Polymorphic microsatellites in the Reeves's pheasant developed by cross-species amplification

Cross-species microsatellite amplification is an effective way of obtaining microsatellite loci for closely related taxa in bird species. The Reeves's pheasant, Syrmaticus reevesii, is a vulnerable species endemic to China. To improve population genetics and parentage analysis studies in this species, we obtained nine polymorphic microsatellite markers, in addition to the nine markers previously isolated, from the cross-species amplification of 52 markers. The number of alleles per locus varied between two and 12 with expected heterozygosity ranging from 0.298 to 0.714 (n = 107). The success rates of simulated paternity tests using CERVUS software improved at different confidence levels after adding these markers to the previous ones.     

Polymorphic microsatellites developed by cross-species amplifications in common pheasant breeds

Genetic variability was analysed in two common breeds of pheasant (Phasianus colchicus L. 1758) by means of cross-species amplifications of microsatellite loci: 154 chicken, Gallus gallus and 32 turkey, Meleagris gallopavo, primers were tested for amplification of pheasant DNA. Thirty-six primers (25 specific for chicken and 11 for turkey) amplified pheasant DNA. Fifteen markers yielded specific products and were tested for polymorphism. Eight of them (55%) were polymorphic, with an average polymorphism of two alleles per locus. Specific polymerase chain reaction (PCR) products were sequenced; repeats were found in 11 of the 15 markers, although only two loci showed the same repeat and could be homologous to chicken ones.     

Polymorphic microsatellites identified by cross-species amplifications in the European Coot Fulica atra

We tested the cross-amplification of 26 microsatellites developed for passerines and an additional three developed for Gallinula species in eight European Coots from two populations. Sixteen microsatellite markers successfully amplified, of which nine were polymorphic with 2–6 alleles (mean 3.7 alleles) and an expected heterozygosity (H _e) ranging from 0.375 to 0.805 (mean H _e = 0.589). On average, we found 2.22 alleles/locus and a mean H _e of 0.440 in one nest, and 2.56 alleles/locus and a mean H _e of 0.494 in the other one. These nine polymorphic markers could be of potential use in studies of genetic variability, population structure and reproductive strategy of European Coots.     

四川雉鹑繁殖习性初报

2005～2008年,在四川省雅江县对四川雉鹑(Tetraophasis szechenyii)的繁殖习性,包括巢、卵、孵卵时间、生长量度和日行为节律进行了初步观察.四川雉鹑同时营树上巢和地面巢,以树上巢为主,占68%(n=25);产卵期集中在4月,正常窝卵数2～5枚(n=9),窝卵孵化率为63.89%(n=12);雌鸟在孵卵期每天离巢1次,离巢平均时间(63.0±22.6)min(n=18),孵卵期24～29 d(n=4);150日龄幼鸟的体重接近成体.在繁殖期,四川雉鹑6:30～7:00时从夜栖树上飞下,行至觅食地觅食,17:00时左右返回夜栖地,19:00～19:30时上树夜栖.     

Isolation and characterization of microsatellites in Primula glaucescens Moretti and their cross-species amplification in P. spectabilis Tratt.

The hexaploid herbaceous species Primula glaucescens Moretti and Primula spectabilis Tratt. (2n = 6x = 66) are two endemics of the Italian Pre-Alps protected by national and international laws. In order to plan conservation strategies for natural populations and to study the influence of the latest glaciation on them we developed a set of microsatellites markers for the endangered Primula species: ten primer pairs allowed analysis of polymorphic disomic loci in P. glaucescens samples and seven of them also amplified polymorphic disomic markers in P. spectabilis. Kirsten Wolff and Giorgio Binelli contributed equally to the paper     

Identification and cross-species amplification of EST derived SSR markers in different bamboo species

The availability of expressed sequence data derived from gene discovery programs became an alternative source of mining simple sequence repeat (SSR) and developing inexpensive genetic markers for the crop improvements. In present study, 10 express sequence tags (EST)-SSR markers were identified from Bambusa oldhamii public sequence data base. Transferability to 25 species of Bambusoideae ranged from 30% to 100%. The number of alleles detected per locus ranged from 2 to 10. All the newly identified SSR markers were found to be moderately to highly polymorphic with an average Polymorphic Information Content (PIC) value of 0.54. As these loci represents transcribed region and recorded high level of cross transferability and reliable amplification across the species, demonstrating the utility of these markers for functional and genetic analyses of bamboo species.     

Development and characterization of microsatellite loci in Chinese medicinal plant Akebia trifoliate ssp. australis and cross-species amplification in closely related taxa

Eleven polymorphic microsatellite loci were isolated and characterized from an AC-enriched genomic library of Akebia trifoliate ssp. australis. The number of allele per locus ranged from 3 to 14. The observed heterozygosity and expected heterozygosity at population level were 0.196–1.000 and 0.522–0.902, respectively. In addition, this set of microsatellites produced robust cross-species amplification in other two related taxa, suggesting these microsatellite markers should provide a useful tool for genetic and conservation studies of the Akebia species.     

Isolation and characterization of microsatellite loci in the endangered tree Berchemiella wilsonii var. pubipetiolata and cross-species amplification in closely related taxa

Twenty-eight polymorphic microsatellite loci were developed and characterized for the endangered tree Berchemiella wilsonii var. pubipetiolata. The observed number of allele ranged from two to seven. The ranges of observed and expected heterozygosity were 0.039–1.000 and 0.038–0.816, respectively. In addition, this set of microsatellites produced robust cross-species amplification in other two endangered taxa: Berchemiella berchemiaefolia and Berchemiella wilsonii, suggesting these microsatellite markers should provide a useful tool for genetic and conservation studies of the globally endangered genus Berchemiella.     

Microsatellite markers for <Emphasis Type="Italic">Pseudopanax</Emphasis> trees and their cross-species amplification in the Araliaceae

In order to study hybridisation, taxonomic boundaries and phylogeography in the genus Pseudopanax (Araliaceae), we developed seven novel microsatellite loci from enriched genomic libraries of P. lessonii and P. crassifolius. These loci were characterised in 16 individuals from a single population of P. crassifolius, and displayed 2–11 alleles per locus. Observed heterozygosities ranged from 0.063 to 0.688. Most loci were polymorphic in the closely related Pseudopanax species, and several loci amplified widely across the Araliaceae.     

Successful amplification of rice chloroplast microsatellites from century-old grass samples from the park grass experiment

We report the successful amplification of microsatellite markers for the chloroplast genome from century-old samples of 2 grasses growing in the Park Grass Experiment (PGE):Anthoxanthum ordoratum andFestuca rubra. This opens the possibility of establishing a long-term genetic time series for the PGE, which began in 1856 and is believed to be the oldest ecological experiment in existence. Although the plant samples used were not originally prepared or stored with molecular analysis in mind, the hexadecyltrimethylammonium bromide (CTAB) method of DNA extraction was successfully used. Obtained DNA was degraded but could be amplified by means of PCR. It produced bands around the expected size for chloroplast microsatellite primers derived from rice. When sequenced, bands showed good homology with sequences from rice chloroplast genomes listed in GenBank (accession No X15901).     

Polymorphic microsatellite DNA amplification customized for chimpanzee paternity testing

DNA segments containing GT/AC dinucleotide repeats in the chimpanzee (Pan troglodytes) genome were screened. Thirteen transformedE. coli colonies were identified with the (GT)₁₀ probe to have chimpanzee DNA fragments containing (GT)_n repeats. These potentially polymorphic (variable n) DNA segments were sequenced. Primers for the polymerase chain reaction (PCR) amplifying these DNA segments were designed. Six pairs of primers yielded polymorphic PCR products. Three of them revealed considerable length polymorphisms and heterozygosities in a group of captive chimpanzees. For studies on chimpanzees in the wild and in captivity, these primers should be useful for paternity testing, for investigating genetic variations, and for improving the genetic maintenance of breeding colonies. The strategy adopted in the present study to obtain PCR primers amplifying polymorphic microsatellite DNA segments may well be applicable to almost all eukaryotic organisms.     

Two multiplexed sets of 21 and 18 microsatellites for Phoxinus phoxinus (L.) and Gobio gobio (L.) developed by cross-species amplification

A total of 68 cyprinid microsatellites were screened for cross-species amplification in order to develop three PCR multiplex kits for the Eurasian minnow (Phoxinus phoxinus L.) and the gudgeon (Gobio gobio L.). The multiplex kits cluster 21 and 18 microsatellites for P. phoxinus and G. gobio, respectively. All loci were evaluated using ~80 individuals sampled from three distinct populations. By multiplexing the markers, we aim to decrease genotyping costs and increase time efficiency. These two microsatellite sets represent a valuable tool for ecological, evolutionary, conservation and management issues.     

Isolation of eight microsatellites loci from the saddled bream, <Emphasis Type="Italic">Oblada melanura</Emphasis> and cross-species amplification in two sea bream species of the genus <Emphasis Type="Italic">Diplodus</Emphasis>

We have developed eight new microsatellite markers for the saddled bream (Oblada melanura) from an enriched genome library protocol. All these loci are polymorphic, with mean allelic diversity of 14.75 (range 3–22), and expected and observed heterozygosities from 0.233 to 0.918 and 0.212 to 0.913, respectively. Cross-species tests in two close relatives of the genus Diplodus (D. sargus and D. vulgaris) revealed successful amplifications at 6 out of 8 loci, with means allele number of 6.67 (range 4–10) and 6.50 (range 4–10), respectively. These results are consistent with the close phylogenetic relationships between the three species, indicating this set of primers might proved useful for studying the levels of genetic diversity and population differentiation in these three species and in other phylogenetically close species of the genus Diplodus and Sparus.     

Characterization of 14 novel microsatellite loci in the endangered <Emphasis Type="Italic">Liriodendron chinense</Emphasis> (Magnoliaceae) and cross-species amplification in closely related taxa

Fourteen polymorphic microsatellite loci were isolated and characterized from an AC-enriched genomic library of Liriodendron chinense (Magnoliaceae). The average allele number of these microsatellites was 5.2 per locus, ranging from three to eight. The observed and expected heterozygosity at population level were 0.07–1.00 and 0.10–0.83, respectively. These sets of microsatellites will be useful for studies of population genetic structure of L. chinense and L. tulipifera as well as to estimate fine-scale gene flow rates.     

Development and characterization of 13 novel microsatellite loci from the flat-headed bat (Tylonycteris pachypus) with cross-species amplification in closely related taxa

13 novel microsatellite loci was isolated using the enriched library method from genomic DNA of the flat-headed bat (Tylonycteris pachypus). These loci showed high levels of genetic polymorphism testing on 54 individuals sampled from Guangxi province, China. The number of observed alleles per locus ranged from 2 to 15. The expected and observed heterozygosity values ranged from 0.170 to 0.900 and from 0.185 to 0.944, respectively. One loci departed significantly from Hardy-Weinberg equilibrium (HWE) after Bonferroni correction and no Linkage disequilibrium was found between any pairs of loci. In addition, these loci were tested in the sister species, Tylonycteris robustula, seven loci amplified successfully and were also polymorphic.     

Isolation of novel microsatellites from Jatropha curcas L. and their cross-species amplification

Jatropha curcas L., a member of the Euphorbiaceae, is widely distributed in different parts of the globe. In the present study, 12 microsatellites were isolated from J. curcas and their cross-species amplification was checked in six species of genus Jatropha. Within J. curcas, observed and expected heterozygosities ranged from 0.94 to 0.54 and from 0.95 to 0.56, respectively. Of the 12 loci, five showed significant deviation from Hardy-Weinberg equilibrium. There was no significant linkage disequilibrium detected between any of the loci. The markers isolated in the present investigation will be useful for assessing the population diversity and genetic structure of J. curcas and also in other species of Jatropha.     

Development of single sequence repeat markers for the ant Aphaenogaster senilis and cross-species amplification in A. iberica, A . gibbosa, A. subterranea and Messor maroccanus

Eight polymorphic microsatellite loci were isolated and characterized for Aphaenogaster senilis, a common ant species distributed in the Western Mediterranean. Characterization of 15 individuals form southern Spain showed moderate to high allelic diversity ranging from 2 to 9 alleles per locus. Cross-species tests on 10 individuals of A. iberica, A. gibbosa, A. subterranea and Messor maroccanus revealed successful amplification for most loci. This set of markers can be useful for population genetic studies and might even prove useful in other phylogenetically close species of the subfamily Myrmicinae.