Molecular evidence on the species status and phylogenetic relationships of Spiranthes parksii, an endangered orchid from Texas

New genetic information on the taxonomic status of an endangered species may have strong implications for its continued protection and that of other listed taxa with questionable species status. While analyzing a more comprehensive project on the molecular phylogeny of North American Spiranthes (Orchidaceae), lack of genetic differentiation between endangered S. parksii, endemic only to Texas, and a common local congener, S. cernua, was discovered. To confirm these results in a comparative context, we focused parsimony analyses on a subset of data from local Spiranthes and the same congeners found elsewhere, expanded by additional local samples of the two focal species. Four DNA sequence segments (two chloroplast, one mitochondrial, one nuclear ribosomal) totaling 3191 base pairs were used separately and together to verify that S. parksii is nested within the same clade as S. cernua, and thus likely to be the same species. Our results concur with another recent investigation using AFLP and microsatellite data that also suggests S. parksii is not unique genetically. Therefore, based on empirical data and the phylogenetic species concept, endangered S. parksii is merely an aberrant local phenotype of and a synonym for S. cernua sensu lato. Our results should be considered during the current review of this orchid’s listing status, despite some recent policy changes on consideration of genetic data after listing.     

Behaviour of Malay peacock pheasant Polyplectron malacense (Aves: Phasianidae)

The breeding behaviour of Malaysian peacock pheasants Polyplectron malacense was studied in West Malaysian rain forest. Males maintained territories, within which some called from cleaned display areas. There were calling interactions between males but no stable dominance hierarchy. Females approached calling males at these display areas for mating. There were year to year variations in males' behaviour, apparently related to the food supply, and it is suggested that food levels may shift the balance between alternative mating strategies.     

物种濒危等级划分与物种保护

介绍了国际与国内濒危物种等级最新标准。探讨了濒危物处等级的划分标准存在的问题和物种的保护优先序。介绍了确定物种保护优先序时的两种不同观点。     

Reproductive biotechnologies for endangered mammalian species

Assisted reproductive techniques (gamete cryopreservation, artificial insemination, embryo transfer, and in vitro fertilization) allow to propagate small fragmented populations of wild endangered species or domestic breeds. There are the best way for producing several offspring from selected genitors in order to avoid inbreeding depression. However, few mammalian species have been well studied for their reproductive biology whereas huge differences have been observed between these species. Furthermore, materials, methods and experimental designs have to be adapted for each case and each limiting factor (wildness, poor quantity of biological material, disparate locations). Genome resource banking is currently arising and the most applied reproductive biotechnology remains artificial insemination. Assisted reproductive techniques currently developed in domestic species (intracytoplasmic sperm injection, nuclear transfer) may offer new opportunities for the propagation of endangered species.     

Molecular evidence for hybridization of species in the genus Gallus except for Gallus varius

A phylogenetic tree for fowl including chicken in the genus Gallus and based on mitochondrial D-loop analysis further supports the hypothesis developed from morphology and progeny production that red junglefowl (RJF) is the direct ancestor of the chicken. The phylogenetic positions of the chicken and the other fowl species in the genus Gallus are of great importance when considering maintenance and improvement of chicken breeds through introgression of genetic variation from wild-type genomes. However, because the phylogenetic analysis based on the DNA sequences is not sufficient to conclude the phylogenetic positions of the fowls in the genus, in the present study, we have determined sequences of whole mitochondrial DNA (mtDNA) and two segments of the nuclear genome (intron 9 of ornithine carbamoyltransferase, and four chicken repeat 1 elements) for the species in the genus Gallus. The phylogenetic analyses based on mtDNA sequences revealed that two grey junglefowls (GyJF) were clustered in a clade with RJFs and chicken, and that one GyJF was located in a remote position close to Ceylon junglefowl (CJF). The analyses based on the nuclear sequences revealed that alleles of GyJFs were alternatively clustered with those of CJF and with those of RJFs and chicken. Alternative clustering of RJF and chicken alleles were also observed. These findings taken together strongly indicate that inter-species hybridizations have occurred between GyJF and RJF/chicken and between GyJF and CJF.     

Molecular evidence for the non-monophyletic status of Naidinae (Annelida, Clitellata, Tubificidae)

Naidinae (former Naididae) is a group of small aquatic clitellate annelids, common worldwide. In this study, we evaluated the phylogenetic status of Naidinae, and examined the phylogenetic relationships within the group. Sequence data from two mitochondrial genes (12S rDNA and 16S rDNA), and one nuclear gene (18S rDNA), were used. Sequences were obtained from 27 naidine species, 24 species from the other tubificid subfamilies, and five outgroup taxa. New sequences (in all 108) as well as GenBank data were used. The data were analysed by parsimony and Bayesian inference. The tree topologies emanating from the different analyses are congruent to a great extent. Naidinae is not found to be monophyletic. The naidine genus Pristina appears to be a derived group within a clade consisting of several genera (Ainudrilus, Epirodrilus, Monopylephorus, and Rhyacodrilus) from another tubificid subfamily, Rhyacodrilinae. These results demonstrate the need for a taxonomic revision: either Ainudrilus, Epirodrilus, Monopylephorus, and Rhyacodrilus should be included within Naidinae, or Pristina should be excluded from this subfamily. Monophyly of four out of six naidine genera represented by more than one species is supported: Chaetogaster, Dero, Paranais, and Pristina, respectively.     

Cryopreservation of semen from endangered pheasants: the first step towards a cryobank for endangered avian species

In order to improve the genetic management of bird species within the European Endangered Programs (EEP), a research project on artificial insemination and cryopreservation of Galliformes semen has been developed. The aim of the program is to create a sperm cryobank for threatened bird species. During this study, semen was collected from 17 pheasant species and specific characteristics of ejaculates were analyzed (volume, sperm concentration, motility, pH). Artificial insemination with fresh semen was performed in nine species and with frozen semen in eight species. Inseminations with frozen and thawed semen were made in 17 species. Viability of fresh and frozen semen was assessed in vitro using double stains, eosin and nigrosin. The effect of pH (7-8.5) on viability of fresh and frozen/thawed spermatozoa was also studied. Chicks hatched in eight and three species after insemination with fresh and frozen/thawed semen, respectively. Species varied widely in semen viability: 1-30% of spermatozoa survived freezing and thawing. There was a negative correlation between the viability of frozen spermatozoa and semen pH. In our experimental conditions, the pH of diluents had no effect on semen viability. However, semen with the highest pH had the lowest quality after freezing and thawing. These experiments demonstrated the feasibility of using a very simple and inexpensive method to achieve artificial insemination and cryopreservation of semen in endangered pheasant species.     

The future of the US endangered species act

The United States Endangered Species Act of J973 (ESA) is the strongest tool for protecting plants and animals in the US and has served as a model of species protection for many other nations. Because the goals of the Act - to conserve all endangered and threatened species in the US and the ecosystems upon which they depend - are widely supported among US environmentalists and biologists, it is commonly believed that these groups offer united support for the legislation. Within the US, however, vigorous debate ensues among conservation biologists as to the effectiveness of the species-oriented approach of the Act.     

Microsporogenesis in the endangered species Cupressus dupreziana A. Camus: evidence for meiotic defects yielding unreduced and abortive pollen

To understand the reproductive biology of Cupressus dupreziana A. Camus (Cupressaceae), a highly endangered Mediterranean conifer, the processes of microsporogenesis and pollen differentiation were investigated cytologically. Pre-meiotic development proved to be similar to the coniferous pattern: the microsporangia differentiated sporogenous tissue in which microsporocytes separated and underwent meiosis. As the meiotic steps proceeded, unexpected irregularities were observed concerning chromosomal and nuclear behaviour. This mainly included: abnormal chromosome segregation and cytokinesis, and nuclear fusion of the meiotic products. The result was the formation, in the same microsporangium, of heterogeneous microspore populations arranged in monads, dyads, triads, tetrads, and polyads, and cytoplasts giving rise to pollen grains of different sizes. This indicates that in C. dupreziana both abortive and unreduced pollen grains are generated. The significance of the finding is discussed in relation to reproductive biology and vulnerability to extinction.     

Semen cryopreservation and the conservation of endangered species

Currently, more than 16,000 plant and animal species are imminently threatened by extinction, often as a direct consequence of anthropogenic influences. One of the measures to halt that process is genetic resource banking. This short review focuses on mammal sperm cryopreservation in combination with assisted reproduction techniques. It summarizes general problems, recent developments, and currently applied protocols and gives an overview of hitherto existing successes of assisted reproduction measures in wild animals in the light of conservation efforts.     

Molecular evidence for cryptic species within Cylicostephanus minutus (Nematoda: Strongylidae)

The nucleotide sequences of the first and second internal transcribed spacers of nuclear ribosomal DNA were determined for adults of Cylicostephanus minutus from different geographical origins. The lengths of first and second internal transcribed spacer sequences ranged from 370 to 372bp and 215 to 216bp, respectively. Pairwise sequence comparisons revealed that some individuals of C. minutus had identical first and second internal transcribed spacer sequences, whereas others differed by 3.0% and 7.4% in their first and second transcribed spacers, respectively. Some individuals with sequence differences originated from the same host. The levels of difference within C. minutus were higher than that between the morphologically distinct species, Cylicostephanus goldi and Cylicostephanus longibursatus (0.8% for the first internal transcribed spacer and 3.8% for the second internal transcribed spacer). The data provide support for the proposal that C. minutus represents a complex of at least two species. In order to study the population genetic structure of C. minutus, a PCR-linked single-strand conformation polymorphism technique was also established.     

Molecular phylogeographic evidence for multiple shifts in habitat preference in the diversification of an amphipod species

An earlier study of ours demonstrated polygenic control of habitat preference between sympatric populations of an estuarine amphipod (Stanhope et al. 1992). Knowledge of the ecological history in this estuary suggested that two new and distinct habitat types (wood debris and Fucus ) had been formed immediately adjacent to each other, in an area which was previously the habitat type common to the rest of the estuary (bank). This suggested the possibility that the ancestral population had been split into two resource specialists (disruptive selection on habitat preference). The genetic relatedness of these three populations (the proposed ancestor and the two proposed descendants) and six others occupying the same three habitat types were investigated on a regional geographic scale, using an extensive set of genomic DNA RFLPs. These data were combined with measures of habitat preference (including genetic tests) in the additional populations. A very strongly supported phylogeographic tree, unequivocally supported a shift in habitat preference in the wood-debris population of this estuary. The data did not, however, support the hypothesis of disruptive selection on the ancestor (bank) with a consequent split into two resource specialists. Instead, it clearly indicated that the occupants of the Fucus habitat type were members of a habitat specific race, and thus their presence in this estuary reflected the expansion of an old resource base. Furthermore, the combined RFLP and habitat preference data, for all nine populations, indicated that the same polygenically based shift in habitat preference that had occurred in the original estuary had occurred independently, in another estuary, 700 km removed, that had similar environmental circumstances.     

A triple-primer PCR method for sexing endangered caprine species

Molecular sexing is becoming an essential technique in understanding the sexual structure and dynamics of natural populations. Herein, we report on a triple-primer PCR method based on the last introns of the ZFX/Y alleles for sex identification in Bovidae, and its successful application to five endangered caprine species. The male samples generated a ~230 bp ZFX-specific fragment and a ~140 bp ZFY-specific fragment, and the female samples only generated the ~230 bp fragment. This method is very sensitive to the Y-linked fragment, thus effectively avoiding false negatives. Genomic DNA extracted from well preserved tissues, non-invasive samples and smoked meat are all usable for analysis with this method.     

Micropropagation of the endangered species Malus niedzwetzkyana for conservation biodiversity in Kazakhstan

In Vitro Cellular & Developmental Biology - Plant - Biodiversity conservation requires advanced and effective ex situ plant propagation techniques. The present study was conducted to optimize...     

Arabia's tallest trees: ecology,distribution and conservation status of the regionally endangered tree species Mimusops laurifolia

Mimusops laurifolia (Forssk.) Friis is the largest tree species in the Arabian Peninsula, and a characteristic species of the threatened southwest Arabian valley forest habitat. In order to understand the distribution and conservation status of M. laurifolia, this paper collates the historical records of M. laurifolia and compares them with recent survey data. This comparison highlights that the abundance of M. laurifolia has dramatically declined at sites that were previously considered important for this species. In Arabia, approximately 100 individuals of M. laurifolia remain at eleven localities. From their size, it is likely that many of these are extremely old trees, but there is currently little regeneration. The main threats to this species are the loss of valley forest habitat through overgrazing and road construction, overexploitation of M. laurifolia for wood as well as climate change. Based on these findings, we recommend that M. laurifolia should be classified as Endangered (EN) in the region on the IUCN red list. We recommend some basic conservation measures and identifies an urgent need for further research on the distribution, propagation and restoration of this species.