DETECTION OF THE PATHOGEN OF VIRAL DISEASE IN SAMBUCUS NIGRA PLANTS

UDC 578.3+582.974 L. Mishchenko, Dr. Sci. Biol, Prof., A. Dunich, PhD Taras Shevchenko National University of Kyiv, Kyiv, Ukraine, O. Taran, PhD National University of Life and Environmental Sciences of Ukraine, Kyiv, Ukraine, L. Glushchenko, PhD Experimental Station of Medicinal Plants of the Institute Agroecology and Environmental of National Academy of Agrarian Sciences of Ukraine, Poltava region, Ukraine

Most reports of elderberry infecting are about Cherry leaf roll virus and carlaviruses. Blueberry scorch virus (BlScV), Elderberry symptomless virus (ElBSV) and several other putative members of the genus Carlavirus (family Betaflexiviridae) have also been reported in elderberry [1,3,4,11,26]. There is report about infecting of Sambucus canadensis plants by filamentous virus which similar on morphological features to carlaviruses [9]. Subsequently, the virus was detected in the Netherlands and was named Elderberry virus A [26]. Recent studies of elderberry samples (Sambucus spp.) from Missouri (USA) showed infecting of these plants with two different viruses, which also belong to the genus Carlavirus [12]. Five novel carlaviruses tentatively named as Elderberry virus A-E (ElVA-ElVE, respectively) were discovered [7,8]. Elderberry carlavirus group 1 (ElVA, ElVB, ElVD)and group 2 (ElVC and ElVE) appear to have emerged from two distinct lineages, containing closely related viruses that infect the same host, indicative of sympatric speciation [Ho et al, 2016]. This, in addition to the recombination analysis, imply that elderberry, along with hop, phlox and potato (respectively infected by Hop latent virus and Hop mosaic virus; Phlox virus B and Phlox virus S; Potato virus P and Potato virus S), are major contributors of the carlaviruses evolution.
Despite the considerable amount of the studies of elderberry viruses in the world and particularly in Europe, such investigations in Ukraine haven't been conducted.
That's why the aim of the research was to obtain the Sambucus nigra plants on the presence of viral diseases.
Materials and methods. For diagnostics of viruses in the plants applied the methods of visual diagnostics, ELISA and transmission electronic microscopy (EM). Contrasting has been made with 2% solution of phosphorus -tungstic acid. Virions are investigated using electron microscope JEM 1230 (JEOL, Japan).
Detection and identification of viruses has been carried out with enzyme-linked immunosorbent assay (DASmodification) using commercial test-systems of firm LOE-WE (Germany). The results of reaction registered on the rider Termo Labsystems Opsis MR(THE USA) with Dynex Revelation Quicklink software at lengths of waves of 405/630 нм. All samples showing values three times higher than the negative controls are assumed as virus positive.
The extinction values (the optical density) of the samples were processed by statistical analysis of Student's criterion, quoted by Lidanski [13]. The confidential intervals were at a significance rate of P ≤ 0.05 of Student's criterion.
Results and discussion. Under observations of wild elderberry plants in Poltava (2015Poltava ( -2016 and Kyiv (2016) regions we detected plants with chlorotic symptoms (a, b) and rolling of leaf tops (c) and twisting up the edges of the leaves (Fig.1). It should be noted that analysis of world scientific literature on elderberry viruses showed that the most common symptom is chlorotic mottling ('blotching') and induced by many viruses [3,20,22]. But the chlorotic foliage symptom is not found in any report.
Filamentous virions 650±50 × 12 nm were found in the elderberry leaves conducting the transmission electron microscopy method. It was marked higher concentration of virions in plants with leaf rolling symptom compared with chlorotic. In addition, earlier PVM and PVY were identified by us in tomatoes with leaf rolling symptoms for the same agroecological conditions (in Poltava and Kiev regions) [16]. Such morphology is characteristic for viruses from the genus Potyvirus (Potyviridae) and Carlavirus (Betaflexiviri-dae). It is known that these genuses have a large number of representatives. So in our research, we settled on potyand carlaviruses that are wide spread in Ukraine and have a wide range of host plants.
Based on the our results and on the data of other scientists we tested elderberry plants with mentioned above symptoms on the presence of carlaviruses (Potato virus M), potyviruses (Potato virus Y, Soybean mosaic virus, Bean yellow mosaic virus) and Alfalfa mosaic virus. Alfalfa mosaic virus was detected in guelder rose (Viburnum), belonging to the same family with the elderberry [3,23].
According to the ELISA results antigens of PVY, PVM, SMV, AMV, and BYMV in the tested elderberry samples were not found (tabl.).