Isolation of microsatellite markers in mungbean,Vigna radiata

A simple and rapid method for isolating microsatellite loci in mungbean, Vigna radiata, based on the 5′‐anchored polymerase chain reaction technique revealed 23 microsatellite loci and six cryptically simple sequence repeats. We report on the characterization of seven polymorphic microsatellite loci in V. radiata. The number of alleles per locus ranged from 2 to 5 while the observed heterozygosity ranged from 0 to 0.9048. These markers should prove useful as tools for detecting genetic variation in mungbean varieties for germplasm management and crossbreeding purposes.     

Isolation,characterization and multiplex genotyping of raccoon tetranucleotide microsatellite loci

We have isolated and characterized 12 polymorphic tetranucleotide microsatellite loci in the raccoon (Procyon lotor). Three multiplex panels comprising the 12 loci were developed and 80 individuals from southeastern and western Ontario were genotyped; allele sizes were assigned without difficulty. One locus isolated was identified as an X‐linked marker. The number of alleles per locus ranged from six to 25 with the average heterozygosity ranging from 0.674 to 0.925. These loci will be used to characterize raccoon population structure across North America, and the data used to further understand the spread of raccoon rabies.     

Isolation and characterization of swift fox tetranucleotide microsatellite loci

Primers have been developed for nine polymorphic tetranucleotide microsatellite loci isolated from the swift fox (Vulpes velox). These can be amplified in three multiplex reactions and all loci can be scored easily and without ambiguity. The number of alleles per locus ranged from three to 14 and the overall heterozygosities ranged from 0.54 to 1.00. All loci were in Hardy‐Weinberg equilibrium, but two loci were in linkage disequilibrium. These loci will be used to monitor endangered swift fox populations on the Canadian prairies.     

New microsatellite markers isolated from mungbean (Vigna radiata (L.) Wilczek)

In this study, we reported the isolation and analysis of new polymorphic microsatellites in mungbean (Vigna radiata (L.) Wilczek). Twelve out of 210 primer pairs screened in 30 mungbean accessions gave polymorphism. The polymorphic markers detected two to three alleles per locus with an average of 2.08. Observed heterozygosity varied from 0 to 0.133, while expected heterozygosity ranged from 0.095 to 0.498. Tests for Hardy-Weinberg equilibrium (HWE) and pairwise linkage disequilibrium of the polymorphic loci revealed that all loci except MB-SSR14 significantly departed from HWE and four pairwise combinations, viz. MB-SSR14 vs. MB-SSR42, MB-SSR42 vs. MB-SSR87, MB-SSR114 vs. MB-SSR121, and MB-SSR175 vs. MB-SSR231 significantly deviated from linkage disequilibrium. The markers are being used to study genetic diversity and genome mapping of mungbean.     

Isolation and characterization of 11 tetranucleotide microsatellite loci in the Egyptian mongoose (Herpestes ichneumon)

We report the isolation of 11 polymorphic tetranucleotide microsatellite loci in the Egyptian mongoose (Herpestes ichneumon). In a sample of 27 individuals, we observed between 4 and 7 alleles per locus and their observed and expected heterozygosities ranged from 0.37 to 0.85 and from 0.44 to 0.79, respectively. All genotypic frequencies conformed to Hardy-Weinberg equilibrium expectations and there were no instances of linkage disequilibrium detected between pairs of loci.     

Isolation of 10 tetranucleotide microsatellite loci in the blackcap (Sylvia atricapilla)

We isolated 10 polymorphic microsatellite DNA loci from the blackcap (Sylvia atricapilla) and optimized them for future studies of population differentiation in populations with different migration strategies in southwestern Germany. The loci were screened for polymorphism using 178 individuals from two populations in Germany and Spain. The primers amplified highly variable loci characterized by two to 19 alleles per locus and their observed and expected heterozygosities range from 0.47 to 0.81 and from 0.50 to 0.91, respectively.     

Isolation of ten tetranucleotide microsatellite loci in the Northern Wheatear (Oenanthe oenanthe)

We isolated 10 polymorphic microsatellite DNA loci from the Northern Wheatear (Oenanthe oenanthe) and optimized these for future studies in population genetics and behavioural ecology. The loci were screened for polymorphism using 107 individuals from one population in Germany. The primers amplified loci with high numbers of alleles ranging from two to 20 alleles per locus.     

Isolation and characterization of seven tetranucleotide microsatellite loci from Atlantic northern bluefin tuna Thunnus thynnus thynnus

Microsatellite‐enriched genomic libraries were obtained from the Atlantic northern bluefin tuna, Thunnus thynnus thynnus and seven tetranucleotide markers were successfully isolated and characterized from this library. These markers were found to have between 1 and 17 alleles in Atlantic northern bluefin tuna and heterozygosity ranged from 0 to 0.85. No deviations from the expectation of Hardy‐Weinburg equilibrium were found for any marker. Several of these markers amplify reliably in other tuna species.     

Characterization of tetranucleotide microsatellite loci in the African Savannah Elephant (Loxodonta africana africana)

Most African elephant (Loxodonta africana africana) populations are isolated and thus threatened by a loss of genetic diversity. As a consequence, genetic analysis of African elephant populations will play an increasing role in their conservation, and microsatellite loci will be an important tool in these analyses. Previously published sets of polymorphic microsatellites developed for African elephants are all dinucleotide repeats, which are prone to typing error. Here, we characterize 11 tetranucleotide microsatellite loci in the African elephant. All loci were polymorphic in 32 faecal samples and two tissue samples from 33 individual African savannah elephants.     

Purification and characterization of Cu-Zn superoxide dismutases from mungbean (Vigna radiata) seedlings

Mungbean contains three isoenzymes of superoxide dismutase designated isoenzyme I, II and III. The two cytosolic superoxide dismutases (I and II) were purified to homogeneity by ammonium sulphate fractionation, ion exchange chromatography on diethylaminoethyl cellulose, gel filtration and preparative polyacrylamide.gel electrophoresis. The molecular weights of isoenzyme I and isoenzyme II were determined to be 33,000 and 31,600 respectively. The subunit molecular weight was approximately 16,000 indicating that the isoenzymes contained two identical subunits. The ultra-violet absorption spectra revealed a maximum at 258–264 nm for the two isoenzymes. Superoxide dismutase I and II were inhibited to different extents by metal chelators. Isoenzyme I was more sensitive to inhibition by cyanide and azide, while isoenzyme II was more susceptible to inhibition by diethyldithiocarbamate ando-phenanthroline. Both the isoenzymes exhibited similar denaturation profiles with heat, guanidinium chloride and urea. The denaturation with urea and guanidinium chloride was reversible. The two copper-zinc enzymes were more stable towards thermal inactivation compared to manganese and iron superoxide dismutases from other sources. The results indicate that the two isoenzymes differ from each other only with respect to charge and sensitivity towards metal chelators.     

Isolation and characterization of seven microsatellite loci in Chioglossa lusitanica (Urodela: Salamandridae)

Seven polymorphic microsatellite markers were isolated and characterized for golden‐striped salamander, Chioglossa lusitanica (Bocage 1864), a salamandrid endemic to the northwestern part of the Iberian Peninsula, from genomic libraries enriched for (GATA)₈ (GACA)₈ (ATG)₉ and (CA)₁₆. These loci were screened in 37–51 individuals from two populations. The number of alleles per locus ranged from five to 19. Heterozygosity ranged from 0.241 to 1.0. The high level of polymorphism revealed by these loci will be extremely useful for the study of population structure and evolutionary history of this species.     

Eight new tetranucleotide microsatellite loci for the agile frog (Rana dalmatina)

We describe eight new polymorphic tetranucleotide microsatellite loci isolated from the agile frog (Rana dalmatina). In 25 individuals from the Nature Reserve Lüneburger Heide (Lower Saxony, Germany), the number of alleles per locus ranged from four to nine and average observed heterozygosities from 69.1% to 80.7%. No evidence for linkage disequilibrium was found and none of the loci showed significant deviation from Hardy-Weinberg expectations. These microsatellite DNA markers are suitable tools for addressing population genetics issues in this endangered species.     

Isolation and characterization of polymorphic tetranucleotide microsatellite loci in the Fire salamander Salamandra salamandra (Amphibia: Caudata)

Ten tetranucleotide and one dinucleotide polymorphic microsatellite loci were cloned and characterized for the Fire salamander (Salamandra salamandra) from 34 populations in Germany. A high genetic diversity (5–22 alleles per locus) and heterozygosity (40.6–95.2%) were observed for these markers. Chord distances for population comparisons of the western evolutionary recolonization lineage in the area near Cologne ranged from 0.139 to 0.366, whereas population comparisons between the western and eastern lineage ranged from 0.541 to 0.670. When compared with classical isolation methods, a sufficient number of polymorphic microsatellites can be obtained for the Fire salamander only from specially enriched sublibraries.     

Isolation and characterization of microsatellite loci in the Caribbean gorgonian Pseudopterogorgia elisabethae

We report on the isolation and characterization of five polymorphic microsatellites in the gorgonian Pseudopterogorgia elisabethae from genomic DNA‐enriched libraries. Forty‐four microsatellites were screened from the libraries with the oligonucleotide probe (CA)₁₂. Five of the screened microsatellites were polymorphic. The levels of polymorphism found in 50 genotyped individuals from a single population suggest that microsatellites are useful tools for the study of genetic variation in gorgonians. These are the first microsatellite loci reported from any gorgonian species.     

Isolation and characterization of microsatellite loci in Tripterygion delaisi

We isolated 49 microsatellite loci from a genomic library of Tripterygion delaisi×anthosoma enriched for CA and GA repeats. Ten loci were screened in 30 individuals with high numbers of alleles per locus (averaging 15.5 ± 2.86) and observed heterozygosity (averaging 0.765 ± 0.052). No deviations from Hardy–Weinberg expectations were detected. These highly polymorphic markers will be useful in determining the spatial patterns of genetic diversity between and within subspecies of Tripterygion delaisi.     

Isolation and characterization of microsatellite loci for storm-petrels

Primers were developed for 10 microsatellite loci for two species of Oceanodroma storm-petrels. Variability was tested in 27 O. castro and 22 O. monteiroi from the Azores, and 24 O. leucorhoa from Norway. At least six loci amplified reliably and were polymorphic in each species. The number of alleles per locus averaged 4.6, and observed heterozygosities averaged 0.41. Most primers also yielded polymerase chain reaction products in O. tethys, O. hornbyi and Pterodroma phaeopygia. These loci are being used to assay population genetic structure in storm-petrels.     

Differences in responses to iron deficiency among various cultivars of mungbean (Vigna radiata (L.) Wilczek)

Ten mungbean cultivars were evaluated for their resistance to iron deficiency in view of chlorosis symptoms, plant growth and seed yield under field conditions on a calcareous soil in Thailand. The KPS2 cultivar was highly susceptible; the KPS1, PSU1 and Pag-asa 1 cultivars were somewhat susceptible; the VC1163B cultivar was moderately tolerant; the CN36, CN60, UT1 and CNM-I cultivars were tolerant; and the CNM8509B cultivar was very tolerant to iron deficiency. Foliar application of a solution of 5 g L^-1 ferrous sulphate was effective in correcting chlorosis that was induced by iron deficiency, and it enhanced both the growth and the yield of susceptible cultivars. Compared with the susceptible cultivar KPS2, the tolerant cultivar UT1 had a greater ability to lower the pH of the nutrient solution in response to iron deficiency. The root-associated Fe³⁺-reduction activity of UT1 that had been grown in -Fe medium was similar to that of the plants grown in +Fe medium when the acidification of the medium occurred. Acidification of the medium in response to iron deficiency might contribute to the efficient solubilization of iron from calcareous soils, and it related more closely to the resistance to iron deficiency than Fe³⁺ reduction by roots in mungbean cultivars.     

Isolation and characterization of nine microsatellite loci in Podarcis bocagei (Squamata: Lacertidae)

Nine dinucleotide microsatellite loci were developed through an enrichment protocol for Bocage's wall lizard, Podarcis bocagei Seoane 1884, a lacertid endemic to the Iberian Peninsula. Nineteen primer pairs were designed and tested. From these, nine loci yielded satisfactory results and were screened on 15–19 individuals. These loci revealed a high level of polymorphism (8–15 alleles) and heterozygosity (0.611–0.947) and will certainly be useful in the study of population structure and evolutionary history of this species.