Genetic diversity and matrilineal structure in Chinese tree shrews inhabiting Kunming,China

Due to their special phylogenetic position in the Euarchontoglires and close affinity to primates,tree shrews have been proposed as an alternative experimental animal to primates in biomedical research.However,the population genetic structure of tree shrews has largely remained unknown and this has hindered the development of tree shrew breeding and selection.Here we sampled 80 Chinese tree shrews（Tupaia belangeri chinensis） in Kunming,China,and analyzed partial mtDNA control region sequence variation.Based on our samples and two published sequences from northern tree shrews（T.belangeri）,we identified 29 substitutions in the mtDNA control region fragment（～ 604 bp） across 82 individuals and defined 13 haplotypes.Seventeen samples were selected for sequencing of the cytochrome b（Cyt b;1134 bp） gene based on control region sequence variation and were analyzed in combination with 34 published sequences to solidify the phylogenetic pattern obtained from control region data.Overall,tree shrews from Kunming have high genetic diversity and present a remarkable long genetic distance to the two reported northern tree shrews outside China.Our results provide some caution when using tree shrews to establish animal models because of this apparent genetic difference.In addition,the high genetic diversity of Chinese tree shrews inhabiting Kunming suggests that systematic genetic investigations should be conducted before establishing an inbred strain for medical and biological research.

Sequence Difference of Mitochondrial DNA Control Region and Genetic Differentiation of Cervinae in China

[Objective] The study aimed at analyzing Genetic Differentiation of Cervinae. [Method] The complete mitochondrial DNA control region of five species of Cervinae was determined by direct DNA sequencing. Homologous sequences of others Cervinae were gained from GenBank, and then analyzed through biology software. [Result] The lengths of their control region were 921-1 072 bp, the nucleotide content of T, A, C, and G was 32.1%, 30.2%, 22.7% and 15.0% separately. Genetic distance of each species ranged from 0.062-0.106, which belonged to level of inter-genus. The molecular phylogenetic tree indicated that red deer, skia deer and White-lipped Deer had close relationship, and they constituted one branch with sambar deer. Eld＇s deer and elk deer constituted the second branch, and hog deer constituted the third branch alone. [Conclusion] Hog deer and fallow deer should be incorporated in Cervus, divergence time is about 1.55-2.60 million years among Cervinae in China.

A Molecular Phylogeny of Macaca Based on Mitochondrial Control Region Sequences

Nucleotide sequences of segments of the mitochondrial control regions were analyzed to infer the phylogenetic relationships among 7 macaques.High nucleotide diversity in Macaca assamensis and relatively low diversity in M.thibetana were found.Based on the ML tree from control regions,species in our study can roughly be sorted into three species groups except for the phylogenetic position of M.fascicularis,i.e.,silenus group,including M.leonina;sinica group,including M.arctoides,M.assamensis,and M.thibetana;and fascicularis group,including M.mulatta and M.cyclopis.A discrepancy between earlier studies (Fooden & Lanyon,1989;Tosi et al,2003a;Deinard & Smith,2001;Evans et al,1999;Hayasaka et al,1996;Morales & Melnick,1998),our result supported the hypothesis that M.fascicularis diverged earlier than M.leonina.Mitochondrial paraphyly in eastern M.mulatta (with respect to M.cyclopis) and eastern M.assamensis (with respect to M.thibetana) were clearly observed in our study.In accordance with the results of Y chromosome,allozyme,nuclear genes and some morphological data (Delson,1980;Fooden & Lanyon,1989;Fooden,1990;Tosi et al,2000,2003a,b;Deinard & Smith,2001),our study on control region sequences supported M.arctoides to be classified into the sinica group.However,this result disagreed with the previous mtDNA studies (Hayasaka et al,1996;Morales & Melnick,1998;Tosi et al,2003a).

A genetic diversity comparison between captive individuals and wild individuals of Elliot’s Pheasant (Syrmaticus ellioti) using mitochondrial DNA

Maintaining genetic diversity is a major issue in conservation biology. In this study, we demonstrate the differences of genetic diversity levels between wild and captive individuals of Elliot’s Pheasant Syrmaticus ellioti. Wild individuals showed a higher genetic diversity level than that of the captive individuals. Nucleotide diversity and haplotype diversity of wild individuals were 0.00628 and 0.993, while those of captive individuals were 0.00150 and 0.584 respectively. Only 3 haplotypes of mtDNA control region sequence were identified among 36 captive individuals, while 16 unique haplotypes were identified among the 17 wild individuals in this study. One captive haplotype was shared by a wild individual from Anhui Province. It is concluded that a low number of founders was the likely reason for the lower level genetic diversity of the captive group. Careful genetic man- agement is suggested for captive populations, particularly of such an endangered species, to maintain genetic variability levels.

The complete mitochondrial genome of the Keeled box turtle Pyxidea mouhotii and phylogenetic analysis of major turtle groups

The complete mitochondrial genome （16,837 bp） from the Keeled box turtle （Pyxidea mouhotii） was determined. The genome content, gene order, and base composition conformed to the consensus vertebrate type mtDNA. However, a remarkable feature was found in this molecule： a large number of （ATTATATC） n direct tandem repeats followed by （TA） n microsatellite at the 3＇ end of the control region （D-loop）, which might be useful as molecular markers for studying population genetics and helpful for species identification and conservation. Besides, to review phylogenetic relationships among major turtle lineages, maximum-likelihood （ML） and Bayesian （BI） analyses were conducted based on concatenated sequences of 13 protein-coding genes from 16 taxa. The resultant ML and BI analyses showed homological topologies, which only differed on the exact placement of Platysternon. Nevertheless, the results strongly supported that 1） Pyxidea mouhotii and Cuora aurocapitata formed a monophyletic clade, whereas Cyclemys atripons was not closer to the Pyxidea-Cuora than to Chinemys reevesii, suggesting that Cyclemys and the Cuora group （containing Pyxidea） may have originated from two ancestors; 2） the Geoemydidae with Testudinidae was a sister group rather than with the Emydidae.

A preliminary investigation on genetic diversity of Sousa chinensis in the Pearl River Estuary and Xiamen of Chinese waters

In this study, the mitochondrial DNA （mtDNA） control region and the mitochondrial cytochrome b gene of stranded Indo-Pacific humpback dolphin （Sousa chinensis） samples from the Pearl River Estuary and Xiamen waters were sequenced and analyzed. The result of mtDNA control region revealed 34 variable sites and four unique haplotypes （named as A, B, C and D） identified among the total samples from these two water areas, and the most common haplotype （A） was shared by 75% of the dolphins sampled from the two water areas. The haplotypic diversity （h） was 0.455 and the nucleodde diversity （π） was 0.0088. The phylogenetic analysis showed that the haplotype A, C, and D were closely related, but the haplotype B （unique for XM01 from Xiamen） was far from the other three. By scanning cytochrome b fragments, two haplotypes （A and B） were identified in these two water areas, and the most common haplotype （A） was shared by 91.67% individuals, while XM01 from Xiamen as the only exception. The data suggest that there is a possibility of gene exchange between the two populations in the Pearl River Estuary and Xiamen waters, and there possibly exists a unique maternal lineage in Xiamen waters.

Broader pattern of tandem repeats in the mitochondrial control region of Perciformes

Perciformes,the largest order of vertebrates with 20 suborders,is the most diverse fish order that dominates vertebrate ocean life.The complete mitochondrial control region(CR) of Trichiurus japonicus(Trichiuridae,Scombroidei) and Pampus sp.(Stromateidae,Stromateoidei) were amplified and sequenced.Together with data from GenBank,the tandem repeats in the mitochondrial CR from 48 species,which covered nine suborders of Perciformes,are reported in this study.The tandem repeats tend to be long in the suborder Percoidei and Stromateoidei.The identical repeats in 21 species of Cichlidae suggest a common origin and have existed before species divergence.Larimichthys crocea shows tandem repeats instead of the typical structure of the central conserved sequence blocks,which was first reported in Perciformes and vertebrates.This might have resulted from interruption of the polymerase activity during the H-strand synthesis.The four broader patterns presented here for the tandem repeats,including those in both the 5' and 3' ends,only in the either 5' or 3' end,and in the central conserved domain of the control region,will be useful for understanding the evolution of species.

Structure of Mitochondrial DNA Control Region of Pholis fangi and Its Phylogenetic Implication

In this study, the entire mitochondrial DNA(mtDNA) control region(CR) of Pholis fangi was amplified via polymerase chain reaction followed by direct sequencing. The length of the mtDNA CR consensus sequence of P. fangi was 853 bp in length. In accordance with the recognition sites as were previously reported in fish species, the mtDNA CR sequence of P. fangi can be divided into 3 domains, i.e., the extended terminal associated sequence(ETAS), the central conserved sequence block(CSB), and the CSB domain. In addition, the following structures were identified in the mtDNA CR sequence of P. fangi: 2 ETASs in the ETAS domain(TAS and cTAS), 6 CSBs in the central CSB domain(CSB-F to CSB-A), and 3 CSBs in the CSB domain(CSB-1 to CSB-3). These demonstrated that the structure of the mtDNA CR of P. fangi was substantially different from those of most other fish species. The mtDNA CR sequence of P. fangi contained one conserved region from 656 bp to 815 bp. Similar to most other fish species, P. fangi has no tandem repeat sequences in its mtDNA CR sequence. Phylogenetic analysis based on the complete mtDNA CR sequences showed that there were no genetic differences within P. fangi populations of the same geographical origin and between P. fangi populations of different geographical origins.

Population genetic structure of the mantis shrimp Oratosquilla oratoria(Crustacea:Squillidae)in the Yellow Sea and East China Sea

The mantis shrimp O ratosquilla oratoria is an ecologically and economically important species in the Western Pacific. In present study, the population genetic structure of O ratosquilla oratoria from the Yellow Sea and East China Sea was examined with mitochondrial DNA control region sequences. In total, 394 samples were collected from 18 locations and 102 haplotypes were obtained. For the Yellow Sea, the overall nucleotide diversity and haplotype diversity were 0.006 9 and 0.946 8, respectively; while across all the East China Sea locations, the overall nucleotide diversity and haplotype diversity were 0.027 94 and 0.979 0, respectively. The results of AMOVA and pairwise F_(ST)(0.145 2, P <0.001) revealed moderate differentiation between the Yellow Sea and East China Sea populations of O. oratoria. However, neither the neighbor-joining tree nor haplotype network showed clades with geographic pattern, which indicated considerable gene flow was existed between the Yellow Sea and East China Sea, and supporting the high larval dispersal ability in this species. Mismatch distribution analysis and neutrality tests suggested that O. oratoria has undergone population expansion event, and the Pleistocene glacial cycles might have an impact on the historical demography of O. oratoria. The genetic information obtained in this study can provide useful information for sustainable improvements for capture fisheries management strategies.

Mitochondrial DNA control region diversity and population structure of Pacific herring(Clupea pallasii)in the Yellow Sea and the Sea of Japan

To investigate the genetic variation and population structure of Pacific herring in the Yellow Sea and the genetic differentiation between the Yellow Sea and the Sea of Japan, fragments of 479-bp mitochondrial DNA control region were sequenced for 110 individuals collected from three different periods in the Yellow Sea and one locality in the Sea of Japan. High haplotype diversity and moderate nucleotide diversity were observed in Pacific herring. AMOVA and exact test of population differentiation showed no significant genetic differentiations among the three populations of the Yellow Sea and suggested the populations can be treated as a single panmictic stock in the Yellow Sea. However, a large and significant genetic differentiation （ФST=0.11; P=0.00） was detected between the populations in the Yellow Sea and the Sea of Japan. The high sea water temperature in the Tsushima Strait was thought a barrier to block the gene exchange between populations of the two sea areas. The neutrality tests and mismatch distribution indicated recent population expansion in Pacific herring.

Limited Heredity Diversity of the Critically Endangered Guizhou Golden Monkeys

Guizhou golden monkey(Rhinopithecus brelichi)was a unique,endangered and endemic primate species in Guizhou Province.It was an isolated population caused by habitat loss and fragmentation due to the human disturbance in the recently 30 years in China,only distributed in Fanjingshan National Natural Reserve,Guizhou Province.To know the background with demonic population structure,we sequenced 867 bp of the mitochondrial DNA D-loop from 312 fresh fecal samples,results showed there 11 haplotypes among these samples,h was 0.517,πwas 0.00413.It indicated that this species had the lowest genetic diversity among four golden monkeys in China and need strengthen the conversation concern for this species immediately.

Directional region control of the thermal fractal diffusion of a space body

We present a directional region control （DRC） model of thermal diffusion fractal growth with active heat diffusion in three-dimensional space. This model can be applied to predict the space body heat fractal growth and study its directional region control. When the nonlinear interference term and the inner heat source term are generalized functions, the relationship between the particle aggregation probability and the interference terms can be obtained using the norm theory. We can then predict the aggregation form of particles in different regions. When the nonlinear interference terms in the model are expressed as a trigonometric function and its composite function, our simulations show that the DRC method of thermal fractal diffusion is effective and has reference value for the directional control of actual fractal growth systems.

Molecular markers for identifying a new selected variety of Pacific white shrimp Litopenaeus vannamei

Selective breeding of the Pacific white shrimp Litopenaeus vannamei during the last decade has produced new varieties exhibiting high growth rates and disease resistance.However,the identification of new varieties of shrimps from their phenotypic characters is difficult.This study introduces a new approach for identifying varieties of shrimps using molecular markers of microsatellites and mitochondrial control region sequences.The method was employed to identify a new selected variety,Kehai No.1(KH-1),from three representative stocks(control group):Zhengda;Tongwei;and a stock collected from Fujian Province,which is now cultured in China's Mainland.By pooled genotyping of KH-1 and the control group,five microsatellites showing differences between KH-1 and the control group were screened out.Individual genotyping data confirmed the results from pooled genotyping.The genotyping data for the five microsatellites were applied to the assignment analysis of the KH-1 group and the control group using the partial Bayesian assignment method in GENECLASS2.By sequencing the mitochondrial control regions of individuals from the KH-1 and control group,four haplotypes were observed in the KH-1 group,whereas14 haplotypes were obtained in the control group.By combining the microsatellite assignment analysis with mitochondrial control region analysis,the average accuracy of identification of individuals in the KH-1group and control group reached 89%.The five selected microsatellite loci and mitochondrial control region sequences were highly polymorphic and could be used to distinguish new selected varieties of L.vannamei from other populations cultured in China.

Comparative Analysis of Mitochondrial Control Region Sequence from Three Flatfish Species(Pleuronectidae)

The 5’-end of the mitochondrial control region sequences of three flatfishes (Pleuronectiformes: Pleuronectidae) were amplified and sequenced. These sequences were compared with those of other three Pleuronectids species retrieved from GenBank. A phylogenetic tree was constructed based on the partial control region sequences. The results of phyloge- netic analysis are consistent with those of conventional systematics. Compared to previous studies, the structure of the 5’-end of mitochondrial control region was analyzed. The terminal associated sequence motif and its complementary motif were i- dentified at the 5’-end of the sequences. A conserved sequence block, named as CM5’d, was identified in the 5’-end of con- trol region sequences in all Pleuronectids. Another central conserved sequence block, named as CSB-F, was detected in the central conserved blocks.

Chaotic motions of the L-mode to H-mode transition model in tokamak

The chaotic dynamics of the transport equation for the L-mode to H-mode near the plasma in a tokamak is studied in detail with the Melnikov method. The transport equations represent a system with external and parametric excitation. The critical curves separating the chaotic regions and nonchaotic regions are presented for the system with periodically external excitation and linear parametric excitation, or cubic parametric excitation, respectively. The results obtained here show that there exist uncontrollable regions in which chaos always take place via heteroclinic bifurcation for the system with linear or cubic parametric excitation. Especially, there exists a controllable frequency, excited at which chaos does not occur via homoclinic bifurcation no matter how large the excitation amplitude is for the system with cubic parametric excitation. Some complicated dynamical behaviors are obtained for this class of systems.

Population Genetic Analysis of Sillago nigrofasciata (Perciformes:Sillaginidae) Along the Coast of China by Sequencing Mitochondrial DNA Control Region

Sillago nigrofasciata, a small to moderate size nearshore species, is newly found along the eastern and southern coasts of China. The present study is carried out in order to analyze the population genetics of the S. nigrofasciata. The control region sequence of mitochondrial DNA revealing 73 haplotypes were obtained from 162 individuals collected at 8 locations along the coast of China. The whole S. nigrofasciata population along the coast of China showed a low nucleotide diversity(0.012) and a high population diversity(haplotype diversity)(0.943), and all the 8 local populations showed low nucleotide diversities(0.014 – 0.001), suggesting the protective measures are effective. The haplotypes of the 8 local populations were widely distributed in haplotype network diagram and neighbor-joining phylogenetic tree, while no branch associating with sampling locations was detected. Recent gene flow analysis showed asymmetric gene exchanges among local populations. The pairwise FST values and unweighted pair-group method with arithmetic mean(UPGMA) tree revealed a certain amount of genetic difference among local populations. Moreover, analysis of molecular variance(AMOVA) reflected genetic differences between hypothetical subdivision groups. Neutral test and mismatch distribution of pairwise nucleotide suggested S. nigrofasciata may have experienced recent population expansion events. The historical geographic events associating with ice age may be the main explanation to the heterogeneity among local populations with short geographic distances, and the homogeneity among local populations with long geographic distances.

Comparative mitochondrial genome analysis of Cynoglossidae(Teleost:Pleuronectiformes)and phylogenetic implications

Generally,a teleostean group(e.g.,family or genus)owns one type or a set of similar mitochondrial gene arrangement.It is interesting,however,that four different types of gene arrangement have been found in the mitochondrial genome(mitogenome)of Cynoglossidae species.So far,the possible mechanisms of mitogenomic gene rearrangement and its potential implications have aroused widespread attention and caused lots of controversy.Here,a total of 21 Cynoglossidae mitogenomes and a newly sequenced mitogenome of Cynoglossus puncticpes(Pleuronectiformes:Cynoglossidae)were compared.The length ranges from 16417 bp to 18369 bp,which is mainly caused by the length heteroplasmy of control region(CR).Further analysis reveals that the difference of tandem repeats acts as a determining factor resulting in the length heterogeneity.Like most gene rearrangements of Cynoglossinae mitogenomes,tRNA-Gln gene encoded by the L-strand has translocated to the H-strand(Q inversion),accompanied by the translocation of CR in C.puncticpes mitogenome.The typical IQM order(tRNA-Ile-Gln-Met)changed to QIM order.Tandem duplication/random loss and mitochondrial recombination were accepted as the most possible models to account for the rearrangements in C.puncticpes mitogenome.Phylogenetic trees showed a strong correlation between the gap spacer in the rearranged QIM area and phylogeny,which provides a fresh idea for phylogenetic studies in future.

Studies on DNA-protein interactions in the upstream regulatory region of the human ε-globin gene promoter

The erythroid- and developmental stage-specific expression of the human ε-globin gene is controlled, in part,by the 5’-flanking DNA sequence of this gene. In the present study, we have used DNA-protein binding assays to identify trans-acting factors which regulate the temporal expression of the human ε-globin gene during development. Using gel mobility shift assays and DNasel footprinting assays, a nuclear protein factor (termed ε-SSF1) in the nuclear extracts from mouse haematopoietic tissues at d 11 and d 13 of gestation was identified. It could specifically bind to the positive control region (between -535 and -453bp) of the human ε-globin gene. We speculated that the E-SSF1 might be an erythroid- and developmental stage-specific activator. In addition, we found another nuclear protein factor (termed ε-R1) in the nuclear extract from mouse fetal liver at d 18 of gestation, which could strongly bind to the silencer region (between -392 and -177bp) of this gene. Therefore, we speculated that the ε-R1 might be an erythroid- and developmental stagespecific repressor. Our data suggest that both ε-SSF1 and ε-R1 might play important roles in developmental regulation of the human ε-globin gene expression during the early embryonic life. On the other hand, we observed that the binding patterns of nuclear proteins from three cell lines (K562, HEL and Raji) to these regulatory regions were partially different. These results suggest that different trans-acting factors in K562, HEL and Raji cells might be responsible for activating or silencing the human ε-globin gene in three different cell lines.

Genetic variation of the small yellow croaker(Larimichthys polyactis)inferred from mitochondrial DNA provides novel insight into the fluctuation of resources

The small yellow croaker(Larimichthys polyactis)belongs to the family Sciaenidae,which is an offshore warm fish species and widely distributed in the western Pacific.In this study,the variation of genetic diversity and genetic differentiation among L.polyactis populations was analyzed by mitochondrial DNA control region.A total of 110 polymorphic sites were checked,which defined 134 haplotypes.High level of haplotype diversity(h=0.993±0.002)was detected in the examined range.Population genetic structure analyse(analysis of molecular variance,Fst)showed there were high gene flow among L.polyactis populations.The result showed that there were relatively high genetic diversity and low genetic differentiation among the Yellow Sea and the East China Sea populations,which can be attributed to diverse habitats,wide distribution range and high mutation rate of control region.Using phylogenetic methods,coalescent analyses(neutrality tests,mismatch distribution analysis,Bayesian skyline analyses)and molecular dating interpreted in conjunction with paleoclimatic and physiographic evidence,we inferred that the genetic make-up of extant populations of L.polyactis was shaped by Pleistocene environmental impacts on the historical demography of this species.Besides,relatively constant genetic diversity and larger effective population size were detected in recent L.polyactis population.The result showed that the fishing policy certainly,such as the summer closed fishing,played a role in protecting resources of L.polyactis.This study can offer a wealth of biological novelties which indicates genetic structure of L.polyactis population and provides the foundation for resources protection and policy setting.

MITOCHONDRIAL DNA POLYMORPHISM IN CONTROL REGION FROM CHINESE YUGU POPULATION

Objective To investigate the mitochondrial DN A sequence polymorphism sites in Chinese YUGU ethnic group and to provide basic da ta used in forensic purpose. Methods Genomic DNA was extracted from the hole blood of 100 unrelated individuals of Chinese YUGU ethnic group by standard chelex-100 method. The sequence polymorphism sites was determined by PCR amplification and direct sequencing. Results 54 polymorphic sites were noted in mtDNA np16091-16418 region, and 46 haplotypes were identifi ed. The genetic diversity was calculated to be 0.9691, and the genetic identity was calculated to be 0.0406. Conclusion There are some particul ar polymorphism sites in Chinese YUGU ethnic group. The results suggest that seq uence polymorphism from np16091-16418 in human mitochondrial DNA can be used as a biological marker for forensic identity.