. Juveniles in groups without deformities did not substantially change lipid content, but the ash content increased. Thus, among four dry diets studied, only Asta starter can be fed effectively and safely to juvenile tench reared intensively under controlled conditions.     

鲤硒缺乏的病理学研究

将健康幼鲤360尾设置3个相同平行处理,每个处理用鱼120尾,随机分为四组,分别投以0 mg/kg、0.15 mg/kg、0.30 mg/kg和0.45mg/kg硒水平的饲料,各试验组的发病率和死亡率与日粮中硒含量高低呈负相关,其发病率分别为46.7%、33.3%、13.3%和0%,死亡率分别为26.7%、16.7%、6.7%和0%。病鱼出现特征性的“瘦背病”和脊柱弯曲等症状。组织学上最突出的变化表现为营养性肌病、营养性肝病、胰腺的变性、坏死等多器官组织的退行性变化。超微结构上,骨骼肌纤维、肝细胞和肾小管上皮细胞线粒体肿胀,嵴断裂,甚至发生溶解,整个线粒体呈囊泡状。硒缺乏幼鲤血液中谷草转氨酶（AST）、谷丙转氨酶（ALT）活性升高,谷胱甘肽过氧化物酶（GSH-Px）、过氧化物歧化酶（SOD）活性降低,丙二醛（MDA）含量升高。     

Quantity, motility and fertility of tench Tinca tinca (L.) sperm in relation to LHRH analogue and carp pituitary treatments

The spermiation of tench males was stimulated with Supergestran containing mammalian LHRHa lecireline at the following doses: 5, 10, 20 and 40 g kg⁻¹ b.w.; then with carp pituitary suspension (CPS) at a dose of 2 mg kg⁻¹ b.w. and with a control of saline physiological solution. The following days, meaning 24, 48 and 72 h after injection, sperm was collected to evaluate volume and the number of sperm per male per kg body weight (B.W.) The percentage of motile sperm and velocity of spermatozoa were measured 48 h after hormonal injection, and 72 h after hormonal injection the sperm was evaluated for fertilization and hatching ability. All 42 males in experimental groups were diploid. Live weight did not differ significantly among experimental groups. The strongest stimulation of spermiation was achieved with LHRHa in dosage of 20 and 40 g kg⁻¹ b.w. and CPS compared to males of the control group and lower dosage of LHRHa. Analysis of variance showed no significant influence of the treatment on the velocity and percentage of motile spermatozoa. The effect of different treatment on the fertilization capacity (the number of spermatozoa per egg was equilibrated) was significant. Significantly the highest quality of sperm collected 72 h after injection expressed by percentage of fertilization and hatching (62–65% fertilization and 61–64% hatching rates, respectively) was found for LHRHa in dosage of 20 and 40 g kg⁻¹ b.w. Significantly the lowest parameters of fertilization and hatching were found for the control group, on the 12% level.     

Kurokura Solution as Immobilizing Medium for Spermatozoa of Tench (Tinca tinca L.)

This study tested KUROKURA solution (Kurokura et al., 1984, Aquaculture 37, 267–274) and its modifications (by increasing NaCl content to 160, 180 and 200 mM) on immobilizing properties for sampling and short-term preservation of potential motility of tench spermatozoa. The immobilizing solution is used because, when collected, the sperm of most samples is contaminated by urine, causing spermatozoa to be of poor quality, with low motilities and velocities (almost 0), thus resulting in a worsened fertilization and hatching rate. Sperm was sampled with a syringe containing an immobilizing solution (IS), allowing an IS:sperm ratio of 2:1, under aerobic conditions at 0–4°C. This sperm solution was stored for 10 h and untreated sperm was collected prior to fertilization as a control. Spermatozoa quality was evaluated for the cell motility and velocity parameters and also for fertilization ability and hatching rate. Results obtained for tench sperm motility, velocity, fertilization and hatching rate showed that only sperm collected in the various immobilizing solutions can be successfully used for artificial insemination and preservation after 10 h at 0–4°C. The best immobilizing solution was found to be KUROKURA 180 (180 mM NaCl, 2.68 mM KCl, 1.36 mM CaCl₂· 2H₂O and 2.38 mM NaHCO₃), giving a fertility and hatching rate of 41%, with no change in rates after 10 h storage of sperm. Control sperm without immobilizing solution showed a fertilization and hatching rate of only 6–7%.     

鲤Rhbdd3蛋白的抗病毒功能

Rhbdd3(Rhomboid domain-containing protein 3)蛋白在哺乳动物天然免疫中发挥了重要作用,但水生动物中rhbdd3基因的确定序列及Rhbdd3蛋白的功能均尚未见报道。为研究鲤(Cyprinus carpio)的Rhbdd3蛋白在鱼类细胞中的功能,探讨其过表达对鱼类病毒感染的影响,本研究通过PCR扩增得到了鲤rhbdd3基因的编码序列,并将其克隆至pCI-neo载体上,构建了真核表达质粒pCI-rhbdd3。pCI-rhbdd3转染鲤上皮瘤细胞EPC(epithelioma papulosum cyprinid)和鲑囊胚细胞CHSE-214(chinook salmon embryo)后利用制备的特异性抗体进行Western blot,检测Rhbdd3蛋白的表达情况,并利用CCK-8试剂检测其过表达对细胞增殖的影响。转染后分别进行鲤春病毒血症病毒(SVCV)和传染性胰腺坏死病毒(IPNV)的感染实验,并利用间接免疫荧光、Western blot和RT-qPCR方法检测Rhbdd3过表达对SVCV和IPNV增殖的影响。结果显示,pCI-rhbdd3转染后Rhbdd3蛋白在EPC和CHSE-214细胞中得到了过表达,且Rhbdd3蛋白的过表达能显著抑制SVCV和IPNV的复制,但不影响两种细胞的正常活性。本研究为鱼类广谱抗病毒药物的开发提供了新的实验依据,也为鱼类抗病毒新品种的培育奠定了重要基础。     

Selective behaviour of a tench, Tinca tinca (L.), stock in different light–substrate combined conditions

Two physical factors light or substrate (fibre glass, artificial vegetation, gravel and mud) were determined in a 24 m long artificial channel to have importance on the distribution of 33 tench 2+, mean size 11.8±0.9 cm S.L. For this purpose in five trials tench could select between the half of the channel occupied by their preferred light intensity (inferior to 10 lux) combined with the less preferred substrate (fibre glass) or the half with the combination of one of the substrates considered and higher light conditions (40 lux). Fish preferred, with a 100% frequency, the optimal light-less preferred substrate when compared with the combinations fibre glass, gravel and mud. When artificial vegetation was the substrate considered, tench selected the combination preferred substrate – non-optimal light intensity with a 98.26±2.68% frequency. This result changed when increasing light intensity up to 150 lux, avoiding the shading effect. We demonstrated that light has a bigger influence than substrate in juvenile tench habitat selection under controlled conditions.     

Top-crossing with evaluation of slaughtering value in common carp (Cyprinus carpio L.) offspring

A three-year test of growth performance andoffspring survival from top-crossing of commoncarp hybrids in low altitude (350 m above thesea level, middle European climate) and in highaltitude (750 m above the sea level, middleEuropean climate) was terminated by assessmentof slaughtering value of edible parts in therespective strains. A recently establishedHungarian synthetic mirror carp strain (HSM)was chosen for testing as maternal strain. TheHSM, as well as wild Amur carp (AC), Ropshacarp (ROP) and Tata carp (TAT) were used aspaternal strains.The evidential strain*altitude interactioneffect was demonstrated as significant forweight of fish (0.0005), fillet without skin(0.0001), index of highbackedness (0.005),index of head length (0.0404),Fulton's coefficient (0.0497),index of widebackedness (0.0315) andgonadosomatic index (0.0082). The interactioneffect was demonstrated as insignificantbetween altitude*sex and betweenstrain*sex.The comparative test of analysis of variance(ANOVA) revealed significantly (P < 0.05)higher weight of fish after killing on lowaltitude (1611.6 g) than on high altitude(1090.3 g), but processed body yield andpercentage of fillet without skin was notsignificantly different (64.0 and 34.2 for lowaltitude, 64.2 and 33.4 for high altitude). Thepercentage of processed body weight andpercentage of fillet without skin in analysisboth altitudes were significantly better infemales (65.5 and 34.6%) than in males (62.5and 33.00%), respectively. The highestsignificant differences in weight of fish afterkilling were found between HSM (890.8 g) andHSM × AC (1283.3 g) in high altitude. In lowaltitude, it was between HSM (1527.2 g) andHSM × AC (1706.9 g) and HSM × ROP (1693.0 g).     

Changes in commercially exploited populations of tench, Tinca tinca (L.), in littoral zones of lakes of northeastern Poland

Commercially exploited tench, Tinca tinca, (L.) populations in 480 lakes in the northeastern Poland were analyzed on the basis of the fishery data from 1949 to 1994. The long-term changes in tench catch were studied in four categories of lakes with respective proportions of littoral zones (<30, 31–60, 61–90 and <90% of total lake area). The analyses were performed with using relative catch (kg ha⁻¹ month⁻¹) of tench. Mean relative catch of tench for 1949–1994 ranged from 0.36 kg ha⁻¹ month⁻¹ in lakes with smallest littoral zone (>30%) to 0.93 kg ha⁻¹ month⁻¹ in lakes with the largest littoral (<90%). Mean relative catch increased with increasing proportions of littoral zones in lakes. However, long-term trends in mean relative catch of tench showed decreasing tendencies in all lakes. The beginning of these tendencies was found about ten years earlier in lakes with smaller littoral (<60%) in comparison to lakes with larger littoral zones (>60%). Mean stocking rates of tench (number of 1-year-old fish per ha of littoral) for 1949–1994 were not significantly different in lakes of studied categories. However, lakes with proportionally less littoral zone had reduced stocking efficiency estimated as number of stocked fish per ha of littoral to require 1 kg ha⁻¹ month⁻¹ of tench.     

Tench (Tinca tinca) broodstock management in breeding station under conditions of pond culture: a review

A basic assumption for carrying out the breeding work is to have well-organised broodstock management. Taking into account the number of populations bred and the limited number of ponds available, it is impossible to avoid rearing several juvenile or adult breeds in mixed stocks. Fish of individual breeds are group-marked with regular renewal of the freeze-branded mark and broodstock adults should be marked individually. To minimise inbreeding or losses in genetic variation, at least 120 fish per strain are reared and when the strain is restored, factorial crosses of at least 15 females and 25 males should be applied. All activities are individually registered in the “Evidence 2003” data-recording software and data can be sorted by the origin of the fish population, of the strain or of individual fish; reproductive and performance parameters are recorded individually also. Further breeding operations with tench comprise the selection after over-wintering and rearing of fish before, during and after the reproductive season.     

Long-term examination of health conditions in monoculture of communally tested amphimictic diploid, diploid gynogenic and triploid tench, Tinca tinca (L.)

Amphimictic diploid (2n), induced triploid (3n) and meiotic gynogenic (MeiG) tench populations were established in 1998 by mass propagation. After separate nursing, fish were group-marked by freeze branding and 200 specimens per population were stocked into an experimental earthen pond for communal test until present. Parasitological and haematological (haemoglobin content Hb, haematocrit PCV and differential leukocyte count) examinations were performed every spring and autumn in the 2+ category, once sex could be registered macroscopically. Altogether 9 genera and/or species of parasites were detected on skin, 7 genera and/or species on gills, 1 genus in eye and 1 species in gut. The highest invasion extensity was recorded in the MeiG tench in the majority of examinations till the age of 4+. The courses of PCV and Hb values for 2n and MeiG populations showed high similarity while those for 3n population were slightly decreased and sex-related differences proved to be insignificant within populations in the majority of examinations. Differential leukocyte counts showed significant differences (p<0.05) of 3n vs. 2n and/or MeiG considering lower proportions of lymphocytes and higher proportions of segmented neutrophils in the majority of cases.     

Results of comparative growing test of all-female and bisexual population in two-year-old common carp (Cyprinus carpio L.)

The aim of this study was to verify whetherall-female carp populations grow faster thanbisexual ones. All-female mirror and scalypopulations were obtained by crossingmasculinizated females (neomales) of Northernmirror carp (M72) with regular females of M72and scaly Ropsa carp (ROP) purebreds,respectively. All-female and bisexual progenyof both mirror and scaly carps were rearedseparately for one year. However, controlgroups of a distinct phenotype of scalinesswere put in ponds with tested fish to make anadjustment for fish weight, which could becaused by environmental influence. Nodifferences were found after eight weeks ofrearing between all-female and bisexualpopulations, but, all-female mirror carppopulation (M72) gained 29.7% more weight(P < 0.0001) by the end of the first year thanthe bisexual (68.5 g ± 15.8 against 52.8 g± 13.9; mean ± S.D., respectively).Before beginning the second growing season,each group was marked and stocked together intriplicate ponds. Corrected weight after secondgrowing season (P < 0.0062) was 9.9% higher inall-female mirror carps (395.9 g ± 98.6)compared to the bisexual (360.2 g ± 89.0).No significant differences were found incorrected weights between scaly all-female andbisexual crossbreds (522.4 g ± 102.4 and518.3 g ± 95.8, respectively). Nodifferences in survival of all-female andbisexual populations were observed.     

锦鲤microRNA-137系统进化、靶基因验证及表达分析

为了探究microRNA-137(miR-137)与锦鲤体色形成的关系,实验对已报道的14种鱼类miR-137前体序列(precursor miR-137, pre-miR-137)进行比对,利用最大似然估计法(Maximum Likelihood Estimate, MLE)构建系统进化树,同时荧光定量PCR (quantitative realtime PCR, qRT-PCR)分析其在体色发生阶段和不同组织中的相对表达水平,并预测其靶基因,随后对靶基因进行GO和KEGG分析,最后通过荧光素酶实验验证miR-137-3p与靶基因的靶向关系,并分析miR-137-3p与靶基因mRNA的表达变化。结果显示,锦鲤与鲤的pre-miR-137序列相似度最高,且pre-miR-137序列在所有鱼类中具有较高的保守性。对miR-137靶基因进行GO和KEGG分析,多个靶基因富集在色素沉积、色素细胞分化等信号通路。定量结果显示,锦鲤出膜后11 d(day post hatching, dph) miR-137-3p表达量达到最高,随后显著降低;miR-137-3p在锦鲤不同组织中均有表达,在眼和肌肉中表达量较高,在皮肤和鳍条等色素细胞存在的组织中也有较高表达,且白色组织表达量显著高于红色组织。荧光素酶实验结果显示,miR-137-3p可结合在mitfa 3′-UTR上并抑制其表达,但对sprb并无显著抑制作用;miR-137-3p与mitfa和sprb在不同发育阶段存在显著负相关,但在组织中不存在相关性。本研究发现,miR-137在鱼类进化过程中高度保守,与锦鲤体色形成具有一定的相关性,且可靶向调控mitfa参与体色的形成,以上结果为进一步探讨miR-137在锦鲤体色形成中的作用提供基础数据。