Phylogeographic structuring and volant mammals: the case of the pallid bat (Antrozous pallidus)

Aim To examine the phylogeographic pattern of a volant mammal at the continental scale. The pallid bat (Antrozous pallidus) was chosen because it ranges across a zone of well‐studied biotic assemblages, namely the warm deserts of North America. Location The western half of North America, with sites in Mexico, the United States, and Canada. Methods PCR amplification and sequencing of the mitochondrial control region was performed on 194 pallid bats from 36 localities. Additional sequences at the cytochrome‐b locus were generated for representatives of each control‐region haplotype. modeltest was used to determine the best set of parameters to describe each data set, which were incorporated into analyses using paup *. Statistical parsimony and measurements of population differentiation (amova , F_ST) were also used to examine patterns of genetic diversity in pallid bats. Results We detected three major lineages in the mitochondrial DNA of pallid bats collected across the species range. These three major clades have completely non‐overlapping geographic ranges. Only 6 of 80 control‐region haplotypes were found at more than a single locality, and sequences at the more conserved cytochrome‐b locus revealed 37 haplotypes. Statistical parsimony generated three unlinked networks that correspond exactly to clades defined by the distance‐based analysis. On average there was c. 2% divergence for the combined mitochondrial sequences within each of the three major clades and c. 7% divergence between each pair of clades. Molecular clocks date divergence between the major clades at more than one million years, on average, using the faster rates, and at more than three million years using more conservative rates of evolution. Main conclusions Divergent haplotypic lineages with allopatric distributions suggest that the pallid bat has responded to evolutionary pressures in a manner consistent with other taxa of the American southwest. These results extend the conclusions of earlier studies that found the genetic structuring of populations of some bat species to show that a widespread volant species may comprise a set of geographically replacing monophyletic lineages. Haplotypes were usually restricted to single localities, and the clade showing geographic affinities to the Sonoran Desert contained greater diversity than did clades to the east and west. While faster molecular clocks would allow for glacial cycles of the Pleistocene as plausible agents of diversification of pallid bats, evidence from co‐distributed taxa suggests support for older events being responsible for the initial divergence among clades.     

The effects of Quaternary glaciations in Patagonia as evidenced by chloroplast DNA phylogeography of Southern beech Nothofagus obliqua

Climatic oscillations during the Quaternary strongly affected the distribution of warm-temperate tree species, which experienced local restrictions into ice-free areas and posterior expansions. To evaluate the impact of these range movements on the genetic structure of populations, we performed a phylogeographical analysis of the species Nothofagus obliqua with chloroplast DNA markers. A total of 27 populations covering the whole natural distribution range were analyzed using polymerase chain reaction-restriction fragment length polymorphism. Diversity (h _T, h _S), allelic richness (r _g), and differentiation among populations for unordered (G _ST) and ordered alleles (N _ST) were calculated. The relationships among haplotypes were evaluated by the construction of a minimum spanning network. The spatial distribution of the genetic variation was analyzed through a Mantel test and with a nested analysis of molecular variance to differentiate between geographic regions. The screening of 11 non-coding regions allowed the identification of 14 haplotypes. A high genetic differentiation was detected (N _ST = 0.875 and G _ST = 0.824) with the existence of phylogeographic structure (p < 0.05). The distribution of the genetic variation was partially explained by the topography of the region when the populations were divided longitudinally into Coastal Mountains, Central Valley, and Andes Mountains (ϕ _RT = 0.093, p = 0.001). In agreement with pollen records, our results support the hypothesis of Coastal refuges since the region harbors high diversity together with older and private haplotypes. Long-lasting persistence of some Coastal populations without contribution to re-colonization is proposed. Additional refuges are also postulated along the Andes and Longitudinal Valley. Survival in multiple glacial refuges is discussed together with the possible migratory routes.     

Maintenance of genetic diversity in Cordia africana Lam., a declining forest tree species in Ethiopia

We assessed genetic variation in a total of 22 populations of the tree species Cordia africana Lam. in Ethiopia and analyzed the country-wide impact of population history, forest disturbance and alteration of land use on extant intraspecific diversity. Amplified Fragment Length Polymorphisms (AFLPs) and chloroplast microsatellite markers were investigated. The analyses of the AFLP data revealed high diversity in all investigated populations: the percentage of polymorphic loci (PPL) ranged from 62.2% to 92.2% and Nei's gene diversity from 0.220 to 0.320 within the populations. The mean PPL and the mean diversity within populations were 85.7% and 0.287, respectively. The analysis of molecular variance revealed a moderate level of differentiation (Φ_ST = 0.07, p < 0.001) among the populations. The Mantel test proved a significant but low correlation (r = 0.31, p < 0.001) between the geographic distance and the genetic differentiation matrices. Only three different cpDNA haplotypes were observed; no more than two haplotypes were found in any population. The dominant haplotype with an overall frequency of 81% was observed in all populations. The level of differentiation among the populations was comparatively low at chloroplast DNA (G _ST = 0.18, R _ST, N _ST = 0.22). The observed patterns and levels of genetic variation within and among the populations indicate that efficient gene flow via pollen and seed is likely to be the main factor contributing to the maintenance of genetic diversity in natural and disturbed conditions.     

Genetic diversity in the mtDNA control region and population structure in the small yellow croaker <Emphasis Type="Italic">Larimichthys polyactis</Emphasis>

The genetic diversity and population genetic structure of the small yellow croaker (Larimichthys polyactis) were investigated. One hundred and fourteen individuals were sampled from 8 localities of the Yellow Sea and the northern East China Sea. Genetic variation in DNA sequences were examined from the first hypervariable region (HVR-1) of the mitochondrial DNA control region. High levels of haplotype diversity (h = 0.98 ± 0.87%) in the HVR-1 region were detected, indicating a high level of genetic diverstiy. A total of 84 polymorphic sites were found, and 87 haplotypes were defined. The pairwise nucleotide differences between samples ranged from 3.83 ± 2.19 to 6.56 ± 3.25. The demographic history of L. polyactis was examined by using neutrality tests and mismatch distribution analysis, which indicated a Pleistocene population expansion at about 49,300–197,000 years. The star burst structure of the minimum spanning tree also suggestted a very recent origin for most haplotypes. Hierarchical molecular variance analysis (AMOVA) and conventional population Fst comparisons revealed no significant genetic structure throughout the examined range, which is inconsistent with previous findings based on the morphological and ecological studies. Long-term dispersal and high gene flow likely have contributed to the genetically homogeneous population structure of the species. The knowledge on genetic diversity and genetic structure will be crucial to establish appropriate fishery management stocks for the species.     

Population genetic structure and evolutionary history of grass carp <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis> in the Yangtze River,China

Ample studies have been conducted to investigate the population genetic structure of grass carp Ctenopharyngodon idella in the Yangtze River, China. However, samples from the upper reaches were not included. In this study, we collected samples from the entire river, including three locations in the upper reaches: Yibin, Banan and Yunyang, two locations in the middle reaches: Shishou and Ruichang and one location in the lower reaches: Hanjiang, and sequenced three mitochondrial coding genes (ND5, ND6 and Cytb) and one control region (i.e., the D-loop). Nineteen haplotypes were observed in grass carp of the Yangtze River through the analysis of combined sequence data sets (around 4428 bp). Haplotype diversity indices (0.6000 ∼ 0.9333) and nucleotide diversity indices (0.0002 ∼ 0.0020) demonstrated low genetic diversity in the Yangtze grass carp. The analysis of molecular variance and the fixation index (F _ST = 0.0202) revealed insignificant genetic difference between samples from different reaches. Two monophyletic lineages of haplotypes were identified, with the lineage A experiencing potential expansion events. Along with previous findings, this study provides a better understanding of genetic diversity and variation of grass carp in the Yangtze River and will be served as an important baseline to evaluate the long-term impact of the Three Gorges Dam and other hydroelectric facilities on fish biodiversity.     

Comparative phylogeography of bagrid catfishes in Taiwan

A phylogeographic analysis of two bagrid catfishes in Taiwan was conducted using sequence data from a portion of the mitochondrial DNA (mtDNA) control region. For Pseudobagrus brevianalis, which is most probably endemic to Taiwan, a total of eight haplotypes were detected in 189 specimens from nine river systems covering its entire distribution range, from northern to central western areas of the island. Obvious genetic differentiation was observed among its populations (average F _ST = 0.753); in particular, the northernmost Tamsui River population was fixed for a single endemic haplotype. Nested clade phylogeographic analysis (NCPA) indicated that the dispersal center of mtDNA was the area around the Touchien River and Holong River, north to the Miaoli Plateau, in northwestern Taiwan, suggesting both northward and southward dispersal in this species. There was no evidence for the validity of P. taiwanensis, the nominal species described from Taiwan, morphologically similar to P. brevianalis. We confirmed that P. adiposalis was distributed discontinuously in three river systems; analysis of 42 specimens from the rivers indicated a total of four haplotypes and population differentiation (average F _ST = 0.876). Fixation into a largely differentiated haplotype in the northernmost Tamsui River population was also found in this species, but different processes for this phylogeographic pattern were implied for the two species. Comparison with P. ussuriensis, a widespread continental species morphologically similar to P. adiposalis, suggested the possibility that P. adiposalis is a group of local populations of P. ussuriensis in Taiwan. Two migration routes vs. random fixation scenarios for the population structure of P. adiposalis are discussed with information on other fishes and the geological history of the island.     

Possible female philopatry of the smooth hammerhead shark Sphyrna zygaena revealed by genetic structure patterns

We assessed the spatial pattern of genetic structure of smooth hammerhead shark Sphyrna zygaena in 10 localities from the Northern Mexican Pacific. A total of 35 haplotypes were identified in 129 sequences of the mtDNA control region. The results showed slight but significant genetic structure among localities (Φ_ST = 0.044, P < 0.001). In addition, the localities with highest number of juveniles were genetically different (Φ_ST = 0.058, P < 0.024), which may be representative of nursery areas. The genetic differentiation pattern can be associated to female philopatry and preference for particular birthing sites. Finally, historical demography shows that S. zygaena populations present a recent demographic expansion that occurred during glacial events in the late Pleistocene to early Holocene.     

Mitochondrial DNA reveals low population differentiation in elongate loach, <Emphasis Type="Italic">Leptobotia elongata</Emphasis> (Bleeker): implications for conservation

Elongate loach (Leptobotia elongata (Bleeker)), an endemic fish species to China, is a famous ornamental freshwater fish. Here, a comparative study of mtDNA control region (D-loop) (835 bp) sequences was performed to analyze its wild population structure and evaluate the genetic diversity for 110 individuals from five locations in the upper reaches of the Yangtze River, China. A total of 49 polymorphic sites and 45 haplotypes yielded high haplotype diversity (h = 0.952), but low nucleotide diversity (π = 0.00454) as that of many fish species. Sequence divergences between haplotypes ranged from 0.0033 ± 0.0011 to 0.0050 ± 0.0012 in intra-groups, and from 0.0037 ± 0.0.0011 to 0.0050 ± 0.0012 between groups. Significant values of Tajima’s D (−1.86383, P < 0.01) and Fu’s F _S (−25.93, P < 0.01), together with uni-modal mismatch distribution, indicated a recent genetic bottleneck or population expansion of the species. Analysis of molecular variance (AMOVA) indicated a small amount of differentiation among groups (1.7%); most of the total variation occurred within groups (98.3%). Also, there was no significant population structure (F _ST = 0.017, P > 0.05), and estimates of gene flows among groups were extremely high (Nm = 28.88), suggesting low genetic divergence between populations in the species. The lack of genetic differentiation among groups is most likely due to the combined gene flow from the downstream movement of eggs and larvae with currents and the upstream or downstream migration of adults throughout the distribution. These groups of L. elongata distributed in upper reaches of the Yangtze River should be considered as a single management unit.     

Streaked horned lark <Emphasis Type="Italic">Eremophila alpestris strigata</Emphasis> has distinct mitochondrial DNA

The Streaked Horned Lark (STHL; Eremophila alpestris strigata) is a federal candidate for listing under the Endangered Species Act. We evaluated the conservation status and level of genetic diversity of the STHL using the complete mitochondrial ND2 gene. We sampled 32 STHLs from the southern Puget Sound region, the Pacific coast, and Whites Island in the Columbia River of Washington, and additional 68 horned larks from Alaska, alpine and eastern Washington, Oregon, California, and Asia (outgroups). Our Maximum Likelihood analysis of 32 haplotypes identified three geographically concordant clades in Pacific coast states: Pacific Northwest (alpine and eastern Washington, Alaska), Pacific Coast (western Washington, California), and Great Basin (eastern Oregon). Each of the three clades was supported by bootstrap values ≥86%. The distance among them varied from 0.72 to 0.79% nucleotide divergence excluding intraclade variation. The relationship among the clades was not resolved. AMOVA also showed significant structuring of haplotypes among the three clades. Differences among clades accounted for 75.7% of sequence variation, differences among localities within clades accounted for 12.1%, and differences among individuals within localities accounted for the remaining 12.2%. Although STHL populations were closely related to the Californian sample, they appeared unique and isolated. All pairwise F _st values involving the STHL samples were significant (except between themselves). STHLs appear to have remarkably low genetic diversity; all 32 STHLs shared the same haplotype. Even with small sample sizes, all other localities had multiple haplotypes. Because the STHL appears to be unique and isolated, and to have little genetic diversity our data suggest it should be a conservation priority.     

Genetic structure analysis of natural <Emphasis Type="Italic">Sargassum muticum</Emphasis> (Fucales,Phaeophyta) populations using RAPD and ISSR markers

Sargassum muticum is important in maintaining the structure and function of littoral ecosystems, and is used in aquaculture and alginate production, however, little is known about its population genetic attributes. In this study, random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were used to investigate the genetic structure of four populations of S. muticum and one outgroup of S. fusiforme (Harv.) Setchell from Shandong peninsula of China. The selected 24 RAPD primers and 19 ISSR primers amplified 164 loci and 122 loci, respectively. Estimates of genetic diversity with different indicators (P%, percentage of polymorphic loci; H, the expected heterozygosity; I, Shannon’s information index) revealed low or moderate level of genetic variations within each S. muticum population, and a high level of genetic differentiations were determined with pairwise unbiased genetic distance (D) and fixation index (F _ST ) among the populations. The Mantel test showed that two types of matrices of D and F _ST were highly correlated whether from RAPD (r = 0.9706, P = 0.009) or ISSR data (r = 0.9161, P = 0.009). Analysis of molecular variance (AMOVA) was conducted to apportion the variations among and within the S. muticum populations. It indicated that variations among populations were higher than those within populations, being 55.82% verse 44.18% by RAPD and 55.21% verse 44.79% by ISSR, respectively. Furthermore, the Mantel test suggested that genetic differentiations among populations were related to the geographical distances (r > 0.6), namely, conformed to the IBD (isolation by distance) model, as expected from UPGMA (unweighted pair group method with arithmetic averages) cluster analysis. On the whole, the high genetic structuring among the four S. muticum populations along the distant locations was clearly indicated in RAPD and ISSR analyses (r > 0.9, P < 0.05) in our study.     

Genetic differentiation and intraspecific structure of Eastern Tropical Pacific spotted dolphins,Stenella attenuata,revealed by DNA analyses

Mitochondrial DNA (mtDNA) control region sequences and microsatellite loci length polymorphisms were used to investigate genetic differentiation in spotted dolphins (Stenella attenuata) in the Eastern Tropical Pacific and to examine the intraspecific structure of the coastal subspecies (Stenella attenuata graffmani). One-hundred and thirty-five animals from several coastal areas and 90 offshore animals were sequenced for 455 bp of the mitochondrial control region, resulting in 112 mtDNA haplotypes. Phylogenetic analyses and the existence of shared haplotypes between the two subspecies suggest recent and/or current gene flow. Analyses using χ², F _ST (based on haplotype frequencies) and Φ_ST values (based on frequencies and genetic distances between haplotypes) yielded statistically significant separation (randomized permutation values P<0.05) among four different coastal populations and between all but one of these and the offshore subspecies (overall F _ST=0.0691). Ninety-one coastal animals from these four geographic populations and 50 offshore animals were genotyped for seven nuclear microsatellite loci. Analysis using F _ST values (based on allelic frequencies) yielded statistically significant separation between most coastal populations and offshore animals, although no coastal populations were distinguished. These results argue for the existence of some genetic isolation between offshore and inshore populations and among some inshore populations, suggesting that these should be treated as separate units for management purposes.     

Genetic diversity and the biogeographical process of Acheilognathus macropterus revealed by sequence variations of mitochondrial cytochrome b gene

In this study, thirty-six individuals of Acheilognathus macropterus were collected from the Heilongjiang River, the Yangtze River, and the Nandujiang River. Partial mitochondrial cytochrome b gene region (636 base pair) was sequenced to these samples and 22 haplotypes were found. With A. chankaensis and A. tokinensis as outgroups, their relationships were analyzed. The p-distances were calculated with Mega software and a molecular phylogenetic tree was constructed using the neighbor-joining (NJ) method. The proportions of main morphological characters were compared as well. P-distances showed that the genetic differences in A. macropterus samples were far smaller than those between these samples and the outgroups. The molecular phylogenetic tree shows that samples with barbels and those without barbels were intermingled. There was no distinctive difference in proportions of morphological characteristics among them. These results suggested that samples with barbels and those without barbels (formally identified as A. taenianalis) are the same species; A. taenianalis is synonymous with A. macropterus. The thirtysix individuals were grouped into five clades and the positions of the samples in the clades were correspondingly grouped within their geographical distributions. Among the five clades, clades 1 and 5 included samples from the Heilongjiang River and Nandujiang River respectively. The samples from the Yangtze River scattered into clades 2, 3, and 4. There were distinctive genetic differences (>5%) among them. Interestingly, the distributions of the 21 samples in these three clades were not correlated to their geographical distributions. It is postulated that these genetic differences were due to the bitterlings’ mating choice mechanism, the prozygotic isolation. The genetic differences between the fish from Nandujiang River and those from the mainland indicated that they were separated early. However, the small genetic differences among the samples and the positions of the fish from the Heilonjiang River in the molecular phylogenetic tree indicate that fish in Heilongjiang River might have dispersed from the Yangtze River to that area much later. __________ Translated from Acta Hydrobiologica Sinica, 2006, 39(2): 134–140 [译自: 水生生物学报]     

Low genetic differentiation among morphologically distinct Labeobarbus species (Teleostei: Cyprinidae) in the Lake Victoria and Albertine basins,Uganda: insights from mitochondrial DNA

Labeobarbus altianalis and L. bynni bynni are hexaploid cyprinid fishes in the genus Labeobarbus. In the Great Lakes region of Africa, these two large-bodied barbs exhibit considerable morphological variations. Their intraspecific classification, currently based on geographical distribution and morphological variation, is of limited application due to the overlapping nature of the water systems. Patterns of genetic variation in the Albertine and Victoria drainage basins in Uganda were assessed using mitochondrial sequences of the control region (470 basepairs) and the cytochrome b gene (627 basepairs). At the control region 36 distinct haplotypes were observed in a total of 177 individuals. Relatively low overall nucleotide diversity (π = 0.010) was observed. Hierarchical analysis of the molecular variance revealed significant differentiation between populations in the total sample (F _ST = 0.172, p < 0.001), among populations within each basin (F _SC = 0.047, p < 0.001), but no significant subdivision among basins (F _CT = 0.131, p > 0.05). Phylogenetic analysis of the control region haplotypes resulted in a star-shaped phylogeny, with no clusters that reflect current taxonomic designation, morphotypes or basins. Phylogenetic analysis of cytochrome b gene sequences resulted in a tree topology similar to that observed for the control region. Thus the morphological characteristics separating the two species are intraspecific variation and the whole sample probably belongs to the same species.     

Population genetic structure of <Emphasis Type="Italic">Sargassum thunbergii</Emphasis> (Fucales,Phaeophyta) detected by RAPD and ISSR markers

Genetic variation of four populations of Sargassum thunbergii (Mert.) O. Kuntze and one outgroup of S. fusiforme (Harv.) Setchell from Shandong peninsula of China was studied with random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers. A total of 28 RAPD primers and 19 ISSR primers were amplified, showing 174 loci and 125 loci, respectively. Calculation of genetic diversity with different indicators (P%, percentage of polymorphic loci; H, the expected heterozygosity; I, Shannon’s information index) revealed low or moderate levels of genetic variations within each S. thunbergii population. High genetic differentiations were determined with pairwise Nei’s unbiased genetic distance (D) and fixation index (F _ST ) between the populations. The Mantel test showed that two types of matrices of D and F _ST were highly correlated, whether from RAPD or ISSR data, r = 0.9310 (P  = 0.008) and 0.9313 (P = 0.009) respectively. Analysis of molecular variance (AMOVA) was used to apportion the variations between and within the S. thunbergii populations. It indicated that the variations among populations were higher than those within populations, being 57.57% versus 42.43% by RAPD and 59.52% versus 40.08% by ISSR, respectively. Furthermore, the Mantel test suggested that the genetic differentiations between the four populations were related to the geographical distances (r > 0.5), i.e., they conformed to the IBD (isolation by distance) model, as expected from UPGMA (unweighted pair group method with arithmetic averages) cluster analysis. As a whole, the high genetic structuring between the four S. thunbergii populations along distant locations was clearly indicated in the RAPD and ISSR analyses (r > 0.8) in our study.     

Genetic differentiation between populations of swimming crab <Emphasis Type="Italic">Portunus trituberculatus</Emphasis> along the coastal waters of the East China Sea

Portunus trituberculatus is a commercially important species widely spread in the East China Sea. Intraspecific variation of the mitochondrial DNA cytochrome oxidase subunit I (mtDNA COI) gene was investigated in 213 individuals from six localities (Changjiang Estuary, Shengsi Islands, Zhoushan Islands, Dongtou Islands, Dinghai Bay, and Quanzhou Bay) ranging from north (31°21′N) to south (24°55′N) coastal waters of the East China Sea. Overall, a total of 27 mtDNA haplotypes and 21 variable sites were detected in the 787 bp segment of COI gene. Analysis of mtDNA COI sequence data revealed that crabs from the six localities were characterized by moderately high haplotypic diversity (h = 0.787 ± 0.026), while sequence divergence values between haplotypes were relatively low (π = 0.00241 ± 0.00098). Each population was characterized by a single most frequent haplotype, shared among all six localities, and a small number of rare ones, typically present in only one or two individuals and representative of a specific population. However, neither the neighbor-joining tree nor the minimum spanning network (MSN) based on the haplotype data exhibited geographical patterns of the six populations. Mismatch distribution analysis of P. trituberculatus individuals sampled from the six localities suggested that sudden population expansion might have occurred in CJ and SS population that might be consistent with over-exploitation of the swimming crab. Analysis of molecular variance (AMOVA) and F _ST statistics showed that significant genetic differentiation existed among the SS, ZS, DT, DH, and QZ populations, suggesting that gene flow might be reduced, even between the geographically close sites, despite the high potential of dispersal. The possible causes of the observed genetic heterogeneity among the P. trituberculatus populations and the potential applications of the mtDNA COI marker in the artificial breeding and fisheries management are discussed. Handling editor: C. Sturmbauer     

Genotype frequency and F ST analysis of polymorphisms in immunoregulatory genes in Chinese and Caucasian populations

Selection and genetic drift can create genetic differences between populations. Cytokines and chemokines play an important role in both hematopoietic development and the inflammatory response. We compared the genotype frequencies of 45 SNPs in 30 cytokine and chemokine genes in two healthy Chinese populations and one Caucasian population. Several SNPs in IL4 had substantial genetic differentiation between the Chinese and Caucasian populations (F _ST ~0.40), and displayed a strikingly different haplotype distribution. To further characterize common genetic variation in worldwide populations at the IL4 locus, we genotyped 9 SNPs at the IL4 gene in the Human Diversity Panel’s (N = 1056) individuals from 52 world geographic regions. We observed low haplotype diversity, yet strikingly different haplotype frequencies between non-African populations, which may indicate different selective pressures on the IL4 gene in different parts of the world. SNPs in CSF2, IL6, IL10, CTLA4, and CX3CR1 showed moderate genetic differentiation between the Chinese and Caucasian populations (0.15 < F _ST < 0.25). These results suggest that there is substantial genetic diversity in immune genes and exploration of SNP associations with immune-related diseases that vary in incidence across these two populations may be warranted.     

Genetic Diversity and Geographical Differentiation of <Emphasis Type="Italic">Desmodium triflorum</Emphasis> (L.) DC. in South China Revealed by AFLP Markers

High levels of genetic variation enable species to adapt to changing environments and provide plant breeders with the raw materials necessary for artificial selection. In the present study, six AFLP primer pairs were used to assess the genetic diversity of Desmodium triflorum (L.) DC. from 12 populations in South China. A high percentage of polymorphic loci (P = 76.16%) and high total gene diversity (H _T = 0.310) were found, indicating that the genetic diversity of D. triflorum is high at the species level. Genetic diversity was also relatively high at the population level (P = 55.85%, H _e = 0.230). The coefficient of gene differentiation among populations (G _ST) was 0.255, indicating that while most genetic diversity resided within populations, there was also considerable differentiation among populations. AMOVA also indicated 24.29% of the total variation to be partitioned among populations (Φ_ST = 0.243). UPGMA clustering analysis based on genetic distances showed that the 12 populations could be separated into three subgroups: an eastern, a western, and a central-southern subgroup. However, a Mantel test revealed no significant correlation (r = 0.286, p = 0.983) between the geographical distances and genetic distances separating these populations; mountain barriers to gene flow and human disturbance may have confounded these correlations. The present study has provided some fundamental genetic data that will be of use in the exploitation of D. triflorum.     

Mitochondrial DNA diversity of <Emphasis Type="Italic">Glyptothorax zanaensis</Emphasis> (Wu et al., 1981) populations in Nu River,China

The hillstream fish, Glyptothorax zanaensis (Wu et al., Oceanol Limnol Sin 12:74–79, 1981) is a freshwater teleost species belonging to the order Siluriformes, endemic to Nu River and Lancang River basin. Because wild stocks of G. zanaensis have declined substantially in recent years, owing to overexploitation, pollution, illegal fishing practices and the development of hydroelectricity in the Nu River, the populations of G. zanaensis have markedly declined in recent years. We studied the mitochondrial DNA cyt b gene to assess the genetic diversity of G. zanaensis in seven tributaries of the Nu River. Seventy variable sites defined 15 haplotypes in 153 individuals. Haplotype diversity and nucleotide diversity were 0.542 ± 0.014 and 0.011 ± 0.007, respectively. The Sunzu River site had the highest haplotype diversity (Hd, 0.893 ± 0.014) and nucleotide diversity ( Pi, 0.010 ± 0.012). Genetic diversity was low in the Dong River, Fengwei River and Mengtong River. Pairwise F _ST values ranged from 0.00 to 1.00. Significant F _ST values were found for Sunzu River sampled sites paired with all other populations and Mengtong river sampled site paired with the others. Furthermore, the Nanpeng River sampled site was significantly differentiated from the Luoming River, Dong River and Fengwei River sampled sites. Such knowledge is essential for developing appropriate policies for the rational management of G. zanaensis.     

Genetic differentiation across eastern Pacific oceanographic barriers in the threatened seahorse <Emphasis Type="Italic">Hippocampus ingens</Emphasis>

Understanding the population structure and evolutionary history of the eastern Pacific seahorse Hippocampus ingens is critical for the effective management of this threatened species. Life history characteristics of H. ingens (site fidelity and brooding of young) may limit gene flow and lead to population differentiation. A recent study analyzing conserved fragments of the mitochondrial cyt b and control region found no population structure. We re-assess this conclusion with a phylogeographic analysis of relationships among 115 individuals of H. ingens over a broader geographic range (San Diego Bay in California, Gulf of California, Central America, Ecuador, and Peru) based on a more variable 428 base pair fragment of the control region. This expanded analysis affirms low overall nucleotide diversity relative to other seahorses (θ_π = 0.004), and shows evidence of a recent bottleneck and population expansion since the middle Pleistocene. AMOVA analysis shows moderate overall population structure (Φ_ST = 0.10, P val = 0.00), and pairwise Φ_ST estimates indicate structure between the Gulf of California and all Pacific coast localities. Knowledge of population structure in H. ingens may improve conservation efforts by identifying evolutionarily important management units, and could determine source regions in the continuing trade of seahorses for traditional Chinese medicine. The level of genetic divergence observed between the Gulf of California and all other localities sampled may distinguish the Gulf as a separate management unit. Additional phylogeographic research with more quickly evolving genetic markers and targeted sampling at the mouth of the Gulf of California is warranted to inform strategies for conservation of this threatened seahorse.     

Comparative chloroplast DNA phylogeography of two tropical pioneer trees, Macaranga gigantea and Macaranga pearsonii (Euphorbiaceae)

Macaranga (Euphorbiaceae) has received much ecological and evolutionary research attention as a genus that includes some of the most conspicuous pioneer trees of Southeast Asian tropical rainforests and because of its manifold associations with ants, including about 30 species that are obligate ant-plants (myrmecophytes). We used sequence data from three chloroplast DNA loci (ccmp5, ccmp6, atpB-rbcL) to assess phylogeographical patterns in species of Macaranga, section Pruinosae, sampled from various regions of Borneo and the Malay Peninsula. Forty-nine chloroplast DNA haplotypes (HT) were identified among 768 specimens from five species, Macaranga gigantea (N = 329; 23 HT), Macaranga pearsonii (N = 347; 21 HT), Macaranga puberula (N = 24; 4 HT), Macaranga hosei (N = 48; 6 HT), and Macaranga pruinosa (N = 20; 5 HT). Forty-one haplotypes were species-specific, whereas eight haplotypes were shared by two, three, or four species and occupied internal positions in a parsimony network. Population genetic parameters based on haplotype frequencies proved to be in a similar range in the non-myrmecophytic M. gigantea and in the ant-associated M. pearsonii, which have overlapping distributions in northern and eastern Borneo. A comparison of G _ST and N _ST values revealed a strong phylogeographic structure in both species, whereas colonization pathways suggested by the network topology were different. Both species exhibited similar levels of haplotypic diversity and moderate to high levels of population differentiation. There were no obvious indications for an influence of the symbiotic ant partners on the population structure of their host plants.