云南青鳉鱼类的分类和分布(鳉形目:青鳉科)

经研究,云南的青鳉鱼类有2种:(1)分布于西双版纳的小青鏘 Oryzias minutillus Smith为中国境内首次记录;(2)另一种是新亚种,定名中华青鳉 Oryzias latipes sinentis subsp.nov.广泛分布于云南及中国其他地区。本新亚种与产自日本的指名亚种日本青鳉 Oryzias latipes latipes Temminck et Schlegel的主要区别在于:本新亚种的第一肋骨连接第二而不是第三脊椎骨上;平均脊椎骨数为29.2±0.6而不是30.3±0.5;胸鳍条多数为9而不是多数为10枚;染色体数目2n=46而不是2n=48。这些差异,明显地表示了由于地理隔离而引起的亚种分化。     

广东淡水鱼类弓背青鳉分类形态性状再描述

食蚊鱼Gambusia affinis的入侵导致弓背青鳉Oryzias curvinotus种群严重下降而趋于濒危。其模式标本产地种群(海南岛那大)已经灭绝,野外种群已多年未见报道。过去,研究者对正模标本的描述存在严重错误且难于复检,现亟需对其分类性状进行再描述及模式标本重建。本文对近期采自广州南沙岛的弓背青鳉孑遗种群进行了分类形态性状再描述,补充了活体及浸制标本彩色照片,并对其雌雄间的形态差异进行了统计分析。结果显示,弓背青鳉区别于其他青鳉鱼类的主要分类检索性状是臀鳍条数目为19.60±0.80(19~22)、体侧纵列鳞数目为28.51±0.67(27~30),定种的关键特征是背鳍-吻端/体长:雄鱼为(80.9±1.3)%、雌鱼为(82.2±1.0)%、全部为(81.6±1.3)%;同时,通过单因素方差分析、主成分分析及判别分析等多元统计方法证实了其种内存在的雌雄异形现象,文中回顾并澄清了有关弓背青鳉的分类形态性状问题。当前亟待加强弓背青鳉保护生物学研究并应采取有力的保护措施尽早对现存种群进行有效保护。     

青鳉(Oryzias latipes)smad3的克隆与表达分析

鱼类性染色体的原始性导致性别决定基因的多样性.高等动物中的单个基因,在鱼类中因为基因组复制而产生多个同源基因,呈遗传多样性.TGF-β家族在硬骨鱼性别决定和性别分化的过程中发挥关键作用.smad3是性腺体细胞衍生因子(gsdf)的下游基因,可将TGF-β信号从细胞表面传递至细胞核.本研究从青鳉精巢和卵巢组织cDNA中克隆了smad3a、smad3b基因的CDS全长序列.蛋白序列的同源性比对和系统发生分析显示,smad3在高等脊椎动物中只有一种,而在鱼类却有两种.RT和定量PCR分析结果显示,smad3在多组织中普遍表达,在精巢中表达高于卵巢.青鳉smad3a和smad3b的表达同性腺分化紧密关联,提示其在性腺雌雄性分化和发育中可能具有重要作用.     

青鳉prdm14的原核表达、多克隆抗体制备及其应用

青鳉(Oryzias latipes)是研究遗传发育和细胞多能性的重要模式鱼类, 为探究prdm14同源基因的潜在作用, 实验将青鳉prmd14经原核表达后制备了兔抗Prdm14多克隆抗体。首先, 将prdm14基因的部分编码区连接到pET32a质粒中, 构建重组表达载体pET32a-prdm14?600。随后将重组载体转化至大肠杆菌(Escherichia coli)Rosetta(DE3), 经异丙基-β-d-硫代半乳糖苷(Isopropyl-β-d-thiogalactoside, IPTG)诱导表达, 获得分子量为60 kD的Prdm14重组蛋白。接着大量诱导蛋白表达并切胶纯化, 免疫家兔(Oryctolagus cuniculus), 6周后获得阳性抗体, 最后通过ELISA和Western blot检测抗体效价及其特异性。结果显示, 在37℃、0.6 mmol/L IPTG、诱导3h的条件下, 可获得Prdm14重组蛋白的高效表达; 制备的兔抗青鳉Prdm14多克隆抗体能够特异性识别青鳉组织中表达的Prdm14蛋白以及在HepG2细胞中过表达的青鳉Prdm14: EGFP融合蛋白。综上所述, 研究首次制备了一种能有效识别青鳉Prdm14的多克隆抗体, 该抗体的获得为后续研究prdm14基因在鱼类多能性干细胞中的作用提供了有力工具。     

一种可高效转染的青鳉肌肉细胞系的建立与应用

在哺乳动物细胞系中转基因的效率相对较高而在鱼类细胞系中相对较低, 为了获得鱼类高效转基因细胞系, 我们从模式鱼类青鳉(Oryzias latipes)中分离和构建了肌肉细胞系OLM。该细胞系类似于纤维细胞, 在28℃ DMEM和10%的胚胎牛血清中培养了超过88代。通过染色体分析, 它属于两倍体并含有48条染色体。在脂质体介导下, 报告基因的转染效率可以达到40%, 在测试的其他鱼类细胞中转染效率最高。此外还构建了稳定转染转基因或者非编码RNA的细胞系。OLM细胞对常见的病毒SVCV、GCRV、SGIV等不敏感。综上所述, 研究在青鳉中建立了一种能高效转染的肌肉细胞系, 它适用于鱼类瞬时和稳定的转基因研究。     

西藏雅鲁藏布江中游发现黄斑褶[鱼兆]

正2016年4月,在西藏雅鲁藏布江中游干流下段的派镇段(29°31′9″N,94°52′12″E,海拔2 897 m),6月在米林段(29°12′1″N,94°5′16″E,海拔2 947 m),2017年7月在里龙段(29°8′32″N,93°54′34″E,海拔2 960 m),使用定置刺网共捕获到15尾疑似黄斑褶(Pseudecheneis sulcatus)的鱼类标本(图1a,b)。2017年4月在雅鲁藏布江下游墨脱段(29°18′10″N,95°16′52″E,海拔682 m)采集到一批黄斑褶标本(图1c,d),并与中游的黄斑褶进行了形态学比较鉴定。     

铜或镉胁迫对海洋青鳉生理生化指标及产卵量的影响

通过研究海水中不同浓度重金属胁迫对海洋青鳉(Oryzias melastigma)生理生化的影响, 探讨与分析各指标间的相互关系, 为海洋重金属污染的生物监测提供参考。按海水水质标准分别设定铜(Cu)和镉(Cd)离子各3个浓度梯度, 测定海洋青鳉中乳酸(LA)、乳酸脱氢酶(LDH)、睾酮(T)、促卵泡素(FSH)、促黄体生成素(LH))及产卵量的动态变化。海洋青鳉中的LA含量随铜胁迫时间的延长而显著降低, 随镉胁迫浓度的增加而升高; LDH活性在铜胁迫下变化不大, 而随镉胁迫时间的延长略有上升; T含量在短期铜胁迫下有所升高, 但随胁迫时间的延长呈显著下降趋势, 却随镉胁迫浓度升高及胁迫时间延长而上升; FSH与LH含量随铜胁迫浓度增加呈显著下降, 随镉胁迫浓度增加呈先降后升趋势, 两者的含量变化与T的动态密切相关。与此同时, 随铜胁迫时间的延长和胁迫浓度的增加, 青鳉的产卵量也呈显著下降趋势。综合表明, 铜或镉胁迫均能显著影响雌性青鳉的LA含量, 增加能量消耗, 进而影响性激素水平, 最终影响到产卵量, 但两类离子的影响存在较大的差异性。因此, 一定浓度的铜或镉胁迫会导致海洋青鳉鱼体内生理生化的变化, 且存在两性差异性, 雌鱼对铜或镉胁迫的响应敏感强, 可选择雌鱼开展相关污染监测。     

青鳉与食蚊鱼竞争排斥的生态形态学解释

生态形态学理论指出,形态相似的物种生态位相似,是导致种间竞争排斥的关键性因素。在鱼类入侵生态学研究中引入生态形态学理论,对于加深理解入侵种与土著种的种间相互关系有着重要意义。本文利用形态分析方法,对广东怀集燕都国家湿地公园入侵种食蚊鱼对土著种鳍斑青鳉和弓背青鳉的影响机制进行研究。结果表明: 弓背青鳉和鳍斑青鳉在研究区域常同域分布,但两者的空间分布格局差异显著。两者形态高度相似,符合生态形态学“形态相似者竞争排斥”理论。与群落其他鱼类相比,食蚊鱼与2种青鳉鱼类形态更为相似。聚类分析发现,食蚊鱼和青鳉鱼类聚合在同一分枝,且其空间生态位的重叠度极低。种群相对密度调查显示,食蚊鱼与青鳉鱼类种群数量呈显著的负相关关系。食蚊鱼的入侵是导致2种青鳉鱼类种群数量明显下降的关键原因。形态特征的相似性能够初步解释食蚊鱼与青鳉鱼类的竞争排斥关系,尚需从不同角度就食蚊鱼入侵对青鳉鱼类的作用机制进行深入探索。     

用模式生物青鳉概观硬骨鱼性别决定及性分化研究进展

鱼类性别决定和性分化呈多元性,既有雌雄同体也有雌雄异体。性腺的雌雄性分化过程受遗传和环境因素(如温度、光照、激素和pH值等)影响,具有可逆可塑性。随着生物技术和基因组学的迅速发展,近年来脊椎动物性别决定和性分化的研究有了重大进展和显著突破。本文通过聚焦青鳉及其他硬骨鱼纲保守存在的dmrt1、gsdf、amh等雄性因子,探讨硬骨鱼普遍存在的雌雄性别可塑可逆信号通路,并介绍了新的基因组编辑和性控育种技术,为单性选育等水产养殖技术的研发提供参考。     

Morphometric and meristic variation in ricefishes,genusOryzias: a comparison with cytogenetic data

Morphometric and meristic data from type and other material of ricefishes of the genusOryzias, particularly onO. latipes andO. curvinotus, were retaken to compare relationships of ricefishes indicated by morphological data with the relationships indicated by the cytogenetic analyses (Uwa, 1986). Along with morphometric and meristic characters traditionally used, two morphometric characters are useful for the classification of this genus: the relative length of the abdominal part and the relative position of the dorsal-fin origin. External anatomical data mostly agree with a classification of ricefishes based on karyology; although, like the cytogenetic data, they are not sufficient to distinguish all species, or to define monophyletic groups.Oryzias latipes is widespread in eastern Asia.Oryzias curvinotus, a name applied to ricefish from Hainan Island, is distributed more widely in southern China, including Hainan, Guangdong, Hong Kong, and Hanoi.     

Karyotypes and geographical distribution of ricefishes from Yunnan,southwestern China

Morphometric and karyotypic studies were made on two species of ricefishes collected from Yunnan, southwestern China.Oryzias latipes from Yunnan had the same morphological and karyological characteristics asO. latipes collected from eastern China. The Yunnan populations had 2n, 46 chromosomes consisting of 3 metacentric, 8 submetacentric, 2 subtelocentric, and 10 acrocentric pairs, the arm number (NF) being 68 (2n=46, NF=68, 3M+8SM+2ST+10A). The karyotype was characterized by having a “large” metacentric pair and nucleolus organizer regions (NORs) on the short arms of a submetacentric pair.Oryzias minutilius from Yunnan had the same morphological characteristics asO. minuiillus from Thailand and Burma, although the karyotype was different from that collected from Thailand. The Yunnan population had 2n, 42 chromosomes consisting of 21 acrocentric pairs, NF being 42 (2n=42, NF=42, 21A). The karyotype was characterized by having NORs at the telomeric regions of an acrocentric pair.Oryzias latipes occurs widely on the eastern Yunnan Plateau where the climate is temperate or subtropical, whereasO. minutilius is found in Xishiangbanna, the southern low mountain areas of Yunnan, where the climate is tropical.     

<Emphasis Type="Italic">Adrianichthys roseni</Emphasis> and <Emphasis Type="Italic">Oryzias nebulosus</Emphasis>, two new ricefishes (Atherinomorpha: Beloniformes: Adrianichthyidae) from Lake Poso,Sulawesi, Indonesia

Two new species of ricefishes or buntingi, Adrianichthys roseni and Oryzias nebulosus, are described from Lake Poso, Sulawesi Tengah, Indonesia, bringing to 12 the number of endemic ricefishes known from Sulawesi. Adrianichthys roseni and its sole congener, A. kruyti, are distinguished from other relatively large-bodied ricefishes, Xenopoecilus, in having orbits that project beyond the dorsal profile of the head; paired preethmoid cartilages (versus single or absent in Xenopoecilus); and 13–16 (versus 8–13) dorsal fin rays. Also, as in A. kruyti, the lower jaw of A. roseni is enclosed in the upper jaw when the mouth is closed; however, the upper jaw is not as large and broadly expanded as in A. kruyti. Adrianichthys roseni differs further from A. kruyti in having fewer scales in a lateral series (approximately 63–65 versus 70–75), attaining a smaller maximum recorded standard length (90mm versus 109mm), and having the lateral process of the pelvic bone in line with the fifth, rather than the eighth, pleural rib. Oryzias nebulosus shares with O. nigrimas, also from Lake Poso, a unique sexual dichromatism in preserved specimens: adult males are dark gray to black, whereas females are a lighter grayish-yellow to brown. Relative to O. nigrimas, O. nebulosus is a small species (maximum recorded SL 33mm versus 51mm), with pelvic fins relatively anterior (lateral process of pelvic bone is in line with the third, fourth, or fifth, modally fourth, rather than the fourth or fifth, modally fifth, pleural rib), dorsal fin relatively anterior (origin above the 12–14th anal fin ray as opposed to the 15–17th anal fin ray), precaudal vertebrae 11–13, modally 12 (versus 13–14, modally 13), and relatively straight, narrow and slender (as opposed to curved, broad, and robust) ossified and cartilaginous portions of the gill arches. Endemic buntingi are threatened by introduction of exotic species, overfishing, and pollution. A conservation plan includes rearing native fishes for local use as ornamental fishes and possible reintroduction.     

Mesozoic origin and ‘out-of-India’ radiation of ricefishes (Adrianichthyidae)

The Indian subcontinent has an origin geologically different from Eurasia, but many terrestrial animal and plant species on it have congeneric or sister species in other parts of Asia, especially in the Southeast. This faunal and floral similarity between India and Southeast Asia is explained by either of the two biogeographic scenarios, ‘into-India’ or ‘out-of-India’. Phylogenies based on complete mitochondrial genomes and five nuclear genes were undertaken for ricefishes (Adrianichthyidae) to examine which of these two biogeographic scenarios fits better. We found that Oryzias setnai, the only adrianichthyid distributed in and endemic to the Western Ghats, a mountain range running parallel to the western coast of the Indian subcontinent, is sister to all other adrianichthyids from eastern India and Southeast–East Asia. Divergence time estimates and ancestral area reconstructions reveal that this western Indian species diverged in the late Mesozoic during the northward drift of the Indian subcontinent. These findings indicate that adrianichthyids dispersed eastward ‘out-of-India’ after the collision of the Indian subcontinent with Eurasia, and subsequently diversified in Southeast–East Asia. A review of geographic distributions of ‘out-of-India’ taxa reveals that they may have largely fuelled or modified the biodiversity of Eurasia.     

Evidence for sympatric speciation in a Wallacean ancient lake

Sympatric speciation has been demonstrated in few empirical case studies, despite intense searches, because of difficulties in testing the criteria for this mode of speciation. Here, we report a possible case of sympatric speciation in ricefishes of the genus Oryzias on Sulawesi, an island of Wallacea. Three species of Oryzias are known to be endemic to Lake Poso, an ancient tectonic lake in central Sulawesi. Phylogenetic analyses using RAD‐seq‐derived single nucleotide polymorphisms (SNPs) revealed that these species are monophyletic. We also found that the three species are morphologically distinguishable and clearly separated by population‐structure analyses based on the SNPs, suggesting that they are reproductively isolated from each other. A mitochondrial DNA chronogram suggested that their speciation events occurred after formation of the tectonic lake, and existence of a historical allopatric phase was not supported by coalescent‐based demographic inference. Demographic inference also suggested introgressive hybridization from an outgroup population. However, differential admixture among the sympatric species was not supported by any statistical tests. These results all concur with criteria necessary to demonstrate sympatric speciation. Ricefishes in this Wallacean lake provide a promising new model system for the study of sympatric speciation.     

Spatial-temporal expression of pum1 and pum2 in medaka Oryzias latipes

Two pumilios, pum1 and pum2, were identified in medaka Oryzias latipes. Oryzias latipes pum1 and pum2 are ubiquitous in the adult tissues but with specific expression in the germ cells of gonads, ovary and testis. Pum1 is expressed in the spermatogonia to spermatocytes whilst pum2 presents in spermatocytes of testis only. Oryzias latipes pum1 and pum2 are maternally supplied RNA with ubiquitous expression in the early stages, and embryonic expression of pum1 and pum2 may begin from early gastrula. Both pum1 and pum2 are expressed in the tissues including brain, eye and trunk, and both are expressed in the gonads after hatching. Taken together, Pum1 and Pum2 may play important roles in embryonic and germ cell development of O. latipes.     

Karyotype evolution and geographical distribution of the Thai-medaka, Oryzias minutillus, in Thailand

The karyotype of Oryzias minutillus was examined with specimens collected from 18 localities in Thailand. Specimens from the south and the northeast had 2n = 42 acrocentric chromosomes; the arm number (NF) was 42 and NORs-chromosomes were acrocentric type (2n = 42, NF = 42, NORs-A). Specimens from the central and the north were characteristic by having 8-12 large metacentric chromosomes (LM-chromosomes). They had 2n = 28–34 chromosomes, and shared the same NF and NORs-chromosomes of submetacentric type (2n = 34-28, NF = 44, NORs-SM). Specimens from the southeast had 2n = 42 or 40 chromosomes. Their karyotypes had the same NF and NORs-chromosomes as those from the central and the north (2n = 40–42, NF = 44, NORs-SM), though they had no, or only one pair of, LM-chromosomes. The karyotype with 42 acrocentric chromosomes seems to be basic for O. minutillus , and consequently those with NORs-SM and LM-chromosomes seem to be caused through pericentric inversion and centric fusion, respectively. We confirmed that the karyotype evolution had occurred in drainage areas of the Mae Nam Chao Phraya and collaterals (the central, north and southeast). On the other hand, the basic karyotype was preserved allopatrically in the peninsula (the south) and the basin of the Mae Nam Mun, a tributary of the Mekong (the northeast).     

Biochemical characterization of the medaka (Oryzias latipes) orthologue for mammalian tissue-type transglutaminase (TG2)

Transglutaminase is an enzyme family responsible for post-translational modification such as protein cross-linking and the attachment of primary amine and/or deamidation of glutamine-residue in proteins. Medaka (Oryzias latipes), a recently established model fish, has similar functional proteins to those characterized in mammals. Previously, we found the apparent orthologues that correspond to human transglutaminases in medaka. In this study, regarding the medaka orthologue of human tissue-type transglutaminase (OlTGT), recombinant protein was expressed in an active form in bacteria cultured at low temperature. Using the recombinant protein, we biochemically characterized the enzymatic activity and also obtained a monoclonal antibody that specifically recognized OlTGT. Immunochemical analysis revealed that OlTGT was not expressed ubiquitously, unlike its mammalian orthologue, but in primarily limited tissues such as the eye, brain, spinal cord, and gas gland.     

Heritability and genetic correlation of abdominal versus caudal vertebral number in the medaka (Actinopterygii: Adrianichthyidae): genetic constraints on evolution of axial patterning?

Variation in the number of abdominal vs. caudal vertebrae is an important source of morphological diversification of fish. It is not clear, however, whether abdominal and caudal regions evolve independently. Regressions of offspring on parents demonstrated substantial additive genetic variation within populations, i.e. heritability, in both abdominal and caudal vertebral numbers of the medaka ( Oryzias latipes ). However, the heritability of caudal vertebrae tended to be smaller than that of abdominal vertebrae in some estimations, suggesting that abdominal and caudal regions are controlled by separate developmental modules. Furthermore, genetic correlation between abdominal and caudal vertebral numbers, estimated using full-sib family means, was negative but weak, supporting independent evolution. In addition, substantial genetic differentiation among populations was demonstrated in abdominal vertebral numbers, but not in caudal numbers. These results support our view that Jordan's rule, a geographical tendency for fish from higher latitudes to have more vertebrae, in this fish reflects local adaptations of abdominal vertebral numbers. In contrast, the low heritability of caudal vertebrae may reflect the intrinsic invariability of genes associated with a change in caudal vertebral numbers. This genetic constraint may have restricted morphological diversification of not only the medaka, but also the Order Beloniformes as a whole.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 867–874.     

河流汞污染对青鳉鱼早期发育的毒性影响

清镇市化工厂是贵州省东门桥河汞污染的重要来源。为了评估这条河流的汞污染程度,本研究调查了东门桥河环境样品中的汞含量和分布情况。河流水体总汞(THg)和甲基汞(MeHg)的平均含量分别为(135.79±47.40)和(0.64±0.58)ng·L-1;生物膜中THg和MeHg的平均含量分别为(3.38±1.91)mg·kg-1和(10.47±0.40)μg·kg^-1;沉积物中THg和MeHg的平均含量分别为(23.21±20.22)mg·kg-1和(32.74±10.96)μg·kg^-1。为了进一步了解这条河流中汞对水生生物的风险,将青鳉鱼胚胎和仔鱼分别暴露于从河中收集的被汞污染的河水、生物膜和沉积物中。青鳉鱼胚胎经15 d暴露后,孵化时间延长、孵化率显著降低,死亡率显著升高,实验暴露还导致胚胎无法破膜、卵黄囊水肿等畸形效应,以及初孵仔鱼不同程度的畸形。青鳉鱼仔鱼经24、48、96 h和21 d暴露后,各暴露组的死亡率和畸形率随着暴露时间的延长而显著升高,且表现出一系列以脊柱弯曲为主的畸形效应。研究表明,东门桥河的汞污染会严重影响青鳉鱼的早期生长发育。