Preliminary investigation on otolith microchemistry of naked carp (Gymnocypris przewalskii) in Lake Qinghai, China

The paper represents the first otolith study on trace element (Sr, Mg) concentrations of naked carp (Gymnocypris przewalskii), an endemic and endangered migratory species in high-altitude Lake Qinghai and its tributaries of China, using electron probe microanalysis (EPMA). Results from life-history transect analyses and X-ray intensity maps suggested that the general reference criterion for discrimination of the salinity habitats of freshwater, brackish and seawater by the otolith Sr:Ca ratio of diadromous fish cannot be used as a reliable indicator for reconstructing the migration history of naked carp between different habitats. On the contrary, Mg concentrations and Mg:Ca ratios were found inconsistent in all otolith samples of G. przewalskii. The otolith regions with higher Mg concentrations (greenish) might coincide with the period that yolk provides nutrition to G. przewalskii. The lower Mg:Ca ratios in most regions of otolith showed that Mg in G. przewalskii might be regulated to that low level during its lifespan in Lake Qinghai. This suggests that the distinct EPMA approach can provide new visualization insight into the ontogenetic and migration patterns of G. przewalskii.     

五条河流青海湖裸鲤的同工酶变异

为全面了解青海湖裸鲤遗传多样性现状,采用水平淀粉凝胶电泳技术分析了青海湖裸鲤[样品采自青海湖的五条支流,分别为泉吉河(QJ)、沙柳河(SL)、哈尔盖河(HEG)、布哈河(BH)以及甘子河(GZ)]的13种同工酶,共记录26个位点,统计了基因频率,计算了其遗传多样性。结果表明:26个基因位点中多态位点6个,分别为Pgm、Hk、Cat、Gdh、Es-t1、S-Mdh,多态位点比例(P0.99)为23.08%,表明青海湖裸鲤的五个群体皆具有较高的遗传多样性水平;分别计算了五个群体的平均杂合度(H)为0.1161(HEG)、0.1178(GZ)、0.1304(SL)、0.1305(BH)、0.1287(QJ),多态位点比例(Ae)分别为1.3112(BH)、1.3069(SL)、1.2542(GZ)、1.2492(HEG)、1.2453(QJ);除泉吉河群体外,其它四个群体的遗传偏离指数均为负数,表明多数群体处于一定程度的杂合子缺失状态;计算了五个群体间的遗传距离以及遗传相似度,结果显示群体间遗传距离与遗传相似度与各个河流的地理位置具有很强的相关性;根据五个群体的遗传距离进行了聚类分析,通过构建系统树图将其基本分为两大种群,甘子河与哈尔盖河两群体为一类群,另外三个群体分为一个类群;计算群体间的基因流(Nm)为7.2013,说明河道之间具有较为广泛的基因交流,并计算了两两群体间的基因流,其中布哈河群体与其它群体间的基因交流都较为广泛。     

Seasonal population dynamics of Neoechinorhynchus qinghaiensis in the carp, Gymnocypris przewalskii przewalskii, from Qinghai Lake, China

Studies on the seasonal population dynamics of Neoechinorhynchus qinghaiensis (Acanthocephala: Neoechinorhynchidae) in its fish host Gymnocypris przewalskii przewalskii in the Qinghai Lake, China, were carried out with samples taken in May 1991, August 1992, November 1992 and February 1993. Prevalences were higher than 44% in all seasons. The mean intensity of infection was above 124 worms per fish. The maximum intensity of worms recovered from a single fish was 1402 in the autumn of 1992. Differences in the mean abundance, mean intensity and prevalence are not statistically significant relative to season and this is likely to be related to the stable temperatures recorded at the bottom of Qinghai Lake. Over-dispersed distributions of N. qinghaiensis in the host population, due to heterogeneity and feeding habits, were observed in all seasons. The size composition of both sexes of N. qinghaiensis showed males to be less than 3.5 mm and females between 0.5 and 4.25 mm, with the main recruitment phase in the worm populations occurring in the autumn, extending through winter and spring with the lowest recruitment occurring in the summer. The maturation and copulation of worms were mainly focused in the summer season. The sex ratio of female to male was both high in winter (1.51:1) and spring (1.48:1). The higher proportion of females and the change in the worm sex ratio in winter can be attributed to the reduced longevity of male worms. As immature male worms exhibit a higher proportion of the worm population than females in all seasons, further studies are needed to determine if such a situation compensates for the shorter life span of males.     

Unusual physiology of scale-less carp,Gymnocypris przewalskii,in Lake Qinghai: a high altitude alkaline saline lake

The scale-less carp (Gymnocypris przewalskii) inhabits Lake Qinghai located on the Qinghai-Tibet plateau (elevation, 3200 m) in western China. The lake waters are alkaline (pH 9.4, titratable alkalinity=30 mmol l(-1)), Mg(2+)-rich (18.7 mmol l(-1)), Ca(2+)-poor (0.30 mmol l(-1)) and saline (9 per thousand ). These fish make annual spawning migrations into freshwater rivers. We investigated the physiology of nitrogen excretion and ionoregulation of fish from the lake and river. Fish from both waters were ammonotelic, although ammonia-N excretion rates were lower in lake fish (175 vs. 344 micromol kg(-1) h(-1), P<0.05) resulting in unusually high levels of ammonia in blood plasma (2.23 vs. 0.32 mmol l(-1)), bile, liver, muscle and brain. Exposure to 0.4 mmol l(-1) total ammonia in lake water ([NH(3)]=0.16 mmol l(-1)) killed fish within 8 h. River fish survived exposure to 1.0 mmol l(-1) total ammonia in river water at pH 8.0 ([NH(3)]=0.023 mmol l(-1)) for 24 h suggesting high ammonia tolerance in lake fish. High glutamate dehydrogenase and glutamine synthetase activities in tissues probably allow the fish to alleviate ammonia toxicity by amino acid accumulation. Neither lake nor river fish relied on urea excretion to remove excess N. Urea-N excretion rates were below 20 micromol kg(-1) h(-1) for both groups, and levels of urea in plasma and tissues were moderate. When exposed to elevated ammonia, urea-N excretion increased slightly (approximately 50 micromol kg(-1) h(-1)) and liver and muscle urea levels increased in the river fish. Plasma ion levels were within the range typical of cyprinids, but river fish had significantly higher plasma [Na(+)] and [Cl(-)] and lower [K(+)] than fish from the lake. During 48-h lake-to-river water transfer, plasma Na(+) and Cl(-) levels rose significantly. Significantly higher Na(+)/K(+)-ATPase activity in the gills of river fish may be related to the higher plasma ion levels. Plasma [Mg(2+)] and [Ca(2+)] were tightly regulated despite the great differences in the lake and river water levels.     

Hydroacoustic study of spatial and temporal distribution of Gymnocypris przewalskii (Kessler, 1876) in Qinghai Lake, China

Gymnocypris przewalskii (Kessler 1876) is an endangered and state-protected rare fish species in Qinghai Lake, China. To further understand the life history and distribution of this fish, five surveys were carried out in Qinghai Lake between 2002–2006. Results of these surveys indicate that fishes were predominantly distributed about 2 m under the surface. In July, significant differences in fish density were found between surface and bottom layers (P = 0.001), and/or between middle and bottom layers (P = 0.025). Fish density was the greatest in the surface layer. In August and October, no significant differences were found between the different layers, but the bottom layer had a greater fish density. Furthermore, there were very large differences among different zones in fish distribution density. Differences in horizontal distribution were not significantly correlated to factors such as water depth and inshore distance, possibly because of very low and uniform fish density. Feeding, changes in water temperature, over-wintering and spawning appeared to influence fish distribution. Hydroacoustic estimates of G. przewalskii biomass in Qinghai Lake increased significantly between 2002 and 2006. We attribute this increase to the management measures put in place to protect this species.     

青海湖裸鲤体外肝胰细胞原代与传代培养

采用组织块移植培养技术,分别用DMEM和RPM11640培养基对青海湖裸鲤肝胰组织细胞进行原代培养。培养48h组织块周围有细胞迁出,并形成生长晕。培养一周可形成单层细胞。对原代培养的单层细胞用胰蛋白酶-EDTA消化后,传代培养至第四代。确立青海湖裸鲤肝胰细胞培养条件为：培养基为DMEM,培养温度为27℃,pH值为7．0—7．5,原代培养血清浓度为20％,传代培养的血清浓度为10％,无需通入CO2和添加细胞生长因子。     

Amplified fragment length polymorphism analysis to identify the genetic structure of the Gymnocypris przewalskii (Kessler, 1876) population from the Qinghai Basin, China

Amplified fragment length polymorphism (AFLP) was used to analyse the genetic structure of 45 individuals of Gymnocypris przewalskii (Kessler, 1876), an endangered and state‐protected rare fish species, from three areas [the Heima (HM), Buha (BH) and Shaliu rivers (SL), all draining into Qinghai Lake]. A total of 563 polymorphic loci were detected. The HM, BH and SL populations have 435, 433 and 391 loci, respectively ( Zhu and Wu, 1975 ), which account for 77.26%, 76.91% and 69.45% of the total number of polymorphic loci of each population, respectively. The Nei indices of genetic diversities (H) of the three populations were calculated to be 0.2869 (HM), 0.2884 (BH) and 0.2663 (SL), respectively. Their Shannon informative indices are 0.4244, 0.4251 and 0.3915, respectively. Research results show that the mean genetic distance between HM and BH is the smallest (0.0511), between BH and SL is the second shortest (0.0608), and between HM and SL is the largest (0.0713), with the mean genetic distance among the three populations being over 0.05. Data mentioned above indicate that the three populations have a certain genetic differentiation. The total genetic diversity (H_t = 0.3045) and the mean value of genetic diversity within the population (H_s = 0.2786) indicate that the variations have mainly come from within the population.     

Micropropagation of an endangered plant species: Coronopus navasii (Brassicaceae)

Plantlets of Coronopus navasii, an endangered species from SE Spain, were successfully regenerated from shoot and root segments excised from young seedlings. Initiation of multiple buds and development of leaves were obtained in MS modified medium plus l mg/l BAP and 0.1 mg/l NAA. Rooting was achieved by transfer of the isolated shoots to fresh MS medium without plant growth regulators. Plant survival of 47% was obtained six weeks after removal from in vitro culture conditions.Abbreviations BAP 6-benzylaminopurine - NAA naphthaleneacetic acid - MS medium see Murashige and Skoog 1962     

Hybridization between native barbless carp (Cyprinus pellegrini) and introduced common carp (C. carpio) in Xingyun Lake, China

Hybridization with introduced fish species is an important threat to native fish species. Here we investigated hybridization between native barbless carp (Cyprinus pellegrini) and introduced common carp (C. carpio) in Xingyun Lake in the Yunnan-Guizhou plateau of China. A total of 203 individuals of Cyprinus from Xingyun Lake were studied by combination of morphological and genetic analyses. Most individuals were strictly intermediate between the two parental species in morphology, strongly suggesting that extensive hybridization has occurred. Bayesian model-based clustering of the genetic data suggests that there are two distinct genetic groups corresponding to barbless and common carp respectively. Many individuals in the two genetic groups showed intermediate morphology, suggesting that both groups actually contain massively introgressed genes. Only two individuals were identified as barbless carp both morphologically and genetically, hinting that this native species is at risk of genetic extinction in Xingyun Lake.     

Local genetic population structure in an endangered plant species, Silene tatarica (Caryophyllaceae)

Genetic substructuring in plant populations may evolve as a consequence of sampling events that occur when the population is founded or regenerated, or if gene dispersal by pollen and seeds is restricted within a population. Silene tatarica is an endangered, perennial plant species growing along periodically disturbed riverbanks in northern Finland. We investigated the mechanism behind the microspatial genetic structure of S. tatarica in four subpopulations using amplified fragment length polymorphism markers. Spatial autocorrelation revealed clear spatial genetic structure in each subpopulation, even though the pattern diminished in older subpopulations. Parentage analysis in an isolated island subpopulation indicated a very low level of selfing and avoidance of breeding between close relatives. The mean estimated pollen dispersal distance (24.10 m; SD = 10.5) was significantly longer and the mean seed dispersal distance (9.07 m; SD = 9.23) was considerably shorter than the mean distance between the individuals (19.20 m; SD = 13.80). The estimated indirect and direct estimates of neighbourhood sizes in this subpopulation were very similar, 32.1 and 37.6, respectively. Our results suggested that the local spatial genetic structure in S. tatarica was attributed merely to the isolation-by-distance process rather than founder effect, and despite free pollen movement across population, restricted seed dispersal maintains local genetic structure in this species.     

Estimating economic values of vegetation restoration with choice experiments: a case study of an endangered species in Lake Kasumigaura, Japan

As the pace of ecosystem restoration increases, more attention has been directed towards understanding public preferences for ecosystem restoration projects. This paper utilizes a choice experiments technique to evaluate the restoration project of an endangered species, Nymphoides peltata (known locally as asaza) in Lake Kasumigaura, Japan. The restoration project evaluated in this paper consists of two attributes: an extinction risk and a lakeshore ecosystem. First, we observe a positive willingness-to-pay for the vegetation restoration project: the mean willingness-to-pay for restoring the endangered specie and avoiding extinction was about JPY6,800 and the mean willingness-to-pay for restoring the covering area to the ideal condition that the species originally inhabited in 1996 was about JPY1,600. Second, we clarify what kinds of individual characteristics affect preferences for restoration projects compared with preferences for the status quo. The results show that respondents who have strong environmental attitudes, visit experience, high income, and are female are more likely to prefer restoration projects to the status quo. Third, we investigate whether knowledge and comprehension of ecological information have any influence on respondents’ value formations. We find that respondents’ comprehension of ecological information is positively related to their willingness-to-pay for the avoidable level of extinction risk attribute, while respondents’ knowledge of ecological terms has no effect. Overall, our findings suggest that the public strongly desires the avoidance of extinction of endangered species, and indicates that researchers should be careful about how ecological information is given.

Cryptic species in an endangered pondweed community (Potamogeton, Potamogetonaceae) revealed by AFLP markers

Cryptic species are morphologically indistinguishable, yet reproductively isolated. Morphological boundaries between species can also be obscured by hybridization and clonality. Determining the roles of reproductive isolation, hybridization, and clonality in morphologically indistinguishable taxa is essential to determining appropriate species-level taxonomic rankings for conservation purposes. The taxonomic status of the endangered Little Aguja pondweed of west Texas, Potamogeton clystocarpus, is uncertain due to a lack of fixed morphological differences between it and two sympatric congeners. Morphology, amplified fragment length polymorphisms (AFLPs), and sequences of the internal transcribed spacer (ITS) region and trnL-F intron and spacer were used to determine the degree of genetic distinctiveness, hybridization and clonality for this rare species. AFLPs indicate that P. clystocarpus is a genetically distinct lineage compared to P. pusillus and P. foliosus. No hybrids involving P. clystocarpus were detected, but two putative hybrids involving P. pusillus and P. foliosus were identified. Clonal growth was only detected in P. pusillus. A combination of morphological and molecular markers was successful in determining the genetic distinctiveness of an endangered cryptic species, Potamogeton clystocarpus. Further sampling in this and adjacent drainages is necessary to assess the degree of endemism of P. clystocarpus and confidently rule out hybridization and clonality in this taxon.     

Five new species of Eimeria Schneider, 1875 from the endangered Tibetan antelope Pantholops hodgsonii (Abel) (Artiodactyla: Bovidae: Caprinae) in the Hoh Xil Nature Reserve Area of Qinghai Province,China

We examined faeces of 76 endangered Tibetan antelopes Pantholops hodgsonii (Abel) in May 2017, from the Hoh Xil Nature Reserve, Qinghai Province, China, and found 62/76 (82%) discharging oöcysts representing five new species of Eimeria Schneider, 1875. Oöcysts of Eimeria pantholopensis n. sp., found in 54/76 (71%) chiru, are subspheroidal/ellipsoidal, 15–22 × 12–19 (18.6 × 16.1) µm, with a length/width (L/W) ratio of 1.0–1.3 (1.2); micropyle cap and 1–3 polar granules are present, but oöcyst residuum is absent. Sporocysts are ovoidal, 7–11 × 4–6 (9.2 × 5.3) µm, with a L/W ratio of 1.6–2.0 (1.7); Stieda body and sporocyst residuum of small, scattered granules are present; each sporozoite contains 2 refractile bodies. Oöcysts of Eimeria wudaoliangensis n. sp. found in 52/76 (68%) chiru, are pyriform, 21–29 × 17–21 (24.9 × 19.0) µm, with a L/W ratio of 1.1–1.5 (1.3); micropyle, micropyle cap and 1–4 polar granules are present, but oöcyst residuum is absent. Sporocysts are ovoidal, 9–13 × 5–8 (11.7 × 6.7) µm, with a L/W ratio of 1.4–2.7 (1.7); Stieda body and sporocyst residuum of disbursed granules are present; sporozoites have a single large refractile body. Oöcysts of Eimeria hodgsonii n. sp. found in 20/76 (26%) chiru, are elongate-ellipsoidal, 25–32 × 18–21 (28.9 × 19.8) µm, with a L/W ratio of 1.2–1.7 (1.5); micropyle, micropyle cap and 1–3 polar granules are present, but oöcyst residuum is absent. Sporocysts are ovoidal, 11–14 × 6–7 (12.3 × 6.8) µm, with a L/W ratio of 1.7–2.1 (1.8); Stieda body and sporocyst residuum as group of large granules lying along the interface between intertwined sporozoites are present; sporozoites have 2 refractile bodies. Oöcysts of Eimeria schalleri n. sp. found in 49/76 (64.5%) chiru, are ellipsoidal, 26–36 × 19–25 (30.4 × 23.2) µm, with a L/W ratio of 1.2–1.5 (1.3); micropyle with micropyle cap and polar granules appearing as many diffuse tiny bodies are present, but oöcyst residuum is absent. Sporocysts are ovoidal, 12–16 × 7–9 (14.2 × 7.8) µm, with a L/W ratio of 1.6–2.1 (1.8); Stieda body and sporocyst residuum are present, the latter as a group of small dispersed granules between intertwined sporozoites; sporozoites with 2 refractile bodies. Oöcysts of Eimeria sui n. sp. found in 4/76 (5%) chiru, are ovoidal, 32–38 × 26–30 (36.6 × 28.6) µm, with a L/W ratio of 1.0–1.4 (1.3); micropyle and micropyle cap and 1–3 polar granules are present, but oöcyst residuum is absent. Sporocysts are ovoidal, 15–18 × 8–10 (16.7 × 8.9) µm, with a L/W ratio of 1.7–2.1 (1.9); Stieda body and sporocyst residuum are present, the latter as a group of dispersed small granules; sporozoites with 2 refractile bodies. Five of 62 faecal samples in which oöcysts were detected (8%) had a single species infection, 13 of 62 (21%) had two species, 28 of 62 (45%) had three species and 16 of 62 (26%) had four species.

     

A new species of Zagrammosoma Ashmead (Hymenoptera,Eulophidae) from Qinghai Province,China

The new species Zagrammosoma dulanense Cao & Zhu, sp. n., from Qinghai Province, China, is described and illustrated. All type specimens were reared from the pupae of Micrurapteryx sophorivora Kuznetzov & Tristan (Lepidoptera: Gracillariidae), a leafmining moth attacking the plant Thermopsis lanceolata R. Br. (Fabaceae). A key to the three known Asian species of Zagrammosoma is provided. All specimens are deposited in the Insect Collection, the Institute of Zoology, Chinese Academy of Sciences, Beijing, China.     

荒漠麻蜥的染色体核型观察

采用常规骨髓细胞制片法,对采自甘肃省民勤县的荒漠麻蜥Eremias przewalskii染色体核型进行分析。荒漠麻蜥染色体核型为2n=38=36I+2m,具18对大型端部着丝粒型和1对微小染色体,属丽斑麻蜥型,与蜥蜴科和麻蜥属的染色体核型一致。系统整理已报道的麻蜥属核型资料,在此基础上探讨麻蜥属的染色体核型特征与演化。     

Development and characterization of microsatellites in Torreya jackii (Taxaceae), an endangered species in China

? Premise of the study: Polymorphic microsatellite loci were developed in Torreya jackii, an endangered species in China, to provide markers for further studies on the genetic diversity of this species. ? Methods and Results: Eight polymorphic loci and one monomorphic locus were developed and characterized in four T. jackii populations (Xianju, Songyang, Pujiang, and Tonglu) from Zhejiang Province, China. The number of alleles per locus ranged from one to eight across 80 T. jackii individuals. At the eight polymorphic loci, the observed heterozygosity ranged from 0.150 to 1.000 and the expected heterozygosity ranged from 0.185 to 0.796. ? Conclusions: The microsatellite loci developed and characterized in this study will facilitate future analyses of the genetic diversity of T. jackii. Such information will aid in designing strategies to conserve this currently endangered species.     

极濒危植物中华水韭屯溪居群调查研究报告

中华水韭（Isoetes sinensisPalmer）为水韭科（Isoetaceae）水韭属（Isoetes）多年生沼泽植物,由于人为干扰日益严重,现分布范围急剧缩小,种群数量稀少,处于濒危状态,已被列为国家一级重点保护野生植物。为科学合理的保护和抢救这一珍稀濒危植物资源提供理论依据,为此对中华水韭屯溪居群的生长环境、面积、株丛数量、水体的pH值、伴生植物种类和生存现状等的调查研究。中华水韭屯溪居群面积500m^2,株丛密度较大,水体pH值为6.55,伴生植物有40余种,其中10种有较高优势度,生存面临多重威胁,保护迫在眉睫。     

Genetic diversity of the endangered species Kirengeshoma palmata (Saxifragaceae) in China

In order to investigate the levels of genetic diversity of the endangered species Kirengeshoma palmata (Saxifragaceae), four extant populations were sampled and analyzed using inter-simple sequence repeats (ISSR) markers. We expected a low genetic diversity level, but our results revealed a high level of intraspecific genetic diversity, probably resulting from this species being in a refuge during the last glaciation (at population level: P = 63.25%, A_e = 1.47, H_E = 0.26 and H_O = 0.37; at species level: P = 79.00%, A = 1.5538, H_T = 0.2586 and H_sp = 0.3104). A low level of genetic differentiation among populations was detected based on Nei's genetic diversity analysis (16.69%) and AMOVA (19.36%). Populations shared high levels of genetic identity. Insect pollination and seed dispersal by wind may have facilitated extensive gene flow and are likely responsible for this present structure of genetic variation.     

Notholca dongtingensis (Rotifera: Monogononta: Brachionidae), a new species from Dongting Lake,China

Notholca dongtingensis n.sp. was found in the second largest lake of China, Dongting Lake. It is related to Notholca labis Gosse, 1887 and N. kozhovi Vassilijewa & Kutikova, 1969. Its main distinguishing taxonomic features are a square-oval lorica, very short anterior spines, anterior lateral spines curving outwards and protrusile posterior margin of the ventral plate. This revised version was published online in July 2006 with corrections to the Cover Date.