In vitro cultivation and biocontrol potential of Botryosphaeria visci against European mistletoe (Viscum album L.)

To improve the biological control of white mistletoe (European mistletoe) additional detailed information on Botryosphaeria visci infection, its basic nutritional requirements, growth, and in vitro growth characteristics is needed. The objectives of this study were to isolate and identify the fungus B. visci associated with Viscum аlbum from Sorbus aucuparia, to provide information on its in vitro introduction and growth characteristics on different media, and to test the pathogenicity of the isolated fungus. To achieve these goals, the morphology of conidiophores from infected parts of mistletoe was evaluated by microscopy. The isolate from fresh collections of V. album was identified as Sphaeropsis visci anamorph of ascomycete Botryosphaeria visci. The morphology of the vegetative mycelium and growth of B. visci varied depending on the media used. The best medium supporting growth and sporulation was oat- meal. Re-infection of European mistletoe in laboratory conditions showed positive results on liquid media, and in field conditions but only after mechanical damage to the mistletoe leaves. Our results expand the knowledge regarding the optimal cultivation of this fungus. This may facilitate further mycological and pathological studies involving B. visci isolates, and the results have a theoretical basis for the implementation of measures for the prevention and control of mistletoe

Assessment of Colobanthus quitensis genetic polymorphism from the Argentine Islands region (maritime Antarctic) by actin, α- and γ-tubulin gene intron analysis

Colobanthus quitensis is one of the two angiosperm plant species commonly spread in the Antarctic. The species has been extensively analyzed at morphological, anatomical and physiological levels, but information regarding its genetic vari-ability remains limited. The aim of the study was to identify molecular genetic differences between C. quitensis populations in one of the Antarctic localities, the Argentine Islands region by estimating the intron length polymorphism of actin, α- and γ-tubulin genes. Samples of C. quitensis from different Antarctic natural populations were collected during the season of the 24th and previous Ukrainian Antarctic expeditions. Total DNA was isolated using the QIAGEN DNeasy Plant Mini Kit. The polymerase chain reaction was carried out with our own degenerate primers. The resulting amplicons were separated and visualized using polyacrylamide gel electrophoresis followed by silver nitrate staining. Molecular genetic analysis of natural populations of C. quitensis was performed using three DNA-marker systems based on the detection of intron length polymor-phism of actin, α- and γ-tubulin genes. A low level of genetic polymorphism of C. quitensis in the studied region by these types of markers was established. By assessing the intron length polymorphism of actin genes of the studied C. quitensis populations it was possible to establish that the populations of Skua Island had unique amplicons characteristic only for this location. This indicates the possibility of further use of the analysis of intron length polymorphism of actin genes for the study of the molecu-lar genetic diversity of the Antarctic pearlwort. At the same time, the results of analysis of the intron length polymorphism of α- and γ-tubulin genes induce selection of more specific primers, taking into account the structure of the C. quitensis genome. C. quitensis in the study region has a low level of genetic variability in intron length polymorphism of actin, α- and γ-tubulin genes. Overall, the results indicate that DNA markers based on gene structure analysis of highly conserved cytoskeletal pro-teins, namely, actin, α- and γ-tubulin, as additional sources of information, can be used for molecular genetic analysis of C. quitensis populations in the Antarctic

Genome-wide identification and evolution of the tubulin gene family in Camelina sativa

Abstract Background Tubulins play crucial roles in numerous fundamental processes of plant development. In flowering plants, tubulins are grouped into α-, β- and γ-subfamilies, while α- and β-tubulins possess a large isotype diversity and gene number variations among different species. This circumstance leads to insufficient recognition of orthologous isotypes and significantly complicates extrapolation of obtained experimental results, and brings difficulties for the identification of particular tubulin isotype function. The aim of this research is to identify and characterize tubulins of an emerging biofuel crop Camelina sativa. Results We report comprehensive identification and characterization of tubulin gene family in C. sativa, including analyses of exon-intron organization, duplicated genes comparison, proper isotype designation, phylogenetic analysis, and expression patterns in different tissues. 17 α-, 34 β- and 6 γ-tubulin genes were identified and assigned to a particular isotype. Recognition of orthologous tubulin isotypes was cross-referred, involving data of phylogeny, synteny analyses and genes allocation on reconstructed genomic blocks of Ancestral Crucifer Karyotype. An investigation of expression patterns of tubulin homeologs revealed the predominant role of N6 (A) and N7 (B) subgenomes in tubulin expression at various developmental stages, contrarily to general the dominance of transcripts of H7 (C) subgenome. Conclusions For the first time a complete set of tubulin gene family members was identified and characterized for allohexaploid C. sativa species. The study demonstrates the comprehensive approach of precise inferring gene orthology. The applied technique allowed not only identifying C. sativa tubulin orthologs in model Arabidopsis species and tracking tubulin gene evolution, but also uncovered that A. thaliana is missing orthologs for several particular isotypes of α- and β-tubulins