Balázs Kolics

Next generation sequencing based development of intron-targeting markers in tetraploid potato and their transferability to other Solanum species.

Intron-targeting (IT) markers were developed from next generation sequencing (NGS) derived transcript sequencing data from the potato cultivar White Lady. The applicability of the IT markers was analyzed in other potato genotypes, and their transferability was studied in other Solanum species: section Archaesolanum (5 species), sect. Solanum (6 species) and a Solanum nigrum population (11 genotypes). Out of 250 randomly chosen transcript sequences, 144 intron harboring loci could be identified for which primer pairs were designed on exons flanking the putative introns. The usefulness of the IT primers was experimentally analyzed on a subset of 40 randomly chosen loci. Statistical analysis of diversity parameters was performed using the ATETRA and POPGENE software packages. By localizing the detected 17 polymorphic loci 11 of the 12 potato chromosomes could be identified. Specificity of the designed IT primers was tested by sequence analysis of amplified IT fragments in a randomly chosen locus. The results revealed the efficiency of NGS derived IT marker development and indicated their utility in diverse molecular analyses including their applicability for cross-species studies.

Description of the song of the bush-cricket Saga rammei Kaltenbach, 1965 (Orthoptera: Tettigoniidae)

A detailed description of the spectrographic and oscillographic features of the male calling song of Saga rammei is presented for the first time. The study is based on Macedonian specimens. The song of S. rammei is a mainly amplitude-modulated signal having wide-band frequency spectrum containing well detectable frequency components between 15 kHz and 80 kHz (intensity maximum between 20–30 kHz). The male song is a sequence of echemes, which are composed of evenly and quickly repeated diplo-hemisyllabic syllables. Echemes begin with a gradually crescending initial part that leads to the main part of the echeme, where the peak amplitude of the syllables is nearly constant. This structure is similar to those described in other European Saga species (S. hellenica, S. campbelli, S. natoliae). However, the results suggest that amongst the European Saga species S. rammei produces the shortest echemes composed of the lowest number of syllables repeated with the highest syllable repetition rate and that the species sings with the shortest echeme repetition period. Those quantitative features make the song of S. rammei distinct and easily recognisable, and therefore could be used by females or males for species recognition.

Re-visiting phylogenetic and taxonomic relationships in the genus Saga (Insecta: Orthoptera).

Twelve of the 13 bushcricket species of the Saga genus are bisexuals and diploids, except the parthenogenetic and tetraploid bush cricket, Saga pedo. Despite a continuous research effort stretching through the 1900s, the taxonomic relationships of the Saga species are still disputed. In this study, our primary aim was to reveal natural relationships of the European Saga species and three of their Asian relatives, with special attention to the problematic taxonomy of two subspecies: S. campbelli campbelli and S. c. gracilis. Following a phylogenetic analysis of eight species, a comprehensive study was carried out on the above three taxa by using acoustic and morphometric approaches in parallel. Our phylogenetic data showed that European Saga species evolved from a monophyletic lineage. The geographical transitional species S. cappadocica was positioned between European and Asian lineages supporting the idea that the European Saga lineage originated phylogeographically from the Asian clade. The above results showed better agreement with the morphological data than with earlier ones based either on karyology or acoustic information only. After reviewing our data, we concluded that Saga pedo has most likely evolved from S. c. gracilis and not from S. rammei or S. ephippigera, as proposed by earlier studies. S. c. gracilis shares the same ITS2 haplotype with S. pedo, indicating that the latter could have evolved from populations of the former, probably through whole genome duplication. Based on acoustic and morphometric differences, we propose to elevate the two subspecies, S. campbelli campbelli and S. c. gracilis, to species level status, as Saga gracilis Kis 1962, and Saga campbelli Uvarov 1921. The present work sets the stage for future genetic and experimental investigations of Saginae and highlights the need for additional comprehensive analysis involving more Asian Saga species.