SUMMARYIn this article, we describe the analysis of over 9000 expressed sequence tags (ESTs) from cDNA libraries obtained from various life cycle stages of Globodera pallida. We have identified over 50 G. pallida effectors from this dataset using bioinformatics analysis, by screening clones in order to identify secreted proteins up-regulated after the onset of parasitism and using in situ hybridization to confirm the expression in pharyngeal gland cells. A substantial gene family encoding G. pallida SPRYSEC proteins has been identified. The expression of these genes is restricted to the dorsal pharyngeal gland cell. Different members of the SPRYSEC family of proteins from G. pallida show different subcellular localization patterns in plants, with some localized to the cytoplasm and others to the nucleus and nucleolus. Differences in subcellular localization may reflect diverse functional roles for each individual protein or, more likely, variety in the compartmentalization of plant proteins targeted by the nematode. Our data are therefore consistent with the suggestion that the SPRYSEC proteins suppress host defences, as suggested previously, and that they achieve this through interaction with a range of host targets.m pp_585 815..828
Multiplex polymerase chain reaction was used to identify the potato cyst nematodes in soil samples from the Ukraine. The results show the occurrence of Globodera pallida in the Uzhhorod region (Zakarpats'ka oblast'), where only G. rostochiensis had been previously reported. In the mixed potato cyst nematode (PCN) populations, G. pallida was less prevalent (2À5%) than G. rostochiensis (95À98%). A phylogenetic analysis based on ribosomal DNA internal transcribed spacer sequences showed that the Ukrainian population of G. pallida had >99% sequence identity with other G. pallida pa2/3 isolates from Europe. This study has demonstrated that polymerase chain reaction-mediated amplification of specific regions of the potato cyst nematode genome is not only highly effective as a species diagnostic tool but is also a sensitive method which can be used for taxonomic purposes with cyst collections which vary in age.Abbreviations: ITS À internal transcribed spacer; PCN À potato cyst nematode; rDNA À ribosomal DNA
A quickscan pest risk analysis for the apple root-knot nematode Meloidogyne mali for the territory of Ukraine was performed. This assessment was initiated in response to the recent (2012/2013) interception of the apple root-knot nematode in the Netherlands and Italy and because of the species inclusion on the EPPO Alert List in 2014. The risk of M. mali introduction, establishment and economic impact in Ukraine was assessed as likely, which proved the need for specific statutory actions to be taken to prevent ingress of the apple root-knot nematode and mitigate its effects in Ukraine. It is stated that the detailed pest risk analysis is required.
Heterodera schachtii Schmidt, 1871 is one of the most economically important pests of sugar beet (Beta vulgaris L.) worldwide. It is also widespread in most sugar beet growing regions in Ukraine causing serious yield reduction and decreasing sugar content of sugar beet in infested fi elds. An advanced parasitic strategy of H. schachtii is employed to support nematode growth, reproduction and harmfulness. In intensive agriculture systems the nematode control measures heavily rely on nematicides and good agricultural practice (crop rota- tion in the fi rst place). But alternative strategies based on nematode resistant sugar beet cultivars and hybrids are required as none of nematicides approved for the open fi eld application are registered in Ukraine. Here we review the achievements and problems of breeding process for H. schachtii resistance and provide the results of national traditional breeding program. Since the beginning of 1980s fi ve sugar beet cultivars (Verchnyatskyi 103, Yaltuschkivska 30, Bilotcerkivska 45, BTs-40 and Yuvileynyi) and seventeen lines partly resistant or toler- ant to H. schachtii have been obtained throughout targeted crossing and progenies assessment in the infested fi elds. The further directions for better utilization of genetic sources for nematode resistance presented in na- tional gene bank collection are emphasized. There is a need for more accurate identifi cation of resistance genes, broader application of reliable molecular markers (suitable for marker-assisted selection of nematode resistant plants in the breeding process) and methods for genetic transformation of plants. Crop cash value and national production capacity should drive the cooperation in this fi eld. Knowledge as well as germplasm exchange are thereby welcomed that can benefi t breeding progress at national and international level.
Following the discovery of Globodera pallida in the Uzhhorod region (Zakarpats'ka oblast') of Ukraine, two populations from this region were further examined. Firstly, their virulence was assessed in relation to two sources of resistance to G. pallida and to other G. pallida populations from Europe and South America. The results showed that the Ukrainian populations were very similar in their virulence to the other European populations. One of the test host genotypes was a new cultivar derived from Solanum tuberosum spp. andigena CPC2802, which proved to be more highly resistant than previously available partially resistant cultivars. The second experiment was a comparison of the mitochondrial Cytochrome B gene from the Ukrainian populations with other European and South American populations. The data indicated that the Ukrainian populations were similar to other European populations.
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