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In Poland, it has been found so far about 260 species of plant and fungi parasitic nematodes. Among the nematodes feeding on plants there are species that have the status of quarantine pests. Before taking the decision to control nematodes, it is necessary to identify the pest species. The identification of nematodes is difficult because of the paucity of easily scorable diagnostic morphological characters. Consequently, molecular identification tools could solve this problem. For molecular identification of species are suitable different PCR-based methods, DNA barcoding and DNA microarrays. The mitochondrial cytochrome oxidase c subunit 1 (COI) gene is one of the most popular markers for species identification across the animal kingdom. Species identification based on COI sequence received name DNA barcoding. DNA microarrays, first created 20 years ago, are well established and able to differentiate hundreds of specimens simultaneously. Microarray technologies, which involve hybridization of short specific probes to target DNA and subsequent detection of the hybridization signal, have been widely used as diagnostic tools.
Characteristic damage caused by feeding leaf nematodes were recorded on sage plants of Compacta erecta group growing in the field in spring 2009. Nematological analysis showed the Aphelenchoides ritzemabosi presence in collected leaves. Seeds were sampled in the autumn from sage plants and then used for seedlings’ production in the next vegetation season. Collected seeds were stored over the winter and then sown into plastic boxes in the glasshouse conditions in February 2010. In April seedlings were replanted into larger pots. They already had symptoms such as weak growth and partial leaf deformations. These were the typical symptoms of damage caused by foliar nematodes which were later found in the field during the vegetation period. The analysis of leaves confirmed A. ritzemabosi presence. Further investigation showed also the presence of nematodes in seeds. In Poland, this was the first record leaf damages caused by A. ritzemabosi on Salvia splendens. Comparison of Aphelenchoides cytochrome oxidase gene subunit I (COI) allowed to design primers for molecular identification of A. ritzemabosi. The primer pair reproducibly amplified the COI region of A. ritzemabosi, generating a 248-bp product using total genomic DNA extracted from individual nematodes.
Analysis of skeletal malformations in embryos that died between day 17 and 21 of incubation obtained from two reproductive flocks of broiler chickens showed exencephaly to be the most common defect. It comprises about 48% of all diagnosed axial skeleton disorders. Analysis of the expression of Hoxa1 and Hoxd3 genes by real time PCR revealed upregulation of Hoxa1 in chicken embryos with exencephaly (16.8-fold higher), when compared with healthy birds. Upregulation of Hoxa1 suggests that this gene may be associated with exencephaly in chickens.
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