A microarray system for Y chromosomal and mitochondrial single nucleotide polymorphism analysis in chimpanzee populations

Chimpanzee populations are diminishing as a consequence of human activities, and as a result this species is now endangered. In the context of conservation programmes, genetic data can add vital information, for instance on the genetic diversity and structure of threatened populations. Single nucleotide polymorphisms (SNP) are biallelic markers that are widely used in human molecular studies and can be implemented in efficient microarray systems. This technology offers the potential of robust, multiplexed SNP genotyping at low reagent cost in other organisms than humans, but it is not commonly used yet in wild population studies. Here, we describe the characterization of new SNPs in Y-chromosomal intronic regions in chimpanzees and also identify SNPs from mitochondrial genes, with the aim of developing a microarray system that permits the simultaneous study of both paternal and maternal lineages. Our system consists of 42 SNPs for the Y chromosome and 45 SNPs for the mitochondrial genome. We demonstrate the applicability of this microarray in a captive population where genotypes accurately reflected its large pedigree. Two wild-living populations were also analysed and the results show that the microarray will be a useful tool alongside microsatellite markers, since it supplies complementary information about population structure and ecology. SNP genotyping using microarray technology, therefore, is a promising approach and may become an essential tool in conservation genetics to help in the management and study of captive and wild-living populations. Moreover, microarrays that combine SNPs from different genomic regions could replace microsatellite typing in the future.     

PERMANENT GENETIC RESOURCES: Characterization of eight microsatellite loci in the woolly mouse opossum, Micoureus paraguayanus, isolated from Micoureus demerarae

Eight novel microsatellite markers were isolated from the woolly mouse opossum from the Amazon Forest in Peru, Micoureus demerarae, using a partial genomic DNA library and an enrichment protocol. These loci were polymorphic in M. demerarae and Micoureus paraguayanus populations from the Atlantic Forest in Brazil with the number of alleles ranging from two to 23. Those eight loci plus another five already described for M. paraguayanus will allow for the evaluation of genetic diversity of populations from the 'Rio Doce' Park, one of the last Atlantic Forest fragments in Minas Gerais state, Brazil.     

Isolation and identification of eight microsatellite loci in the Cherokee darter (Etheostoma scotti) and their variability in other members of the genera Etheostoma, Ammocrypta, and Percina

The Cherokee darter Etheostoma scotti is a federally threatened fish endemic to the Etowah River system of northwest Georgia. In order to analyse the population structure and genetic diversity of this fish, eight tetranucleotide microsatellite genetic markers were developed. The marker set was applied to 13 additional darter species to test cross-species amplification and polymorphism. Successful amplification was obtained for all eight loci in each of the 13 other species of darters, with between seven and eight polymorphic loci per species.     

Microsatellite markers for eastern hemlock (Tsuga canadensis)

We describe polymerase chain reaction primer pairs and reaction conditions for amplification of 15 microsatellite loci from eastern hemlock (Tsuga canadensis). The primers were tested on 23 individuals from a natural population in southwestern North Carolina, USA. These primers yielded an average of 5.9 alleles per locus (range of 2-14), an average observed heterozygosity of 0.45 (range 0.14-0.73), and an average polymorphic information content of 0.54 (range 0.28-0.86). In addition, eight of the primer pairs were found to amplify microsatellite loci in one or more additional species of Tsuga.     

Isolation and characterization of 12 dinucleotide microsatellites in the European eel, Anguilla anguilla L., and tests of amplification in other species of eels

Twelve polymorphic dinucelotide microsatellites in the freshwater eel Anguilla anguilla L. were isolated and characterized. Genetic diversity was assessed in eels from Lake Constance, Germany. Allele numbers ranged from five to 26 per locus with observed heterozygosities between 0.125 and 0.875. A portion of locus AangCT77 aligns with a transcribed region of the zebrafish gene crystallin beta B2. Cross-species amplification of most markers was possible for nine other Anguilla eel species. The newly developed primer pairs will facilitate population and conservation genetic studies in order to refine the understanding of the subtle population genetic structure typical of eels, and to identify interspecies admixture due to global trade.     

Development of polymorphic microsatellite markers for the New Zealand black stilt (Himantopus novaezelandiae) and cross-amplification in the pied stilt (Himantopus himantopus leucocephalus)

Eight polymorphic microsatellite primer pairs were developed for the critically endangered New Zealand black stilt, Himantopus novaezelandiae, representing the first microsatellite markers available for birds in the family Recurvirostridae. The number of alleles ranged from two to four per locus. Observed and expected heterozygosities ranged from 0.30 to 0.80 and from 0.37 to 0.70, respectively. All eight loci were polymorphic in the related species Himantopus himantopus leucocephalus, indicating these primer pairs may be useful for additional taxa in the globally distributed genus Himantopus.     

Development of a new vitrification solution, VSL, and its application to the cryopreservation of gentian axillary buds

Vitrification methods are convenient for cryopreserving plant specimens, as the specimens are plunged directly into liquid nitrogen (LN) from ambient temperatures. However, tissues and species with poor survival are still not uncommon. The development of vitrification solutions with high survival that cover a range of materials is important. We attempted to develop new vitrification solutions using bromegrass cells and found that VSL, comprising 20% (w/v) glycerol, 30% (w/v) ethylene glycol, 5% (w/v) sucrose, 10% (w/v) DMSO and 10 mM CaCl₂, gave the highest survival following cryopreservation, as determined by fluorescein diacetate staining. However, the cryopreserved cells showed little regrowth, for unknown reasons. To check its applicability, VSL was used to cryopreserve gentian axillary buds and the performance was compared with those of conventional vitrification solutions. Excised gentian stem segments with axillary buds (shoot apices) were two-step precultured with sucrose to induce osmotic tolerance prior to cryopreservation. Gentian axillary buds cryopreserved using VSL following the appropriate preculturing approach exhibited 78% survival (determined by the regrowth capacity), which was comparable to PVS2 and PVS1 and far better than PVS3. VSL had a wider optimal incubation time (20–45 min) than PVS2 and was more suitable for cryopreserving gentian buds. The optimal duration of the first step of the preculture was 7–11 days, and preculturing with sucrose and glucose gave a much higher survival than fructose and maltose. VSL was able to vitrify during cooling to LN temperatures, as glass transition and devitrification points were detected in the warming profiles from differential scanning calorimetry. VSL and its derivative, VSL+, seem to have the potential to be good alternatives to PVS2 for the cryopreservation of some materials, as exemplified by gentian buds. Mitsuteru Suzuki, Pramod Tandon and Masaya Ishikawa contributed equally to the work.     

Differences of ecological functions inside and outside the wetland nature reserves in Sanjiang Plain, China

Conservation effectiveness of wetland nature reserves is determined by both the management intensity inside the reserves and the hydrological status outside the reserves. Therefore, differences of ecological functions inside and outside the reserves are an integrated indictor for assessing conservation effectiveness. Based on the land use map created from Landsat-TM satellite image and 1:50000 Digital Elevation Model (DEM) data in the study area, the catchments that belong to the wetland reserves were determined as their hydrological sensitive zones by SWAT (Soil and Water Assessment Tool) hydrological model. The ecological function indices of wetland reserves and their corresponding hydrological sensitive zones were calculated through expert consultation and value assessment on wetland ecosystem service functions. The results showed that the ecological functions of national wetland reserves were better than those of local reserves in general. However, the wetland ecological functions of hydrological sensitive zones of the former were not always better than those of the latter. Meanwhile, clustering analysis showed that the wetland ecological functions of several adjacent reserves in Nongjiang-Bielahonghe watershed were similar. But correlation analysis found that there existed a remarkable positive correlation between the wetland ecological function indices of local reserves and their hydrological sensitive zones.     

达赉湖自然保护区疣鼻天鹅繁殖行为初步观察

2006年8～11月和2007年4～8月,在内蒙古达赉湖国家级自然保护区对疣鼻天鹅(Cygnus olor)的繁殖数量、繁殖行为及迁飞时间进行研究.结果显示,疣鼻天鹅通常4月初迁来,10月底迁离,居留期长达196 d左右(n=3).4月下旬开始求偶,时间一般为上午进行,没有固定求偶地点,每次求偶所需时间平均172 s(n=8).主要依靠炫耀行为来保卫和标记领域.2006与2007年的繁殖种群数量均为12只(6对),2006年育成幼鸟数分别为4、2、4、3、6、3只;2007年仅有3对繁殖成功,幼鸟数分别为4、4、5只.     

江西齐云山自然保护区两栖爬行动物资源调查与区系分析

为掌握江西齐云山自然保护区野生动物资源现状,促进自然保护区的有效管理和建设,作者于2007年1月10～19日、5月16～24日、7月26日～8月8日,前后3次对该保护区两柄爬行动物资源进行了实地考察.调查结果显示,齐云山保护区现已记录72种两栖爬行动物,隶属5目18科.其中两栖纲2目7科24种,爬行纲3目11科48种.齐云山保护区动物地理区划属东洋界华中区东部丘陵平原亚区,其区系组成以60种东洋界种类占明显优势,其余12种均为广布种,无古北界物种分布.大眼斜鳞蛇福建亚种(Pseudoxenodon macrops fukienensis)和寒露林蛙(Rana hanluica)为江西省两栖爬行动物新纪录,福建钝头蛇(Pareas stanleyi)为江西省赣南地区爬行动物新纪录.齐云山保护区是海南棱蜥後(Tropidophorus hainanus)和福建华珊瑚蛇(福建丽纹蛇)(Sinomicrurus kettoggi)在江西省的第二个发现地,是海南闪鳞蛇(Xenopeltis hainanensis)在江西省的第三个采集地.通过平均动物区系相似性分析发现,齐云山地理位置独特,是华中区和华南区物种重要的扩散过渡地域.最后对齐云山保护区的保护与管理工作提出了建议.