Effect of α-cypermethrin and pirimiphos-methyl on wing morphology of Tribolium castaneum (Herbst) and T. confusum Jacquelin du Val: a comparative study.

Tribolium castaneum (Herbst) and Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae) are widespread and serious pests of stored products. Various insecticides are applied aiming to effectively manage both species. Here, two insecticides are tested, the pyrethroid α-cypermethrin and the organophosphate pirimiphos-methyl, hypothesizing that they can lead to morphological changes in the certain body parts of the adult offspring of treated T. castaneum and T. confusum parental female adults. For this purpose, the geometric morphometric method to the elytra and hindwings was applied. Both males and females were included in the analysis. The results showed that adult individuals of T. confusum showed higher tolerance to both insecticides compared to T. castaneum adults. This finding is reflected in analyses of both pairs of wings in T. confusum where changes in shape were negligible. The hindwings of T. castaneum experienced deformations to both insecticides. More significant changes in wing shape were observed in the α-cypermethrin treatment compared to pirimiphos-methyl. In the case of T. castaneum, even the shortest exposure to insecticides (5 min) is enough to provoke shape changes in the hindwings. Deformities in offspring, caused after treatment of their parents with insecticides, could moderate the frequency of insecticidal applications in storages.

Sub-Lethal Effects of Pirimiphos-Methyl Are Expressed to Different Levels in Wings of Three Stored-Product Coleopterans: A Geometric Morphometrics Investigation.

Tenebrio molitor L. (Coleoptera: Tenebrionidae), Prostephanus truncatus (Horn), and Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae) are noxious insect pests of grains in storages. Pirimiphos-methyl is widely used to protect grains at the post-harvest stage. However, the sub-lethal impact of this active ingredient on the offspring of all three coleopterans remains unknown. Thus, mated females of each species were exposed separately to pirimiphos-methyl at short exposures (30 min, 3, 5, 8, 16, 24, and 36 h), where the elytra and hindwings of the adult progeny were analyzed with the geometric morphometrics method. Males and females of all species were incorporated into the analysis. The results revealed variability among species. Tenebrio molitor was the most sensitive among three species, displaying significant deformations in the elytra and hindwings. Males had more conspicuous morphological changes than females. Prostephanus truncatus hindwings exhibited deformities after 36 h of exposure to pirimiphos-methyl. In contrast, R. dominica offspring were not affected by pirimiphos-methyl. In light of our findings, organophosphorus insecticides may cause variable sub-lethal effects to stored-product insects. This issue may lead to different insecticidal treatments according to the targeted stored-product species.

Genetic Variability in Balkan Paleoendemic Resurrection Plants Ramonda serbica and R. nathaliae Across Their Range and in the Zone of Sympatry.

The genus Ramonda includes three Paleoendemic and Tertiary relict species that survived in refugial habitats of the Balkan Peninsula (R. nathaliae and R. serbica) and the Iberian Peninsula (R. myconi). They are all "resurrection plants," a rare phenomenon among flowering plants in Europe. Ramonda myconi and R. nathaliae are diploids (2n = 2x = 48), while R. serbica is a hexaploid (2n = 6x = 144). The two Balkan species occur in sympatry in only two localities in eastern Serbia, where tetraploid potential hybrids (2n = 4x = 96) were found. This observation raised questions about the existence of gene flow between the two species and, more generally, about the evolutionary processes shaping their genetic diversity. To address this question, genetic markers (AFLP) and an estimate of genome size variation were used in a much larger sample and at a larger geographic scale than previously. The combination of AFLP markers and genome size results suggested ongoing processes of interspecific and interploidy hybridization in the two sites of sympatry. The data also showed that interspecific gene flow was strictly confined to sympatry. Elsewhere, both Ramonda species were characterized by low genetic diversity within populations and high population differentiation. This is consistent with the fact that the two species are highly fragmented into small and isolated populations, likely a consequence of their postglacial history. Within sympatry, enormous variability in cytotypes was observed, exceeding most reported cases of mixed ploidy in complex plant species (from 2x to >8x). The AFLP profiles of non-canonical ploidy levels indicated a diversity of origin pathways and that backcrosses probably occur between tetraploid interspecific hybrids and parental species. The question arises whether this diversity of cytotypes corresponds to a transient situation. If not, the question arises as to the genetic and ecological mechanisms that allow this diversity to be maintained over time.

Strategies of accumulation of potentially toxic elements in Minuartia recurva and M. bulgarica.

The aim of this study was to determine the concentrations of potentially toxic elements in soil samples and plant tissues of Minuartia recurva and M. bulgarica, predominantly or exclusively calcifuge species. Biological concentration (BCs) and translocation factors (TFs) were used to evaluate their accumulation potential. Considerable differences were observed between M. recurva and M. bulgarica assessions in terms of accumulation strategies of potentially toxic elements (PTEs). In M. recurva, most of the elements analyzed (Mn, Cu, Zn, Cd, and Co) were transported to the shoot, whereas in M. bulgarica, these elements remained predominantly in the roots. The Cu concentrations in the shoot samples of M. recurva from an abandoned iron-copper mine at Mt. Kopaonik were clearly above the notional hyperaccumulation threshold, characterizing this species as a possible Cu hyperaccumulator. Additionally, strong accumulation potential for Cr, Ni, Zn, Pb, and Cd was observed in M. recurva assessions, but without significant accumulation due to the low concentrations of these elements in the soils. The strong accumulation capacity and the different strategies in tolerance to PTEs indicate a potential of the two species for an application in phytoremediation: M. recurva for phytoextraction and M. bulgarica for phytostabilization.

Changes in the wing shape and size in Drosophila melanogaster treated with food grade titanium dioxide nanoparticles (E171) - A multigenerational study.

Drosophila is among the most commonly used models for toxicity assessment of different types of nanoparticles. This study aims to examine the effects of a constant exposure to the low concentration of human food grade titanium dioxide nanoparticles (TiO2 E171) on Drosophila melanogaster wing morphology over multiple generations. Subsequently, the Geometric Morphometrics Analysis was employed to examine possible changes in the wing shape and size of the treated flies. The treatment resulted in the diminishment but not a disruption in the sexual dimorphism in wings. Consequently, the female flies were clearly separated from the male flies by the differences in wing morphology as in the control group. A splitting by generations was overly similar within the control and the treatment, but it was slightly more pronounced in the treatment. However, the observed generational differences seemed mostly random between generations, irrespective of the treatment. Specifically, the treated groups displayed slightly higher splitting by generations in females than in males. Regardless of the generation, the results show a clear splitting by the differences in the wing shape between the treated flies and the flies from control. The mean value of centroid size, which refers to the wing size, of both female and male wings was smaller in the treatment when compared to the control. The overall effect of TiO2 was to induce significant difference in Drosophila wing morphology but it did not alter the general wing morphology pattern. Therefore, the change in the wings occurred only within the normally allowed wing variation.

Does the exposure of parental female adults of the invasive Trogoderma granarium Everts to pirimiphos-methyl on concrete affect the morphology of their adult progeny? A geometric morphometric approach.

Insecticidal applications may result to morphological deformations upon exposed insects or their offspring production. In the present study, we tested whether pirimiphos-methyl can induce deformities to wings of progeny production of the invasive khapra beetle, Trogoderma granarium (Coleoptera: Dermestidae) when its parental female adults have been treated with this organophosphorus active ingredient. For that purpose, we analysed both elytra and hindwings of both sexes of T. granarium progeny production by using the geometric morphometrics method. Our results showed that the wings of progeny of the pirimiphos-methyl-treated T. granarium parental female adult individuals suffered certain changes in their usual shape depending on size. Deformations occurred on both pairs of wings, but changes were more noticeable on the hindwings. A longer than 5-h exposure of parental female adults to pirimiphos-methyl, resulted in progeny with more deformed wings than in those individuals emerged after the exposure of their parental female adults in shorter periods on the toxicant. Generally, wings of both sexes were sensitive to pirimiphos-methyl, distinguishing the control group from the insecticidal treatments. The existence of deformed adults could be a useful indicator of earlier insecticidal applications as surface treatments and/or grain protectants in the storage facilities.

Assessment of trace element accumulation potential of Noccaea kovatsii from ultramafics of Bosnia and Herzegovina and Serbia.

In this work, we present the results of the investigation of trace elements (Fe, Mg, Ni, Zn, Cu, Cr, Co, Cd, Pb) accumulation potential of Noccaea kovatsii (Heuff.) F. K. Mey., from the Balkan Peninsula. The study included eight populations from ultramafic soils, six from Bosnia and Herzegovina, and two from Serbia. Principal component analysis (PCA) was used to reveal relationships of elements in soil, and Pearson's correlation coefficients for analysing associations of available quantities of elements in soil and those in roots and shoots of N. kovatsii. Uptake and translocation efficiency was assessed by using bioconcentration (BCF) and translocation factors (TF). All the analysed populations of N. kovatsii emerged as strong Ni accumulators, with the highest shoot concentrations of 12,505 mg kg-1. Even thought contents of Zn in plant tissues of N. kovatsii were under the hyperaccumulation level (602 mg kg-1 and 1120 mg kg-1 respectively), BCF was up to 667, indicating that certain surveyed populations have strong accumulative potential for this element.

Resurrection plants of the genus Ramonda: prospective survival strategies - unlock further capacity of adaptation, or embark on the path of evolution?

Paleoendemic species of the monophyletic genus Ramonda (R. myconi, R. serbica and R.~nathaliae) are the remnants of the Tertiary tropical and subtropical flora in Europe. They are the rare resurrection plants of Northern Hemisphere temperate zone. Ramonda serbica and R. nathaliae are chorologically differentiated in the Balkan Peninsula and occupy similar habitats in calcareous, northward slopes in canyons and mountainsides. They remain well-hydrated during spring, late autumn and even in winter. In summer and early autumn when plants are subjected to drought and thermal stress, their desiccation tolerance comes into operation and they fall into anabiosis. Investigations revealed the permanent presence of ubiquitine and its conjugates, high amounts of oxalic acid and proline. Both species are homoiochlorophyllous. It enables them to rapidly resume photosynthesis upon rehydration, but also makes them susceptible to reactive oxygen species formation. Dehydration induces activation of antioxidative enzymes (ascorbate peroxidase, glutathione reductase, polyphenol oxidase), increase in amounts of AsA and GSH, phenolic acids, dehydrins, sucrose, and inorganic ions. Plasma membranes, characterized by high amount of cholesterol, are subjected to decrease in membrane fluidity mostly on account of increased level of lipid saturation. Cytogenetic analysis revealed that R. nathaliae is a diploid (2n = 48) and probably evolutionary older species, while R. serbica is a hexaploid (2n = 144). Two species live together in only two localities forming hybrid individuals (2n = 96). Polyploidization is the major evolutionary mechanism in the genus Ramonda that together with hybridization ability indicates that these relict species which have preserved an ancient survival strategy are not the evolutionary "dead end."The species of the genus Ramonda are promising sources of data important for understanding the complex strategy of resurrection plants' survival, appraised through a prism of their evolutionary and adaptive potential for multiple environmental stresses.