Genetic structure of the common terrestrial pulmonate snail,Cryptozona siamensis (Pfeiffer, 1856), in Thailand

Allozyme variation was examined in 432 specimens of the pulmonate snail, Cryptozona siamensis, from 24 localities across three regions of Thailand plus an additional population from Malaysia. Although this snail is a cosmopolitan species it was predicted that C. siamensis should exhibit a large degree of genetic subdivision across these regions due to a low level of inter-population gene flow. To test this hypothesis, 10 enzyme systems were screened using horizontal starch gel electrophoresis, from which 14 presumed allozyme loci were scored and 13 found to be polymorphic. For these 13 polymorphic loci the expected heterozygosity levels were moderate (H_exp = 0.036–0.183, mean = 0.101). Genetic differentiation among the samples was low, with a low F_ST value (0.254) and genetic distance (D = 0.000–0.124, mean = 0.034) and no significant isolation by distance (IBD) across the whole sampled range of Thailand or Thailand plus Malaysia. However, an IBD pattern in the population substructure in the central Thailand region was detected. The low F_ST and moderate H_exp values in various populations of C. siamensis suggests a high degree of gene flow/migration between populations across the three geographic regions of Thailand (northern, central and southern) as well as the sampled population in Malaysia, perhaps associated with the anthropochoric effect of snail dispersal. The apparent IBD in the central Thailand region could be the consequence of local dynamics though this region.     

Mitochondrial genetic variation and population structure of the striped snakehead,Channa striata in Malaysia and Sumatra,Indonesia

We explored the genetic diversity and structure of the striped snakehead (Channa striata) across Malaysia and Sumatra (Indonesia) using the partial mtDNA CO1 gene. Twenty five populations (n = 345) were assayed and subdivided into six regions, following the physiogeographical barriers. Populations Sega (SG), Tanjung Tambutan (TR), Kajang (KJ) and Linggi (LG) are highly diversified (Hd: 0.484–0.762, π: 0.0033–0.0059) which could serve as candidates for a selective breeding programme. The only population that contributed to the total allelic richness is Takengon (CS) as it is highly differentiated from other populations and genetically variable within population. We detected two major phylogenies: 1) northwest Peninsular Malaysia and 2) all other regions of Malaysia and Sumatra, Indonesia. They are products of the physical restriction to gene flow between the two lineages by the Bintang Mountain Range. A total of 92.4% of the population pairwise comparison F_ST showed significant structuring, yet several geographically distant populations showed a close genetic relationship. The discrepancy is due to ancient population dispersal and human-mediated translocation. These major findings provide an important base study for initiating a selective breeding program. The high population genetic diversity requires independent conservation as they contain most of the total diversity in this area.     

Genetic Diversity of a Tropical Tree Species,Shorea leprosula Miq. (Dipterocarpaceae), in Malaysia: Implications for Conservation of Genetic Resources and Tree Improvement1

Genetic diversity and population genetic structure of Shorea leprosula was investigated using seven natural populations distributed throughout Peninsular Malaysia and one natural population from Borneo. The mean population and species level genetic diversity were exceptionally high (H_e= 0.369 ± 0.025 and 0.406 ± 0.070, respectively). Heterozygosity varied among populations, ranging from 0.326 to 0.400, with the highest values found in the populations from central Peninsular Malaysia. Correlations among ecological factors (longitude, latitude, and annual rainfall) were not significant (P > 0.05), indicating that these ecological variables were not responsible for the observed genetic differences among populations. The Bangi adult population exhibited a higher level of observed heterozygosity but lower fixation indices in comparison to its seedling population. All other seedling populations also showed positive fixation indices (f), indicating a general excess of homozygotes. This also may suggest selection against homozygotes between the seedling and adult stages. A low level of population differentiation was detected (G_ST= 0.117 with the Lambir population and GST= 0.085 without the Lambir population). Furthermore, gene flow (N_m) between populations was not significantly correlated with geographical distances for the populations within Peninsular Malaysia. Cluster analysis also did not reflect geographical proximity and gave little insight into the genetic relatedness of the populations. This may indicate that the populations sampled are part of a continuous population with fragmentation having occurred in the recent past.     

Phylogeographic Pattern of the Striped Snakehead,Channa striata in Sundaland: Ancient River Connectivity,Geographical and Anthropogenic Singnatures

A phylogeographic study of an economically important freshwater fish, the striped snakehead, Channa striata in Sundaland was carried out using data from mtDNA ND5 gene target to elucidate genetic patterning. Templates obtained from a total of 280 individuals representing 24 sampling sites revealed 27 putative haplotypes. Three distinct genetic lineages were apparent; 1)northwest Peninsular Malaysia, 2)southern Peninsular, east Peninsular, Sumatra and SW (western Sarawak) and 3) central west Peninsular and Malaysian Borneo (except SW). Genetic structuring between lineages showed a significant signature of natural geographical barriers that have been acting as effective dividers between these populations. However, genetic propinquity between the SW and southern Peninsular and east Peninsular Malaysia populations was taken as evidence of ancient river connectivity between these regions during the Pleistocene epoch. Alternatively, close genetic relationship between central west Peninsular Malaysia and Malaysian Borneo populations implied anthropogenic activities. Further, haplotype sharing between the east Peninsular Malaysia and Sumatra populations revealed extraordinary migration ability of C. striata (>500 km) through ancient connectivity. These results provide interesting insights into the historical and contemporary landscape arrangement in shaping genetic patterns of freshwater species in Sundaland.     

Anadromy and heterogenous population of a tropical shad Tenualosa ilisha in Malaysia,as revealed by otolith microchemistry and molecular evidence

Tenualosa ilisha was found recently in the Perak River in western Peninsular Malaysia. Molecular phylogenetic and haplotype network analyses suggest that T. ilisha has two genetically distinct populations/groups: (i) Peninsular Malaysia (Malaysia population), and (ii) Peninsular Malaysia, Thailand, India and Bangladesh (Indian Ocean population). The results also suggest that the T ilisha population in Peninsular Malaysia is genetically heterogeneous with a typical anadromous migration pattern.     

Genetic diversity and population structure of Schizopygopsis younghusbandi Regan in the Yarlung Tsangpo River inferred from mitochondrial DNA sequence analysis

The genetic diversity and population structure of Schizopygopsis younghusbandi from six sites in the middle reach of the Yarlung Tsangpo River were examined based on mitochondrial DNA cytochrome b (Cyt b) gene and control region (D-loop) sequences. 1141 bp of complete Cyt b sequences and 737 bp of partial D-loop sequences for 153 individuals of S. younghusbandi were obtained by using PCR amplification and sequencing. The results showed that S. younghusbandi populations had high haplotype diversity and low nucleotide diversity, and the population genetic diversity of this species was at a low level. Analysis of molecular variance revealed that most genetic variation occurred within populations, and that genetic differentiation was at low or moderate levels. The network of haplotypes based on Cyt b gene showed that there were two groups within the examined populations. Neutrality tests and mismatch distribution analysis suggested that this species might have undergone a population expansion and the expansion time was estimated to be 0.25–0.46 Ma BP. All these results would be crucial to establish scientific strategies for effective conservation and sustainable exploitation of wild resources of S. younghusbandi.     

PCR-RFLP analysis of mitochondrial DNA cytochrome b gene among Haruan (Channa striatus) in Malaysia

Haruan (Channa striatus) is in great demand in the Malaysian domestic fish market. In the present study, mtDNA cyt b was used to investigate genetic variation of C. striatus among populations in Peninsular Malaysia. The overall population of C. striatus demonstrated a high level of haplotype diversity (h) and a low-to-moderate level of nucleotide diversity (π). Analysis of molecular variance (AMOVA) results showed a significantly different genetic differentiation among 6 populations (F_ST = 0.37566, P = 0.01). Gene flow (Nm) was high and ranged from 0.32469 to infinity (∞). No significant relationship between genetic distance and geographic distance was detected. A UPGMA tree based on the distance matrix of net interpopulation nucleotide divergence (d_A) and haplotype network of mtDNA cyt b revealed that C. striatus is divided into 2 major clades. The neutrality and mismatch distribution tests for all populations suggested that C. striatus in the study areas had undergone population expansion. The estimated time of population expansion in the mtDNA cyt b of C. striatus populations occurred 0.72-6.19 million years ago. Genetic diversity of mtDNA cyt b and population structure among Haruan populations in Peninsular Malaysia will be useful in fisheries management for standardization for Good Agriculture Practices (GAP) in fish-farming technology, as well as providing the basis for Good Manufacturing Practices (GMP).     

Phenotypic profile of Aedes albopictus (Skuse) exposed to pyrethroid-based mat vaporizers and underlying detoxification mechanisms: A statewide report in Selangor,Malaysia

This study examines the biological efficacy of four mosquito mat vaporizers each containing different active ingredients: prallethrin with PBO, dimefluthrin, prallethrin, and d-allethrin. The glass chamber assay was used to evaluate their efficacy on Aedes albopictus (Skuse) (Diptera: Culicidae) from nine districts in Selangor, Malaysia. Aedes albopictus exhibited different knockdown rates, with 50% knockdown times, KT₅₀, varying from 1.19 to 2.00 min, 1.22 to 2.20 min, 1.39 to 5.85 min, and 1.39 to 1.92 min for prallethrin with PBO, dimefluthrin, prallethrin and d-allethrin, respectively. In general, all populations of Ae. albopictus were completely knocked down after exposure to all active ingredients except Hulu Selangor population, which showed 96.00% knockdown against d-allethrin. On the contrary, mortality rates were observed from 84.00–100.00%, 84.00–100.00%, 90.67–100.00% and 90.67–100.00% in populations tested with prallethrin with PBO, dimefluthrin, prallethrin and d-allethrin, respectively. Moreover, significant correlations between mortality rates of prallethrin with PBO vs dimefluthrin (r = 0.836, P = 0.003), prallethrin with PBO vs prallethrin (r = 0.760, P = 0.011), and prallethrin vs d-allethrin (r = 0.694, P = 0.026) were also observed, suggesting cross-resistance among pyrethroids. d-allethrin was found to be high in insecticidal activity, followed by prallethrin, prallethrin with PBO, and dimefluthrin. In consistent with mortality due to insecticide exposure, elevated levels of enzyme activities were also demonstrated in Sabak Bernam, Hulu Selangor, Gombak, Petaling, Hulu Langat and Klang populations.     

Ranging behaviour,activity, habitat use,and morphology of the Malay civet (Viverra tangalunga) on Peninsular Malaysia and comparison with studies on Borneo and Sulawesi

The Malay civet (Viverra tangalunga) is found throughout Southeast Asia, but there have been very few studies on its natural history and ecology. We present here the results from the first radio telemetry study of this species on Peninsular Malaysia and compared our findings to similar studies on Borneo and Sulawesi. From August 2004 to August 2006, we captured 11 Malay civets in Krau Wildlife Reserve and radio-tracked seven adults (four males and three females). The mean weight of males (6.6 kg) was significantly higher than females (5.8 kg). Both sexes on Peninsular Malaysia were larger than their counterparts on Borneo and Sulawesi. There was no significant difference between the mean sizes of male and female home ranges on Peninsular Malaysia; the mean home range size for both sexes was 143 ha (95% MCP), which was larger than the mean range size recorded on Borneo and Sulawesi. We found that the Malay civet is a solitary, territorial species on Peninsular Malaysia: mean range overlap was 15% for males and 0% for females, and the home range of each male overlapped one or two females. Malay civets were mainly nocturnal, with some periods of rest during the night; daytime rest sites were within dense ground cover. Lowland forest is an essential habitat for this species; although Malay civets were sometimes found in plantations, they did not venture far from forested habitat.     

First discovery of Pleistocene orangutan (Pongo sp.) fossils in Peninsular Malaysia: Biogeographic and paleoenvironmental implications

Nine isolated fossil Pongo teeth from two cave sites in Peninsular Malaysia are reported. These are the first fossil Pongo specimens recorded in Peninsular Malaysia and represent significant southward extensions of the ancient Southeast Asian continental range of fossil Pongo during two key periods of the Quaternary. These new records from Peninsular Malaysia show that ancestral Pongo successfully passed the major biogeographical divide between mainland continental Southeast Asia and the Sunda subregion before 500 ka (thousand years ago).     

Genetic diversity and population structure of the Asian green mussel Perna viridis in South China Sea based on microsatellite markers

The green mussel, Perna viridis is ecologically and economically important in the coastal region of the South China Sea. Determining its population genetic structure at this fine geographic scale will help sustainable management of natural stocks. In this study, we examined the genetic diversity and population genetic structure of P. viridis from four locations in the South China Sea (n = 45–48) using nine microsatellite loci. The results showed moderate levels of genetic diversity in all four samples (mean A = 13.222–14.000, mean A_e = 7.092–7.571, mean A_r = 12.894–13.746, mean H_o = 0.596–0.656, mean H_e = 0.690–0.733) and a large effective population size estimate for the pooled sample (total N_e estimates = infinity, 95% CI = 1869.0-infinity). We did not detect any sign of recent bottleneck events in P. viridis populations in the South China Sea. The conventional and a model-based analysis reveal low, non-significant genetic divergence among the four samples (F_ST = − 0.001–0.005, P > 0.05/6). The results obtained from this study can provide valuable genetic information for the conservation and fishery management of P. viridis by retaining the high N_e estimates.     

Trio under threat: can we secure the future of rhinos,elephants and tigers in Malaysia?

Three of Malaysia’s endangered large mammal species are experiencing contrasting futures. Populations of the Sumatran rhino (Dicerorhinus sumatrensis) have dwindled to critically low numbers in Peninsular Malaysia (current estimates need to be revised) and the state of Sabah (less than 40 individuals estimated). In the latter region, a bold intervention involving the translocation of isolated rhinos is being developed to concentrate them into a protected area to improve reproduction success rates. For the Asian elephant (Elephas maximus), recently established baselines for Peninsular Malaysia (0.09 elephants/km² estimated from one site) and Sabah (between 0.56 and 2.15 elephants/km² estimated from four sites) seem to indicate globally significant populations based on dung count surveys. Similar surveys are required to monitor elephant population trends at these sites and to determine baselines elsewhere. The population status of the Malayan tiger (Panthera tigris jacksoni) in Peninsular Malaysia, however, remains uncertain as only a couple of scientifically defensible camera-trapping surveys (1.66 and 2.59 tigers/100 km² estimated from two sites) have been conducted to date. As conservation resources are limited, it may be prudent to focus tiger monitoring and protection efforts in priority areas identified by the National Tiger Action Plan for Malaysia. Apart from reviewing the conservation status of rhinos, elephants and tigers and threats facing them, we highlight existing and novel conservation initiatives, policies and frameworks that can help secure the long-term future of these iconic species in Malaysia.     

An observation of the loss of genetic variability in prenant’s schizothoracin,Schizothorax prenanti,inhabiting a plateau lake

Prenant’s schizothoracin (Schizothorax prenanti) was found naturally distributed in a plateau lake (Lake Wuxu) in upper reaches of the River Jiulong. The genetic diversities of the populations from L. Wuxu and R. Jiulong were investigated according to analysis of mtDNA control region and compared against three other reported river populations. The L. Wuxu population showed lower haplotype (h = 0.74) and nucleotide (π = 0.001) diversities compared to the R. Jiulong population (h = 0.87; π = 0.012); the lake population had suffered from a bottleneck, which might contribute to its low genetic diversity; the lake population significantly differentiated from the river populations, and significant gene flow was hardly found between river populations and L. Wuxu population, suggesting that there is a strong population structure between the lake and river populations. The L. Wuxu population was ecologically isolated from the R. Jiulong population, while the relatively low genetic variability had not affected its viability.     

Genetic diversity of a dominant species Stipa bungeana and its conservation strategy in the Loess Plateau of China

In order to investigate the genetic diversity and population structure of Stipa bungeana under the background of grassland utilization and grazing exclusion, ten S. bungeana populations were selected from different steppe type in the Loess Plateau of China. Sequence-related amplified polymorphism (SRAP) marker was used to assess the genetic diversity. Fifteen SRAP primer combinations generated a total of 482 amplification bands, 418 (86.72%) were polymorphic bands. A relatively high level of genetic diversity (PPB = 89.80%, h = 0.1972, H = 0.3154) was detected at the species level, but the genetic diversity was low at the population level (PPB = 17.01–33.33%, h = 0.0438–0.0967, H = 0.0361–0.1447). AMOVA analysis revealed a high level of genetic differentiation among populations (Φ_ST = 0.6757), and a limited among-population gene flow (N_m = 0.1200). There was no significant correlation between genetic distance and geographic distance by Mantel test (r = 0.1126, P = 0.204). Conservation implications were proposed for S. bungeana.     

Genetic diversity in Mentha cervina based on morphological traits,essential oils profile and ISSRs markers

Morphological, phytochemical and genetic differences were studied to evaluate the level and distribution of diversity in twelve populations of the Portuguese endangered medicinal plant Mentha cervina L. Morphological variation was correlated with ecological conditions at the site of origin. Pulegone was the major essential oils compound in all of the populations collected at full flowering (68–83%), in different growing conditions (51–82%), and for all the developmental stages studied (47–82%). Although clusters were defined, the analysis revealed a high chemical correlation among all populations (S_corr ≥ 0.95%). Inter-simple sequence repeats markers were used to assess the population structure and genetic variation. Populations exhibited a relatively low genetic diversity (PPB = 14.3–64.6%, H_e = 0.051–0.222, I = 0.076–0.332), with high structuring between them (G_ST = 0.51). However, the genetic diversity at species level was relatively high (PPB = 97.7%; H_e = 0.320). The levels and patterns of genetic diversity were assumed to result largely from a combination of evolutionary history and its unique biological traits, such as breeding system, clonal growth, low capacity of dispersion and habitat fragmentation. The relatively low genetic diversity in the populations analyzed indicates that the maintenance of their evolutionary potential is at risk if population sizes are maintained and if there is no protection of the habitats.     

Genetic diversity and population structure analysis of Qinghai-Tibetan plateau schizothoracine fish (Gymnocypris dobula) based on mtDNA D-loop sequences

Gymnocypris dobula is a commercially important fishery species and mostly distributed in the freshwater of Tibetan plateau. In this study, genetic diversity and intraspecific population differentiation were examined by using mitochondrial DNA D-loop sequences in 97 individuals sampled from three localities (Pali, Lasa and Yanghu) in the Tibetan plateau. Two hundred and fourteen polymorphic sites and 50 haplotypes were defined among the three localities. Genetic diversity analysis showed that the highest genetic diversity level was found in Pali population. Phylogenetic relationships analysis results indicated that closer phylogenetic relationships were found between the Yanghu and Lasa populations. Genetic population differentiation analysis indicated that the majority of variation (84.91%) was attributed to variations among populations and the largest differentiation was found between Pali and Lasa localities (F_st = 0.874, Nm = 0.036). Moreover, the historical demographic events were assessed by implementing the mismatch distribution analysis, Fajima's D test and Fu's Fs test. The results indicated that the Pali population had undergone a demographic expansion, possibly within the last 0.163 MYA (Million Years Ago). Our study firstly identified the population genetic structure of the G. dobula, which could be helpful for artificial breeding, fishery stock identification and resource conservation for this species.     

Genetic diversity and population structure of the red stingray,Dasyatis akajei inferred by AFLP marker

To examine population genetic diversity and variation of the red stingray Dasyatis akajei, samples from 1 freshwater region and 6 coastal localities within its distribution range were analyzed by using amplified fragment length polymorphism (AFLP) technology. A total of 207 loci were identified by 4 primer combinations from 87 individuals, 174 of which were polymorphic (84.1%). A high level of Nei's gene diversity was observed with the overall value of 0.230 ± 0.179. No significant genealogical clusters associated with sampling sites were revealed on the UPGMA tree. Both analysis of molecular variance (AMOVA, Φ_ST = 0.085, P = 0.00) and pairwise F_ST indicated significant genetic differentiation among four marine samples. However, no particular genetic differentiation was detected between marine and the limited sampling freshwater populations (AMOVA, Φ_CT = 0.056, P > 0.05). Except for the TZ vs. WZ (5.193), the gene flow estimates (N_m) demonstrated the effective immigrants were 1.918-2.976, suggesting low level dispersal between pairwise marine populations. Species-specific habits (demersal and sluggish habits) are probably responsible for the population structuring.     

Host-associated genetic differentiation in rice grasshopper, Oxya japonica, on wild vs. cultivated rice

Rice grasshopper, Oxya japonica, is one of the most important pests in south China, mainly inhabiting fields of wild rice (Oryza rufipogon) and cultivated rice (Oryza sativa). In this study, we used AFLP marker to investigate the genetic diversity and population structure of rice grasshoppers collected from south China, with emphasis on testing the hypothesis that there was significant genetic differentiation among grasshopper populations associated with different hosts (i.e. wild vs. cultivated rice). Seven populations consisting of 104 individuals were sampled from Hainan Island and the mainland of south China. Eight primer combinations produced 564 reliable bands, of which 563 were polymorphic. O. japonica showed considerable genetic variation at population level, with gene diversity (H_E) ranging from 0.1103 to 0.2035. Genetic diversity were studied on seven populations, and generally three populations from wild rice had higher levels of genetic diversity (H_E = 0.1635) than the other four populations feeding on cultivated rice (H_E = 0.1327). We observed high population differentiation, with F_st ranging from 0.4172 to 0.7652 among the seven populations. However, Mantel test detected no significant correlation between genetic distance and geographical distance (r = 0.3541; p = 0.0689). By contrast, we found significant genetic differentiation between groups collected from different hosts. These data suggested that the anthropogenic activity in cultivated rice fields (i.e. pesticides, fertilization and cultivation) could have played an important role in shaping the genetic structure of O. japonica.     

Fragmented habitat drives significant genetic divergence in the Chinese endemic plant,Urophysa henryi (Ranuculaceae)

Urophysa henryi (Oliv.) Ulbr., endemic to China with small populations, is known as a medicinal plant. In this study, ISSR markers were used to assess the genetic diversity and population structure throughout its entire distribution areas. Twelve primers revealed high genetic diversity at the species level (PPB = 95.6%; H = 0.3441; I = 0.5111), as well as high level of genetic differentiation (F_ST = 0.659, p < 0.001; G_ST = 0.677) and restricted gene flow (Nm = 0.239) among populations. According to the UPGMA and PCoA analysis, the 9 populations were clustered into three main groups, which were roughly in accordance with their geographical regions. In addition, a significant correlation between the genetic difference and geographic distances among populations was detected from the IBD analysis (r = 0.516, p = 0.003). These results indicated that the habitat heterogeneity and physical barriers play important roles in the modern distribution pattern and population divergence of U. henryi. However, human activities have posed serious threat to its living environment and continued survival. It is necessary to adopt some measures to restrict anthropogenic disturbances and preserve the existing populations.