Background: The rapid increase in the amount of protein and DNA sequence information available has become almost overwhelming to researchers. So much information is now accessible that high-quality, functional gene analysis and categorization has become a major goal for many laboratories. To aid in this categorization, there is a need for non-commercial software that is able to both align sequences and also calculate pairwise levels of similarity/identity.

BackgroundAnthocyanin pigments aid in reproduction and provide ultraviolet protection to land plants. We have examined the phylogenetic relationships among the five primary enzymes responsible for producing anthocyanin pigment in its three major forms. Dihydroflavonol 4-reductase (DFR), anthocyanidin synthase (ANS), Flavonoid 3’glucosyltransferase (F3GT), flavonoid 3’hydroxylase (F3’H), and flavonoid 3’5’ hydroxylase (F3’5’H) are responsible for the final steps in anthocyanin pigment production.ResultsWe were interested in how conserved the anthocyanin pathway genes may be among land plants, and evolutionarily how far back into the plant lineage anthocyanin production may be traced. The DFR, ANS, F3GT, and F3’H genes date back 450 million years to the first land plants. Mosses, spike mosses, and ferns express these four products, although there is no evidence of sequence orthologues for these genes in algae. Additionally, F3’5’H is not evident in organisms that predated gymnosperms.ConclusionOur findings support the hypothesis that “blue” anthocyanin pigments did not evolve until 300-350 mya along with the gymnosperms, although the “red” anthocyanin pigments may be as ancient as the mosses (~450 mya).Electronic supplementary materialThe online version of this article (doi:10.1186/1999-3110-55-10) contains supplementary material, which is available to authorized users.

Within Barnegat Bay, New Jersey, Zostera marina populations have declined by 62% over the last 20 years, and restoration efforts have met with mixed success. We have completed a microsatellite-based genetic investigation of eight populations of Z. marina within Barnegat Bay to determine whether the genetic stock origins of the plants used in management projects may affect restoration success. Additionally, we assessed the genetic diversity of Z. marina in Barnegat Bay to better understand its population structure. Clonal diversity ranged from 0.70 to 0.95 for the populations studied. Individually, Barnegat Bay populations are not genetically diverse, and there is also little divergence among populations. The Atlantic populations had mean Hobs values (0.20-0.34) that were far lower than the Hexp values (0.69-0.83). Also, the F IS values in all of the eastern populations indicate a surfeit of homozygotes over heterozygotes, suggesting a low degree of outcrossing in the Barnegat Bay populations. Six of the ten populations studied (Ham Island, Manahawkin Bay, Shelter Island, Marsh Elder, Harvey Cedar Sedge, and Long Island) show evidence of historical bottlenecks. Mean estimated F ST values would suggest that most alleles are undergoing moderate genetic differentiation, with values that range from 0.06 to 0.13. Oyster Creek and Sedge Island demonstrate the largest estimated effective population sizes and may be the most appropriate populations for use in future eelgrass restoration projects.

The ILR1-like family of hydrolase genes was initially isolated in Arabidopsis thaliana and is thought to help regulate levels of free indole-3-acetic-acid.We have investigated how this family has evolved in dicotyledon, monocotyledon and gymnosperm species by employing the GenBank and TIGR databases to retrieve orthologous genes. The relationships among these sequences were assessed employing phylogenomic analyses to examine molecular evolution and phylogeny. The members of the ILR1-like family analysed were ILL1, ILL2, ILL3, ILL6, ILR1 and IAR3. Present evidence suggests that IAR3 has undergone the least evolution and is most conserved. This conclusion is based on IAR3 having the largest number of total interspecific orthologues, orthologous species and unique orthologues. Although less conserved than IAR3, DNA and protein sequence analyses of ILL1 and ILR1 suggest high conservation. Based on this conservation, IAR3, ILL1 and ILR1 may have had major roles in the physiological evolution of ‘higher’ plants. ILL3 is least conserved, with the fewest orthologous species and orthologues. The monocotyledonous orthologues for most family-members examined have evolved into two separate molecular clades from dicotyledons, indicating active evolutionary change. The monocotyledon clades are: (a) those possessing a putative endoplasmic reticulum localizing signal; and (b) those that are putative cytoplasmic hydrolases. IAR3, ILL1 and ILL6 are all highly orthologous to a gene in the gymnosperm Pinus taeda, indicating an ancient enzymatic activity. No orthologues could be detected in Chlamydomonas, moss and fern databases.

BackgroundThe acquisition of high-quality DNA for use in phylogenetic and molecular population genetic studies is a primary concern for evolutionary and genetic researchers. Many non-destructive DNA sampling methods have been developed and are used with a variety of taxa in applications ranging from genetic stock assessment to molecular forensics.ResultsThe authors have developed a field sampling method for obtaining high-quality DNA from sunfish (Lepomis) and other freshwater fish that employs a variation on the buccal swab method and results in the collection of DNA suitable for PCR amplification and polymorphism analysis. Additionally, since the circumstances of storage are always a concern for field biologists, the authors have tested the potential storage conditions of swabbed samples and whether those conditions affect DNA extraction and PCR amplification. It was found that samples stored at room temperature in the dark for over 200 days could still yield DNA suitable for PCR amplification and polymorphism detection.ConclusionThese findings suggest that valuable molecular genetic data may be obtained from tissues that have not been treated or stored under optimal field conditions. Furthermore, it is clear that the lack of adequately low temperatures during transport and long term storage should not be a barrier to anyone wishing to engage in field-based molecular genetic research.

Zostera marina (eelgrass) can be found in the North Atlantic on the coast of Europe and on the east and west coasts of North America. Over the last 30 years, this once robust species has been reduced to sparse patchy populations due to disease and anthropogenic effects. In order to better understand the consequences of this devastation on the population genetics of the species, we have analyzed the population structure of western Atlantic Z. marina, employing microsatellite DNA polymorphisms. Although high fixation index values suggest moderate genetic differentiation among most of the Z. marina sites, population diversity was low. This lack of diversity was supported by a general dearth of observable heterozygotes in these sites; mean observed heterozygosity values (0.14-0.46) were lower than the mean expected heterozygosity values (0.57-0.81). Additionally, the mean F(IS) (coefficient of local inbreeding) values in these sites were positive, again indicating a surfeit of homozygotes. Allelic richness suggests that Chesapeake Bay has the greatest internal genetic diversity of the sites studied. Inbreeding seems prevalent in these American populations, suggesting possible reproductive fitness problems in the future. There is evidence of demographic bottlenecking and particularly low genetic diversity in Long Island. Northern Maine had the highest effective population size, suggesting a possible use in future restoration projects.

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Within Barnegat Bay, New Jersey, eelgrass (Zostera marina) populations have declined by 62 % over the last 20 years. To better understand the consequences of this devastation, we have previously employed microsatellite DNA polymorphisms to analyze the population structure of Z.marina within Barnegat Bay, as well as along the eastern United States seaboard. We have restored populations of Z. marina in Barnegat Bay over the last 10 years to help assess the best planting conditions and ecotypes that might be used in long‐term restoration strategies. In this study, we examined the genetic health of the restored populations compared to that of the donor eelgrass populations within the bay. Using microsatellites, we can identify which parental founding ecotypes survived the restoration process over multiple generations. The frequency of observed heterozygotes, although higher than in the natural populations, still indicates reduced levels of diversity and connectivity. The inbreeding frequency is high in the restored populations, but lower than what is seen in the native populations. All restored populations have effective population values >50, suggesting a high probability of survival in the short term.