Biogeography of land snail genus Acusta (Gastropoda: Camaenidae): Diversification on East Asian islands.

East Asia has highly diverse and endemic biota due to its complex geological and climatic history and its diversified topography. The continental and insular distributions of land snail genus Acusta in East Asia provide a good opportunity to compare the evolutionary processes in this group under different biogeographical conditions. In this study, we inferred the evolutionary history of the land snail genus Acusta by a molecular phylogeny and investigated how the palaeogeographic events shaped species diversity and the distribution of the Acusta genus within the island arc. A concatenated dataset generated from sequences of one nuclear (ITS2) and two mitochondrial (16S, COI) gene fragments, include most of nominal taxa of the genus, four related species and one outgroup. We constructed the phylogeny and the evolutionary history of the genus through maximum-likelihood and Bayesian inference methods, using a Bayesian molecular clock and ancestral range estimation. Our results suggested that currently recognized species in Acusta are polyphyletic. The traditionally accepted concept of the affinity of Acusta and Bradybaena is not supported. The hypothesis of colonization via land bridges during the Pleistocene glaciations for the biota of East Asian islands is not supported. Instead, the origin and diversification of the genus Acusta was dated to the late Miocene-Pliocene from an area around North and Northeast China to South China and East Asian islands Three major evolutionary lineages were identified. Two of the major lineages demonstrate distinct evolutionary histories, as sympatric speciation is the major speciation process for the continental clade, while the insular clade originated from founder events. Taiwan functioned as an important source of diversification for species on the East Asian islands possibly through passive dispersal of different mechanisms. The sea level fluctuations caused by the Pleistocene glacial cycles play a role in the subsequent dispersion and diversification of species of the continental clade, such as the more recent range expansion of A. redfieldi from South China to Taiwan and Japan.

Systematic revision of the Taiwanese genus Kurixalus members with a description of two new endemic species (Anura, Rhacophoridae).

Two new species of rhacophorid tree frog were identified in Taiwan. In both new taxa, derived reproductive characteristics of laying eggs in tree holes and oophagous tadpoles are shared with Kurixalus eiffingeri, but they are divergent from each other in molecular genetics, mating calls, and tadpole and adult morphology. The morphological characteristics and the molecular phylogenetic evidence support the hypothesis that the two new species, Kurixalus berylliniris sp. n. and Kurixalus wangi sp. n., are both monophyletic lineages.

A molecular forensic method for identifying species composition of processed marine mammal meats.

We used universal primers designed for the cytochrome oxidase I (CO I) sequence of the order Cetacea and the family Phocidae to prove that meat fritters sold in Taiwan contained meat from two seal, six cetacean, and one pig species. The sequence information for CO I obtained in this study was limited and population genetics data for the eight sampled marine mammalian species was insufficient to deduce where these marine mammals were hunted. Regardless of the geographic origins of the marine mammal flesh, sale and consumption of marine mammals in Taiwan violates the Wildlife Conservation Act. This study provides PCR primers that could enable government testing of suspect meats to curtail the illegal trade in marine mammal products.

Complete mitogenomes of two Puntius in Taiwan: P. semifasciolatus and P. snyderi (Cypriniformes: Cyprinidae).

We sequenced the complete mitochondrial genome of two spotted barbs native to Taiwan: Puntius semifasciolatus and Puntius snyderi. The complete mitochondrial genomes are 16,594 and 16,578 bp in size, respectively. Both of them contain 37 genes coding for 13 proteins, 2 rRNAs, 22 tRNAs, and 1 control region. They share the same gene arrangement pattern that was identical with most vertebrates. Nucleotide sequence divergence (K2P distance) between the two whole mitochondrial genomes was 7.63%. These two spotted barbs show very close relationship based on the comparison of the characters of their mitochondrial genomes.

Complete mitochondrial genome of the Kikuchi's minnow Aphyocypris kikuchii (Teleostei, Cyprinidae).

We have sequenced the complete mitochondrial genome of the Kikuchi's minnow, Aphyocypris kikuchii (Oshima 1919), which is an endemic species to Taiwan. The complete mitochondrial genome is 16,601 bp in size, containing 37 genes coding for 13 proteins, 2 rRNAs, 22 tRNAs, and 1 control region. It has the typical vertebrate mitochondrial gene arrangement. The sequence information could play an important role in resolving the conflict on its current taxonomic position and preservation of genetic resources for helping conservation of the endangered species.

Complete mitochondrial genome of the Moltrecht's minnow, Aphyocypris moltrechti (Teleostei, Cyprinidae), in comparison with A. kikuchii.

We sequenced the complete mitochondrial genome of the Moltrecht's minnow, Aphyocypris moltrechti, which is known as an endemic species to Taiwan. The complete mitochondrial genome is 16,617 bp in size, consisting of 37 genes coding for 13 proteins, 2 rRNAs, 22 tRNAs, and 1 control region. Its gene arrangement pattern was identical with most vertebrates. We compared the mitochondrial genome of A. moltrechti with that of the Kikuchi's minnow, Aphyocypris kikuchii, which had been considered closely related to A. moltrechti within a same genus. Nucleotide sequence divergence (K2P distance) between the two whole mitochondrial genomes was 11.62%. The detailed comparison between the mitochondrial genomes of two species was done.

Complete mitochondrial genome of the Chinese rasbora Rasbora steineri (Teleostei, Cyprinidae).

We describe the complete mitochondrial genome sequence of the Chinese rasbora, Rasbora steineri, which is a small cyprinid distributed in southeastern Asia. The circle genome (16,530 bp) consists of 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes and 1 control region. It has the typical vertebrate mitochondrial gene arrangement.

High-throughput avian molecular sexing by SYBR green-based real-time PCR combined with melting curve analysis.

BACKGROUND: Combination of CHD (chromo-helicase-DNA binding protein)-specific polymerase chain reaction (PCR) with electrophoresis (PCR/electrophoresis) is the most common avian molecular sexing technique but it is lab-intensive and gel-required. Gender determination often fails when the difference in length between the PCR products of CHD-Z and CHD-W genes is too short to be resolved. RESULTS: Here, we are the first to introduce a PCR-melting curve analysis (PCR/MCA) to identify the gender of birds by genomic DNA, which is gel-free, quick, and inexpensive. Spilornis cheela hoya (S. c. hoya) and Pycnonotus sinensis (P. sinensis) were used to illustrate this novel molecular sexing technique. The difference in the length of CHD genes in S. c. hoya and P. sinensis is 13-, and 52-bp, respectively. Using Griffiths' P2/P8 primers, molecular sexing failed both in PCR/electrophoresis of S. c. hoya and in PCR/MCA of S. c. hoya and P. sinensis. In contrast, we redesigned sex-specific primers to yield 185- and 112-bp PCR products for the CHD-Z and CHD-W genes of S. c. hoya, respectively, using PCR/MCA. Using this specific primer set, at least 13 samples of S. c. hoya were examined simultaneously and the Tm peaks of CHD-Z and CHD-W PCR products were distinguished. CONCLUSION: In this study, we introduced a high-throughput avian molecular sexing technique and successfully applied it to two species. This new method holds a great potential for use in high throughput sexing of other avian species, as well.