线粒体控制区在鱼类种内遗传分化中的意义

线粒体DNA（mtDNA）作为分子标记已被广泛应用于各物种系统发生的研究。mtDNA控制区序列（D-Loop）以其较高的突变积累对于研究物种种内的遗传分化具有重要价值。鱼类是脊椎动物中最原始但在种属数量上又最占优势的类群,其物种繁多,分布广泛,起源复杂,研究其系统发生历来是令人饶有兴趣的课题。D-Loop在研究鱼类种内遗传分化中具有多方面的重要意义。近年来,已有越来越多的研究工作将D-Loop作为分子标记来探讨各种鱼类的种内遗传分化,并且获得了许多有启发性的结果。青海湖是我国内陆最大的咸水湖,湖中主要鱼类为青海湖裸鲤（Gymnocypris przewalskii）,D-Loop分析初步结果显示青海湖及其周围河流中的裸鲤似乎没有新的种内遗传分化现象。      

盐碱胁迫对青海湖裸鲤耗氧率、血浆渗透浓度和离子浓度的影响

采用呼吸生理和血液生理的方法,探讨了性成熟青海湖裸鲤(Gymnocypris przewalskii)对盐度、碳酸盐碱度的适应性以及青海湖裸鲤在盐碱胁迫下的渗透调节和离子调节规律。将裸鲤从淡水突变到盐水(盐度14.01±0.01)、碱水(碳酸盐碱度33.03±0.01mmol·L-1)和青海湖水(盐度12.12±0.02、碳酸盐碱度29.07±0.09mmol·L-1)中,测定了在不同水质胁迫下裸鲤耗氧率、血浆渗透压和离子浓度的变化。结果表明:与淡水对照组比较,呼吸耗氧率除了盐度组在3h有显著性升高(P<0.05),其他实验组96h内均未出现显著性变化(P>0.05);裸鲤的窒息点为0.14～0.17mg·L-1,各实验组窒息点均没有显著性差异(P>0.05);碱度组血浆的渗透浓度和Na+、Cl-、Ca2+离子浓度在96h内未出现变化(P>0.05),盐度组和青海湖水组在所测时间段有显著的升高(P<0.05),并且随着胁迫时间的延长,在72h达到峰值。各实验组血浆K+浓度变化没有血浆Na+、Cl-明显,K+浓度有增高的趋势,且都在72h达到了峰值。青海湖裸鲤具有较强的盐碱耐受性,能通过渗透调节和离子调控适应高盐碱环境,而消耗能量较少。     

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。     

The youngest split in sympatric schizothoracine fish (Cyprinidae) is shaped by ecological adaptations in a Tibetan Plateau glacier lake

Although new empirical evidence shows that sympatric speciation has occurred in some species, there are few indisputable model organisms for this process of speciation. The two subspecies ( Gymnocypris eckloni eckloni and G . e . scoliostomus ) of the schizothoracine Gymnocypris fish species complex from a small glacier lake in the Tibetan Plateau, Lake Sunmcuo, fit several of the key characteristics of the sympatric speciation model. We used combined mitochondrial control region sequences and the cytochrome b gene (1894 bp) to address the phylogenetics and population genetics of 232 specimens of G . e . eckloni and G . e . scoliostomus , as well as all of its closely related sister species. We found that: (i) a total of four old lineages were uncovered in the widespread G . e . eckloni , of which only one was shown to be shared with all G . e . scoliostomus individuals and (ii) the new subspecies ( G . e . scoliostomus ) evolved in Lake Sunmcuo from the ancestral G . e . eckloni population within approximately 0.057 Ma. These two taxa of the species complex are morphologically distinct, and reproductive isolation is further suggested. Ecological disruptive selection based on morphological traits (e.g. mouth cleft characters) and food utilization may be a mechanism of incipient speciation of two sympatric populations within Lake Sunmcuo. This study provides the first genetic evidence for sympatric speciation in the schizothoracine fish.     

青海湖裸鲤寄生舌状绦虫的空间格局研究

舌状绦虫裂头蚴只寄生在体长小于２２０ｍｍ的青海湖裸鲤中,其种群在宿主种群中呈聚集分布。其聚集分布的强度随寄生虫种群平均密度的增加而降低。由于舌状绦虫在宿主体腔生长,产生明显的空间拥挤效应,可能导致宿主死亡或被食鱼鸟类捕食而转移到终末宿主中。在体长小于１２０ｍｍ的宿主中,密度依赖的死亡过程可能是使聚集强度降低的原因;而体长１４０－２００ｍｍ的鱼中,则是非密度依赖的全死过程使聚集强度增加．     

布哈河青海湖裸鲤鱼苗鱼卵的时空分布研究

2008年7月1日到10月7日,使用水文过河缆道及圆锥网接滤过式集苗器进行采样,对青海湖布哈河鱼类早期资源的时空分布进行了研究,以期了解青海湖裸鲤的自然繁殖状况,为其资源监测及生态保护提供基础性资料。结果表明:布哈河中鱼类早期资源种类单一,主要为青海湖裸鲤,采样期间有多次苗汛,主要苗汛出现于7月份,9月上旬仅有小规模苗汛;鱼苗平均密度3.17ind.·m^-3,最高达54ind.·m^-3;晚上鱼苗密度(22:00-22:10)与下午鱼苗密度(14:00-14:10)有显著性差异(df=98,P<0.05);青海湖裸鲤的鱼卵在上层平均密度为0.075ind.·m^-3,下层平均密度0.175ind.·m^-3,两者亦存在显著性差异(df=10,P<0.05);鱼苗的漂流运动主要集中在晚上;总体上看,鱼苗主要分布在河流上层,有近岸分布的趋势,而鱼卵则主要分布在河流下层,接近河流中央位置;鱼苗断面系数为0.43,鱼卵断面系数为2.32。     

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.     

沙柳河青海湖裸鲤早期资源发生量及时空分布

为了掌握青海湖裸鲤(Gymnocypris przewalksii)资源补充状况, 于2019年5月26日至9月2日逐日在青海湖第二大支流沙柳河开展了青海湖裸鲤早期资源丰度时空变化特征的调查研究, 共采集青海湖裸鲤鱼卵3386粒, 仔鱼4690尾。调查发现, 亲鱼自5月底开始洄游, 鱼卵6月初出现, 6月底至7月初达到高峰, 随后逐渐下降, 至8月中旬基本消失。仔鱼数量自7月初呈波动式上升, 8月达到高峰, 9月初逐渐消失。经估算, 沙柳河鱼卵径流量为25.58×106粒, 仔鱼径流量为62.00×106尾。鱼卵仔鱼空间分布为从河口往上丰度依次递减, 断面水平分布为右岸>左岸>中心。Kruskal-Wallis检验表明, 昼夜鱼卵丰度存在显著差异(P<0.05); 昼夜仔鱼丰度存在极显著性差异(P<0.01), 鱼卵仔鱼漂流高峰期均集中在夜间。鱼卵丰度与流速呈显著正相关(P<0.05), 仔鱼丰度与流速呈极显著正相关(P<0.01), 与径流量日上涨率呈极显著正相关(P<0.01)。研究报道了沙柳河青海湖裸鲤早期资源现状, 填补了该水域青海湖裸鲤早期资源研究的空缺, 为青海湖裸鲤上溯亲本资源量和入湖幼鱼资源量估算提供了数据支撑, 可为青海湖裸鲤资源管理与保护提供科学依据。     

青海湖裸鲤不同年龄鉴定材料的年轮特征

通过各种年龄鉴定材料比较研究了青海湖裸鲤的年轮特征。微耳石和星耳石可用于鉴定年龄,而矢耳石易碎,不适合鉴定年龄。4种年龄鉴定材料对青海湖裸鲤年龄的判别能力为:背鳍条>微耳石>臀鳞>脊椎骨。对8龄以下个体,用背鳍条磨片鉴定年龄效果较佳;对8龄以上个体,臀鳞、背鳍条磨片和脊椎骨上的年轮计数明显低于微耳石磨片,微耳石磨片是高龄青海湖裸鲤较为可靠的年龄鉴定材料。       

黄河裸裂尻鱼群体遗传结构和Cyt b序列变异

测定了来自黄河上游和柴达木盆地托索湖的裸裂尻鱼共16个个体的Cytb基因全序列(1141bp),探讨了种群结构和遗传多样性。用MEGA2.1软件分析了碱基组成和序列变异;以青海湖裸鲤、花斑裸鲤和极边扁咽齿鱼为外类群,用PAUP*4.0b10程序构建了单倍型NJ树;用Arlequin Ver.2000程序计算了群体间遗传变异值(Fst)和Nm值以及群体分化概率值。结果显示,来自柴达木水系托索湖的裸裂尻鱼没有形成单系群,Fst=0.204(P0.05),Nm=1.95。初步判断,黄河和柴达木水系托索湖的裸裂尻鱼未显著分化,支持将柴达木裸裂尻鱼(Schizopygopsis kessleri)归并入黄河裸裂尻鱼(Schizopygopsis pylzovi)的形态学结果。两种群核苷酸多样度()分别为0.0012和0.0026,表现为较低水平。根据校正的分子钟推测,黄河和托索湖裸裂尻鱼群体分歧时间为距今7万年左右的更新世末期,结合地理分布的资料和古地质事件,对黄河裸裂尻鱼群体分布水系间的历史联系进行了分析。       

人工模拟条件下青海湖裸鲤自然繁殖环境条件需求研究

为对青海湖裸鲤(Gymnocypris przewalskii)的自然繁殖需求条件进行定量研究, 实验结合青海湖裸鲤自然产卵场原位生境调查, 通过室内人工模拟构建产卵环境诱发野生亲鱼自然繁殖, 解答可控环境中青海湖裸鲤自然繁殖的环境需求。结果表明, 青海湖裸鲤自然繁殖发生与水温、水深、流速、光照及底质因素密切相关, 其中卵石河床质较细砂等底质环境可显著提升自然交配诱导率及繁殖效果; 在合适的底质条件下, 青海湖裸鲤自然繁殖的适宜条件: 水深为0.15—0.2 m、流速为0.2—0.4 m/s、水温为10.8—14.3℃; 15D﹕9L比例的光周期有利于促进青海湖裸鲤自然繁殖的发生。在人工模拟环境中, 水深≥0.45 m、流速≥0.8 m/s、温度≥17℃或≤6℃、全光照等环境中未观测到自然繁殖活动的发生, 这些被认为是其自然繁殖环境的限制条件。研究阐明了可控环境中青海湖裸鲤野生亲鱼自发产卵繁殖的环境需求, 构建人工模拟产卵环境技术, 为自然产卵场生境调查评估、改进人工繁育模式提供技术支撑和新思路。     

6个地区青海湖裸鲤肌肉营养成分分析

采用常规生化分析方法,测定青海湖6个不同地区的青海湖裸鲤(Gymnocypris przew alskii)肌肉主要营养成分,对其营养价值进行评价。结果表明,淡水青海湖裸鲤肌肉中脂肪含量显著大于(P<0.05)其他5个地区,蛋白含量显著大于(P<0.05)黑马河青海湖裸鲤肌肉的蛋白含量,6个地区青海湖裸鲤肌肉营养成分存在一定的地区差异。总体上,青海湖裸鲤肌肉(鲜样)中粗蛋白含量为17.26%~18.87%;粗脂肪含量为1.51%~3.37%;灰分含量为1.42%~1.61%;18种氨基酸总量15.48%~17.16%,其中除色氨酸外7种人体必需氨基酸总量6.38%~7.05%,4种鲜味氨基酸总量5.82%~6.49%。青海湖裸鲤肌肉中蛋白质含量较高,氨基酸种类齐全,必需氨基酸与鲜味氨基酸含量较高,尤其是淡水青海湖裸鲤品质更佳,淡水的生态环境可能更适合青海湖裸鲤的生长。     

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.     

青海湖裸鲤自然产卵场的生境特征及无人机遥测判别——以泉吉河为例

自然产卵场是维持物种延续最关键的栖息地,青海湖裸鲤（Gymnocypris przewalskii）的自然繁殖栖息地状况和生境特征尚缺乏详细的定量研究。以青海湖入湖河流泉吉河为例,在平水期采用现场调查和无人机遥测的方法对青海湖裸鲤的自然产卵场分布及生境状况进行调查,确定其产卵场生境特征参数,建立基于无人机遥测识别产卵场的方法并进行复核验证。结果表明：泉吉河河道形态可分为弯曲型、分汊型和顺直型等三种典型河型,青海湖裸鲤自然产卵场主要分布在弯曲型和分汊型河道中,分汊型河道几乎100%都有产卵场分布,弯曲型河道有70%为产卵场;产卵场河道平均长度（135.13±61.13） m,平均宽度（30.01±17.51） m,平均曲折度1.09±0.07;平均面积（4586.6±4201.61） m²;产卵场常位于缓水浅滩处,平均水深（0.19±0.10） m （范围：0.03-0.44m）,平均流速（0.24±0.20） m/s （范围：0.01-0.81m/s）,河床质为含有沙粒的卵石（粒径：163-256mm）底质。河道形态、沙洲分布、水深特征等特征参数可作为无人机遥测识别产卵场的判断条件,并实现验证成功。研究结果可为开展整个流域的青海湖裸鲤自然产卵场现状评估及保护对策制定提供技术支撑。     

青海湖裸鲤生活史类型的研究

用模糊聚类法定量研究了青海湖裸鲤的生活史类型,以平衡产量模式进行了验证,并用世代生物量法探讨了其合理捕捞强度与捕捞年龄.结果表明：青海湖裸鲤属比较典型的k-选择类型鱼类;青海湖裸鲤平衡产量曲线与达氏鳇的平衡产量曲线非常相似;青海湖裸鲤的临界年龄为15.67龄,整个世代生物量在11～16龄达到最大;建议对青海湖裸鲤进行捕捞的最小捕捞年龄为12龄,捕捞强度不超过0.1.     

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.     

AFLP and RAPD analyses of genetic diversity of wild and/or weedy Guizotia (Asteraceae) from Ethiopia

Amplified fragment length polymorphism (AFLP) and random amplified polymorphic DNA (RAPD) markers were used to provide estimates of the comparative genetic variation within and among populations of various Guizotia taxa with the goal of conserving and utilizing their genetic diversity. The percentage of polymorphic loci (P(S)) ranged from 28.5%-90% (AFLP) and 85.6%-99.6% (RAPD). The overall gene diversity estimate () has shown slight variation among taxa ranging from 0.32-0.37 (AFLP) and from 0.22 to 0.28 (RAPD). The within population diversity of "Chelelu" and "Ketcha" was found to be unexpectedly high. Both parameters used to estimate population differentiation (G(ST) and F(ST)) revealed the highest population differentiation G. zavattarii in followed by G. arborescens. Genetic variation among populations within a taxon was highly significant for all the five taxa as revealed by AMOVA (P<0.0001). The need for immediate conservation activities for G. arborescens and G. zavattarii, and factors that contribute to the existing genetic variability and population genetic structures are discussed.     

Length–weight relationships of three endemic fish species from the upper Yellow River,China

Length–weight relationships (LWRs) of three Chinese endemic fish species (Gymnocypris eckloni eckloni Herzenstein, 1891, Platypharodon extremus Herzenstein, 1891 and Triplophysa pseudoscleroptera (Zhu & Wu, 1981)) were analyzed, collecting from the upper Yellow River (Xunhua and Guide Counties, Qinghai Province). Totally 347 specimens monthly collected by gill nets with 40 mm mesh size and traps with 5 m length from August to October 2014, were used to estimate the relationship parameters. The allometric factor b values ranged from 2.96 in T. pseudoscleroptera to 3.33 in G. eckloni eckloni. The condition factor a values varies between 0.0063 and 0.0175 in three species. Three WLRs of fishes distributed in the upper Yellow River were firstly reported and will fill in some of the empty spaces on endemic species in the Fishbase.     

A population genetics study of Anopheles darlingi (Diptera: Culicidae) from Colombia based on random amplified polymorphic DNA-polymerase chain reaction and amplified fragment lenght polymorphism markers

The genetic variation and population structure of three populations of Anopheles darlingi from Colombia were studied using random amplified polymorphic markers (RAPDs) and amplified fragment length polymorphism markers (AFLPs). Six RAPD primers produced 46 polymorphic fragments, while two AFLP primer combinations produced 197 polymorphic fragments from 71 DNA samples. Both of the evaluated genetic markers showed the presence of gene flow, suggesting that Colombian An. darlingi populations are in panmixia. Average genetic diversity, estimated from observed heterozygosity, was 0.374 (RAPD) and 0.309 (AFLP). RAPD and AFLP markers showed little evidence of geographic separation between eastern and western populations; however, the F ST values showed high gene flow between the two western populations (RAPD: F ST = 0.029; Nm: 8.5; AFLP: F ST = 0.051; Nm: 4.7). According to molecular variance analysis (AMOVA), the genetic distance between populations was significant (RAPD:phiST = 0.084; AFLP:phiST = 0.229, P < 0.001). The F ST distances and AMOVAs using AFLP loci support the differentiation of the Guyana biogeographic province population from those of the Chocó-Magdalena. In this last region, Chocó and Córdoba populations showed the highest genetic flow.     

青海湖裸鲤体腔寄生蠕虫群落研究

青海湖裸鲤体腔为三种寄生蠕虫寄生,即裂头绦虫裂头蚴,舌状绦虫裂头蚴和对盲囊缄虫的三期幼虫,这几种蠕虫不同大小的青海湖裸鲤中数量差异很大,将其看作一个组分群落,并根据其发生数量进行模糊聚类和极点排序,结果将其分为二个亚组分群落,即体长小于１５０ｍｍ的青海湖裸鲤体腔蠕虫亚组分群落和体长大于１５０ｍｍ的个体的体腔蠕虫亚组分群落。这与宿主的行为和食性改变有密切关系,舌状绦虫与另两种蠕虫间有显著负协调关系。