The food plant preferences of Phratora vitellinae (Coleoptera: Chrysomelinae)

The specialisation of the beetles from the Petite Camargue on Salix nigricans was confirmed by laboratory food-plant trials. When S. nigricans is not present in the choice offered, other Salix species are accepted and the following ranking, in decreasing order of preference, can be established: S. nigricans, S. purpurea, Populus nigra, P. tremula, S. alba, s. caprea and S. cinerea. S. nigricans leaves are the richest in salicin of the 4 most acceptable plant species. The 3 least acceptable species have the undersurface of their leaves covered with trichomes.The food-plant preferences of 2 populations from central Europe and 2 from Belgium differ both in the field and in the laboratory. The level of dietary specialisation also differs from population to population. The populations originating from localities (Oignie in Belgium and the Petite Camargue in central Europe) where their favorite food-plants in the field and in the laboratory (respectively P. tremula and S. nigricans) are abundant, show clearer preferences than the populations from localities (Grammont in Belgium and Herrliberg in central Europe) where these plants are scarcer or even absent, due to human influence.The Salicaceae tested, as seen by the more specialised beetles of Oignie and the Petite Camargue, include both a strongly preferred plant (P. tremula or S. nigricans) and two or three strongly disliked species (S. alba, S. caprea and S. cinerea). for these beetles, laboratory preferences and field observations of host plant species are in agreement. It is interesting that S. nigricans, although not present in Belgium, is the second most preferred food of the Belgian beetles in laboratory trials. Similarly, P. tremula, the preferred food of the Belgian beetles, is well accepted by the central European beetles.For the less specialised beetles of Grammont and Herrliberg, the same Salicaceae include strongly disliked speicies but no single strongly preferred species. In the Herrliberg population, with rather poorly defined preferences, preference for the favorite species of the more specific population from the same geographical area (Petite Camargue) can readily be produced in the laboratory by conditioning.     

Influence of phenolglucosides and trichome density on the distribution of insects herbivores on willows

The effects of both trichome density and phenolglucoside content of leaves of 76 willow hybrids (Salix alba x fragilis) were measured to estimate their influence on the distribution of Phratora vitellinae (L.), Plagiodera versicolora Baly (Coleoptera: Chrysomelidae) and Pontania proxima (Lepeletier 1823) (Hymenoptera: Tenthredinidae) in a nursery at Gramont, Belgium.The willows showed differences in their phenolglucoside content and pilosity of leaves and are classified on these basis into four groups by a clustering method. Correlations and multiple regressions showed that these chemical and physical characteristics are good predictors of the abundance of insects. First, the abundance of larvae of Ph. vitellinae, adults of Pl. versicolora and galls of P. proxima is correlated positively with a high phenolglucoside content and a low pilosity of the leaves. Secondly, the distribution of adults of Ph. vitellinae and of larvae of Pl. versicolora is influenced by neither the chemical nor the physical leaves characteristics studied.     

Beetle assemblages on willow trees: do phenolic glycosides matter?

Herbivorous beetles were sampled in central Slovakia and in the Tatra Mountains of northern Slovakia from seven different Salix species which are partly characterized by smooth leaves containing phenolic glycosides and partly by hairy leaves containing tannins. The collection included about 8000 individuals representing 129 species. Of the 129 species, 77 species are able to use the willows as hosts; the remaining 52 ‘tourist’ species comprise less than 3% of the collected individuals. The data on species richness and abundance did not generally support the ‘feeding specialization’ hypothesis. The proportion of specialized (= monophagous and oligophagous) beetles feeding on willows of both morphological and biochemical groups was in the same range. Host plants of the two groups could support high diversity and high evenness values, even when leaf characteristics and plant chemicals largely influenced species assemblage. The region in which the willow trees grow had a considerable impact on host plant use. Generalist beetle species predominated in central Slovakia. By contrast, in the Tatra Mountains, specialist feeders which are able to use phenolic glycosides to their advantage were predominant. The number of species and the total density of individuals collected from willows containing phenolic glycosides (S. fragilis and S. purpurea) did not usually vary between the two regions. In contrast, fewer species and individuals were found in the Tatra Mountains when they settled on willow species containing tannins (S. caprea and S. cinerea). Also, the phylogenetic status of host plants affected species assemblages. In central Slovakia willow species of the subgenus Vetrix (S. purpurea, S. caprea and S. cinerea) generally showed a higher beetle diversity (H_s) than species of the subgenus Salix (S. fragilis, S. alba and S. triandra), although both subgenera comprise species of both morphological and biochemical groups. Furthermore, when the analysis was restricted to beetles of central Slovakia, which should be most adapted to their host plants (i.e. catkin feeders and phyllophages in the adult and larval stage), the phylogenetic status was found to be more important than any single leaf character measured.     

Chlorogenic acid as an antiherbivore defence of willows against leaf beetles

In the leaves of 13 Finnish willow species, the content of a phenolic, chlorogenic acid, was found to vary from 0 up to 18 mg g^–1 D.W. Effects of pure chlorogenic acid on insect feeding behaviour were tested using four common leaf beetle species which are in the field mainly found on willows with low-chlorogenic acid leaves. One species, Lochmaea capreae L., was invariably deterred by pure chlorogenic acid applied in naturally occurring concentrations on the willow leaves. Accordingly, in 2-choice laboratory feeding trials L. capreae was found to prefer low-chlorogenic acid leaves of four willow species over high-chlorogenic acid leaves of Salix pentandra L. and S. myrsinifolia Salisb. When presented on the leaves of S. phylicifolia L, pure chlorogenic acid inhibited also the feeding by Phratora polaris Sp.-Schn. Instead, chlorogenic acid had no significant effect on Ph. polaris when it was presented on the leaves of another willow S. cinerea L. In laboratory, Ph. polaris did not show general preference for willow species with low chlorogenic acid content in their leaves. Thus, the response of Ph. polaris to chlorogenic acid seems to depend on the plant species. Apparently variation in other traits such as leaf hairyness may easily override the potential effect of chlorogenic acid content on Ph. polaris. To two other leaf beetle species, Galerucella lineola F. and Plagiodera versicolora Laich., chlorogenic acid is an ineffective deterrent even at unnaturally high concentrations. In laboratory, G. lineola and P. versicolora did not prefer willows with low chlorogenic acid content in their leaves. Thus, among four studied leaf beetle species, only L. capreae seems to be clearly affected by this phenolic. Therefore, overall importance of chlorogenic acid as a defence against willow-feeding leaf beetles appears to be very limited.     

Intra- and interspecific effects of larval secretions in some chrysomelids (Coleoptera)

The larval secretions of the Chrysomelinae (Coleoptera, Chrysomelidae) are generally considered to be chemical defense substances against predators and parasites. The experiments presented in this paper provide evidence that the range of activity of these larval secretions also extends to interactions between different conspecific developmental stages and between competing phytophagous species. Four chrysomeline species were tested: Gastrophysa viridula De Geer, Phaedon cochleariae (F.), Plagiodera versicolora (Laich.), and Phratora vitellinae (L.). In the latter species, we did not test the intraspecific effect of the natural larval secretion, but the activity of its purchasable main component, salicylaldehyde (synthetic larval secretion). In G. viridula, Ph. cochleariae, and Ph. vitellinae the (synthetic) larval secretions act as oviposition deterrents against conspecific females. An investigation of the persistence of the oviposition-deterring activity of the larval secretion in G. viridula revealed that the deterrent effect is lost after 20 to 26 hours. In each of the four tested species the (synthetic) larval secretion deters feeding of conspecific adults. Since P. versicolora and Ph. vitellinae may occur on the same host plant (e.g. the willow Salix fragilis L.), the interspecific effect of the larval secretion between competing phytophagous species was examined in these two willow leaf beetles. The larval secretions of both species and salicylaldehyde, respectively, act as interspecific repellent and feeding deterrent against adults of the other species.

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Zusammenfassung Die Sekrete der Larven der Chrysomelinae (Fam. Chrysomelidae) werden im allgemeinen als chemische Abwehrstoffe gegen Prädatoren und Parasiten betrachtet. Die hier dargestellten Untersuchungen zeigen, daß die Wirkung der Larvensekrete nicht nur auf die Feindabwehr begrenzt ist. Das Wirkungsspektrum umfaßt darüber hinaus auch Interaktionen zwischen verschiedenen Entwicklungsstadien einer Art bzw. zwischen konkurrierenden phytophagen Arten. Folgende Chrysomelinenarten wurden untersucht: Gastrophysa viridula De Geer, Phaedon cochleariae (F.), Phratora vitellinae (L.) und Plagiodera versicolora (Laich.). Da das Larvensekret von Ph. vitellinae Salicylaldehyd als Hauptkomponente enthält, wurde hier-wenn nicht anders erwähnt-die Wirkung von synthetischem Salicylaldehyd getestet. Bei G. viridula, Ph. cochleariae und Ph. vitellinae wirkt das Larvensekret bzw. die authentische synthetische Substanz als hochwirksames Eiablage-Deterrens auf konspezifische Weibchen. Die eiablagehemmende Wirkung des Larvensekretes von P. versicolora wurde nicht getestet. Eine Untersuchung der Wirkungsdauer der eiblagehemmenden Aktivität des Larvensekretes von G. viridula ergab, daß die Hemmwirkung nach 20 bis 26 Stunden nicht mehr nachweisbar ist. Bei allen oben genannten vier Arten hemmt das. Larvensekret bzw. dessen authentische synthetische Substanz den Fraß von Adulten derselben Art. Diese fraßhemmende Aktivität ist bei G. viridula, Ph. cochleariae und Ph. vitellinae mindestens 18 Stunden wirksam. Das Larvensekret von P. versicolora zeigte bei 18-stündiger Versuchsdauer keine fraßhemmende Wirkung auf Adulte derselben Art; erst bei Reduktion der Versuchsdauer auf drei Stunden konnte hier eine signifikant fraßhemmende Wirkung nachgewiesen werden. P. versicolora und Ph. vitellinae können als Weidenblattkäfer an derselben Wirtspflanze auftreten. Deshalb wurde auch die interspezifische Wirkung der Larvensekrete dieser beiden Chrysomelinenarten geprüft. Das Larvensekret von Ph. vitellinae sowie auch dessen authentische synthetische Substanz (Salicylaldehyd) wirkt auf adulte P. versicolora als Repellent und Fraßhemmstoff. Ebenso wirkt umgekehrt das Larvensekret von P. versicolora abschreckend und fraßhemmend auf adulte Ph. vitellinae.

     

Biological diversity of Salix taxa in Cu,Pb and Zn phytoextraction from soil

The aim of the study was to estimate the efficiency of copper (Cu), lead (Pb) and zinc (Zn) phytoextraction by 145 Salix taxa cultivated in an area affected by industrial activity. Survivability and biomass of plants were also analyzed. The highest Cu, Pb and Zn content in shoots was 33.38 ± 2.91 (S. purpurea × viminalis 8), 24.64 ± 1.97 (S. fragilis 1) and 58.99 ± 4.30 (S. eriocephala 7) mg kg^?1 dry weight, respectively. In the case of unwashed leaves, the highest content of these metals was 135.06 ± 8.14 (S. purpurea 26), 67.98 ± 5.27 (S. purpurea 45) and 142.56 ± 12.69 (S. alba × triandra 2) mg kg^?1 dw, while in washed leaves it was 106.02 ± 11.12 (S. purpurea 45), 55.06 ± 5.75 (S. purpurea 45) and 122.87 ± 12.33 (S. alba × triandra 2) mg kg^?1 dw, respectively. The differences between the highest and lowest values for Cu, Pb and Zn were 545%, 20500% and 535% in shoots; 2692%, 2560% and 7500% in unwashed leaves; and 3286%, 2221% and 6950% in washed leaves, respectively. S. acutifolia was able to effectively accumulate all three metals jointly, producing shoots that were well developed in both length and diameter when compared with the other tested willows—an ability that would suggest its high suitability for practical application.     

The presence or absence of phenolglycosides in Salix (Salicaceae) leaves and the level of dietary specialisation of some of their herbivorous insects

Summary European Salix species fall into at least 2 groups on the nature of the secondary compounds in their leaves. Some species such as S. nigricans, S. purpurea and S. fragilis contain phenolglycosides in their leaves. Additionally, there are species such as S. alba, S. caprea and S. cinerea with leaves containing no phenolglycosides.Using published food plant list for the weevils, the sawflies and the caterpillars of the British moth, I tried to see if the presence or absence of phenolglycosides in the willow leaves is related to the degree of dietary specialisation of the insects feeding on these leaves.The results show that the Salix with phenolglycosides tend to be the food of the specialised herbivores and to be avoided by generalist. Conversely, Salix without phenolglycosides tend to be eaten by more generalist insects and are avoided by the more specialised.Moreover, the faunas of the different Salix species with phenolglycosides in their leaves are more similar to each other than to the faunas of the Salix species having no phenolglycosides in their leaves.     

Host‐plant quality adaptively affects the diapause threshold: evidence from leaf beetles in willow plantations

1. Voltinism of herbivorous insects can vary depending on environmental conditions. The leaf beetle Phratora vulgatissima L. is univoltine in Sweden but will sometimes initiate a second generation in short‐rotation coppice (SRC) willow plantations. 2. The study investigated whether increased voltinism by P. vulgatissima in plantations can be explained by (i) rapid life‐cycle development allowing two generations, or (ii) postponed diapause induction on coppiced willows. 3. In the field, no difference was found in the phenology or development of first‐generation broods between plantations (S. viminalis) and natural willow habitats (S. cinerea). However, the induction of diapause occurred 1–2 weeks later in SRC willow plantations. 4. Laboratory experiments indicated no genetic difference in the critical day‐length for diapause induction between beetles originating from plantations and natural habitats. Development time was unaffected by host‐plant quality but critical day‐length was prolonged by almost an hour when the beetles were reared on a non‐preferred willow species (S. phylicifolia). When reared on new leaves from re‐sprouting shoots of recently coppiced willow plants, diapause incidence was significantly less than when the beetles were reared on mature leaves from uncoppiced plants. 5. The study suggests that P. vulgatissima has a plastic diapause threshold influenced by host‐plant quality. The use of host‐plant quality as a diapause‐inducing stimulus is likely to be adaptive in cases where food resources are unpredictable, such as when new host‐plant tissue is produced after a disturbance. SRC willows may allow two beetle generations due to longer growing seasons of coppiced plants that grow vigorously.     

Nutrient levels in malformed and healthy tissues of mango (Mangifera indica L.)

Samples of malformed and healthy panicles of mango (Mangifera indica L.) as well as leaves and shoots bearing them were collected at different stages of development (fully swollen buds, bud inception, fully grown panicles prior to full bloom and at full bloom) over two consecutive years and were analysed for their macro- and micronutrient status. In addition, malformed and healthy seedlings were collected and analysed. Malformed panicles were found to be significantly higher in N at all the developmental stages except at bud inception. Phosphorus and K also tended to accumulate in malformed panicles at later stages of their development. In general, malformed panicles exhibited lower levels of P, K and Ca than healthy panicles. The differences in levels of Mg and S in malformed and healthy panicles were not significant. All micronutrients were in much lower concentrations in malformed panicles except for Mn which appears to accumulate in malformed panicles particularly at the early stages of development. The leaves on the shoots bearing malformed panicles also showed a tendency to accumulate N, while P, Mg and S were always higher in leaves on shoots bearing healthy panicles. The leaves on shoots bearing healthy panicles had lower levels of Fe, Cu and Mn, whereas levels of Zn and B tended to be higher in leaves on shoots bearing malformed panicles. The nutrient concentration differences between the two kinds of shoots were generally nonsignificant (P=0.05), except for K and S which were significantly lower in shoots bearing malformed panicles. The shoots bearing malformed panicles showed significantly (P=0.05) higher levels of almost all nutrients compared with shoots bearing healthy panicles. Vegetative malformation was found to be associated significantly (p=0.05) with higher amounts of all nutrients except Ca which was significantly higher in healthy seedlings. The present study, therefore, seems to point to lower Ca as one of the pre-disposing factors causing malformation in mango.A part of Ph.D. thesis of the senior author.A part of Ph.D. thesis of the senior author.     

Phenolic glucosides as feeding cues for willow-feeding leaf beetles

The effects of individual phenolic glucosides and total glucoside fractions on the feeding behaviour of three willowfeeding leaf beetles (Coleoptera: Chrysomelidae) were tested in the laboratory. Feeding preferences of the tested leaf beetles were strongly influenced by certain phenolic glucosides which are typical secondary compounds of willows (Salicaceae:Salix). Two of the tested leaf beetles,Galerucella lineola andLochmaea capreae showed rather similar responses to glucoside treatments. Both of them were strongly stimulated by total glucoside fractions fromSalix triandra and by its major glucoside salidroside. The third species,Phatora vitellinae, was attracted most by the fractions fromS. myrsinifolia andS. pentandra, and by two related salicylate glucosides, tremulacin and salicortin. Food selection pattern of the tested beetles in the laboratory concords fairly well with their distributions in the field and with the occurrence of phenolic glucosides in their host willows. Phenolic glucoside extracts stimulated more feeding than individual pure glucosides. This indicates that different compounds have synergistic effects in the feeding behaviour of leaf beetles. Our results clearly show that willow leaf beetles select their food based on phenolic glucosides of their host plants.     

Post hibernation dispersal of three leaf-eating beetles (Coleoptera: Chrysomelidae) colonising cultivated willows and poplars

1 We studied the spring dispersal of three common chrysomelids, from overwintering habitats into cultivated willow and poplar coppices at four sites in southern England over 2 years. 2 Adult Galerucella lineola, Phratora vulgatissima and P. vitellinae overwintered under the bark of mature trees or in other niches that simulated this habitat, within a few hundred metres of the coppice plantation. Relatively few beetles remained in the coppice fields during the winter. 3 Phratora vitellinae at the poplar sites emerged several weeks later than G. lineola and P. vulgatissima at the willow sites, reflecting the later leafing of poplar compared to willow. For all species, dispersal was by flight, with most activity during warm periods. Dispersal continued for several weeks for the willow- feeding species but was shorter for P. vitellinae. 4 All three species initially colonized the edge of the coppice field. Typically, 80% or more of the beetles colonising a plantation were within 8 m of the edge. Both Phratora spp. accumulated in the plantation edge zone for several weeks before leaving this area and colonising the crop interior. 5 The patterns of dispersal and colonization identified by this study may facilitate chrysomelid management practices in infested short rotation coppice that avoid the need for insecticide applications over the entire plantation. Keywords Chrysomelidae, Galerucella lineola, Phratora vitellinae, Phratora vulgatissima, poplar, short rotation coppice, willow, winter dispersal.     

Leaf beetle grazing does not induce willow trichome defence in the coppicing willow Salix viminalis

Abstract 1 Willows are frequently attacked and defoliated by adult leaf beetles (Phratora vulgatissima L.) early in the season and the plants are then attacked again when new larvae emerge. The native willow Salix cinerea has previously been shown to respond to adult grazing by producing new leaves with an increased trichome density. Subsequent larval feeding was reduced on new leaves. This type of induced plant response may reduce insect damage and could potentially be utilized for plant protection in agricultural systems. 2 Here, we investigated if the willow species most commonly used for biomass production in short rotation coppice, Salix viminalis, also responds to adult beetle grazing by increasing trichome density. Larval performance and feeding behaviour on plants previously exposed to adult beetles was compared with that on undefoliated control plants in a greenhouse. 3 We found an overall decrease in trichome density within all the plants (i.e. trichome density was lower on new leaves compared to that for older basal leaves on S. viminalis). However, leaves of beetle defoliated plants had a higher trichome density compared to control plants. Larval growth and feeding was not affected by this difference between treatments. Larvae appeared to remove trichomes when feeding on S. viminalis, a behaviour that might explain the lack of difference between treatments.     

Susceptibility of willow clones (Salix spp.) to herbivory by Phyllodecta vulgatissima (L.) and Galerucella lineola (Fab.) (Coleoptera, Chrysomelidae)

Levels of damage by mixed natural infestations of the leaf-feeding chrysomelid beetles, Phyllodecta vulgatissima (L.) (the blue willow beetle) and Galerucella lineola (Fab.) (the brown willow beetle), were determined in replicated field plots of 24 Salix clones at Long Ashton (Bristol, UK) during 1993–94. Over the same period, the host plant preferences of both chrysomelids were investigated in a standard multiple-choice laboratory procedure, where beetles were enclosed in Petri dishes with leaf discs cut from young pot-grown trees propagated from shoot cuttings taken from 20 of the 24 willow clones represented in the field study. The laboratory experiments indicated that P. vulgatissima and G. lineola had similar host plant preferences in the range of willows examined (r >0.85). In both field and laboratory, the least preferred Salix clones and hybrids were those of 5. eriocephala, followed by S. purpurea, S. burjatica, S. dasyclados and S. triandra. Clones of S. eriocephala and S. purpurea were frequently rejected altogether in laboratory tests. Most preferred were clones of S. viminalis and several hybrids of S. viminalis, S. aurita, S. caprea and S. cinerea. These results substantiate the reports that P. vulgatissima and G. lineola are deterred from feeding on willows which have relatively high concentrations of phenolic (salicylate) glucosides in the leaves. The least preferred willows, particularly S. eriocephala, S. purpurea and S. burjatica, could be of great potential value in plant breeding for resistance to these willow beetle pests.     

Temporal change in the location of egg-laying by a bud-galling sawfly,Euura mucronata,on growing shoots of Salix cinerea

Summary Temporal change in the egg-laying behaviour of Euura mucronata was studied on growing shoots of Salix cinerea. Many new shoots arise from each previous year's shoot of S. cinerea; they grow very rapidly at the beginning of the oviposition period of E. mucronata, but their growth pattern changes dramatically during the 3 weeks of oviposition. When oviposition commences, all new shoots are equally long, and E. mucronata females lay eggs on them randomly; but toward the end of oviposition, eggs are laid on the longest of the new shoots, which originate on the distal part of the previous year's shoot. By the end of oviposition, the mean proportion of ovipositor borings that had resulted in egg-laying was 65.8% on distal shoots and declined linearly to 25% on shoots arising from the base of the previous year's shoot. The proportion of attacked buds from which larvae emerged declined non-linearly from 13.8% on the distal shoot to 3.5% on the basal shoots. The position of the shoot accounted for 67% and 73% of the variance in success of egg-laying and larval development, respectively.     

Exploration of meaningful subspecies definitions of the carnivorous Northern Pitcher Plant (Sarracenia purpurea), throughout its geographic distribution

The carnivorous wetland plant, Sarracenia purpurea (the northern pitcher plant) is native to eastern and Midwestern North America. This species is abundant within its habitat but suitable habitat is increasingly scarce, raising interest in S. purpurea restoration and conservation. Complicating conservation planning, two controversial subspecies of S. purpurea are historically defined primarily on morphologic traits: S. purpurea subsp. purpurea distributed north of Maryland and S. purpurea subsp. venosa distributed south of Maryland. S. purpurea is also found in three distinct habitat types defined by substrate: acidic Sphagnum peatlands, acidic sandy savannahs, and alkaline marl wetlands. In species level studies, S. purpurea leaves have exhibited morphological plasticity in response to environmental variability, bringing into question the validity of subspecies definitions based on morphology alone. This study examined morphologic and genetic variation throughout S. purpurea’s natural distribution, encompassing both the traditional subspecies and three unique habitat types testing the validity of traditional subspecies definitions. Genetic analysis indicated possible ecological significance of considering a new grouping of S. purpurea populations into Midwest, N. East Coast and S. East Coast populations (AMOVA % variation = 13.34, P = 0.0078) based on genetic differentiation. Morphological variation in leaf shape measurements supported this division as well as indicating plasticity associated with environmental variables. This study conservatively suggests that new, geographical area conservation units may be a more important conservation unit for preserving S. purpurea genetic variation and morphological plasticity than traditional subspecies definitions.     

Plant phenology-mediated indirect effects: The gall midge opens the phenological window wider for a leaf beetle

We examined whether larvae of the gall midge Rabdophaga rigidae (Diptera: Cecidomyiidae) can modify the seasonal dynamics of the density of a leaf beetle, Plagiodera versicolora (Coleoptera: Chrysomelidae), by modifying the leaf flushing phenology of its host willow species, Salix serissaefolia and Salix eriocarpa (Salicaceae). To test this, we conducted field observations and a laboratory experiment. The field observations demonstrated that the leaf flushing phenology of the willows and the seasonal dynamics of the beetle density differed between shoots with stem galls and shoots without them. On galled shoots of both willow species, secondary shoot growth and secondary leaf production were promoted; consequently, leaf production showed a bimodal pattern and leaf production periods were 1 to 2 months longer than on non‐galled shoots. The adult beetle density on galled shoots was thus enhanced late in the season, and was found to change seasonally, synchronizing with the production of new leaves on the host willow species. From the results of our laboratory experiment, we attributed this synchrony between adult beetle density and willow leaf flush to beetles’ preference to eat new leaves rather than old. Indeed, beetles consumed five times more of the young leaves when they were fed both young and old leaves. These results indicate that stem galls indirectly enhance the adult beetle density by enhancing food quality and quantity late in the beetle‐feeding season. We therefore conclude that midge galls widen the phenological window for leaf beetles by extending the willows’ leaf flush periods.     

Behavioral responses of a leaf beetle to injury-related changes in its salicaceous host

Summary Adults of the leaf beetle Plagiodera versicolora Laich, avoided previously injured shoots of Salix alba Tristis in favor of nearby uninjured ones. The response was rapid and persisted for five days of study. During brief periods of observation, the vast majority of beetles engaged in behaviors other than locomotion such as feeding and resting. Furthermore, the proportions of beetles walking were similar on injured and uninjured shoots. It appears that under these experimental conditions previous injury did not greatly increase searching behavior of the beetles. Leaves on previously injured shoots received less subsequent herbivory than uninjured ones. Findings of these experiments are consistent with patterns of herbivory observed in the field and known reproductive responses of beetles to diets of injured leaves. These results provide new information toward understanding relatively low levels of herbivory on individual willow leaves associated with the feeding of P. versicolora.     

Aster Yellows Subgroup 16SrI‐C Phytoplasma in Rhododendron hybridum

Symptoms of unknown aetiology on Rhododendron hybridum cv. Cunningham's White were observed in the Czech Republic in 2010. The infected plant had malformed leaves, with irregular shaped edges, mosaic, leaf tip necrosis and multiple axillary shoots with smaller leaves. Transmission electron microscopy showed phytoplasma‐like bodies in phloem cells of the symptomatic plant. Phytoplasma presence was confirmed by polymerase chain reaction using phytoplasma‐specific, universal and group‐specific primer pairs. Restriction fragment length polymorphism analysis of 16S rDNA enabled classification of the detected phytoplasma into the aster yellows subgroup I‐C. Sequence analysis of the 16S‐23S ribosomal operon of the amplified phytoplasma genome from the infected rhododendron plant (1724 bp) confirmed the closest relationship with the Czech Echinacea purpurea phyllody phytoplasma. These data suggest Rhododendron hybridum is a new host for the aster yellows phytoplasma subgroup 16SrI‐C in the Czech Republic and worldwide.     

Total Phenols,Flavonoids and Antioxidant Activity in <i>Annona muricata</i> and <i>Annona purpurea</i> Callus Culture

Callus cultures of Annona muricata and Annona purpurea were induced in Murashige and Skoog (MS) medium supplemented with different concentrations of 1-naphthylacetic acid (NAA), 6-benzyladenine (BA) and 2,4-dichlorophenoxyacetic acid (2,4-D) utilized hypocotyls with explant. The highest percentage of callus formation was the treatment supplemented with 3 mg L^-1 NAA for A. muricata (100%) while for A. purpurea in lower percentage (75%). BA stimulated the formation of shoots in all the evaluated concentrations, being the concentration of 2 mg L^-1 the one that induced the greater formation of shoots for A. muricata (23 shoots/explant) and A. purpurea (28 shoots/explant). The content of total phenols, flavonoids and antioxidant activity was measured in the callus obtained from both species. The results showed that a higher content of total phenols was quantified in callus of A. purpurea (27.8 mg g^-1 dw) compared to A. muricata (23.2 mg g^-1 dw). The highest content of total flavonoids was observed in the callus of A. purpurea (8.0 μg g^-1 dw). Antioxidant activity was determined by the 2,2-diphenyl-1-picrylhydracil radical assay. The concentration required for 50% inhibition (IC50) of the 2,2-diphenyl-1-picrylhydracil radicals were 4.22 μg mL^-1 in methanolic extracts of callus of A. muricata, while in extracts of callus of A. purpurea was 2.86 μg mL^-1, in both cases was greater than that found for leaves. Callus culture of the species studied in this work represents an alternative for the production of natural antioxidants.     

Agaricus bisporus compost improves the potential of Salix purpurea × viminalis hybrid for copper accumulation

The aim of the study was to determine the ability of spent mushroom compost (SMC) from the production of Agaricus bisporus (A. bisporus) to stimulate the growth and efficiency of copper (Cu) accumulation by Salix purpurea × viminalis hybrid. Roots, shoots and leaves were analysed in terms of total Cu content and selected biometric parameters. Due to the absence of information regarding the physiological response of the studied plant, low molecular weight organic acids (LMWOAs), phenolic compounds and salicylic acid (SA) contents were investigated. The obtained results clearly demonstrate the effectiveness (usefulness) of SMC in promoting the growth and stimulation of Cu accumulation by the studied Salix taxon. The highest Cu content in roots and shoots was found at the 10% SMC addition (507±22 and 380±11 mg kg^?1 DW, respectively), while there was a reduction of the content in leaves and young shoots (109±8 and 124±7 mg kg^?1 DW, respectively). In terms of physiological response, lowered secretion of LMWOAs, biosynthesis of phenolic compounds and SA, as well as accumulation of soluble sugars in Salix leaves was observed with SMC addition. Simultaneously, an elevation of the total phenolic content in leaves of plants cultivated with SMC was observed, considered as antioxidant biomolecules.