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.     

cDNA cloning and expression of erythropoietin in the plateau zokor （Myospalax baileyi） from the Qinghai-Tibet Plateau

All organisms respond to variation in their environments and manage environmental stress through metabolic adjustments.The plateau zokor(Myospalax baileyi) is an endemic and keystone subterranean rodent species that inhabits the Qinghai-Tibet Plateau between 2800 and 4200 m above sea level.It is a hypoxic-tolerant mammal with a high ratio of oxygen utilization that enables it to cope with its harsh surroundings.To explore the molecular mechanism of altitude acclimatization of the plateau zokor,we cloned the zokor erythropoietin(Epo) gene and used real-time PCR to compare Epo mRNA levels in zokors inhabiting 16 different altitudes.The full-length zokor Epo open reading frame was 579 bp that encoded a precursor peptide of 192 amino acids with a signal peptide of 26 residues.The Epo gene of the plateau zokor was 81%-95% homologous to that of human,mouse,rat,root vole and the Golan Heights blind mole rat,with the highest homology(95%) to species of the genus Spalax.Epo mRNA was detected mainly in the zokor kidney and spleen among 8 selected tissues.The level of Epo mRNA increased in the liver and kidney with increases in altitude.The increase in the kidney was 5 times that in the liver.Remarkably,expression of Epo mRNA in the kidney of zokors living at the highest altitude(4268 m) was 12-fold higher than that of zokors living at the lowest(2492 m) altitude.These findings provide essential information for understanding the possible role of Epo in adaptation to hypoxia in the plateau zokor.     

Phylogeny and divergence of Chinese Angiopteridaceae based on chloroplast DNA sequence data （rbcL and trnL-F）

Marattioid ferns are an ancient lineage of primitive vascular plants that first appeared in the middle Carboniferous. Extant members are almost exclusively restricted to tropical regions, and the spe- cies-rich family Angiopteridaceae are limited in their distribution to the eastern hemisphere; relation- ships within the group are currently vague. Here the phylogenetic relationship between Angiopteris Hoffm. and Archangiopteris Christ et Gies. was evaluated based on the sequence analysis of chloro- plast rbcL gene and trnL-F intergenic spacer with MEGA2 and MrBayes v3.0b4. On the basis of the phy- logenetic pattern and fossil record, we further estimated the divergence time for the two genera. The phy- logenetic trees revealed that all species of Angiopteris and Archangiopteris in this study formed a monophyletic group with strong statistical support, but the relationship between the two genera re- mained unresolved based on individual sequence analysis. On the other hand, the sequence analyses of combined data set revealed that Archangiopteris species diverged first, indicating that Archangiop- teris may not be a direct derivative as traditionally assumed. The clade of Angiopteris and Archan- giopteris appears to have diversified in the late Oligocene (≈26 Ma) based on the molecular estimate. Thus, the evolutionary history of extant Angiopteris and Archangiopteris has been characterized by ancient origin and recent diversification, and these groups are not relic and endangered lineages as traditionally considered.     

Quantitative Assessment on Endangerment Degree of Rare Animal and Plant Species in Tibet, China

It is of profound theoretical and practical significance to study endangerment status of rare species in Tibet. Index system is firstly set down for quantitative assessment of rare animal and plant species, then endangerment degree of wildlife under special state protection are calculated, which is expressed by value E. The results reveal that Yunnan snub-nosed monkey （Rhinopithecus bieti）, Himalayan tahr （Hemitragus jemlahicus） and Gigantic Cypress （Cupressus gigantea） have the highest E respectively in animals and plants. According to value E, all species are categorized into 4 ranks： critically endangered（0.6-0.8）, endangered （0.4- 0.6）, vulnerable （0.2-0.4） and lower risk （ E ≤ 0.2）. By comparison of the first five animals and plants of the highest E, each sub-ecozone bears a distinct identity.     

The role of different cues in the brain mechanism on visual spatial attention

The visual spatial attention mechanism in the brain was studied in 16 young subjects through the visual search paradigm of precue-target by the event-related potential (ERP) technique, with the attentive ranges cued by different scales of Chinese character and region cues. The results showed that the response time for Chinese character cues was much longer than that for region cues especially for small region cues. With the exterior interferences, the target stimuli recognition under region cues was much quicker than that under Chinese character cues. Compared with that under region cues, targets under Chinese character cues could lead to increase of the posterior P1, decrease of the N1 and increase of the P2. It should also be noted that the differences between region cues and Chinese character cues were affected by the interference types. Under exterior interferences, no significant difference was found between region cues and Chinese character cues; however, it was not the case under the interior interferences. Considering the difference between the exterior interferences and the interior interferences, we could conclude that with the increase of difficulty in target recognition there was obvious difference in the consumption of anterior frontal resources by target stimuli under the two kinds of cues.     

Expression and function of a new angiogenic factor AA98 target molecule at the maternal-embryonic boundary of rhesus monkey

The target molecule of monoclonal antibody AA98 (AA for short) is a new vascular endothelial cell related factor and plays a role in angiogenesis as indicated by the previous data. To investigate its role in angiogenesis and placentation in primate, we examined its expression in the implantation sites on D17, 19, 28 and 34 of gestation in rhesus monkey by immunohistochemistry and Western immunoblot. Western blot analysis showed that the primary antibody used in this study was specific for its epitope. AA protein was mainly expressed in small blood vessels and in some cytotrophoblast cells. The AA staining was found mainly in the endothelial cells and vascular small muscle. This observation supported the AA‘s role in angiogenesis. AA was spatio-temporarily expressed in cytotrophoblasts: weak in proliferating trophoblast within cell column and endovascular trophoblast, strong in trophoblastic subpopulation within the basal plate and vascular trophoblast; AA staining within the basal plate was down-regulated during early placentation. The shift of AA98 expression in extravillous trophoblasts suggestes a role of this new factor during the course of cytotrophoblast metastasis and spiral artery remodeling. The spatio-temporarily expression indicats that AA98 could be also used as a trophoblast cellular marker to characterize the acquisition of a vascular endothelial and invasive phenotype.     

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.     

Cloning, sequencing and expression analysis of cDNA encoding a constitutive heat shock protein 70 （HSC70） in Fenneropenaeus chinensis

The cDNA encoding hsc70 of Chinese shrimp Fenneropenaeus chinensis was cloned from hepatopancreas by RT-PCR based on its EST sequence. The full length cDNA of 2090 bp contained an open reading frame of 1956 nucleotides and partial 5‘- and 3‘-untranslated region(5‘- and 3‘-UTR). PCR amplification and sequencing analysis showed the existence of introns in the region of 1--547 bp, but they did not exist in the region of 548--2090 bp of hsc70 cDNA. When the deduced 652 amino acid sequence of HSC70 was compared with the members of HSP70 family from other organisms, the results showed 85.9% similarity with HSC71 from Oncorhynchus mykiss and HSC70 from Homo sapiens. It also exhibited 85.8% similarity with HSP70 from Mus musculu and 85.4% with HSC70 from Manduca sexta. Expression analysis showed that hsc70 mRNA was espressed constitutively in hepatopancreas, muscle, eyestalks, haemocytes, heart, ovary, intestine and gills in Fenneropenaeus chinensis. No difference could be detected on hsc70 mRNA level in muscle between heat-shocked and control animals.     

鲇和大口鲇同工酶比较研究

采用聚丙烯酰胺垂直平板凝胶电泳技术，对鲇和大口鲇的肌肉、心脏、肝脏、肾脏、脑、眼等6种组织器官乳酸脱氢酶（LDH）和酯酶（EST）同工酶进行了比较研究。结果表明：2种鲇的同种组织中乳酸脱氢酶（LDH）同工酶的酶谱相同，但在不同组织中表现出明显的组织特异性；而2种鲇的酯酶（EST）同工酶谱表现出明显的组织特异性和物种特异性。笔者还对2种鲇的亲缘关系进行了探讨。     

四川泸沽湖湿地冬季水鸟调查研究

鸟类多样性是生物多样性的重要组成部分,研究泸沽湖湿地的鸟类多样性对湿地生态环境保护具有重要意义。2014—2016年冬季,对四川泸沽湖湿地水鸟进行了全面调查和系统整理,结果显示:共记录水鸟42种,隶属6目11科,其中,留鸟10种,冬候鸟32种;东洋界6种,古北界34种,广布种2种;国家Ⅰ类保护鸟类2种,占物种总数的4.76%;国家Ⅱ类保护鸟类2种,占物种总数的4.76%,四川省保护鸟类5种,占11.90%;极危(CR)物种1种,近危(NT)物种2种,低危(LC)物种34种。根据记录到的水鸟种类和数量,对其群落特征及其季节变化进行了分析,同时结合泸沽湖湿地的特点,对四川泸沽湖湿地水鸟的组成、种群数量及多样性等进行了分析,发现四川泸沽湖湿地水鸟多样性呈下降趋势,主要原因为人为干扰过大。     

中华鲟囊胚细胞的长期保存及其克隆胚胎发育观察

为保存濒危鱼类中华鲟种质资源,在0、-20、-80、-196℃4种不同的温度下对其囊胚细胞进行了长期保存试验,细胞存活期分别为10d、30d和2年,在液氮中(-196℃)保存2年后仍有60.7%存活率;用冷冻复苏后的细胞进行核移植,并对克隆胚胎的发育进行了观察.共移植417枚卵,获得2尾克隆幼鲟,成功率为0.48%,表明长期冷冻保存的囊胚细胞仍具有发育全能性,通过克隆技术能获得存活个体.此项技术为濒危物种的保护开辟了新途径.     

刺猬和三种游蛇科动物冬眠期和活动期乳酸脱氢酶同工酶的比较

采用聚丙烯酰胺凝胶电泳技术，对刺猬（Erinaceus dealbatus）及游蛇科的黑眉锦蛇（Elaphe taeniura）、红点锦蛇（E.rufodorsata）和赤链蛇（Dinodon rufozonatum）等在冬眠期和活动期骨骼肌、心肌、肝等的乳酸脱氢酶（lactate dehydrogenase,LDH）同工酶谱进行了分析，并用比色法测定了以上各种组织的LDH活性。结果表明：LDH同工酶的电泳图谱只有红点锦蛇的肝和骨骼肌中存在活动期和冬眠期的明显差异，在其余动物两个时期的组织中未表现出明显变化；刺猬的肝脏LDH活性在冬眠期显著地高于活动期，而其余的动物组织冬眠期LDH活性均显著低于活动期；三种蛇类组织LDH4均未见表达。     

基于微卫星的喀斯特地区长梗吊石苣苔和吊石苣苔的遗传多样性研究

长梗吊石苣苔Lysionotus longipedunculatus (W.T. Wang) W.T. Wang是我国吊石苣苔属中的一个比较罕见的物种,目前濒危等级被评估为极危CR Blac (ⅰ,ⅱ,ⅴ),而同属的吊石苣苔L. pauciflorus Maxim.分布范围非常广泛、生境变化多样,濒危等级为无危LC。本研究通过筛选出16个微卫星位点对长梗吊石苣苔的两个居群进行遗传多样性研究,并对比相近地区的吊石苣苔居群,比较研究导致长梗吊石苣苔濒危的原因。结果显示二者的遗传多样性都比较低,有较高自交现象,但吊石苣苔具有明显的走茎,可以有效地克隆繁殖,相较之下,长梗吊石苣苔较弱的克隆繁殖能力可能是导致其濒危的原因之一。     

濒危植物翅果油树的研究进展及其开发前景

翅果油树为我国特有的濒危植物,已列为国家二级重点保护树种。本文比较系统地综述了翅果油树的种质资源、生态学特性、植物生物学和生物化学以及资源开发和应用前景。同时对翅果油树濒危的可能原因进行了分析,并提出了保护措施。