Inefficient photosynthesis in the Mediterranean orchid Limodorum abortivum is mirrored by specific association to ectomycorrhizal Russulaceae

Among European Neottieae, Limodorum abortivum is a common Mediterranean orchid. It forms small populations with a patchy distribution in woodlands, and is characterized by much reduced leaves, suggesting a partial mycoheterotrophy. We have investigated both the photosynthetic abilities of L. abortivum adult plants and the diversity of mycorrhizal fungi in Limodorum plants growing in different environments and plant communities (coniferous and broadleaf forests) over a wide geographical and altitudinal range. Despite the presence of photosynthetic pigments, CO2 fixation was found to be insufficient to compensate for respiration in adult plants. Fungal diversity was assessed by morphological and molecular methods in L. abortivum as well as in the related rare species Limodorum trabutianum and Limodorum brulloi. Phylogenetic analyses of the fungal internal transcribed spacer (ITS) sequences, obtained from root samples of about 80 plants, revealed a tendency to associate predominantly with fungal symbionts of the genus Russula. Based on sequence similarities with known species, most root endophytes could be ascribed to the species complex encompassing Russula delica, Russula chloroides, and Russula brevipes. Few sequences clustered in separate groups nested within Russula, a genus of ectomycorrhizal fungi. The morphotypes of ectomycorrhizal root tips of surrounding trees yielded sequences similar or identical to those obtained from L. abortivum. These results demonstrate that Limodorum species with inefficient photosynthesis specifically associate with ectomycorrhizal fungi, and appear to have adopted a nutrition strategy similar to that known from achlorophyllous orchids.     

Characterization of 14 tetranucleotide microsatellite loci derived from Pacific herring

Spawning aggregations of Pacific herring (Clupea pallasi) often exhibit significant interannual variation in allele frequencies of neutral gene markers. We isolated 14 tetranucleotide microsatellites to examine hypothetical processes that may produce this unique genetic signal. We developed and tested primer pairs for each locus and then estimated locus variability in samples (n = 60) from two populations. The number of alleles per locus ranged from five to 49. The expected heterozygosity across loci and populations ranged from 0.20 to 0.96. These microsatellites will be useful for estimating genetic variation in herring on a fine geographical scale.     

Isolation and characterization of microsatellite loci in Epidendrum puniceoluteum, an endemic orchid from the Atlantic Rainforest

Epidendrum puniceoluteum is an endemic orchid of Atlantic Rainforest, restricted to few populations only due to the destruction and fragmentation of its native habitat. Here, we report on the development of 10 microsatellite markers isolated from this orchid species. Genetic variability was characterized in two distant populations from Brazil coast. The number of alleles observed for each locus ranged from two to 12 and with an average of 6.4 alleles per locus. These microsatellites should be valuable tools for studying both fine-scale genetic structure of scattered E. puniceoluteum population and patterns will be useful genetic markers for other closely related taxa.     

Novel microsatellite loci in the grass snake (Natrix natrix) and cross-amplification in the dice snake (Natrix tessellata)

Six novel polymorphic microsatellite loci are presented for the grass snake (Natrix natrix), a species with declining populations in many regions. The number of alleles per locus ranged from two to seven. Four dice snake (Natrix tessellata) microsatellites were polymorphic in the grass snake with three to four alleles. At two loci, the expected heterozygosity differed significantly from observed heterozygosity. Cross-amplification of the grass snake markers in the dice snake showed two polymorphic microsatellites with two and four alleles.     

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 ten tetranucleotide microsatellite loci in rock carp, <Emphasis Type="Italic">Procypris rabaudi</Emphasis> (Tchang)

Ten tetranucleotide microsatellite loci were isolated from GATA-enriched library of rock carp (Procypris rabaudi). The loci were tested in 25 individuals from four populations. The number of alleles per locus ranged from 3 to 6. Polymorphism information content (PIC) ranged from 0.460 to 0.788. Observed and expected heterozygosities ranged from 0.375 to 0.739 and from 0.531 to 0.833, respectively. These microsatellites will facilitate the study of population genetics of rock carp.     

Development of polymorphic microsatellite markers for Dalbergia nigra (Papilionoideae), an endangered tree from the Brazilian Atlantic Forest

Dalbergia nigra is an endangered tree restricted to the Brazilian Atlantic Forest. Ten polymorphic microsatellite loci were isolated and characterized in 47 trees from two populations. The total number of alleles per locus ranged from three to 12 alleles. The levels of observed and expected heterozygosities ranged from 0.304 to 0.740 and from 0.278 to 0.872, respectively. Significant deviations from Hardy-Weinberg were detected for only three loci in each population. No pair of loci exhibited significant linkage disequilibrium. These microsatellites provide an efficient tool to investigate genetic structure in forest remnants with the purpose of conservation of this species.     

Isolation and characterization of microsatellite loci from the coppertail skink (Ctenotus taeniolatus)

We report on the isolation and characterization of eight microsatellite markers for the coppertail skink (Ctenotus taeniolatus). From a sample of 27 individuals, the number of alleles per locus ranged from seven to 13 and observed and expected heterozygosities ranged from 0.714 to 0.818 and from 0.738 to 0.899, respectively. These loci will be used to assess anthropogenic disturbance on coppertail skink populations. These are the first microsatellites reported for the genus Ctenotus, despite being the most speciose genus of lizards in Australia.     

Isolation and characterization of 36 polymorphic microsatellite markers in Schima superba (Theaceae)

? Premise of the study: Our objective was to develop microsatellite markers to investigate the level of genetic diversity within and among populations in a dominant evergreen broad-leaved tree, Schima superba, in southern China. ? Methods and Results: Thirty-six microsatellite markers were developed and showed polymorphism in three populations. The number of alleles per locus ranged from six to 34, with an average of 19. The observed and expected heterozygosities ranged from 0.242 to 1.000 and from 0.504 to 0.945, respectively. ? Conclusions: The developed microsatellites will be useful for studying genetic diversity and population structure in S. superba.     

Genetic variation in populations of the threatened seagrass Halophila beccarii (Hydrocharitaceae)

Halophila beccarii (Hydrocharitaceae), a small monoecious seagrass, has been listed as a threatened species. In this study, a total of 106 samples were collected from four Chinese populations located at the northern limit of its distribution range. Using six polymorphic microsatellites, we found low genetic variation in this species, in which the mean number of alleles per locus was 2.8, and 16 multi-locus genotypes were revealed. In the four populations, the mean number of alleles per locus ranged from 1.2 to 2, one to eight genotypes were found, and clonal diversity ranged from 0 to 0.23; observed and expected heterozygosity ranged from 0.17 to 0.40 and from 0.09 to 0.24, respectively. Strong genetic differentiation was found among the populations, and the standardized fixation index (F′_ST) was 0.787. Species traits (i.e., clonal growth) and bottleneck effects due to drastic population fluctuation may contribute to the observed low genetic variation.     

Microsatellites in the Karoo scrub-robin, Cercotrichas coryphaeus (Passeriformes: Muscicapinae): isolation and characterization of 13 autosomal and two sex-linked loci

Fifteen polymorphic microsatellites were developed for the Karoo scrub-robin, Cercotrichas coryphaeus. Here we describe and characterize microsatellite variation of 13 autosomal loci and two Z-linked loci in 42 individuals from two distinct South African populations. The number of alleles per locus varied from three to 13 and values of observed heterozygosity ranged from 0.318 to 0.900. These loci will be used to test hypotheses relating to fine-scale social structure and mating strategies in this cooperatively breeding species.     

Polymorphic microsatellites for the study of fragmented populations of Pitcairnia geyskesii L. B. Smith (Bromeliaceae), a specific saxicolous species of inselbergs in French Guiana

Pitcairnia geyskesii is a saxicolous bromeliad endemic to the granitic inselbergs of French Guiana and Surinam. Seven polymorphic microsatellite markers were characterized to investigate the polymorphism among individuals sampled on five inselbergs in French Guiana. The total number of alleles per locus varied from four to 14. Mean observed heterozygosities at each microsatellite locus ranged from 0.114 to 0.629. This set of microsatellites offers an efficient tool for more detailed investigations on the genetic structure of fragmented populations living on such a singular and insular habitat within tropical rainforest.     

Isolation and characterization of microsatellites in the bird-pollinated, autohexaploid, Eremophila glabra ssp. glabra (R.Br. (Ostenf.)) (Myoporaceae), an Australian endemic plant

Thirty-eight microsatellite loci were developed for the bird pollinated, autohexaploid, Eremophila glabra ssp. glabra. A genomic library was screened with dinucleotide and trinucleotide sequence repeats. Polymorphism ranged from one to 21 alleles per locus. Twenty-four loci exhibited null alleles, based on patterns of inheritance between maternal and progeny phenotypes. Cross-species amplification of nine Eremophila species was successful for most primers, indicating wide applicability across the genus. These microsatellites will be used to study the gene flow patterns of fragmented populations of E. glabra ssp. glabra.     

Isolation and characterization of six microsatellites in the Mexican beaded lizard Heloderma horridum

Six polymorphic microsatellite loci were isolated and characterized in the threatened Mexican beaded lizard, Heloderma horridum. The number of alleles per locus ranged from three to 12, with observed heterozygosity estimates ranging from 0.00 to 0.77, and expected heterozygosity estimates ranging from 0.00 to 0.73. These microsatellites will provide a valuable tool for the investigation of the genetic variation and structure of both wild and captive H. horridum populations.     

Polymorphic microsatellite loci for population studies of the razor clam, <Emphasis Type="Italic">Sinonovacula constricta</Emphasis>

The razor clam (Sinonovacula constricta) is an important aquacultured bivalve in China. The natural populations of this species are decreasing quickly. To facilitate studies on genetic diversity and population structure of wild populations, microsatellites were isolated from a CA enriched genomic library. Eight microsatellite loci were polymorphic in 30 individuals from Chongming in Shanghai, China. The number of alleles per polymorphic locus varied from 6 to 13 and the values of observed heterozygosity and expected heterozygosity ranged from 0.350 to 1.000 and from 0.602 to 0.902, respectively. These microsatellites are being used in studying population differentiation and genetic diversity for effective conservation and management genetic resources of S. constricta.     

Development of 10 polymorphic microsatellite markers from herbicide‐bleached tissues of the brooding pocilloporid coral Seriatopora hystrix

Here we report the isolation of 44 microsatellites from the brooding, pocilloporid coral, Seriatopora hystrix, developed from a partial genomic DNA library using a repeat enrichment protocol. A further eight previously published microsatellites were also tested; five of these were developed for S. hystrix, whereas three were isolated from corals of the closely related genus Pocillopora. Out of these, we incorporated nine and 10 primer pairs into two multiplex reactions that reliably amplified polymorphic microsatellites in populations from the west and the east coast of Australia, respectively. Number of alleles ranged from three to 22 per locus.     

Characterization of long transcribed microsatellites in Helianthus annuus (Asteraceae)

? Premise of the study: Research on the evolutionary role of exonic microsatellites currently lacks an understanding of the evolutionary pressures that promote or limit their expansion. Contrasting levels of variability and genetic structures at anonymous and transcribed microsatellite loci of varying lengths are likely to provide useful insights regarding the relative strength of selection acting on different classes of microsatellites. We have developed primers for long transcribed microsatellites in Helianthus annuus to make these comparisons. ? Methods and Results: Eight relatively long microsatellites from sequences in the expressed sequence tag database of H. annuus were characterized. A total of 63 individuals from three populations in Kansas were genotyped. The number of alleles per locus ranged from four to 11 with an average observed heterozygosity of 0.723. ? Conclusions: Our study has generated suitable tools for studying the population genetics of long transcribed microsatellites that are potentially influenced by selection.     

Development of microsatellite markers for Dimorphandra mollis (Leguminosae), a widespread tree from the Brazilian cerrado

? Premise of the study: Microsatellite markers were developed for Dimorphandra mollis (Leguminosae), a widespread tree in the Brazilian cerrado (a savanna-like vegetation). ? Methods and Results: Microsatellite markers were developed from an enriched library. The analyses of polymorphism were based on 56 individuals from three populations. Nine microsatellite loci were polymorphic, with the number of alleles per locus ranging from three to 10 across populations. The observed and expected heterozygosities per locus and population ranged from 0.062 to 0.850 and from 0.062 to 0.832, respectively. ? Conclusions: These microsatellites provide an efficient tool for population genetics studies and will be used to assess the genetic diversity and spatial genetic structure of D. mollis.     

Polymorphic microsatellites for the study of Aconitum napellus L. (Ranunculaceae), a rare species in France

Aconitum napellus is a perennial European Ranunculaceae that is currently decreasing in France. Six polymorphic microsatellite markers were characterized to study the polymorphism among individuals sampled from six populations in France. The total number of alleles per locus varied from two to four. Mean observed heterozygosities at each microsatellite locus ranged from 0.05 to 0.19. This set of microsatellites offers an efficient tool for more detailed investigations on population genetic structure of a rare species that can reproduce both sexually and by vegetative spread.     

Identification and characterization of microsatellite loci in Intsia palembanica (Leguminosae), a valuable tropical timber species

We present 25 polymorphic microsatellite loci for Intsia palembanica, a highly valued timber species in the Indo-Malayan region, Australia and western Pacific islands. Microsatellite loci were tested for polymorphism across a total of 76 individuals from three natural populations from Irian Jaya, Indonesia. The average number of alleles for these microsatellites was 12.1 per locus, ranging from four to 19. The observed heterozygosity within the natural populations ranged from 0.01 to 0.96. The markers will enable us to evaluate the spatial-temporal population genetic structure and gene flow dynamics among populations. They can also be used for tracking and tracing wood from legally logged concessions.