DNA barcoding is a useful tool for species identification using standardized genomic DNA fragments. The genus Amentotaxus, consisting of five or six species, is confined to South China, Northeast India, Laos, and Vietnam. All species have been assessed as globally or nationally threatened. However, there is uncertainty about the number of species involved, especially in the border areas of southern China, northern Vietnam, and Laos. We selected five DNA barcodes (rbcL, matK, trnH-psbA, trnL-F, and internal transcribed spacer (ITS)) to evaluate their discrimination ability in this genus, and to investigate the current taxonomy of Amentotaxus. Our results indicate that all the selected barcoding regions showed a high level of universality for PCR and sequencing. When six species are recognized, the nuclear ribosomal DNA region ITS and the chloroplast DNA region trnL-F used on their own provided the highest identification success (60%). Two barcode combinations that included either ITS or trnL-F had the same species discrimination ability. Combinations using additional barcodes did not improve the species identification success. When only five species are recognized, with A. hatuyenensis T. H. Nguyen treated as a synonym of A. yunnanensis H. L. Li, the discrimination rate rises to 100%. Our results also indicate that recent collections from Yunnan province, China, Lao Cai province, Vietnam, and Laos may represent a potential new species. The findings from this study will be very useful for the formulation of appropriate conservation strategies for threatened Amentotaxus species in national and trans-boundary regions.
Mugil cephalus sensu lato is a globally distributed complex of cryptic species whose distribution range and evolutionary history remains largely unknown. In the North West (NW) Pacific three species have been identified genetically among fish described morphologically as M. cephalus. Their distribution ranges are largely parapatric and has been proposed to mirror different thermal preferences. To date, few samples have been analyzed from South China Sea, which limits inferences on the evolutionary history of the species complex. We sampled fish identified morphologically as M. cephalus along Vietnamese shores and characterized them using the sequence polymorphism of two mitochondrial genes, the cytochrome oxidase I and cytochrome b. This demonstrated that all three species described in the NW Pacific are present in both northern and southern Vietnamese waters. Although the difference in species abundance reflects those observed in the NW Pacific, no phylogeographic pattern was revealed. In addition, no population structure was observed whatever the species or the distribution range considered, which indicates a significant level of gene flow that maintains genetic homogeneity of the three species. It is also conceivable that each species experienced a recent population expansion from a single ancestral population. Finally we suggest that if the cold waters of the NW Pacific present a physiologic challenge leading to the almost parapatric distribution of the three species, then it is likely that the warm surface temperatures of the South China Sea negate this barrier.
Steinernema robustispiculum n. sp. (Rhabditida: Steinernematidae) was isolated from woodland in Chumomray National Park, Sason, Sathay, Kontum, Vietnam. Its morphology, morphometrics, cross-hybridisation and the ITS-rDNA sequence analysis revealed that S. robustispiculum clearly differs from other known Steinernema spp. As in the cases of S. intermedium (Poinar, 1985), S. robustispiculum has very robust spicules, but it can be distinguished by the longer tail of the infective juvenile, lower E%, shorter spicules, the shape of the spicules, the number of genital papillae in the caudal region and the presence of a mucron on the male tail. S. robustispiculum has a lateral field resembling that of S. sangi Phan, Nguyen & Moens, 2001, but can be distinguished by a higher E%, higher D%, smaller length to width ratio of the spicules and the morphology of both the spicule head (manubrium) and the dorsal lobe of the spicule. The morphometrics of infective juveniles of S. robustispiculum are similar to those of S. monticolum Stock, Choo & Kaya, 1997; these species can be distingusihed by the position of the excretory pore, the smaller length to width ratio of the spicules, and the length and morphology of the spicule head (manubrium). The phylogenetic relationships within Steinernema Travassos, 1927, including the newly sequenced Vietnamese species S. robustispiculum n. sp., S. loci Phan, Nguyen & Moens, 2001, S. thanhi Phan, Nguyen & Moens, 2001 and S. sangi, are presented based on analyses of the ITS-rDNA. The ITS RFLP profiles obtained from 17 different restriction enzymes are also presented.
The endangered Dipterocarpus dyeri is distributed in forests of southeast Vietnam, which is expected to have low genetic diversity and high genetic population structure due to habitat loss and overexploitation. To provide valuable information for conservation measures, nine polymorphic microsatellites were used to analyze 151 adult trees from six populations, representing the natural range of D. dyeri in Vietnam. Contrary to the expectation, this species has relatively high genetic diversity within populations, low genetic differentiation among populations, and suggesting high gene flow. Results of bottleneck tests showed evidence of a bottleneck in the TaKou population. Analysis of molecular variance showed a high genetic variation within populations compared to among populations. Bayesian analysis also indicated two genetic lineages related to geographical distances. These results highlight the importance of conserving the genetic resources of D. dyeri and will provide guidelines for species conservation in Vietnam.
Steinernema ashiuense sp. n. was collected by a Galleria baiting method from grassland along the riverbank of the Yura river, near the Ashiu Research Forest Station of Kyoto University. The new species is characterised by an infective juvenile body length of 768 (720-800) µm, lateral field with five equally developed ridges (i.e., six lines or incisures), head smooth lacking horn-like structures, excretory pore located at level of middle of pharynx, hyaline portion short (ca one-third of tail length). Males have 50-65 µm long, broad, slightly yellowish spicules; spicule length / spicule width = 4 (3.5-4.4), and two subventral and one subdorsal pair of genital papillae in the postcloacal region. The analysis of ITS-rDNA sequence placed S. ashiuense sp. n. in the 'feltiae-kraussei-oregonense' group in the clade containing S. robustispiculum and S. monticolum. It can be distinguished from these species by morphological characters of the infective juveniles and males.
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