Isolation and characterization of microsatellite loci in the Brazilian orchid <Emphasis Type=Italic>Epidendrum fulgens</Emphasis>

Epidendrum fulgens has a patchy distribution along the Atlantic Rainforest in the Brazilian coast, due to the destruction of its native habitat. Here, we report on both the development of nine new microsatellite markers isolated from this species and the characterization of their allele variability in two distant and unrelated populations. The number of alleles observed for each locus ranged from 2 to 17 with an average of 6.4 alleles per locus. These microsatellites should be valuable tools for studying the effect of habitat fragmentation on the genetic structure of E. fulgens populations.     

Isolation and characterization of twenty-one polymorphic microsatellite loci in the Tibetan macaque (<Emphasis Type=Italic>Macaca thibetana</Emphasis>)

Twenty-one microsatellite loci were isolated from AC-enriched library of Tibetan macaque (Macaca thibetana). The number of alleles at the 21 microsatellite loci ranged from 8 to 15, with an average of 12.2 per locus. Polymorphism information content (PIC) ranged from 0.805 to 0.910 with an average of 0.873. The observed and expected heterozygosities ranged from 0.208 to 0.792 and from 0.843 to 0.938, respectively. These microsatellite loci will be useful for future studies that relate to the genetic diversity and population structure of Tibetan macaque.     

Isolation and characterization of twelve polymorphic microsatellite loci in the buff-throated partridge (<Emphasis Type=Italic>Tetraophasis szechenyii</Emphasis>)

Twelve polymorphic microsatellite markers were isolated from an AC-enriched genomic library of Buff-throated partridge (Tetraophasis szechenyii). The allele number of these loci ranged from three to 13 (average 7.75 per locus) in tested individuals. Polymorphism information content ranged from 0.532 to 0.882 with an average of 0.721. Average observed and expected heterozygosities were 0.559 (range from 0.240 to 0.792) and 0.772 (range from 0.621 to 0.910), respectively. These microsatellite markers will be useful for the assessment of genetic diversity, relatedness identification of family and analysis of population structure in wild buff-throated partridge.     

Isolation and characterization of microsatellite loci for the jack mackerel (<Emphasis Type=Italic>Trachurus murphyi</Emphasis> Nichols, 1920)

Eight microsatellite loci were developed for the jack mackerel (Trachurus murphyi), a fish of significant commercial importance in the Southeast Pacific. Genetic variation at these loci was examined in 15 samples from the locality of Talcahuano (Chile). All eight were highly polymorphic, with a number of alleles per locus ranging from 4 to 22 and an observed heterozygosity from 0.429 to 1. These markers will be useful to address issues of population genetics, ecology, conservation and fisheries management related to that species.     

Isolation of polymorphic microsatellite markers from <Emphasis Type=Italic>Przewalskia tangutica</Emphasis> (Solanaceae)

Microsatellite-containing regions were isolated and characterized in Przewalskia tangutica Maxim (Solanaceae), an endemic and endangered species to the Qinghai-Tibetan Plateau of China. An enrichment protocol yielded 200 positive clones. We designed primers to amplify 29 unique microsatellites, 12 of which amplified cleanly and were polymorphic. A survey of 17 individuals showed that these loci are highly variable with the number of alleles ranging from 3 to 12, and expected heterozygosity ranged from 0.2929 to 0.4947. Those markers will be useful for studies of population structure and intraspecific variation in P. tangutica.     

<Emphasis Type=Italic>In vitro</Emphasis> propagation of ten threatened species of <Emphasis Type=Italic>Mammillaria</Emphasis> (Cactaceae)

Mammillaria species are the most numerous within Cactaceae family, and some of them are threatened with extinction as a result of human activities. In this work, results of in vitro propagation are presented for ten Mammillaria species, testing 20 combinations of indole-3-acetic acid (IAA) and kinetin. Best results on shoot formation were obtained using kinetin at two levels: 27.9 and 46.5 μM. All IAA levels tested were able to induce de novo shoot formation in M. bocasana, M. densispina, M. hahniana, M. hutchisoniana, M. orcutii, M. pectinifera, M. perbella, M. picta, M. rhodantha, and M. zephyranthoides. Depending on the IAA level tested, four responding groups were observed concerning their highest shoot-formation number. For all species, the highest average of shoot formation was achieved with 5.7:46.5 or 11.4:46.5 μM IAA/kinetin, yielding 4.8 and 4.7 shoots per explant, respectively, in 60 d. Rooting of regenerated shoots was achieved by leaving the explants in their shoot-induction medium or transferring them to half-strength MS medium. Hardening of regenerated plants was successfully achieved by planting them in peat moss substrate after a desiccation treatment at room temperature for 3 d.     

Development and characterization of 12 compound microsatellite markers in <Emphasis Type=Italic>Platypus </Emphasis><Emphasis Type=Italic>quercivorus</Emphasis> (Murayama) (Coleoptera: Platypodidae)

Twelve compound microsatellite loci were isolated from the ambrosia beetle Platypus quercivorus. Every locus was polymorphic among 50 individuals from two localities, with two to six alleles per locus, without linkage disequilibrium. The observed and expected heterozygosities ranged from 0.140 to 0.740 and from 0.255 to 0.731, respectively. These markers will be available for population genetic studies and parentage analysis of this beetle.     

Efficient separation and purification of allophycocyanin from <Emphasis Type=Italic>Spirulina</Emphasis> (<Emphasis Type=Italic>Arthrospira</Emphasis>) <Emphasis Type=Italic>platensis</Emphasis>

Allophycocyanin (APC) is a minor component of phycobiliproteins in cyanobacteria and red algae. This paper describes a simple and inexpensive extracting method for isolating APC from Spirulina (Arthrospira) platensis with high efficiency. The crude phycobiliprotein extract was pretreated by ammonium sulfate fractionation. Then, by adding hydroxylapatite into crude phycobiliprotein extract dissolved in 20 mM phosphate buffer (pH 7.0), APC was selectively adsorbed by hydroxylapatite but C-phycocyanin (C-PC) was not. The hydroxylapatite was collected and APC was extracted from the crude phycobiliprotein extract. Then, the enriched APC was washed off from the hydroxylapatite using 100 mM phosphate buffer (pH 7.0). In this simple extracting method it was easy to remove C-PC and isolate APC in large amounts. The absorbance ratio A ₆₅₀/A ₂₈₀ of extracted APC reached 2.0. The recovery yield was 70%, representing 4.61 mg · g⁻¹ wet weight. The extracted APC could be further purified by a simple anion-exchange chromatography with a pH gradient from 5.6 to 4.0. The absorbance ratio A ₆₅₀/A ₂₈₀ of the purified APC reached 5.0, and the overall recovery yield was 43%, representing 2.83 mg · g⁻¹ wet weight. Its purity was confirmed by native polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate-PAGE.     

Analysis of microsatellite DNA of rainbow trout (<Emphasis Type=Italic>Parasalmo</Emphasis> (<Emphasis Type=Italic>Oncorhynchus</Emphasis>) <Emphasis Type=Italic>mykiss</Emphasis>) of Kamchatka: Selection of loci and optimization of the method

The variation of a sample of rainbow trout (Parasalmo (Oncorhynchus) mykiss) from natural populations of several rivers of the Kamchatka Peninsula with respect to 43 microsatellite DNA loci has been studied. These loci were earlier used for analysis of Asian populations of closely related salmonids. Ten of them may be regarded as markers and seen promising for further studies on intraspecific relationships of rainbow trout of Kamchatka. Their use in studies on more numerous samples from different localities and populations of Parasalmo (O.) mykiss in the Asian part of the species range will ensure efficient population genetic analysis of the Kamchatka population group of this species.     

Morphological and molecular characterization of two ITS groups of <Emphasis Type=Italic>Erysiphe</Emphasis> (Erysiphales) occurring on <Emphasis Type=Italic>Syringa</Emphasis> and <Emphasis Type=Italic>Ligustrum</Emphasis> (Oleaceae)

ITS sequences determined for 53 Erysiphe specimens on Syringa and Ligustrum collected in Europe, East Asia, and North and South America were divided into two ITS groups, S and K types. Phylogenetic analysis showed that these two ITS types do not share a common ancestor and form separate clades. The K type on Ligustrum was identified as Erysiphe ligustri based on the three-dimensional branching pattern of appendages. Morphological observations showed that there are some morphological differences—pigmentation of appendages and number of ascospores per ascus—between the S and K types on Syringa. Based on these morphological observations, the S and K types on Syringa were identified as E. syringae and E. syringae-japonicae, respectively. The recent abundant production of chasmothecia by lilac powdery mildew in Europe was caused by E. syringae-japonicae introduced from East Asia. DNA sequence analyses of the rDNA ITS region and the 28S rDNA, tub2, CYP51, and Chs1 genes did not support an interspecific hybrid origin for E. syringae-japonicae. Haplotype analysis suggested that E. syringae originated in North America and independently migrated to East Asia and Europe/South America.