CABINET MINISTRY OF UKRAINE NATIONAL UNIVERSITY OF LIFE AND ENVIRONMENTAL SCIENCES OF UKRAINE Ecological monitoring of lucerne mosaic virus in agricultural landscapes of NULE’s enterprise “Agronomic research station” Author: E.E. Gladun Supervisors: Prof. Dr. Agricultural Sci. V.М. Chaika PhD. in Agriculture, Associate Prof. І.О. Antipov

Aim and goals of master’s thesis Inspection of lucerna plantation to identify infection of mosaic virus. Diagnostic, identification and revealing of pathways of lucerne mosaic viruses (LMV) distribution in agrarian cenoses. To achieve the aim following goals were determined: 1. To provide visual diagnostic of lucerne planting to reveal affection by lucerne mosaic viruses (LMV). 2. To account host weeds of lucerne virus infections. To calculate amount of host weeds of lucerne virus infections among plantation. 3. To provide LMV diagnostic using methods of electronic microscop and immunoenzyme analisis. 4. To reveal ecological pathways of (LMV) distribution in agrarian cenoses.

Alfalfa mosaic virus Symptoms: - rugosity of leaves; - top necrosis; - light-green mosaic; - yellow mosaic. Lucerne mosaic virus

Experimental sites are situated at territory of NULE’s enterprise “Velykosnitinske agronomic educational-research station of O. V. Muzichenka” Lucerne The object of our study was lucerne plants. Bush is high, semi- straight, leaf is green, open-type, petals are licht-violet.

Visual diagnostic of LMV Edge necrosis on lucerne leaves A plant with symptoms of dot mosaic

Affected by viral chlorosis lucern plant (left), healthy plant (right)

Symptoms, revealed on lucerne plants №SymptomAmount % from general amount 1Rolling leaves Viral chlorosis31,6 3Mosaics94,8 4Dwarf plants42,1 5 Total plants with symptoms 3116,6 6 Plants without symptoms 15683,4 7Inspected plants187100

Species composition and presence of weeds on lucerne plantation № Species of weed Spec. on m 2 % from genera l amoun t 1 Setaria glauca 2851,8 2 Echinochloa crusgalli35,5 3 Orache (Chenopodium album) Amaranth (Amaranthus retroflexu) 712,9 5 Wheat grass (Agropуron repens L.) 2 3,7 6 Sow-thistle Circium arvensis 4 7,4 7 Genral amount

Results of electronic microscopical study of lucerne samples Bacilloid virions of lucerne mosaic virus (size 250х60 nm)

Results of lucerne samples, method of IEA № of sample Indexes of optical density, wave length 405 nm RepetitionMean ,4110,4210,416 20,4060,4130,454 0,424 30,4090,4120,421 0, ,4200,401 0,412 50,4140,4020,419 0,412 60,4230,4170,416 0,419 71,7211,7311,750 1,734 80,4130,4220,414 0,416 90,4110,4190,421 0, ,4080,4130,404 0,408 11(PK)1,9631,9211,944 1,943 12(NK)0,4030,4110,415 0,410

CONCLUSIONS 1. Visual diagnostics of lucern plantation has beem provided to assess affection by lucerne mosaic virus. As a result of investigation four types of symptomes were revealed: rolling leaves 8% from general amount of inspected plants, viral chlorosis – 1,6%, intravein, dot, wave, circle, edge mosaics – 4,8%, dwarfness – 2,1%. Rolling leaves and mosaics occured the most often. 2. As a result of inspection host plants were revealed: (Setaria glauca), (Echinochloa crusgalli),(Chenopodium album), (Amaranthus retroflexus),(Circium arvensis), (Agropуron repens L.). Sensitive to LMV species were between these weeds, they may be affected by virus, accumulate it and transmite this virus: sow- thistle, amaranth, orache. Following symptoms were observed: chlorosis in sow- thistle, rolling and mosaics in amaranth, chlorosis and mosaics in orache.

3. During observation time it has been revealed that lucern plants, affected by LMV, loose natural resistance and got affected by different deceases more intensely. 4. LMV diagnostic using electronic microscopy method was performed. It has been shown that virus, causing symptoms belongs to Alfamovirus group, it has bacilloid morphology and size of particles 250х60 nm. 5. As a result of immunoenzyme analysis presence of LMV in sample # 7 was detected in optical density 1,734 nm.

RECOMMENDATIONS After analysis of conclusions mentioned above it can be summarised that LMV is distributes in agrocenoses of NULE’s enterprise “Velikosnitinske educational-research station” both on cultural (in that case has economic value) and wild-growing plants. As a result of infection decrease of productivity is evident, to prevent distribution of LMV is possible in case of : 1.Using sanated planting material, which has genetical resistance to viral agents. 2. Following to high culture of land cultivating. 3. Regulation of vectos quantity and weed the fields, especially destruction of that weeds that are reservoirs of LMV. 4.Providing of total control concerning viral bearing of lucern planting, especially those, which would be used as a seeds.

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