Isolation and characterization of microsatellite markers in Perilla frutescens Brit

We isolated and characterized 13 polymorphic microsatellite primers from Perilla frutescens Brit. var. frutescens by using a modified method that involves one‐way PCR amplification with single primer prior to enrichment with an ‘oligo hook’. The efficiency of this procedure for isolating unique microsatellite sequences was approximately 77%. The number of alleles per microsatellite locus ranged from three to 10 with an average of 6.5 alleles per locus while fragment size varied from 156 to 298 bp. The observed and expected heterozygosities ranged from 0.52 to 0.86 and 0.52 to 0.89, respectively. These newly isolated microsatellite markers are expected to provide valuable resources for different genetic studies currently underway in our Perilla genome research program.     

CONSUMERS REPORT PREFERENCES WHEN THEY SHOULD NOT: A CROSS-CULTURAL STUDY

If a chi‐squared analysis were to be performed to determine whether preferences were significant in a paired preference test, the appropriate expected frequencies in the analysis would represent those that would occur should the consumers have no preference. One way of determining these ‘no preference’frequencies, for a particular test situation, would be to note the preference responses obtained when the stimuli were putatively identical. Over 2000 consumers were given paired preference tests in which the stimuli were putatively identical. Response conditions and consumer groups were varied and the proportions of preference and no preference responses were noted. In a preliminary experiment, further research was seen to be justified when for putatively identical stimuli, judges did not exclusively express lack of preference; many expressed a preference for one or other of the stimuli. Further experiments were conducted using a written response condition and putatively identical potato chips (potato crisps) as stimuli. Using a single ‘no preference’option, variation in the placement of this option at either first, second or third place on the response sheet had no significant effect on the response frequencies. The proportion of ‘no preference’responses was approximately 30% in all cases. A previously reported 40‐20‐40 distribution was not always confirmed. The experiment was repeated with Korean consumers; these had fewer ‘no preference’responses. Deriving preferences from hedonic scales, rather than having judges respond to preference options, increased the proportion of ‘no preference’responses, with American judges still having more than Korean judges. Yet there are logical objections to extracting preference data from hedonic scales. Increasing the number of ‘no preference’options to two or three, had the effect of increasing the number of ‘no preference’responses up to as much as approximately 60%. Extending the results to Koreans, using two ‘no preference’options, it was seen that only the judges in an anonymous response condition had significantly fewer ‘no preference’responses than Americans. The use of these response frequencies as expected frequencies in chi squared analyses was illustrated, after adjustments for counterbalancing.     

Complex phylogeographic patterns in the freshwater alga Synura provide new insights into ubiquity vs. endemism in microbial eukaryotes

The global distribution, abundance, and diversity of microscopic freshwater algae demonstrate an ability to overcome significant barriers such as dry land and oceans by exploiting a range of biotic and abiotic colonization vectors. If these vectors are considered unlimited and colonization occurs in proportion to population size, then globally ubiquitous distributions are predicted to arise. This model contrasts with observations that many freshwater microalgal taxa possess true biogeographies. Here, using a concatenated multigene data set, we study the phylogeography of the freshwater heterokont alga Synura petersenii sensu lato. Our results suggest that this Synura morphotaxon contains both cosmopolitan and regionally endemic cryptic species, co‐occurring in some cases, and masked by a common ultrastructural morphology. Phylogenies based on both proteins (seven protein‐coding plastid and mitochondrial genes) and DNA (nine genes including ITS and 18S rDNA) reveal pronounced biogeographic delineations within phylotypes of this cryptic species complex while retaining one clade that is globally distributed. Relaxed molecular clock calculations, constrained by fossil records, suggest that the genus Synura is considerably older than currently proposed. The availability of tectonically relevant geological time (10⁷–10⁸ years) has enabled the development of the observed, complex biogeographic patterns. Our comprehensive analysis of freshwater algal biogeography suggests that neither ubiquity nor endemism wholly explains global patterns of microbial eukaryote distribution and that processes of dispersal remain poorly understood.     

Estrogen and Androgen Target Cells in the Brain of Fishes, Reptiles and Birds: Phylogeny and Ontogeny

The cellular and subcellular distribution of 3H estradiol inthe forebrain of goldfish, platyfish, lizard, turtle, and ringdove was studied by thaw-mount autoradiography. The topographicdistribution of estrogen target cells in these non-mammalianspecies corresponds to the general pattern established earlierin our laboratory for mammals. In all non-mammalians studied,accumulations of estrogen target cells exist in preoptic-septal,central hypothalamic and thalamic areas. While allocorticaldistribution of estrogen target cells is present in the avianand reptilian species, it appears to be absent in the teleosts.Differences in brain evolution probably account for it. Thisphylogenetically delayed acquisition of telencephalic steroidtarget sites seems paralleled by an ontogenetic delay, as observedin the chick embryo: early embryonic appearance of estrogenuptake (receptors) in preoptic and central hypothalamic regionsand late appearance of estrogen receptors in the amygdala. Androgendistribution in the forebrain of the dove and lizard overlapswith the estrogen distribution with some exceptions.     

Sibling species in the Chrysis ignita complex: molecular,morphological and trophic differentiation of Baltic species,with a description of two new cryptic species (Hymenoptera: Chrysididae)

Cryptic species complexes cause major challenges for taxonomists and alter understanding of species diversity. In Northern Europe, the Chrysis ignita species group is one such complex with numerous sympatric sibling species. The objective of this paper is to assess the taxonomy of 15 species from this group using three different approaches: molecular, morphological and trophic differentiation. The analysed set of molecular markers included a 7400‐bp‐long sequence of the mitochondrial genome covering complete sequences of CO1, CO2, ATP8, ATP6, CO3, ND3, 16S and 12S rRNA, nine tRNAs and a partial sequence of CytB, as well as a 3880‐bp‐long sequence of the nuclear DNA covering a part of 18S rRNA, the ITS1, 5.8S rRNA, ITS2 and a part of 28S rRNA. Discrete diagnostic characters of each species sequence were retrieved using the Characteristic Attribute Organisation System algorithm and a molecular identification key was compiled. The study revealed a higher evolutionary rate of the genes ATP8, ATP6, CO3, ND3 and CytB compared to that of CO1, CO2 and 16S; the studied nuclear markers demonstrated a lower evolutionary rate than the mitochondrial markers. A consensus tree compiled based on the combined mtDNA and nuclear markers with a strongly supported topology resolved the position of the C. schencki – C. parietis sp.n. clade as sister to the C. ignita – C. impressa clade and supported the monophyly of the C. angustula – C. longula clade. We compiled a morphometric species identification key applying linear discriminant equations. The trophic differentiation was assessed using data on host preferences of ten Chrysis species reared from trap‐nests; the analysis demonstrated that most of them are specialists exploiting a single or a few taxonomically related host species. In most cases, all three approaches supported the distinct status of the included species. Moreover, two previously undescribed species were consistently supported by the molecular methods. Therefore, we describe these as new, namely C. horridula sp.n. and C. parietis sp.n. Only C. mediata and C. solida were not clearly distinguished using the molecular phylogeny reconstruction methods. However, based on distinctive niche divergence, the presence of molecular characters and morphometric differences, we consider them as phylogenetically young but distinct species. In view of the weak morphological and molecular differentiation, the widely overlapping distribution areas and often similar habitat preferences and the trophic specialization, the C. ignita complex presents a possible model for studies of sympatric cryptic speciation. This published work has been registered in ZooBank, http://zoobank.org/urn:lsid:zoobank.org:pub:1EBAF0E1‐5FB7‐4CF4‐A595‐C11982448360 .