No abstract

In the north Adriatic basin, a morphologically and genetically distinct lineage of grayling is found, designated as the Adriatic grayling. In Slovenia, the Adriatic grayling is restricted to the Soča river system, where it is critically endangered. The most pertinent threat is stocking with nonnative, highly divergent Sava (Danubian) drainage stock, and this activity has been going on for more than four decades. The present study was designed to characterise the genetic structure of the Adriatic grayling in Slovenia, with particular emphasis on estimating the degree of introgression with nonindigenous stocked grayling. We analysed polymorphism at 154 microsatellite loci in samples representing grayling from the Adriatic and Danubian drainage stock. A relatively high number (12) of alleles, diagnostic for the Adriatic grayling, were identified. However, a correspondence analysis based on individual multilocus genotypes also revealed that there is no distinctive Adriatic group but rather a dispersed multitude of individuals that cannot be unambiguously distinguished from the more homogenous Danubian population. A Bayesian analysis of individual admixture coefficients confirmed this pattern and revealed extensive introgression between the Adriatic grayling and stocked grayling of Danubian origin. Average individual admixture coefficients showed that only between 50 and 60% of the original gene pools remained, and only few non-introgressed indigenous individuals could be identified. Microsatellite-based individual admixture analysis appear to be an important tool for identifying remaining non-introgressed indigenous individuals that could be used for restoring the original populations.

Genetic variation in the southern green stink bug Nezara viridula (Linnaeus) from 11 geographically separated sampling locations (Slovenia, France, Greece, Italy, Madeira, Japan, Guadeloupe, Galapagos, California, Brazil and Botswana) was studied by sequencing 16S and 28S rDNA, cytochrome b and cytochrome c oxidase subunit I gene fragments and random amplified polymorphic DNA (RAPD) analysis. Sequencing revealed 11 distinct haplotypes clustering into lineages A, B and C. Lineage C was characteristic for a single analysed specimen from Botswana. Lineage B was detected in Japan, and it probably arose in Asia. Haplotypes of European and American specimens belonged to lineage A; specimens from France, Slovenia, Madeira and Brazil shared highly similar haplotypes (>99%) from subgroup A1, while all the specimens from Greece, California, Galapagos and Guadeloupe shared a haplotype from subgroup A2. RAPD data were more variable but consistent with mtDNA sequences, revealing the same clustering. They separated the Botswanian specimen from Japanese specimens and from a group of more closely related specimens from Europe and America. Sequence and RAPD results both support the African origin of N. viridula, followed by dispersal to Asia (lineage B) and, more recently, by expansion to Europe and America (lineage A). RAPD analysis revealed two highly supported subgroups in Japan, congruent with mtDNA lineages A2 and B, suggesting multiple colonization of Japan. Invariant sequences at the 28S rDNA combined with other results do not support the hypothesis that cryptic (sibling) species exist within the populations investigated in this study.

-The genetic structure of Apis mellifera carnica bee from Slovenia, collected from 269 localities of ten Slovenian districts, was assessed by mitochondrial and nuclear DNA analyses. The level of genetic variability within and among districts was low. All of the samples were fixed for one newly found mtDNA haplotype of the C phylogenetic lineage, designated as C2C. A low level of variability was observed for all microsatellite loci, showing a very homogenous structure of the Carniolan bee population.

-In order to illuminate the phylogeography of brown trout (Salmo trutta) populations in the Balkan state of Serbia, the 561 bp 5'-end of mtDNA control region of 101 individuals originating from upland tributaries of the Danubian, Aegean and Adriatic drainages were sequenced and compared to corresponding brown trout sequences obtained in previous studies. Among 15 haplotypes found, 14 were considered native, representing the Danubian and Adriatic lineages of the brown trout, while one haplotype (ATcs1), found only in two individuals originating from two stocked rivers, corresponded to the Atlantic lineage and was considered introduced. Native haplotypes exhibited a strong geographic pattern of distribution: the Danubian haplotypes were strictly confined to the Danubian drainage, while the Adriatic haplotypes dominated in the Aegean and Adriatic drainages; most of the total molecular variance (69%) was attributed to differences among the drainages. Phylogenetic reconstruction, supplemented with seven haplotypes newly described in this study, suggested a sister position of the Atlantic-Danubian and Adriatic-Mediterranean-marmoratus ("southern") phylogenetic group, and pointed to the existence of a distinct clade, detected within the "southern" group. The data obtained confirmed our expectation of the existence of high genetic diversity in Balkan trout populations, and we recommend more widespread surveys covering trout stocks from the region.phylogeography / Salmo trutta / mitochondrial DNA / control region / Balkan Peninsula

Analysis of both uni‐(two mtDNA gene sequences) and bi‐parentally (seven microsatellite loci) inherited genetic markers, together with analysis of 40 morphological characters, described Salmo ohridanus as a highly divergent member of the genus Salmo. Based on comparative substitution rate differences at the cytochrome b gene, and a rough estimated age of the Salmo trutta complex (i.e. at least 2 million years), the S. ohridanus and Salmo obtusirostris clade probably split from a common ancestor of brown trout Salmo trutta >4 million years ago, overlapping with minimum age estimates of the formation of Europe's oldest freshwater habitat, Lake Ohrid. Comparative analysis with Lake Ohrid brown trout (known regionally as Salmo letnica), supported the notion that these fish have more recently colonized the lake and phylogenetically belong to the Adriatic lineage of brown trout. It is further suggested that species‐specific saturation in the mtDNA control region underestimated the divergence between S. ohridanus and S. trutta. Evidence of rare hybridization between S. ohridanus and Lake Ohrid brown trout was seen at both mtDNA and microsatellite markers, but there was no support for extensive introgression.

Salmothymus obtusirostris (soft‐muzzled trout) is endemic to the South Adriatic drainage. Owing to its unusual appearance, which resembles both trout and grayling, it has been initially classified as a separate genus. However, this classification is ambiguous and has never been firmly established. We have studied mtDNA (control region and cytochrome b gene) and nuclear DNA (a part of LDH C*1 gene) variation between soft‐muzzled trout from the upper part of the River Neretva, Bosnia and Herzegovina, and other salmonid representatives in order to examine how the current classification is congruent with molecular data. On the basis of sequence identity of mtDNA control region among several genera (i.e. Salmo, Oncorhynchus, Salvelinus, Acantholingua, Brachymystax, Thymallus and Coregonus) a close relationship between Salmothymus, Salmo and Acantholingua was established. Phylogenetic analysis on a combined data set of mitochondrial and nuclear DNA, supported by 100% bootstraping, indicated that S. obtusirostris and A. ohridana are sister taxa which exhibit a closer relationship to S. trutta than to S. salar. This finding refutes the current classification, which recognizes S. obtusirostris as separate genus, and instead suggests its reclassification on the species level as Salmo obtusirostris. © 2002 The Linnean Society of London, Biological Journal of the Linnean Society, 2002, 77, 399–411.

The northern part of the peri-Adriatic river system is inhabited by a native grayling population. In comparison with the Danubian population most of the Adriatic population is regarded as morphologically distinct and therefore recognized as racially differentiated. DNA sequence analysis of the 394-bp fragment of the mtDNA control region and 363-bp fragment of the cytochrome b gene region from grayling specimens of the Adriatic, Danubian and Atlantic river systems revealed nine composite mtDNA haplotypes. The genetic divergence of the Adriatic and the Danubian-Atlantic haplotypes ranged from 2.52 to 3.1O0/o and from 3.38 to 3.92% in the cytochrome b gene and control region, respectively. On the basis of sequence data an attempt was made to elucidate the phylogeography of the Adriatic population. The molecular data indicate an ancient monophyletic origin and also confirm its distinct position within the Thymallus thymallus species, in spite of the fact that stocking with the Danubian type occurred in the Adriatic river system. Therefore the Adriatic population deserves special attention. As a first step in its preservation, its formal distinction by using the name Adriatic grayling is proposed. 0 2001 The Linnean Society of London ADDITIONAL KEY WORDS: mitochondria1 DNAcontrol regioncytochrome b gene, phylogeography, conservation biology.