中国扬子鳄保护存在的主要问题及管理对策研究

在调查分析扬子鳄野生及饲养种群保护现状的基础上,提出了当前中国扬子鳄保护存在的主要问题:野生扬子鳄栖息地岛屿化严重及自然保护区管理体制的缺陷;饲养种群基础设施规模的局限及没有建立完整谱系资料等。同时提出了相应的管理对策:改变保护区的管理体制,恢复扬子鳄丘陵人工湿地型栖息地及建立扬子鳄成片滩涂型湿地栖息地;加大对宣城扬子鳄繁殖研究中心基础设施建设的投入,加快饲养种群的遗传谱系建设,开展扬子鳄种群生存力分析及有效种群大小评估。     

淮河乌鳢线粒体DNA控制区结构分析及遗传多样性研究

利用线粒体DNA控制区(D-loop)序列分析淮河淮滨段、凤台段、蚌埠段、洪泽湖的野生乌鳢(Ophicephalus argus)种群遗传结构及种群历史.结果表明,在790bp的同源序列中,4个种群共检测到变异位点22个,占全部序列的2.78%,84个个体共检测到33种单倍型;4个种群的平均单倍型多样性(h)、核苷酸多样性(Pi)分别为0.956 8、0.0038,表明淮河野生乌鳢种群的遗传多样性水平较高.4个种群间的遗传分化指数Fst为0.046 0,仅3.10%的变异来自种群间(AMOVA分析),基因交流值为10.3696,种群间K2-P遗传距离为0.003～0.005,从而显示乌鳢种群间没有发生明显的地理分化.NJ树揭示4个种群的个体组成2个谱系,但这2个谱系与地理分布并不相关.中性检验、错配分析和Network网络亲缘关系分析皆表明乌鳢鱼种群有过种群扩张,扩张时间约在末次冰期早期,距今51.8ka BP～74.6ka BP.     

扬子鳄DMRT1基因DM保守区的克隆及序列分析

DMRT1基因是DMRT基因家族的一个重要成员,在动物性别决定过程中发挥重要作用.本文参照人DMRT1基因DM保守区及有关文献,设计了一对简并引物,扩增了扬子鳄的DMRT1基因,并对扩增产物进行了亚克隆与测序.结果在雌雄扬子鳄个体中各获得一个预期的基因片段,长度为140bp,序列无性别差异性,命名为扬子鳄AsDMRT1基因.序列同源性分析表明,扬子鳄DMRT1基因DM盒区核苷酸序列与人和鼠相应基因的同源性分别为87%、86%;编码的氨基酸序列与人和鼠相应基因编码序列的同源性分别为95%、97%.充分显示了DMRT1基因在进化上的高度保守性.     

扬子鳄卵的人工孵化及幼鳄饲养与越冬的初步研究

关于扬子鳄(Alligatot Sinensis)卵的人工孵化及幼鳄饲养与越冬的问题,国内以往未曾见到有这方面的资料报导。近年来扬子鳄种群数量日益减少,主要原因之一,是产卵季节母鳄产下的卵常遭产区群众及小孩破坏(捣毁巢卵),致使种群得不到补充之故。为了探讨对鳄卵进行人工孵化及幼鳄饲养与越冬问题,我们曾于1976年8月至1979年7月,先后在皖南宣城采集二窝鳄卵,在室内做了人工孵化及幼鳄饲养与越冬的实验,均已获得成功。这对今后进行扬子鳄的人工繁殖,以及对其种群数量的恢复和发展,均具有一定的现实意义。此外,对动物志的编写工作,也有一定的参考价值。现将初步结果报告如下,仅供参考。     

帘蛤科两种经济贝类种群的ITS-1序列遗传多样性分析

利用核糖体DNA转录间隔区ITS-1序列对硬壳蛤(Mercenaria mercenaria)的养殖群体和青蛤(Cyclinasinensis)的野生种群遗传多样性进行了分析.经过PCR扩增、连接与测序后,得到硬壳蛤ITS-1的长度为718～724 bp,青蛤的为665～683 bp,序列用ClustalX1.83多重比对后,在725 bp组成的同源片段中有714个碱基序列为保守位点,通过DNAsp软件分析,该种群的核酸多态性指数Pi为0.00302,每位点Eta为0.00304,并检测出4个单倍型(Haplotype)序列.青蛤在由695 bp组成的同源片段中有644个碱基序列为保守位点,该种群的Pi为0.03071,每位点Eta为0.0378,共测出6个单倍型(Haplotype)序列.利用PAUP4.10软件计算出每个种群内单倍型序列间的相对遗传距离,其中,硬壳蛤的遗传距离仅为0.0014～0.0056,说明其DNA遗传多样性的丰度较低;而野生青蛤种群的遗传距离为0.0036～0.0105,说明其遗传多样性比较丰富.文章还初步讨论了双壳类遗传多样性变动的因素及种质保护等问题.     

浙江建立两个保护扬子鳄基地

扬子鳄是我国特产的珍稀动物,是目前生活在地球上的鳄类中的“临危种”.1973年联合国把它列为世界保护和禁运动物. 扬子鳄分布的面积越来越小,目前仅生活在安徽、浙江、江苏三省交界的部分水域地区.为了挽救这种动物,浙江省建立了两个扬子鳄保护饲养基地:一个在吉安县灵峰寺,一个在长兴县管埭公社尹家大队.这一带河流纵横交错,大小鱼塘星     

几种鹤类线粒体DNA的差异和亲缘关系分析

采用聚合酶链式反应,从丹顶鹤(G.joponensis♂)、白枕鹤(G.vipio♀)、灰鹤(G.grus)及丹顶鹤♂和白枕鹤♀的4个杂交子代中分别扩增出线粒体DNA细胞色素b基因,并测定出细胞色素b碱基序列,它们之间的序列差异在0.1%～7.2%之间.序列分析表明几种鹤类细胞色素b基因序列的碱基变化主要是转换和颠换,转换的比率大于颠换;绘制几种鹤类细胞色素b之间的亲缘关系进化树表明丹顶鹤和灰鹤的进化距离更近,4个杂交子代与母本白枕鹤的亲缘关系更近.     

陕西秦巴山区中华蜜蜂线粒体DNA的遗传多样性

为研究陕西秦巴山区中华蜜蜂遗传多样性,对陕西秦巴山区20个地理种群的200群中华蜜蜂线粒体DNA tRNAleu~COⅡ基因片段进行测序分析。结果显示:线粒体DNA基因片段长度为349bp,其中包括90bp非编码区和259bp的COⅡ部分序列,A+T含量大于85%,表现出A+T偏倚性;共检测到25个变异位点和29种单倍型,单倍型多样度为0.680±0.037,核苷酸多样度为0.002 94±0.000 25;聚类分析发现陕西秦巴山区中华蜜蜂各地理种群间未发生明显遗传分化,与北京、吉林、甘肃等地及日本东方蜜蜂亲缘关系也较近,而与印度东方蜜蜂亲缘关系较远。     

桃抗病基因同源序列的克隆与分析

根据已克隆的STK类抗病基因的保守区设计简并引物对4个不同桃的基因组DNA进行PCR扩增,将获得的扩增片段的回收产物,应用pGEM-T easy裁体试剂盒进行了目的基因片段的克隆.随机挑取若干单克隆经PCR鉴定确认后,进行测序,共获得11个各不相同的片段序列.氨基酸序列分析中,有6个片段能推导出完整的氨基酸序列.除了两侧都基本具有STK类抗病基因产物的保守结构域.     

疣尾蜥虎线粒体基因组全序列及其基因组成

测定了疣尾蜥虎(Hemidactylus frenatus) 的线粒体基因组全序列.序列全长为16 891 bp, 包括13 个蛋白质编码基因、2 个rRNA 基因和22 个tRNA 基因.除大部分基因由重链编码外,仅1个蛋白编码基因和8个tRNA基因由轻链编码.碱基组成的偏好与其他脊椎动物线粒体DNA接近.没有发现长的基因间隔区,说明该基因组的结构十分紧凑.基因组的组成与典型脊椎动物的相近,即没有发现重排、基因或控制区的重复等在其它有鳞类动物中出现过的异常特征.除单性生殖的壁虎外,现有的壁虎类线粒体基因组在基因含量和顺序上是一致的.作为蜥虎属线粒体基因组全序列的惟一代表,该序列有望在有鳞类系统发生的推断上发挥一定的作用.     

Testing reintroduction as a conservation strategy for the critically endangered Chinese alligator: Movements and home range of released captive individuals

The Chinese alligator (Alligator sinensis) is considered the most critically endangered crocodilian as a result of the near total loss of its habitat and its extremely small and fragmented wild populations. Plans for population recovery lie mostly with wetland res- toration and the reintroduction of captive-reared animals. We carried out a first-trial release of 3 adult Chinese alligators (1♂, 2♀) into a pond at the Hongxing conservation site, Xuancheng, southern Anhui Province; the animals were radio-tracked from May to October in 2003. We hypothesized that after a period of adaptation, the alligators would establish definable home ranges. Two (1♂, 1♀) of the 3 alligators were monitored for the whole of the tracking period. The male had an annual home-range size of 7.61 hm2, and the female 4.00 hm2. Water temperature and pond water level were two important factors influencing the alligators’ distributions, and daily movements. The radio-tracked alligators had overlapping home ranges, which notably included the one substantial island in the pond; that island is the only known nesting site of the local native wild alligators. Aggressive interactions between the released alligators and native wild alligators were observed during the breeding season around this island. All the three reintroduced alligators survived the winter of 2003 and were alive in the same pond in 2008. We concluded that the Hongxing conservation site provided a suitable habitat for the reintroduced alligators. However, the low water level in the pond resulting from farmland irrigation in August and September can be a substantial threat to the alligators’ survival. Therefore, regu- lations on irrigation in summer and autumn are needed to balance the water needs of the alligators and agriculture.     

影响野生扬子鳄生存的环境因素分析

通过对被保护物种生存环境的分析是探知该物种灭绝和种群数量下降的重要手段．2002年9月间，作者在参加南陵县野生扬子鳄栖息地生境的考察工作的基础上，以安徽省南陵县野生扬子鳄生存环境作为对象，对野生扬子鳄的生存的环境因素进行分析评价．结果表明，影响野生扬子鳄生存环境质量的主要因素为食物、隐蔽条件和水体以及人的活动．在安徽省南陵县二个官方指定的保护点中长乐点尽管食物、隐蔽条件和水三大生境基本要素都较为理想，但因为没有足够的栖息地因此这里并不适合野生扬子鳄种群的生存．比较而言，如对楂林点野生扬子鳄的生存环境进行一定的人工修饰，此地应该有着较为合适的生存环境．     

野生扬子鳄栖息地土壤金属元素含量初步研究

2002年9—11月，作者在皖南地区对扬子鳄栖息地进行了质量评估，主要是对现有野生鳄的栖息地以及曾经有鳄分布的栖息地共20样点土壤中的Cd、Pb、Zn、Cu、Fe、Ca、Mg等7种金属元素含量进行测定，对重金属元素的数据分析，采用我国土壤元素背景值作为评价标准，每个样点的污染程度以多因子评价指数为指标，发现曾经有鳄分布的样地土壤中的重金属元素含量较其它样地的重金属元素含量高，野生扬子鳄对于栖息地的无机环境是有一定的依赖性的，人类的生产、生活不但影响了野生扬子鳄的生物环境，也影响和改变了野生扬子鳄的无机环境。     

Complete mitochondrial DNA sequence of Chinese alligator,Alligator sinensis, and phylogeny of crocodiles

The 16746-neueleotide (nt) sequence of mitochondrial DNA(mtDNA) of Chinese alligator,Alligator sinensis,was determined using the Long-PCR and primer walking methods.As is typical in vertebrates,the mtDNA encodes 13 proteins,2 rDNA,22 tRNA genes,and a noncoding control region.The Composition of bases is respectively 29.43%A,24.59%T,14.86%G,31.12% C.The gene arrangement differs from the common vertebrate gene arrangement,but is similar to that of other crocodiles,DNA sequence data from 12S rRNA,168 rRNA,protein-coding genes and combined sequence data were used to reconstruct the phylogeny of reptiles with the MP and ML methods.With this large data set and an appropriate range of outgroup taxa,the authors demonstrate that Chinese alligator is most closely related to American alligator among three crocodilian species,which suppors the traditional viewpoint.According to the branch lengths of ML tree from the combined data set,the primary divergence between Alligator and Caiman genus was dated at about 74.9 Ma,the split between Chinese alligator and American alligator was dated at 50.9Ma.     

扬子鳄（Alligator sinensis）眼球形态学参数的测量

本文对11条扬子鳄的眼球的各种形态学参数进行了测量。结果表明,扬子鳄眼球形状呈稍扁圆球形;眼球大小与其它鳄类相似,且随年龄增长而增长;角膜在眼球的覆盖面比昼行哺乳类大,比水生哺乳类小;晶状体与角膜比值较小,这表明扬子鳄眼球聚焦能力较差;另外,角膜、巩膜和晶状体的生长关系表明,角膜轴径比巩膜轴径生长快,角膜屈光度随年龄增长而加大,因而成鳄较幼鳄近视。     

Establishment and cryopreservation of liver, heart and muscle cell lines derived from the Chinese alligator （Alligator sinensis）

The Chinese alligator, Alligator sinensis, is a critically endangered species. A conservation project of gene resources for an endangered species first involves the preservation of organs, tissues, gametes, genomic DNA libraries and cell lines. The present study is the first to establish and cryopreserve cell lines of liver, heart and muscle tissues from the Chinese alligator. The study revealed that there was a large discrepancy in cell migration time in primary cultures among liver (11-12 d), heart (13-14 d) and muscle (17-18 d) tissue pieces. The differences in time in primary cell culture suggested that it was relatively easy to build visceral-derived cell lines for reptiles. Biological analysis showed that the population doubling time for thawed cells was approxi- mately 36 h. Karyotyping revealed that the frequency of Chinese alligator cells showing chromosome number as 2n=32 was 88.6%-93.4%. Chinese alligator cell lines established here provide a vital resource for research and are likely to be useful for protection of this rare and critically endangered species. Furthermore, the establishment of these methods may supply technical and theoretical support for preserving genetic resources at the cellular level for other reptile species.     

The taxonomic status ofMoschus moschiferus anhuiensis

Since the musk deer distributed in the Dabie Mountains, Anhui was defined asMoschus moschiferus anhuiensis, its status has been disputed based on researches of morphology and ecology. This study further probed into its taxonomic status with principal components analysis (PCA) on skull measurements and mtDNA cytochrome b sequences (367 bp) analysis so as to clear its status. It is concluded that the figure and skull in Anhui musk deer are different from other species, and DNA divergence between it and other species is 6.24%–7.90%, which belongs to inter-specific difference. Thus, the study defines this musk deer as one distinct species,Moschus anhuiensis (Wang, Huet al.)     

MHC II DRB variation and trans-species polymorphism in the golden snub-nosed monkey (Rhinopithecus roxellana)

Genetic variation is generally believed to be important in studying endangered species’ adaptive potential.Early studies assessed genetic diversity using nearly neutral markers,such as microsatellite loci and mitochondrial DNA(mtDNA),which are very informative for phylogenetic and phylogeographic reconstructions.However,the variation at these loci cannot provide direct information on selective processes involving the interaction of individuals with their environment,or on the capability to resist continuously evolving pathogens and parasites.The importance of genetic diversity at informative adaptive markers,such as major histocompatibility complex(MHC) genes,is increasingly being realized,especially in endangered,isolated species.Small population size and isolation make the golden snub-nosed monkey(Rhinopithecus roxellana) particularly susceptible to genetic variation losses through inbreeding and restricted gene flow.In this study,we compared the genetic variation and population structure of microsatellites,mtDNA,and the most relevant adaptive region of the MHC II-DRB genes in the golden snub-nosed monkey.We examined three Chinese R.roxellana populations and found the same variation patterns in all gene regions,with the population from Shennongjia population,Hubei Province,showing the lowest polymorphism among three populations.Genetic drift that outweighed balancing selection and the founder effect in these populations may explain the similar genetic variation pattern found in these neutral and adaptive genes.     

扬子鳄肌肉组织成份测试和营养评价

古医籍早有记载，鳄肉可食。根据江苏省理化测试中心和南京大学现代化分析中心的测定，本文在国内首次发表了鳄肉中所含脂肪、蛋白质、氨基酸、维生素、宏量及微量元素的全部资料，说明鳄肉营养丰富，胜过猪、牛、羊、鸡、鸭肉，并具有海洋生物特有的ＤＨＡ（脑黄金）。对进一步研究怀开发我省特产扬子鳄有重要意义。     

扬子鳄幼鳄身体量度间的回归参数分析

扬子鳄头部和躯干随年龄增加按一定比速增长.据目前研究,躯干和头部长度之间具有较显著的相关性.扬子鳄幼体的4个身体量度数据和14个头部形态学参数被测量和记录下来.头部量度与身体量度间的数学关系通过回归分析提取出来.体重值采取log转化,再与体长进行回归分析,得到了直线方程.回归方程为由头部参数来估计出扬子鳄幼体体长与体重等数据提供了可行的途径.