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Botanicals, such as essential oils (EO) and their compounds, are considered a viable eco-friendly alternative to synthetic insecticides, which threaten human health and ecosystem functioning. In the present study, we explored the potential use of two EO compounds, trans-anethole (phenylpropanoid) and S-(+)-carvone (monoterpene ketone), against gypsy moth larvae (GML), a serious pest of deciduous forests and orchards. GML feeding, survival, molting, and nutritional physiology were assessed at different compound concentrations and compared with the effects of the commercial botanical product NeemAzal®-T/S (neem). The impact of botanicals on GML feeding was assessed by the leaf-dipping method and showed the highest antifeeding activity of neem in the no-choice assay. GML that were offered a choice were deterred by anethole and attracted by low concentrations of carvone and neem. Ingestion of botanicals was more effective in inducing mortality and reducing molting than residual contact exposure. Anethole and carvone were better toxicants but worse growth regulators than neem. Assessing nutritional indices revealed reduced growth, consumption, and food utilization in larvae fed on botanical-supplemented diets. The highest metabolic cost of food processing was recorded in carvone-fed larvae, which exhibited a negative growth rate. The results suggest that anethole and carvone might be used as control agents against GML.

The Colorado potato beetle (CPB) is a serious pest of economically important Solanaceae species. The use of essential oil compounds in pest management has been proposed as an alternative to harmful chemical insecticides that disturb human health and ecosystem functioning. We examined the antifeedant activity of three concentrations (0.125%, 0.25% and 0.5%) of pure camphor and a thujone-camphor mixture against 3rd instar larvae and adults. Their efficacy was evaluated according to the degree of leaf damage and avoidance of treated leaves by the CPB. Treatment of potato leaves significantly reduced leaf damage compared to the control. Leaf protection increased at higher concentrations of the examined compounds. Camphor was more effective against larvae and the thujone-camphor mixture was more effective against adults. Additionally, adults moved faster towards the control leaf disc in the two-choice olfactometer assay if an alternative disc was treated with a thujone-camphor mixture, whereas larvae responded similarly to the two potential repellents. However, after contact with the leaf disc treated with the highest compound concentration, the larvae escaped faster from the thujone-camphor mixture than from pure camphor. In conclusion, both examined compounds are promising eco-friendly antifeedants, but their efficacy depends on the developmental stage of the beetle, compound type and applied concentration.

Acanthoscelides obtectus Say (Coleoptera: Chrysomelidae: Bruchinae), is one of the most important pests of the common bean Phaseolus vulgaris L. Without appropriate management it may cause significant seed loss in storages. In search for means of environmentally safe and effective protection of beans we assessed biological activity of thymol, an oxygenated monoterpene present in essential oils of many aromatic plants. We studied contact toxicity of thymol on bean seeds and its effects on adult longevity and emergence in F1 generation. Furthermore, we determined acetylcholinesterase (AChE), superoxide dismutase (SOD), catalase (CAT), mixed-function oxidase (MFO), carboxylesterases (CarE) and glutathione S-transferase (GST) activities in response to 24 h exposure of beetles to sublethal and lethal thymol concentrations. Our results showed that thymol decreased adult survival, longevity and percentage of adult emergence. Higher median lethal concentration (LC50) was recorded in females indicating their higher tolerance comparing to males. Overall, activities of SOD, CAT and CarE increased at sublethal and MFO increased at both sublethal and lethal thymol concentrations. On the other hand, GST and AChE activities decreased along with the increase in thymol concentrations from sublethal (1/5 of LC50, 1/2 of LC50) to lethal (LC50). Enzyme responses to the presence of thymol on bean seed were sex-specific. In the control group females had lower CarE and higher SOD, CAT and GST activity than males. In treatment groups, females had much higher CAT activity and much lower CarE activity than males. Our results contribute to deeper understanding of physiological mechanisms underlying thymol toxicity and tolerance which should be taken into account in future formulation of a thymol-based insecticide.

The Colorado potato beetle, Leptinotarsa decemlineata (Say), is one of the most destructive pest species to have developed resistance to most chemical insecticides. We determined the composition and evaluated the potential of Tanacetum parthenium L. and Tanacetum vulgare L. (Asteraceae family) essential oil (EO) application as an alternative eco-friendly control strategy against L. decemlineata. We assessed the antifeedant activity for L. decemlineata larvae and adults by estimating the damage to potato leaves treated with three concentrations of EOs dissolved in ethanol (0.125, 0.25 and 0.5%). Results showed that T. parthenium EO was more effective against larvae, and T. vulgare was more effective against adults. In an olfactometer assay, the time required to choose an untreated leaf disc did not depend on the Tanacetum species, or life stage examined. However, the concentration of EO exhibited a significant effect on the behaviour of both developmental stages. At higher EO concentrations, both third instar larvae and adults require less time to choose an untreated leaf disc. Additionally, T. parthenium EO provoked more rapid movement away from the treated leaf disc than T. vulgare, especially at the highest concentration. Successful modification of L. decemlineata behaviour by the two Tanacetum oils suggests that they possess the potential for use in potato protection.

The gypsy moth (Lymantria dispar L. (Lepidoptera: Erebidae)) is a serious pest of hardwood forests. In the search for an environmentally safe means of its control, we assessed the impact of different concentrations of essential oils (EOs) from the seeds of three Apiaceae plants (anise Pimpinella anisum, dill Anethum graveolens, and fennel Foeniculum vulgare) on behavior, mortality, molting and nutritional physiology of gypsy moth larvae (GML). EOs efficacy was compared with commercial insecticide NeemAzal(R)-T/S (neem). The main compounds in the Eos were trans-anethole in anise; carvone, limonene, and alpha-phellandrene in dill; and trans-anethole and fenchone in fennel seed. At 1% EOs concentration, anise and fennel were better antifeedants and all three EOs were more toxic than neem. Neem was superior in delaying 2nd to 3rd larval molting. In the 4th instar, 0.5%, anise and fennel EOs decreased relative consumption rate more than neem, whereas all three EOs were more effective in reducing growth rate, approximate digestibility and efficiency of conversion of food into body mass leading to higher metabolic costs to GML. Decrease in consumption and metabolic parameters compared to control GML confirmed that adverse effects of the EOs stem from both pre- and post-ingestive mechanisms. The results indicate the potential of three EOs to be used for gypsy moth control.

U radu je ispitano delovanje rastvora etarskih ulja majčine dušice, ruzmarina i bosiljka i njihovih odabranih komponenata timola, α-pinena, 1,8-cineola i linalola na odrasle jedinke pasuljevog žiška (Acanthoscelides obtectus Say)..., In this research the effect of solutions of thyme, rosemary and basil essential oils and their selected constituents thymol, α-pinene, 1,8-cineole and linalool was tested against the bean weevil adults (Acanthoscelides obtectus Say)...

Gypsy moth, Lymantria dispar L. (Lepidoptera: Erebidae) feeds on a large number of tree species, while ash, Fraxinus spp. (Lamiales: Oleaceae) species are considered resistant and are only sporadically eaten. To assess the conditions under which late instar gypsy moth larvae (GML) can temporarily use non-host common ash ( CA) (F. excelsior L.), and to evaluate their ability to recover from ingestion of this toxic food, we determined the relative growth rate, the relative consumption rate and the amount of produced feces in different laboratory feeding trials. Our report is the first to show that under specific circumstances, the resources acquired after short-term consumption of CA leaves can be utilized for larval growth. We varied the intensity of density and starvation stress prior to feeding on CA leaves. We observed that after moderate stress a group of GML was temporarily capable of coping with CA leaves. Although observed growth and consumption were much lower on CA than on the optimal host oak, Quercus cerris L. (Fagales: Fagaceae), CA-oak-switched larvae showed the ability to recover from short-term use of a toxic non-host foliage. This suggests that feeding on CA might enable GML to survive under conditions of food shortage.

Simple Summary Compounds of botanical origin, including essential oils (EOs), which coevolved as plant defense mechanism against herbivores and pathogens have been proposed as a promising strategy for post-harvest control of stored product insects. Despite several drawbacks, such as low stability, phytotoxicity and food odor changes at high concentrations, EOs are believed to be safe for human and environmental health and thus have an advantage comparing to conventional insecticides. The present study was aimed to examine acute toxicity and effects of thyme (Thymus vulgaris) EO on longevity, behavior and physiology of the bean weevil (Acanthoscelides obtectus), a major pest of stored bean seeds. We found that exposure to thyme oil reduced adult survival and longevity and induced damages to lipids and proteins in a concentration-dependent manner. Sublethal EO concentrations also deterred female egg laying and strongly inhibited adult emergence. Efficacy of such low EO concentrations in suppression of adult emergence implies that thyme EO might be cost-effective and environmentally low risk botanical insecticide for bean seed protection against A. obtectus. The bean weevil Acanthoscelides obtectus Say (Coleoptera: Chrysomelidae: Bruchinae) can cause significant losses in production of its primary host common bean Phaseolus vulgaris L. To avoid bean protection with environmentally risky chemical insecticides and provide sustainable and safe production of food, new pest management methods based on natural compounds are investigated. In the present study, we evaluated protective potential of the essential oil (EO) from the common thyme Thymus vulgaris L. applied on bean seeds. We assessed residual contact toxicity of thyme EO and its effects on A. obtectus longevity, oviposition and adult emergence. Furthermore, to elucidate the role of oxidative stress in thyme EO toxicity, we estimated the levels of oxidatively damaged proteins and lipids, as well as the level of thiol

The development of "green" alternatives to chemical pesticides could play a crucial role in integrated pest management (IPM). Their use is considered either as a substitution for or in addition to hazardous synthetic products. We analysed the influence of three concentrations of tansy (Tanacetum vulgare L.) essential oil (EO), previously characterised by GC-MS, on the survival and moulting of the 2nd instar and the nutritional indices of the 4th instar gypsy moth (Lymantria dispar L.) larvae. In a residual contact toxicity assessment, the exposure to tansy EO caused low mortality ( lt 10%) while larval development was significantly slowed down, i.e., the percentage of larvae that moulted into the 3rd instar was reduced. On the other hand, when tansy EO was incorporated into the diet digestive toxicity assay), high mortality and a lack of moulting after 120 h of eating were recorded for the highest applied concentration of EO. During 48 h of feeding on EO-supplemented food at concentrations of 0.5 and 1% (v/v), the relative growth rate (RGR) of the 4th instar larvae significantly decreased, which can be explained by a significant reduction of the relative consumption rate (RCR) and significantly or marginally significantly lower efficiency of conversion of ingested food into insect biomass (ECI). Although the RCR was also reduced with the lowest applied EO concentration (0.1%), the ECI was not affected which meant the RGR was as high as it was for the control larvae. ECI changes, when two higher EO concentrations were applied, were due to a reduction in the efficiency of conversion of digested food into biomass (ECD), while approximate digestibility was unaffected by the presence of EO in the food. Our results on the significant negative effects of tansy EO on gypsy moth larval survival, development time, and nutritional physiology suggest that it could be considered in future designs for botanical insecticides for gypsy moth control.

The bean weevil (Acanthoscelides obtectus, Say) is a serious pest of stored bean seeds. Bean weevil control relies heavily on the use of synthetic insecticides. In the search for a sustainable alternative, the residual contact toxicity and anti-oviposition activity of thyme (Thymus vulgaris L.), rosemary (Rosmarinus officinalis L.) and basil (Ocimum basilicum L.) essential oils as well as their dominant components (thymol, alpha-pinene, 1,8-cineole and linalool) were tested against A. obtectus adults. Out of the seven tested botanicals, T. vulgaris oil, thymol and linalool exhibited the highest toxic potential (>90% mortality). Females were less susceptible than males. The insecticidal activity of these botanicals was much greater when they were applied on glass compared to direct application to the bean. All tested botanicals reduced oviposition by bean weevil females. T. vulgaris oil, thymol and a-pinene also deterred bean weevil oviposition, as revealed by a two-choice test. Our research shows that T. vulgaris oil and thymol are promising and sustainable alternatives to synthetic pesticides for protecting stored beans against the bean weevil.

The bean weevil (Acanthoscelides obtectus, Say) is a serious pest of stored bean seeds. Bean weevil control relies heavily on the use of synthetic insecticides. In the search for a sustainable alternative, the residual contact toxicity and anti-oviposition activity of thyme (Thymus vulgaris L.), rosemary (Rosmarinus officinalis L.) and basil (Ocimum basilicum L.) essential oils as well as their dominant components (thymol, alpha-pinene, 1,8-cineole and linalool) were tested against A. obtectus adults. Out of the seven tested botanicals, T. vulgaris oil, thymol and linalool exhibited the highest toxic potential (>90% mortality). Females were less susceptible than males. The insecticidal activity of these botanicals was much greater when they were applied on glass compared to direct application to the bean. All tested botanicals reduced oviposition by bean weevil females. T. vulgaris oil, thymol and a-pinene also deterred bean weevil oviposition, as revealed by a two-choice test. Our research shows that T. vulgaris oil and thymol are promising and sustainable alternatives to synthetic pesticides for protecting stored beans against the bean weevil.

Larvae of gypsy moth (Lymantria dispar L.) are major defoliators of deciduous forests and urban environment. The biological means of regulation of moth population density were incorporated along with conventional treatment in the integrative management concept. The effects of tansy (Tanacetum vulgare L.) essential oil (EO) to gypsy moths were monitored through the effects of residual contact and digestive toxicity on the mortality and development in the second instar larvae, but also through the growth and feeding indices of the fourth instar larvae. Although EO presence didn’t cause residual contact toxicity or significantly affect larval mortality, tested oil notably elongated developmental time by delaying the molting moment and reduced percentage of molted larvae. On the other hand, EO digestion caused significant mortality in the second instar larvae (72% compared to control, at the end of experiment). Digestion of food enriched with tansy oil decreased the efficiency of the diet, and led to reduced mass gain, and therefore induced the delay of the molting moment, or completely stopped molting into the third instar larvae. Besides that, EO reduced the daily mass gain (0.06 mg mg-1 day-1 compared to 0.3 mg mg-1 day-1 in control) and the food consumption rate (0.61 mg mg-1 day-1 compared to 1.23 mg mg-1 day-1 in control) in the fourth instar larvae and caused decrease of the conversion rate of digested food to larvae body mass (10.62% compared to 26.05% in control). Despite that, food assimilation rate to larvae body mass was not significantly decreased, which moderated negative effects of EO digestion as well as reduced food intake.

For long-term protection of stored products there is a growing demand to replace chemical insecticides due to their effects on human health and environmental safety. Particulate materials, such as inert dusts and various submicron and nanomaterials have been extensively tested as viable alternatives. This is the first study on the insecticidal impact of alumina powder (alpha-Al2O3) on the bruchid pest, Acanthoscelides obtectus (Say) (Coleoptera: Chrysomelidae: Bruchinae). By altering the fuel to oxidant molar ratio (F/O: 0.5, 0.8, 1.2) in an autocombustion reaction we synthesized alumina powders with specific surface area and particle size varying from the nano- to micron scale. It was found that particle morphology influenced survival and progeny number of A. obtectus. The order of powders from low to high efficacy in reducing beetle performance (F/O-0.8 lt F/O-0.5 lt F/O-1.2) correlated well with increase in surface area, pore volume and diameter, and decrease in particle size. Survival was also affected by time of exposure, the applied dose and sex. The estimated median lethal concentration of the most efficient powder F/O-1.2 was significantly lower in males (LC50 = 330.4 ppm) than in females (LC50 = 409.6 ppm). Our results suggest that alumina powder can be considered for seed protection against A. obtectus, particularly during long-term storage, as it is cost effective, exerts limited toxicity to humans and demands no repetitive use like conventional pesticides.

Ključna pretpostavka evolucione teorije starenja jeste da opadanje uzrasno-specifičnog preživljavanja organizama predstavlja rezultat smanjenja intenziteta prirodne selekcije nakon početka reprodukcije. U osnovi evolucije starenja nalaze se dva populaciono-genetička mehanizma: akumulacija mutacija (AM) i antagonistička plejotropija (AP). U skladu sa pretpostavkama evolucione teorije starenja, kod eksperimentalnih populacija pasuljevog žiška (Acanthoscelides obtectus) selektovanih više od 170 generacija za ranu (E režim) ili kasnu (L režim) reprodukciju, uočena je divergencija u dužini života i drugim preadultnim i adultnim osobinama životne istorije. Evolucija veće dužine života u L režimu u saglasnosti je sa pretpostavkama AP modela, dok u evoluciji kraćeg života E žižaka važnu ulogu ima akumulacija mutacija. Tretiranjem E i L žižaka pesticidima (parakvat i tebufenpirad) koji generišu slobodne radikale, potvrđena je pozitivna korelacija između dužine života i otpornosti na oksidativni stres. Budući da mitohondrija predstavlja glavno mesto sinteze energije i slobodnih radikala, u ovoj tezi je istraženo da li je uzrasno-specifična selekcija oblikovala varijabilnost mtDNK, nDNK i njihovih epistatičkih interakcija. Genotipizacijom mitohondrijskog COI gena i mikrosatelitskih lokusa pokazana je efikasnost uzrasno-specifične selekcije u međupopulacionoj divergenciji mitohondrijskog i jedarnog genskog pula kod pasuljevog žiška. Odvijanje procesa mito-jedarne koevolucije analizirano je korišćenjem mito-jedarnih linija u kojima su E i L mitohondrijski genomi kombinovani sa E i L jedarnim genomima. Pokazano je da su linije sa narušenim mito-jedarnim interakcijama imale niže aktivnosti mitohondrijskih kompleksa elektron transportnog lanca i lošiju preadultnu performansu., According to the evolutionary theory of ageing, age-specific decline in survival is the result of decreasing intensity of natural selection after the onset of reproduction. The evolution of ageing may be explained by two different, but not mutually exclusive, genetic mechanisms: antagonistic pleiotropy (AP) and mutation accumulation (MA). In accordance with evolutionary theory of ageing, two sets of the seed beetle (Acanthoscelides obtectus) experimental lines selected for more than 170 generations for early (E regime) or late (L regime) reproduction, show divergence in longevity and ageing patterns, as well as in other preadult and adult life history traits. Evolution of long life in L regime is predominantly based on antagonistic pleiotropy gene effects, while mutation accumulation plays important role in evolution of short life in E regime. By exposure of E and L beetles to two pesticides (paraquat and tebufenpyrad) that generate free radicals, it was confirmed that extended longevity in L beetles (especially in females) was associated with higher oxidative stress resistance. Additionally, since mitochondria are main site of free radical and energy production, effects of age-specific laboratory selection on mtDNA, nDNA and their epistatic interaction were assessed. Genotyping of mitochondrial COI gene and nuclear microsatellite loci showed that selection in E and L regimes led to divergent evolution of mtDNA and nDNA gene pools. Mitonuclear coevolution was investigated using mitonuclear introgression lines in which E and L mitochondrial genomes were expressed in both E and L nuclear background. It was found that lines with disrupted mitonuclear interactions had lower activity of the mitochondrial electron transport chain complexes and lower preadult performance.

Ključna pretpostavka evolucione teorije starenja jeste da opadanje uzrasno-specifičnog preživljavanja organizama predstavlja rezultat smanjenja intenziteta prirodne selekcije nakon početka reprodukcije. U osnovi evolucije starenja nalaze se dva populaciono-genetička mehanizma: akumulacija mutacija (AM) i antagonistička plejotropija (AP). U skladu sa pretpostavkama evolucione teorije starenja, kod eksperimentalnih populacija pasuljevog žiška (Acanthoscelides obtectus) selektovanih više od 170 generacija za ranu (E režim) ili kasnu (L režim) reprodukciju, uočena je divergencija u dužini života i drugim preadultnim i adultnim osobinama životne istorije. Evolucija veće dužine života u L režimu u saglasnosti je sa pretpostavkama AP modela, dok u evoluciji kraćeg života E žižaka važnu ulogu ima akumulacija mutacija. Tretiranjem E i L žižaka pesticidima (parakvat i tebufenpirad) koji generišu slobodne radikale, potvrđena je pozitivna korelacija između dužine života i otpornosti na oksidativni stres. Budući da mitohondrija predstavlja glavno mesto sinteze energije i slobodnih radikala, u ovoj tezi je istraženo da li je uzrasno-specifična selekcija oblikovala varijabilnost mtDNK, nDNK i njihovih epistatičkih interakcija. Genotipizacijom mitohondrijskog COI gena i mikrosatelitskih lokusa pokazana je efikasnost uzrasno-specifične selekcije u međupopulacionoj divergenciji mitohondrijskog i jedarnog genskog pula kod pasuljevog žiška. Odvijanje procesa mito-jedarne koevolucije analizirano je korišćenjem mito-jedarnih linija u kojima su E i L mitohondrijski genomi kombinovani sa E i L jedarnim genomima. Pokazano je da su linije sa narušenim mito-jedarnim interakcijama imale niže aktivnosti mitohondrijskih kompleksa elektron transportnog lanca i lošiju preadultnu performansu., According to the evolutionary theory of ageing, age-specific decline in survival is the result of decreasing intensity of natural selection after the onset of reproduction. The evolution of ageing may be explained by two different, but not mutually exclusive, genetic mechanisms: antagonistic pleiotropy (AP) and mutation accumulation (MA). In accordance with evolutionary theory of ageing, two sets of the seed beetle (Acanthoscelides obtectus) experimental lines selected for more than 170 generations for early (E regime) or late (L regime) reproduction, show divergence in longevity and ageing patterns, as well as in other preadult and adult life history traits. Evolution of long life in L regime is predominantly based on antagonistic pleiotropy gene effects, while mutation accumulation plays important role in evolution of short life in E regime. By exposure of E and L beetles to two pesticides (paraquat and tebufenpyrad) that generate free radicals, it was confirmed that extended longevity in L beetles (especially in females) was associated with higher oxidative stress resistance. Additionally, since mitochondria are main site of free radical and energy production, effects of age-specific laboratory selection on mtDNA, nDNA and their epistatic interaction were assessed. Genotyping of mitochondrial COI gene and nuclear microsatellite loci showed that selection in E and L regimes led to divergent evolution of mtDNA and nDNA gene pools. Mitonuclear coevolution was investigated using mitonuclear introgression lines in which E and L mitochondrial genomes were expressed in both E and L nuclear background. It was found that lines with disrupted mitonuclear interactions had lower activity of the mitochondrial electron transport chain complexes and lower preadult performance.

U radu je ispitivano delovanje alkoholnih rastvora etarskih ulja anisa, mirođije i morača i njihovih dominantnih komponenata trans-anetola i karvona na gusenice gubara (Lymantria dispar L.). Antifidna aktivnost etarskih ulja i njihovih dominantnih komponenata primenjenih u koncentracijama 0.1, 0.5 i 1.0 % ispitivana je u dva odvojena ogleda, ogledu bez izbora i ogledu sa izborom. Ispitivanje je vršeno na gusenicama drugog stupnja, a rezultati ogleda su očitavani nakon 48 časova od početka ogleda. Rezidualna kontaktna i digestivna toksičnost etarskih ulja anisa, mirođije i morača i njihovih dominantnih komponenata primenjenih u koncentracijama 0.05, 0.1, 0.25, 0.5 i 1.0 %, ispitivana je na gusenicama gubara drugog stupnja. Pored mortaliteta gusenica, praćen je i uticaj navedenih toksičnosti na presvlačenje gusenica iz drugog u treći larveni stupanj. Ogledi u kojima je ispitivan uticaj toksičnosti ispitivanih etarskih ulja i nihovih dominantnih komponenata na mortalitet i zaustavljanje presvlačenja gusenica trajali su 120 časova, a rezultati su očitavani na svaka 24 časa. Delovanje ispitivanih etarskih ulja i njihovih dominantnih komponenata, primenjenih u koncentracijama 0.1, 0.25 i 0.5 %, na indekse rasta i ishrane gusenica gubara četvrtog stupnjsa sagledavano je u ogledima za ispitivanja njihovog uticaja na relativnu brzine rasta gusenica (RGR), relativnu brzinu konzumacije hrane gusenica (RCR), efikasnost asimilacije hrane kod gusenica (AD) i efikasnosti konverzije unete i svarene hrane kod gusenica (ECI i ECD). Rezultati ogleda su očitavani nakon 48 časova od početka ogleda. Za poređenje rezultata je korišćeno biološko sredstvo NeemAzal (standard). Statistička obrada podataka izvršena je uz pomoć softverskog paketa Statistica 7.0. (StatSoft, Inc). Analizom dobijenih rezultata utvrđeno je da dobru antifidnu aktivnost poseduju etarsko ulje anisa i njegova dominantna komponenta trans-anetol primenjeni u koncentraciji 1.0 %. Rezidualna kontaktna toksičnost ispitivani, In this work the influence of ethanol solutions of essential oils of anise, fennel and dill and their dominant components trans-anethole and carvone on gypsy moth larvae (Lymantria dispar L.) was tested. Antifeedant activity of essential oils and their dominant components applied in concentrantions 0.1, 0.5 i 1.0 % was tested in two separate tests, no choice and choice feeding ones. The testing was done on the second instar and the results of the experiments were recorded 48 h after the beginning of the experiment. Residual concant and digestive toxicity of essential oils of anise, fennel and dill and their dominant components applied in concentrations 0.05, 0.1, 0.25, 0.5 i 1.0 % were tested on the second instar gypsy moth larvae. The influence of mentioned toxicity on moulting from the second to the third instar was followed, too. The influence of toxicity of essential oils and their dominant components on mortality and disturbing moulting was evaluated after 24, 48, 72, 96 and 120 h after the beginning of the experiment. The influence of tested essential oils and their dominant components applied in concentrations 0.1, 0.25, 0.5 % on growth and feeding indices of the fourth insters was tested in the asseys about their influence on relative growth rate (RGR), relative consuption rate (RCR), aproximate digestibility (AD) and efficiency of conversation of ingested food (ECI), as well as efficiency of conversation of digested food (ECD). The results were recorded 48 h after the beginning of the experiment. Botanical standard NeemAzal was used for comparing results. Statistical data proccesing was done by softer Statistica 7.0. The analyzying of the results confirmed that essential oil of anise and its dominant component tras-anethole applied in concentration 1.0 % possess good antifeedant activity. Residual contact toksicity of tested essential oils and their dominant components as well as their influence on disturbing moulting was not satisfied. Tested essential oil

Fenotipska plastičnost, kao univerzalno svojstvo svih organizama, predstavlja sposobnost jednog genotipa da u različitim uslovima životne sredine razvije različite fenotipske vrednosti osobina. Ovaj fenomen predstavlja važan mehanizam fenotipskog variranja insekatа i nalazi se u osnovi formiranja različitih pravaca evolucionih promena njihovih populacija uzrokujući adaptivnu divergenciju i posledičnu međupopulacionu reproduktivnu izolaciju. Od posebne važnosti je i uloga fenotipske plastičnosti u procesima ekološke specijacije, kada ekološki zasnovana divergentna selekcija između životnih sredina ograničava protok gena između populacija. Koristeći pogodnosti laboratorijske evolucije, ova studija proučava osobine životne istorije, njihova međusobna ograničenja i obrasce plastičnih odgovora, populaciono-ekološke i selekcione obrasce kao i promenu reproduktivnog ponašanja kod dve grupe laboratorijskih populacija fitofagnog insekta, Acanthoscelides obtectus, koje su evoluirale na različitim biljkama domaćinima (pasulj, Phaseolus vulgaris i naut, Cicer arietinum). Populacije selektovane na različitim biljkama pokazuju jasne razlike u životnim strategijama i stepenu plastičnosti osobina životne istorije i ponašanja. Načelno, populacije selektovane na pasulju imaju izraženiju sposobnost plastičnog odgovora na variranje identiteta biljke domaćina u poređenju sa populacijama selektovanim na nautu. Selekcioni režimi pokazuju i umeren stepen reproduktivne izolacije čime se ističe značaj divergentne selekcije u početnim fazama procesa nastanka novih vrsta., Phenotypic plasticity, a universal feature of all organisms, represents genotypes’ ability to produce alternative phenotypes in different environmental conditions. In insects, this phenomenon represents an important mechanism of phenotypic variation and it is fundamental for the establishment of different evolutionary trajectories of their populations causing adaptive divergence and consequential interpopulation reproductive isolation. Phenotypic plasticity is of particular importance during the process of ecological speciation, when ecologically based divergent selection between environments reduces gene flow among populations. Using the benefits of laboratory evolution, this study investigates life history traits, their trade-offs and plasticity patterns, population dynamics and selection patterns, as well as changes in reproductive behaviour, in two sets of laboratory populations of a phytophagous insect, Acanthoscelides obtectus, evolved on different host plants (white beans, Phaseolus vulgaris and chickpeas, Cicer arietinum). Populations selected on different plant hosts demonstrate distinct life history strategies and plasticity levels of life history and behavioural traits. Generally, populations selected on beans demonstrate prominent ability to plastically respond to host plant variation compared to chickpea selected populations. Selection regimes demonstrate moderate level of reproductive isolation emphasizing the importance of divergent selection in the first phases of the speciation.

Ključna pretpostavka evolucione teorije starenja jeste da opadanje uzrasno-specifičnog preživljavanja organizama predstavlja rezultat smanjenja intenziteta prirodne selekcije nakon početka reprodukcije. U osnovi evolucije starenja nalaze se dva populaciono-genetička mehanizma: akumulacija mutacija (AM) i antagonistička plejotropija (AP). U skladu sa pretpostavkama evolucione teorije starenja, kod eksperimentalnih populacija pasuljevog žiška (Acanthoscelides obtectus) selektovanih više od 170 generacija za ranu (E režim) ili kasnu (L režim) reprodukciju, uočena je divergencija u dužini života i drugim preadultnim i adultnim osobinama životne istorije. Evolucija veće dužine života u L režimu u saglasnosti je sa pretpostavkama AP modela, dok u evoluciji kraćeg života E žižaka važnu ulogu ima akumulacija mutacija. Tretiranjem E i L žižaka pesticidima (parakvat i tebufenpirad) koji generišu slobodne radikale, potvrđena je pozitivna korelacija između dužine života i otpornosti na oksidativni stres. Budući da mitohondrija predstavlja glavno mesto sinteze energije i slobodnih radikala, u ovoj tezi je istraženo da li je uzrasno-specifična selekcija oblikovala varijabilnost mtDNK, nDNK i njihovih epistatičkih interakcija. Genotipizacijom mitohondrijskog COI gena i mikrosatelitskih lokusa pokazana je efikasnost uzrasno-specifične selekcije u međupopulacionoj divergenciji mitohondrijskog i jedarnog genskog pula kod pasuljevog žiška. Odvijanje procesa mito-jedarne koevolucije analizirano je korišćenjem mito-jedarnih linija u kojima su E i L mitohondrijski genomi kombinovani sa E i L jedarnim genomima. Pokazano je da su linije sa narušenim mito-jedarnim interakcijama imale niže aktivnosti mitohondrijskih kompleksa elektron transportnog lanca i lošiju preadultnu performansu., According to the evolutionary theory of ageing, age-specific decline in survival is the result of decreasing intensity of natural selection after the onset of reproduction. The evolution of ageing may be explained by two different, but not mutually exclusive, genetic mechanisms: antagonistic pleiotropy (AP) and mutation accumulation (MA). In accordance with evolutionary theory of ageing, two sets of the seed beetle (Acanthoscelides obtectus) experimental lines selected for more than 170 generations for early (E regime) or late (L regime) reproduction, show divergence in longevity and ageing patterns, as well as in other preadult and adult life history traits. Evolution of long life in L regime is predominantly based on antagonistic pleiotropy gene effects, while mutation accumulation plays important role in evolution of short life in E regime. By exposure of E and L beetles to two pesticides (paraquat and tebufenpyrad) that generate free radicals, it was confirmed that extended longevity in L beetles (especially in females) was associated with higher oxidative stress resistance. Additionally, since mitochondria are main site of free radical and energy production, effects of age-specific laboratory selection on mtDNA, nDNA and their epistatic interaction were assessed. Genotyping of mitochondrial COI gene and nuclear microsatellite loci showed that selection in E and L regimes led to divergent evolution of mtDNA and nDNA gene pools. Mitonuclear coevolution was investigated using mitonuclear introgression lines in which E and L mitochondrial genomes were expressed in both E and L nuclear background. It was found that lines with disrupted mitonuclear interactions had lower activity of the mitochondrial electron transport chain complexes and lower preadult performance.

Fenotipska plastičnost, kao univerzalno svojstvo svih organizama, predstavlja sposobnost jednog genotipa da u različitim uslovima životne sredine razvije različite fenotipske vrednosti osobina. Ovaj fenomen predstavlja važan mehanizam fenotipskog variranja insekatа i nalazi se u osnovi formiranja različitih pravaca evolucionih promena njihovih populacija uzrokujući adaptivnu divergenciju i posledičnu međupopulacionu reproduktivnu izolaciju. Od posebne važnosti je i uloga fenotipske plastičnosti u procesima ekološke specijacije, kada ekološki zasnovana divergentna selekcija između životnih sredina ograničava protok gena između populacija. Koristeći pogodnosti laboratorijske evolucije, ova studija proučava osobine životne istorije, njihova međusobna ograničenja i obrasce plastičnih odgovora, populaciono-ekološke i selekcione obrasce kao i promenu reproduktivnog ponašanja kod dve grupe laboratorijskih populacija fitofagnog insekta, Acanthoscelides obtectus, koje su evoluirale na različitim biljkama domaćinima (pasulj, Phaseolus vulgaris i naut, Cicer arietinum). Populacije selektovane na različitim biljkama pokazuju jasne razlike u životnim strategijama i stepenu plastičnosti osobina životne istorije i ponašanja. Načelno, populacije selektovane na pasulju imaju izraženiju sposobnost plastičnog odgovora na variranje identiteta biljke domaćina u poređenju sa populacijama selektovanim na nautu. Selekcioni režimi pokazuju i umeren stepen reproduktivne izolacije čime se ističe značaj divergentne selekcije u početnim fazama procesa nastanka novih vrsta., Phenotypic plasticity, a universal feature of all organisms, represents genotypes’ ability to produce alternative phenotypes in different environmental conditions. In insects, this phenomenon represents an important mechanism of phenotypic variation and it is fundamental for the establishment of different evolutionary trajectories of their populations causing adaptive divergence and consequential interpopulation reproductive isolation. Phenotypic plasticity is of particular importance during the process of ecological speciation, when ecologically based divergent selection between environments reduces gene flow among populations. Using the benefits of laboratory evolution, this study investigates life history traits, their trade-offs and plasticity patterns, population dynamics and selection patterns, as well as changes in reproductive behaviour, in two sets of laboratory populations of a phytophagous insect, Acanthoscelides obtectus, evolved on different host plants (white beans, Phaseolus vulgaris and chickpeas, Cicer arietinum). Populations selected on different plant hosts demonstrate distinct life history strategies and plasticity levels of life history and behavioural traits. Generally, populations selected on beans demonstrate prominent ability to plastically respond to host plant variation compared to chickpea selected populations. Selection regimes demonstrate moderate level of reproductive isolation emphasizing the importance of divergent selection in the first phases of the speciation.

Larvae of the gypsy moth, Lymantria dispar L. (Lepidoptera: Lymantriidae), a generalist species, frequently encounter spatial and temporal variations in diet quality. Such variation favoured the evolution of high behavioural and physiological plasticity which, depending on forest stand composition, enables more or less successful exploitation of the environment. Even in mixed oak stands, a suitable habitat, interspecific and intraspecific host quality variation may provoke significant variation in gypsy moth performance and, consequently, defoliation severity. To elucidate the insufficiently explored relationship between gypsy moth and oaks (Fagaceae), we carried out reciprocal switches between Turkey oaks (Quercus cerris L.) and less nutritious Hungarian oaks (Quercus frainetto Ten.) (TH and HT groups), under controlled laboratory conditions, and compared larval performance between the switched larvae and larvae continuously fed on either Turkey oak (TT) or Hungarian oak (HH). We found that larval traits were most strongly affected by among-tree variation in oak quality and identity of the host consumed during the fourth instar. Switching from Turkey to Hungarian oak (TH) led to a longer period of feeding, decrease of mass gain, growth, and consumption rate, lower efficiency of food use and nutrient conversion, and increase of protease and amylase activities. Larvae exposed to the reverse switch (HT) attained values of these traits characteristic for TT larvae. It appeared that the lower growth in the TH group than in the TT group was caused by both behavioural (consumption, pre-ingestive) and metabolic (post-digestive) effects from consuming oaks. Multivariate analyses of growth, consumption, and efficiency of food use revealed that early diet experience influenced the sensitivity of themost examined traits to less suitableHungarian oaks, suggesting the development of behavioural and physiological adjustments. Our results indicate that lower risks of defoliation b

U radu je ispitivano delovanje alkoholnih rastvora etarskih ulja anisa, mirođije i morača i njihovih dominantnih komponenata trans-anetola i karvona na gusenice gubara (Lymantria dispar L.). Antifidna aktivnost etarskih ulja i njihovih dominantnih komponenata primenjenih u koncentracijama 0.1, 0.5 i 1.0 % ispitivana je u dva odvojena ogleda, ogledu bez izbora i ogledu sa izborom. Ispitivanje je vršeno na gusenicama drugog stupnja, a rezultati ogleda su očitavani nakon 48 časova od početka ogleda. Rezidualna kontaktna i digestivna toksičnost etarskih ulja anisa, mirođije i morača i njihovih dominantnih komponenata primenjenih u koncentracijama 0.05, 0.1, 0.25, 0.5 i 1.0 %, ispitivana je na gusenicama gubara drugog stupnja. Pored mortaliteta gusenica, praćen je i uticaj navedenih toksičnosti na presvlačenje gusenica iz drugog u treći larveni stupanj. Ogledi u kojima je ispitivan uticaj toksičnosti ispitivanih etarskih ulja i nihovih dominantnih komponenata na mortalitet i zaustavljanje presvlačenja gusenica trajali su 120 časova, a rezultati su očitavani na svaka 24 časa. Delovanje ispitivanih etarskih ulja i njihovih dominantnih komponenata, primenjenih u koncentracijama 0.1, 0.25 i 0.5 %, na indekse rasta i ishrane gusenica gubara četvrtog stupnjsa sagledavano je u ogledima za ispitivanja njihovog uticaja na relativnu brzine rasta gusenica (RGR), relativnu brzinu konzumacije hrane gusenica (RCR), efikasnost asimilacije hrane kod gusenica (AD) i efikasnosti konverzije unete i svarene hrane kod gusenica (ECI i ECD). Rezultati ogleda su očitavani nakon 48 časova od početka ogleda. Za poređenje rezultata je korišćeno biološko sredstvo NeemAzal (standard). Statistička obrada podataka izvršena je uz pomoć softverskog paketa Statistica 7.0. (StatSoft, Inc). Analizom dobijenih rezultata utvrđeno je da dobru antifidnu aktivnost poseduju etarsko ulje anisa i njegova dominantna komponenta trans-anetol primenjeni u koncentraciji 1.0 %. Rezidualna kontaktna toksičnost ispitivani, In this work the influence of ethanol solutions of essential oils of anise, fennel and dill and their dominant components trans-anethole and carvone on gypsy moth larvae (Lymantria dispar L.) was tested. Antifeedant activity of essential oils and their dominant components applied in concentrantions 0.1, 0.5 i 1.0 % was tested in two separate tests, no choice and choice feeding ones. The testing was done on the second instar and the results of the experiments were recorded 48 h after the beginning of the experiment. Residual concant and digestive toxicity of essential oils of anise, fennel and dill and their dominant components applied in concentrations 0.05, 0.1, 0.25, 0.5 i 1.0 % were tested on the second instar gypsy moth larvae. The influence of mentioned toxicity on moulting from the second to the third instar was followed, too. The influence of toxicity of essential oils and their dominant components on mortality and disturbing moulting was evaluated after 24, 48, 72, 96 and 120 h after the beginning of the experiment. The influence of tested essential oils and their dominant components applied in concentrations 0.1, 0.25, 0.5 % on growth and feeding indices of the fourth insters was tested in the asseys about their influence on relative growth rate (RGR), relative consuption rate (RCR), aproximate digestibility (AD) and efficiency of conversation of ingested food (ECI), as well as efficiency of conversation of digested food (ECD). The results were recorded 48 h after the beginning of the experiment. Botanical standard NeemAzal was used for comparing results. Statistical data proccesing was done by softer Statistica 7.0. The analyzying of the results confirmed that essential oil of anise and its dominant component tras-anethole applied in concentration 1.0 % possess good antifeedant activity. Residual contact toksicity of tested essential oils and their dominant components as well as their influence on disturbing moulting was not satisfied. Tested essential oil

Kadmijum je jedan od najpotentnijih polutanata koji dospeva u životnu sredinu, iz prirodnih i/ili antropogenih izvora, i ulazi u žive sisteme preko lanaca ishrane, ostvarujući svoj toksični efekat na svim nivoima biološke organizacije. U ovom radu ispitivan je uticaj kadmijuma u koncentracijama od 10 (C1), 30 (C2) i 50 μg Cd/g (C3) suve hrane na komponente adaptivne vrednosti, heritabilnost u širem smislu, genetičke i fenotipske korelacije, fenotipsku plastičnost, cenu plastičnosti, i na obim i pravce delovanja selekcije, kao i uticaj 50 μg Cd/g suve hrane na sistem antioksidativne zaštite kod gubara tokom larvenog razvića, razvića lutke i adultnog perioda. Gubar, Lymantria dispar L., je polifagna, invazivna vrsta koja zbog svog štetnog delovanja na šumske ekosisteme spada u grupu organizama od posebnog interesa za istraživanja fizioloških i evolucionogenetičkih odgovora koji utiču na promenu brojnosti populacije ove vrste. Kadmijum dovodi do produžetka razvića kod mlađih larvenih stupnjeva, do smanjenja mase lutke kod oba pola i skraćivanja razvića lutke i života adultnih ženki. Polovi se značajno razlikuju u trajanju pojedinih faza larvenog razvića, naročito u C3 grupi. Za većinu ispitivanih osobina postoji značajan nivo heritabilnosti u širem smislu. Pri tome, prisustvo kadmijuma u C3 grupi dovodi do značajnog povećanja heritabilnosti trajanja razvića larvi četvrtog stupnja i kod mužjaka za trajanje larvenog razvića do 5. stupnja u odnosu na kontrolu i u odnosu na ženke. Značajna varijabilnost fenotipske plastičnosti postoji kod svih mlađih larvenih stupnjeva u svim grupama, i kod oba pola za pojedine osobine u okviru različitih sredina ukazujući na potencijal evolucije adaptivnog plastičnog odgovora u stresnim uslovima sredine..., Cadmium is one of the most potent pollutants which enters the environment through natural and/or anthropological sources and into the living organisms though the food chain thus realizing its toxic effect in all levels of biological organization. This work examines cadmium effects at 10 (C1), 30 (C2) and 50 μg Cd/g (C3) concentrations of dry food on the fitness-related traits, broad-sense heritability, genetic and phenotypic correlations, phenotypic plasticity, cost of plasticity and on direction of selection force. Also, this work examines the effects of 50 μg Cd/g of dry food on antioxidative defense system in gypsy moth during larval development, pupal development and the adult period. Lymantria dispar L. gypsy moth is a polyphagous, invasive species which, due to its detrimental influence on forest ecosystems, is considered an organism of particular interest for researches of physiological and evolutionary-genetic responses which can influence population dynamics of this species. Cadmium prolongs the development duration in early larval instars, it also reduces larval mass in both genders and larval development and life expectancy in adult females. Duration of certain larval instars is significantly different between genders, especially in C3 group. Most of the examined traits showed a significant level of broad-sense heritability. Thus, the presence of cadmium in C3 group significantly increases the heritability of the fourth instar duration and the heritability of development duration of male larvae from hutching until the 5th instar in comparison with control and female larvae, respectively. Significant variability of phenotypic plasticity is present in all of the early instars regardless of the group, as well as in both genders for certain traits depending on different environments which points to the potential of the evolution of adaptive plastic response in stressful environments. An index of phenotypic plasticity according to Cheplik and Li’s method shows

Temperatura i kvalitet hrane utiču na performansu larvi gubara, Lymantria dispar L. nezavisno ili u međusobnoj interakciji. Strategije preživljavanja larvi gubara u temperaturno kao i nutritivno heterogenoj sredini obuhvataju različite tipove reverzibilne i ireverzibilne fenotipske plastičnosti, koje preko uticaja na usvajanje i raspodelu resursa utiču na osobine životne istorije i rezistentnost prema ekstremnim uslovima životne sredine. U cilju ispitivanja efekata temperature i kvaliteta hrane, tj. sadržaja proteina i ugljenih hidrata u hrani na komponente adaptivne vrednosti, kao i ekspresiju genetičke varijabilnosti, larve gubara su izložene delovanju tri različite temperature (suboptimalna, optimalna i supraoptimalna) i 4 kombinacije hranljivog sastava dijete, koje su se međusobno razlikovale kako u ukupnom sadržaju proteina i ugljenih hidrata, tako i u njihovom međusobnom odnosu. U istim eksperimentalnim uslovima ispitivana je uloga procesa varenja, odnosno aktivnosti digestivnih enzima u usklađivanju odnosa i količine unetih nutienata sa potrebama organizma na različitim temperaturama. Takođe, ispitan je uticaj nutritivne vrednosti i balansiranosti hrane na senzitivnost gubara prema stresnim temperaturama. Nepovoljne temperature i nizak sadržaj proteina u hrani, kao i disbalans proteina u odnosu na ugljene hidrate, smanjuju performansu larvi gubara. Uticaji temperature i kvaliteta hrane na komponente adaptivne vrednosti: preživljavanje, trajanje razvića, masu i relativnu brzinu rasta, uglavnom su međusobno nezavisni. Pokazano je da povišena temperatura smanjuje preživljavanje i trajanje razvića larvi ali dovodi do povećanja relativne brzine rasta. Nutritivni sastav hrane nije uticao na preživljavanje, ali je nizak sadržaj proteina u hrani dovodio do produžavanja razvića, smanjenja mase i relativne brzine rasta larvi. Relativna brzina rasta larvi je bila manja i pri visokom sadržaju ugljenih hidrata u hrani, dok je smanjenje mase larvi na hrani sa niskim sadrža, Temperature and food quality affect the performance of gypsy moth larvae Lymantria dispar L. independently or in an interaction with each other. Survival strategies of gypsy moth larvae in temperature and nutritionally heterogeneous environments include various types of reversible and irreversible phenotypic plasticity, which due to the effect of uptake and distribution of resources affect the life-history traits and resistance to extreme environmental conditions. In order to investigate the direct and interactive effects of temperature and food quality on fitness components, as well as the expression of genetic variation, gypsy moth larvae were exposed to three different temperatures (suboptimal, optimal and supraoptimal) and 4 sets of nutrient composition of the diet, which differed in protein and carbohydrate content. Under the same experimental conditions, the role of digestion and digestive enzyme activity in adjusting nutrient quantity and ratio with organism needs at different temperatures was investigated. Also, it was investigated the effect of nutritional value of the food on sensitivity of gypsy moth larvae to stressful temperatures. An adverse temperature and low protein content in food, as well as an imbalance of protein compared to carbohydrates, reduced performance of gypsy moth larvae. Effects of temperature and food quality on fitness components - survival, developmental time, larval weight and relative growth rate were mainly independent. It has been shown that elevated temperature reduces survival and duration of development, but leads to an increase of the relative growth rate. Nutritional composition of food had no effect on survival, but the low protein content led to prolonged developmental time, reduced larval weight and relative growth rate of gypsy moth larvae. The relative growth rate of larvae was lower if carbohydrate content in food was high, while larval weight reduction was greater if protein content was low and the carbohydrate conte

Kombinovanje ili “slaganje” različitih gena u transgenim biljkama radi postizanja uspešnije kontrole patogena i štetočina i/ili većeg prinosa predstavlja jednu od glavnih oblasti istraživanja savremene biotehnologije. Orizacistatini I i II (OCI i OCII), proteinazni inhibitori različitih specifičnosti, pokazali su potencijal u kontroli štetočina koje koriste cisteinske proteinaze za digestiju proteina. Da bi se pojačao njihov inhibitorni potencijal i, eventualno, povećala efikasnost ovih inhibitora u kontroli štetočina, oba cistatina su koeksprimirana u transformisanim biljkama tri sorte krompira. “Slaganje” orizacistatinskih gena kod sorti Dragačevka i Dezire ostvareno je postupkom ko-transformacije i zabeležena je frekvenca kointegracije OCI i OCII gena od 20-22%. Kod sorte Jelica sekvencijalna re-transformacija se pokazala kao efikasniji pristup: frekvenca integracije OCII gena nakon re-transformacije OCI-transformisane linije iznosila je 91%. Istovremeno, “slaganje” dva orizacistatnska gena, bilo postupkom ko- ili re-transformacije, postignuto je upotrebom nptII gena kao jedinog selekcionog markera. Ekspresija OCI i OCII gena indukovana povređivanjem i akumulacija biološki aktivnih rekombinantnih OCI i OCII proteina potvrđena je kod svih analiziranih OCI/OCII transformisanih linija krompira. OCI/OCII linije krompira nisu ispoljavale značajna odstupanja od normalnog fenotipa, što ukazuje na nizak nivo somaklonalnih varijacija i odsustvo uticaja rekombinantnih OCI i OCII na metabolizam biljke domaćina. Iako nije uticala na preživljavanje, ishrana larvi krompirove zlatice (Leptinotarsa decemlineata Say) listovima krompira koji eksprimiraju oba orizacistatina imala je značajan uticaj na različite osobine performanse rasta i razvića larvi. Larve hranjene transformisanim listovima su se presvlačile ranije, i tokom L2 i L3 stupnja uvećavale masu do 29,7% brže i konzumirale listove do 29,1% brže u odnosu na one hranjene netransformisanim listovima. Istovremeno, larve na, The combination or stacking different genes in transgenic plants to achieve disease and pest control and/or higher crop yield is one of a major method of contemporary biotechnology. Oryzacystatins I and II (OCI and OCII), inhibitors with different specificity, show potential in controlling pests that utilize cysteine proteinases for protein digestion. To strengthen this inhibitory range and, possibly, achieve an additive effect in the overall efficiency of these proteins against pests, both cystatins were co-expressed in three potato cultivars. Oryzacystatin genes pyramiding in Dragačevka and Desiree cultivars were achieved by co-transformation with OCI and OCII genes co-integration frequency of 20-22%. For Jelica cultivar sequential re-transformation was more efficient approach: OCII gene integration frequency following re-transformation of an OCI-expressing line was 91%. Additionally, pyramiding of different oryzacystatin genes, by co- or re-transformation approach, were achieved using the nptII gene as the only selection marker. Wounding induction of OCI and OCII gene transcripts and accumulation of biologically active OCI and OCII recombinant proteins was confirmed in all analyzed OCI/OCII transformed lines. OCI/OCII potato lines did not exhibit morphological abnormalities, indicating low level of somaclonal variation or interference of the recombinant OCI or OCII with host plant metabolism. In the absence of significant mortality, feeding Colorado potato beetle larvae (Leptinotarsa decemlineata Say) on OCI/OCII-expressing foliage had an impact on various aspects of the growth and developmental performances of larvae. Larvae feeding on transformed potato leaves tended to molt earlier and, especially during L2-L3 stages, gain weight up to 29.7% faster and consume leaf material up to 29.1% faster, compared to those on untransformed foliage. Larvae on OCI/OCII foliage were also reach maximum weight gained three days earlier and slow down earlier in preparation for

Temperatura i kvalitet hrane utiču na performansu larvi gubara, Lymantria dispar L. nezavisno ili u međusobnoj interakciji. Strategije preživljavanja larvi gubara u temperaturno kao i nutritivno heterogenoj sredini obuhvataju različite tipove reverzibilne i ireverzibilne fenotipske plastičnosti, koje preko uticaja na usvajanje i raspodelu resursa utiču na osobine životne istorije i rezistentnost prema ekstremnim uslovima životne sredine. U cilju ispitivanja efekata temperature i kvaliteta hrane, tj. sadržaja proteina i ugljenih hidrata u hrani na komponente adaptivne vrednosti, kao i ekspresiju genetičke varijabilnosti, larve gubara su izložene delovanju tri različite temperature (suboptimalna, optimalna i supraoptimalna) i 4 kombinacije hranljivog sastava dijete, koje su se međusobno razlikovale kako u ukupnom sadržaju proteina i ugljenih hidrata, tako i u njihovom međusobnom odnosu. U istim eksperimentalnim uslovima ispitivana je uloga procesa varenja, odnosno aktivnosti digestivnih enzima u usklađivanju odnosa i količine unetih nutienata sa potrebama organizma na različitim temperaturama. Takođe, ispitan je uticaj nutritivne vrednosti i balansiranosti hrane na senzitivnost gubara prema stresnim temperaturama. Nepovoljne temperature i nizak sadržaj proteina u hrani, kao i disbalans proteina u odnosu na ugljene hidrate, smanjuju performansu larvi gubara. Uticaji temperature i kvaliteta hrane na komponente adaptivne vrednosti: preživljavanje, trajanje razvića, masu i relativnu brzinu rasta, uglavnom su međusobno nezavisni. Pokazano je da povišena temperatura smanjuje preživljavanje i trajanje razvića larvi ali dovodi do povećanja relativne brzine rasta. Nutritivni sastav hrane nije uticao na preživljavanje, ali je nizak sadržaj proteina u hrani dovodio do produžavanja razvića, smanjenja mase i relativne brzine rasta larvi. Relativna brzina rasta larvi je bila manja i pri visokom sadržaju ugljenih hidrata u hrani, dok je smanjenje mase larvi na hrani sa niskim sadrža, Temperature and food quality affect the performance of gypsy moth larvae Lymantria dispar L. independently or in an interaction with each other. Survival strategies of gypsy moth larvae in temperature and nutritionally heterogeneous environments include various types of reversible and irreversible phenotypic plasticity, which due to the effect of uptake and distribution of resources affect the life-history traits and resistance to extreme environmental conditions. In order to investigate the direct and interactive effects of temperature and food quality on fitness components, as well as the expression of genetic variation, gypsy moth larvae were exposed to three different temperatures (suboptimal, optimal and supraoptimal) and 4 sets of nutrient composition of the diet, which differed in protein and carbohydrate content. Under the same experimental conditions, the role of digestion and digestive enzyme activity in adjusting nutrient quantity and ratio with organism needs at different temperatures was investigated. Also, it was investigated the effect of nutritional value of the food on sensitivity of gypsy moth larvae to stressful temperatures. An adverse temperature and low protein content in food, as well as an imbalance of protein compared to carbohydrates, reduced performance of gypsy moth larvae. Effects of temperature and food quality on fitness components - survival, developmental time, larval weight and relative growth rate were mainly independent. It has been shown that elevated temperature reduces survival and duration of development, but leads to an increase of the relative growth rate. Nutritional composition of food had no effect on survival, but the low protein content led to prolonged developmental time, reduced larval weight and relative growth rate of gypsy moth larvae. The relative growth rate of larvae was lower if carbohydrate content in food was high, while larval weight reduction was greater if protein content was low and the carbohydrate conte

Gubar (Lymantria dispar L.) spada u red najinvazivnijih polifagnih vrsta insekata, koja naseljava šume, voćnjake i urbana staništa. U ovom radu je kod larvi gubara trećeg dana četvrtog larvenog stupnja, ispitivan akutni i hroničan uticaj dve koncentracije kadmijuma (10 i 30 μgCd/g suve hrane), kao i oporavak od istih, na različite osobine larvi: komponente adaptivne vrednosti (mortalitet, trajanje prvog stupnja, trajanje razvića do ulaska u četvrti larveni stupanj, masa i relativna brzina rasta larvi), aktivnosti enzima srednjeg creva (proteaza, tripsin, leucin aminopeptidaza, α i β-glikozidaza, esteraza i glutation-S-transferaza), kao i procenat mase srednjeg creva u odnosu na masu larve. Kod svih ispitivanih enzima, osim kod GSTa, prisutan je trend pada aktivnosti na kadmijumu u odnosu na kontrolu. Hronična izloženost kadmijumu nije pokazala značajan uticaj na mortalitet prvog stupnja, kao i ukupan mortalitet, masa larvi se smanjuje pri dugotrajnim tretmanima, dok se u toku akutnog tretmana brzina rasta larvi povećava. Analize varijanse su pokazale da se kod proteolitičkih enzima aktivnost menja u zavisnosti od koncentracije metala. Između proteolitičkih enzima na svim tretmanima najverovatnije postoji neka vrsta zajedničke regulacije sinteze ili sekrecije u prisustvu kadmijuma. Regulacioni mehanizmi β-glikozidaze i GSTa se razlikuju u odnosu na ostale proučavane enzime. Zbog ograničenih energetskih resursa postoji veliki broj negativnih korelacija između mase i enzima. Znak korelacija između α i β-glikozidaza se menja na stresu u odnosu na kontrolu. Na različitim tretmanima kadmijumom plastičnost enzima se menja. Pošto se indeks fenotipske plastičnosti ne menja u toku delovanja kadmijuma, promena osobina adaptivne vrednosti ili njihova plastičnost, se ne mogu uzeti kao dobri indikatori prisustva kadmijuma. Zbog velike osetljivosti leucin aminopeptidaze i esteraze u toku akutnog stresa, postojanja korelacija sa velikim brojem digestivnih enzima, velike plastičnost, The Gypsy moth (Lymantria dispar L.) represents one of the most invasive polyphagous insect species, which inhabits forests, orchards and even urban habitats. We investigated the acute and chronic effects of two cadmium concentrations (10 and 30 μgCd/g dry food), as well as the recovery from chronic stress on various traits of 4th instar gypsy moth larvae: fitness components (mortality, duration of development of the first larval instar, duration of development from hatching to molting into the 4th instar, larval mass and relative growth rate), midgut enzyme activities (activity of protease, trypsin, leucine aminopeptidase, α and β-glucosidase, esterase and glutathione S-transferase), as well as the percentage of the midgut mass in relation to the larval mass. All examined enzymes, with the exception of the glutathione S-transferase, showed a decrease of activity in relation to control. Larval mass decreased during the long-term treatment, while the acute treatment resulted in the growth rate increase. The analysis of variance showed that the activity of proteolytic enzymes depended on metal concentration. Significant broad-sense heritability was discovered after short-term exposure to cadmium. Regulation mechanisms of β-glucosidase and glutathione S-transferase differ in relation to the rest of the examined enzymes. There are a large number of negative correlations between larval mass and enzymes due to the limited energy resources. The correlation sign between α and β- glucosidases changes during the stress conditions in relation to the control. During different cadmium treatments plasticity of enzymes is changed. Changes of life history traits or their plasticity cannot be good indicators of cadmium presence. Due to a great sensitivity of leucine aminopeptidase and esterase during the acute stress, correlated response to a large number of digestive enzymes, great plasticity in stressful conditions, these two enzymes, with further research, could be potential biom

Uticaj različitog inteziteta svetlosti, kao jednog od abiotičkih faktora životne sredine, na nivo stabilnosti razvića i morfološku varijabilnost osobina cveta I. pumila ispitivan je na jedinkama populacija koje naseljavaju dva prirodna staništa različitiog inteziteta i kvaliteta raspoložive svetlosti (otvoreno i zasenčeno stanište), kao i na jedinkama populacija poreklom sa otvorenog i zasenčenog staništa, koje su bile izložene tretmanima visokog i niskog svetlosnog inteziteta svetlosti u eksperimentalnim uslovima. Osnovna hipoteza je bila da pod uticajem suboptimalnog intenziteta svetlosti dolazi do smanjenja stabilnosti razvića cvetnih organa što se može utvrditi na osnovu stepena asimetrije radijalno i bilateralno simetričnih cvetnih organa. Stepen fluktuirajuće asimetrije i radijalne asimetrije kao mere nestabilnosti razvića jedinke ili populacije, zavisi od intenziteta delovanja biotičkih i abiotičkih stresora, a pošto je stepen asimetrije negativno korelisan sa stabilnošću razvića, a pozitivno sa razvojnim šumom, na osnovu toga treba očekivati veće vrednosti indeksa asimetrije, tj. veću nestabilnost razvića u stresnijim sredinskim uslovima. Veličina i obrazac stabilnosti razvića osobina cveta I.pumila su se razlikovali u zavisnosti od analizirane osobine, kao i od evolucione istorije istraživanih populacija. Prosečne vrednosti indeksa radijalne asimetrije su bile više u zasenčenom nego u otvorenom staništu, ukazujući da je zasenčeno stanište verovatno stresnija sredina. Populacije poreklom sa otvorenog i zasenčenog staništa su se razlikovale u srednjoj vrednosti indeksa radijalne asimetrije osobina cveta u okviru svakog od svetlosnih tretmana. Fluktuirajuća asimetrija, kao mera bilateralne simetrije osobina cveta I.pumila, imala je veće vrednosti u prirodnim staništima sa visokim svetlosnim intezitetom u poređenju sa niskim, na osnovu čega bi se moglo zaključiti da je otvoreno stanište stresnije za ovaj tip osobina cveta. Aktuelni svetlosni uslovi u alternativ, The main goal of thesis was to examine the influence of an abiotic environmental factor, such as light intensity, on the developmental stability level and morphological variability of I. pumila floral traits. The individuals, from natural habitat populations which located different light condition habitats (exposed and shaded one), as well as the individuals of populations derived from natural habitat but raised in experimental light conditions of low and high intensity, were used. The basic hypothesis was whether suboptimal light intensity decreased developmental stability of floral traits. Since the level of fluctuating and radial asymmetry is in negative correlation with developmental stability, but is positively correlated with developmental noise, the larger values of asymmetry index are to be expected in more stressful conditions. The developmental stability size and pattern of I. pumila floral traits differed depending on analyzed trait, as well as on the evolutionary history of analyzed populations. The radial asymmetry index showed higher average values in shaded habitat comparing to the exposed one, meaning that the exposed location is more stressful environment for development of the identical replicas of different floral organs. Populations derived from open and shaded habitat differed in an average value of radial asymmetry index of floral traits inside each light intensity treatments, but didn’t differ in fluctuating asymmetry level.Statistically significant interaction between treatment and population, that we got for the radial asymmetry of the standard length and all the indices of fluctuating asymmetry, indicate that the effect of the light intensity on the sensitivity of developmental stability of Iris pumila floral traits depends on the population origin: whether they originated from an open or a shaded habitat. The actual light regimes in different habitats create statistically significant differences in phenotypic and genetic correlation matric

Uticaj različitog inteziteta svetlosti, kao jednog od abiotičkih faktoraživotne sredine, na nivo stabilnosti razvića i morfološku varijabilnostosobina cveta I. pumila ispitivan je na jedinkama populacija kojenaseljavaju dva prirodna staništa različitiog inteziteta i kvalitetaraspoložive svetlosti (otvoreno i zasenčeno stanište), kao i na jedinkamapopulacija poreklom sa otvorenog i zasenčenog staništa, koje su bileizložene tretmanima visokog i niskog svetlosnog inteziteta svetlosti ueksperimentalnim uslovima. Osnovna hipoteza je bila da pod uticajemsuboptimalnog intenziteta svetlosti dolazi do smanjenja stabilnostirazvića cvetnih organa što se može utvrditi na osnovu stepena asimetrijeradijalno i bilateralno simetričnih cvetnih organa. Stepen fluktuirajućeasimetrije i radijalne asimetrije kao mere nestabilnosti razvića jedinke ilipopulacije, zavisi od intenziteta delovanja biotičkih i abiotičkih stresora,a pošto je stepen asimetrije negativno korelisan sa stabilnošću razvića, apozitivno sa razvojnim šumom, na osnovu toga treba očekivati većevrednosti indeksa asimetrije, tj. veću nestabilnost razvića u stresnijimsredinskim uslovima. Veličina i obrazac stabilnosti razvića osobinacveta I.pumila su se razlikovali u zavisnosti od analizirane osobine, kao iod evolucione istorije istraživanih populacija. Prosečne vrednostiindeksa radijalne asimetrije su bile više u zasenčenom nego u otvorenomstaništu, ukazujući da je zasenčeno stanište verovatno stresnija sredina.Populacije poreklom sa otvorenog i zasenčenog staništa su se razlikovaleu srednjoj vrednosti indeksa radijalne asimetrije osobina cveta u okvirusvakog od svetlosnih tretmana. Fluktuirajuća asimetrija, kao merabilateralne simetrije osobina cveta I.pumila, imala je veće vrednosti uprirodnim staništima sa visokim svetlosnim intezitetom u poređenju saniskim, na osnovu čega bi se moglo zaključiti da je otvoreno staništestresnije za ovaj tip osobina cveta. Aktuelni svetlosni uslovi ualternativnim svetlosnim staništima su, The main goal of thesis was to examine the influence of an abioticenvironmental factor, such as light intensity, on the developmentalstability level and morphological variability of I. pumila floral traits. Theindividuals, from natural habitat populations which located differentlight condition habitats (exposed and shaded one), as well as theindividuals of populations derived from natural habitat but raised inexperimental light conditions of low and high intensity, were used. Thebasic hypothesis was whether suboptimal light intensity decreaseddevelopmental stability of floral traits. Since the level of fluctuating andradial asymmetry is in negative correlation with developmental stability,but is positively correlated with developmental noise, the larger valuesof asymmetry index are to be expected in more stressful conditions. Thedevelopmental stability size and pattern of I. pumila floral traits differeddepending on analyzed trait, as well as on the evolutionary history ofanalyzed populations. The radial asymmetry index showed higheraverage values in shaded habitat comparing to the exposed one, meaningthat the exposed location is more stressful environment for developmentof the identical replicas of different floral organs. Populations derivedfrom open and shaded habitat differed in an average value of radialasymmetry index of floral traits inside each light intensity treatments,but didn’t differ in fluctuating asymmetry level.Statistically significantinteraction between treatment and population, that we got for the radialasymmetry of the standard length and all the indices of fluctuatingasymmetry, indicate that the effect of the light intensity on the sensitivityof developmental stability of Iris pumila floral traits depends on thepopulation origin: whether they originated from an open or a shadedhabitat. The actual light regimes in different habitats create statisticallysignificant differences in phenotypic and genetic correlation matrices’structures of morphologi