Effect of variation in photoperiodic response on diapause induction and developmental time in the willow leaf beetle, Plagiodera versicolora

The willow leaf beetle, Plagiodera versicolora (Coleoptera: Chrysomelidae) overwinters in adult diapause. In this study, the photoperiodic responses for diapause induction and developmental time were examined in the Ishikari (Hokkaido, Japan) population of P. versicolora. All females entered reproductive diapause under short daylength (L10:D14), but 31.7% of females did not enter diapause under long daylength (L16:D8). The developmental time from oviposition to adult emergence was significantly longer at L10:D14 than that at L16:D8. Norm of reaction curves illustrated variation among families in the photoperiodic responses for diapause induction and for developmental time. ANOVA indicated significant family × photoperiod interactions in the developmental time. At L16:D8, developmental time was positively correlated with the incidence of diapause in females. This means that a female having a longer developmental time tends to have a longer critical photoperiod. Such variation may be maintained by differences in selection pressures on the growth rate and the critical photoperiod for diapause induction between univoltine and bivoltine genotypes because Ishikari is located in a transitional area between populations with univoltine and bivoltine life cycles.     

Effects of leaf toughness on mandibular wear of the leaf beetle, Plagiodera versicolora

Abstract. 1. The effects of leaf toughness on mandibular wear of the leaf beetle Plagiodera versicolora Laich. (Coleoptera: Chrysomelidae) were evaluated by feeding adult beetles young (tender) and old (tough) leaves of Salix babylonica and S. alba 'Tristis'.
2. Tough leaves erode the cutting surface of beetle jaws more so than tender leaves.
3. Beetles with worn jaws consumed leaves at a slower rate than ones with less mandibular wear.
4. Because rates of leaf consumption and egg production are positively correlated, increased mandibular wear may reduce beetle fecundity.
5. These results support the belief that leaf toughness may act as a potent defence affecting morphology, feeding behaviour, and ultimately spatial and temporal patterns of herbivores.     

Spatio-temporal abundance of flushing leaves shapes host selection in the willow leaf beetle, Plagiodera versicolora

In the willow leaf beetle, Plagiodera versicolora (Laicharting) (Coleoptera: Chrysomelidae), food resources available for adults are severely restricted by leaf toughness, which increases with age. Nevertheless, females require their own food almost all their life in order to produce eggs. In this paper, we have focused our attention on the spatio-temporal abundance of flushing leaves and have examined its effect on host-plant selection by adults among four co-occurring willow species ( Salix chaenomeloides , Salix eriocarpa , Salix integra , and Salix serissaefolia ) (Salicaceae) by field observations and experiments at two spatial scales. Among the various factors associated with this, the amount of new leaf production contributed maximally to variation in adult abundance. By conducting two experiments, we confirmed that the adults preferentially flew towards willow trees with abundant flushing leaves. Furthermore, we detected substantial seasonal changes in new leaf abundance and realized fecundity in the field, and a strong positive correlation was observed between them. Availability of adult food resources limited the reproductive performance of adults, particularly in mid-summer when only S. serissaefolia produced a few new leaves. These results supported the substantial effect of new leaf abundance on adult abundance in the field. Thus, we concluded that adult feeding is a critical factor that shapes the host-plant selection of P. versicolora and determines its seasonal occurrence through the dispersal and settlement of adults.     

柳蓝叶甲的生物学特性室内观察

在实验室条件下(27±0.5℃,12 D:12 L)研究了柳蓝叶甲Plagiodera versicolora的生物学特性。结果表明,幼虫每隔2~3 d蜕皮1次,共蜕皮2次。成虫产卵前期4 d,世代周期17 d。产卵具有一定的周期性,约10 d为1个周期。叶龄对幼虫的生长发育和成虫的产卵行为有极显著的影响。当幼虫以成熟叶为食料时,其生长发育速率明显减慢,各龄期延长,蛹重较轻;成虫以成熟叶或老叶为食料时产卵几乎完全被抑制,而以幼叶为食料时几乎每天均能产卵。此外,柳蓝叶甲产出卵表面化合物对产卵行为有极为显著的影响。卵表面的有机提取物对成虫产卵具有显著的引诱作用;相反,卵表面的水提取物对产卵有一定的驱避作用。     

Nitrogen homeostasis in a willow leaf beetle, Plagiodera versicolora, is independent of host plant quality

While foliar nitrogen (N) content of host plants depends on environmental conditions, N content of herbivorous insects may remain relatively constant due to homeostasis. However, it is unknown to what extent insects can maintain their body elemental composition against natural variation in host plant quality. The present study examined the performance and N content of a willow leaf beetle, Plagiodera versicolora Laicharting (Coleoptera: Chrysomelidae), when fed leaves of host willow, Salix eriocarpa Franchet et Savatier (Salicaceae), with varying nutritional status. Water content, toughness, and N content of willow leaves varied seasonally, and they affected performance of the leaf beetle. The leaf beetle achieved high performance when fed young leaves. On the other hand, the N content of the leaf beetle changed little, and it was independent of that of willow leaves, indicating strong N homeostasis of the leaf beetle. We discussed the function of N homeostasis in herbivorous insects in tritrophic level interactions.     

Strong material benefits and no longevity costs of multiple mating in an extremely polyandrous leaf beetle, Chrysochus cobaltinus (Coleoptera: Chrysomelidae)

It is well established that females of many species exhibitpolyandry. Although such behavior often increases female fitnessby augmenting fecundity or enhancing the genetic diversity andvigor of their offspring, it often reduces female longevity.It has been argued that trade-offs between these costs and benefitsshould limit the degree to which females remate. However, theexistence of highly polyandrous species suggests substantialpolyandry benefits and/or minimal costs in some systems. Femalesof the leaf beetle, Chrysochus cobaltinus, are extremely polyandrous,providing an opportunity to examine the factors influencingthe evolution of such behaviors. We compared the fecundity andlongevity of singly mated females, females that mated multipletimes with the same male, and females that mated multiple timeswith different males. Compared with females in the single matingtreatment, females in both multiple mating treatments exhibiteda significant reduction in latency to oviposition and, due toan increase in daily egg production, significant increases inlifetime fecundity. This difference diminished as the time sincelast mating increased. There were no differences in fecunditybetween the 2 multiple mating treatments, indicating that mateidentity does not influence the material benefits of multiplemating. Surprisingly, female longevity did not differ amongtreatments. The pronounced fecundity benefits that females gainfrom multiple mating, coupled with a lack of longevity costs,apparently explains the extreme polyandry in this species. Inaddition, the existence of material fitness benefits via conspecificmatings raises the intriguing possibility that in a C. cobaltinus–Chrysochusauratus hybrid zone, heterospecific matings may confer similarbenefits to Chrysochus females.     

Factors contributing to the seasonal occurrence of the willow leaf beetle Plagiodera versicolora

We conducted a 3-year study of a natural population of the willow leaf beetle Plagiodera versicolora (Coleoptera: Chrysomelidae) on a river bank of the Inukami River, Shiga, central Japan, where four willow species (Salix chaenomeloides, S. eriocarpa, S. integra, and S. serissaefolia) occur sympatrically. Our survey showed that: (1) at the study site, the abundance of P. versicolora greatly varied among years and among willow species; (2) adult abundance changed seasonally with species-specific patterns on different willow species; and (3) the dispersal-settlement of adults had the most pronounced effects on the seasonal population growth rate of P. versicolora. Factors affecting these results were discussed.     

Mating behavior of the cabbage beetle, Colaphellus bowringi (Coleoptera: Chrysomelidae)

Abstract  Circadian mating rhythms, mating frequency, mating duration, and the effect of mating duration on fecundity and fertility in the cabbage beetle, Colaphellus bowringi were investigated in the laboratory. Mating occurred throughout the 24-h cycle but the majority of copulations occurred in the photophase with two apparent peaks, one at 8:00 and another at 16:00. Mating frequency observations for 10 consecutive days indicated that pre-mating period of C. bowringi was about 4 days, and pairs mated an average of 5 times per day and an average of 40 times during the first 10 days. There was a negative correlation between mating frequency and mating duration during the consecutive mating. The mean duration of the first copulation (136.24 ± 4.62 min) was significantly longer than those of the second (57.87 ± 2.03 min), third (53.05 ± 2.05 min) and fourth copulation (30.86 ± 2.98 min). Fecundity showed a slight increase with increasing mating duration but no significant difference among treatments. However, fertility was significantly influenced by the mating duration in this species. Mating of 20-min duration did not produce viable eggs. The mean percentage of fertile eggs with completed mating duration (204.43 ± 18.96 min, 56.75% fertile eggs) was significantly higher than those with 60 min (39.55%) and 30 min (17.91%) mating duration, suggesting that the longer mating duration might be associated with transfer of more sperm that are used to increase the fertility of eggs.     

Soil nutrients and water availability interact to influence willow growth and chemistry but not leaf beetle performance

We investigated the effects of soil nutrient and water availability on the growth and chemistry of the silky willow (Salix sericea Marshall), and on the performance of the imported willow leaf beetle (Plagiodera versicolora Laichartig). Our major aims were to determine whether there are nutrient–water interactions on plant traits and whether this leads to parallel interactions for herbivore performance. We used a 2 × 3 fully factorial design, which consisted of high and low nutrient treatments crossed with dry, field capacity, and flooded water treatments. We found that nutrient additions increased plant growth, but only in field capacity and flooded conditions (nutrient–water interaction). Leaf nitrogen content also depended on the interaction between soil nutrients and water: nutrient addition resulted in a larger increase in foliar nitrogen in the field capacity treatment than in the flooded and dry treatments. Of the two phenolic glycosides measured, salicortin and 2′‐cinnamoylsalicortin, only one was affected by the treatments. 2′‐cinnamoylsalicortin concentration was lower in the high nutrient–dry treatment compared with the other treatments. In contrast to plant responses, there were no interactions found for larval or pupal weight or development time. Nutrient addition led to an increase in female pupal weight, and foliar N was positively correlated with female pupal weight and negatively correlated with female development time. In addition, leaf water was positively correlated with female development time. The lack of interactions for insect performance may stem from the small absolute differences in foliar nitrogen content associated with the interaction between the nutrients and water. Taken together, our results suggest that nutrient–water interactions influence plant traits that are potentially important for insect performance (leaf nitrogen and water), but these interactions do not produce parallel interactions in beetle performance.     

Effects of Leaf Age on Oviposition and on Offspring Fitness in the Imported Willow Leaf Beetle Plagiodera versicolora (Coleoptera: Chrysomelidae)

Imported willow leaf beetles Plagiodera versicolora oviposit on willow leaves, and both larvae and adults feed on the leaves. In the field, eggs were found on leaves near the center of branchlets, and the number of eggs per cluster was independent of the leaf area and position. However, in the laboratory, females chose young leaves over old leaves, for both oviposition and feeding and choice did not rely on information on relative position or size of leaves. Developing on young versus old leaves may provide both advantages and disadvantages. In the laboratory, larvae developed more quickly and attained a greater adult weight when fed young versus old leaves, perhaps because of increased mandibular wear of larvae fed old leaves. However, in the field, survival of eggs was lower on young versus old leaves. In the laboratory, rates of cannibalism and survivorship to adulthood did not differ on young versus old leaves.     

Endemism patterns in the Italian leaf beetle fauna (Coleoptera,Chrysomelidae)

In this contribution the results of a zoogeographical analysis, carried out on the 123 endemic leaf beetle species (Coleoptera: Chrysomelidae) occurring in Italy and its immediately adjacent regions, are reported. To assess the level of faunistic similarity among the different geographic regions studied, a cluster analysis was performed, based on the endemic component. This was done by calculating the Baroni Urbani & Buser’s similarity index (BUB). Finally, a parsimony analysis of endemicity (PAE) was used to identify the most important areas of endemism in Italy.     

Reproductive and subsocial behaviour in the ovoviviparous leaf beetle Gonioctena sibirica (Coleoptera: Chrysomelidae)

Abstract.

• 1 In the spring, females of the leaf beetle Gonioctena sibirica deposited larvae on the ventral surface of growing young leaves situated on the apical position of shoots of the willow Salix bakko.
• 2 The parent females remained with the larvae usually on the underside of the basal part of leaves, facing toward the base of shoots. When other arthropods approached, the females temporarily moved towards these intruders, showing aggressive behaviour such as swinging the body or stamping the legs. Many females remained with their larvae until the larvae grew into the final (fourth) instar. No female produced an additional brood in the field.
• 3 Broods from which parent females were experimentally removed suffered higher mortality than those in which females were left intact. Arthropods such as spiders and ants were observed preying on the larvae. In contrast, the survivorship of broods from which females were removed and intruders were excluded with a sticky substance applied to the base of twigs was not different from that of control broods. These results demonstrate that the main mortality factor of offspring is pedestrian arthropod predators and females physically repel the predators.
• 4 Potentially alternative reproductive strategies, such as producing a large number of offspring by iteroparity and/or larger brood(s) with less or no care, seem to be inhibited in G.sibirica by larval dependence on growing young leaves which are temporally limited and by ovoviviparity which may have limited brood size.

     

Repeated mating with the same male increases female longevity and fecundity in a polyandrous leaf beetle Galerucella birmanica (Coleoptera: Chrysomelidae)

Female multiple mating (polyandry) is widespread across Insecta, even if mating can be costly to females. To explain the evolution and maintenance of polyandry, several hypotheses, mainly focusing on the material (direct) and/or the genetic (indirect) benefits, have been proposed and empirically tested in many species. Considering only the direct benefits, repeatedly‐mated females are expected to exhibit the same fitness as multiply‐mated females under the same mating frequency. In the present study, we compare the fitness of females received monandrous repeated mating (MM) and polyandrous multiple mating (PM) in a polyandrous leaf beetle Galerucella birmanica and assess female mate preference with regard to polyandry or monandry. Our data indicate that the longevity and the egg‐laying duration of MM females are significantly longer than that of PM females. MM females produce significantly more hatched eggs than PM females over their lifetime under the same mating frequency, which results from the high hatching rate of eggs produced by MM females. PM females mated with novel virgin males in the second mating suffer decreased longevity and lifetime fecundity compared with PM females mated with novel mated males in the second mating. Once‐mated females are more likely to re‐mate with familiar males than novel males. By contrast to expectations, the results of the present study suggest that repeated mating provides females with more direct benefits than multiple mating in G. birmanica, and females prefer to re‐mate with familiar males. The possible causes of this finding are discussed.     

Characterization and chromosome location of satellite DNA in the leaf beetle Chrysolina americana (Coleoptera, Chrysomelidae)

This paper is the first record of the satellite DNA of the specialized phytophagous genus Chrysolina. The satellite DNA of Chrysolina americana is organized in a tandem repeat of monomers 189 bp long, has a A + T content of 59.6 % and presents direct and inverted internal repeats. Restriction analysis of the total DNA with methylation sensitive enzymes suggests that this repetitive DNA is undermethylated. In situ hybridization with a biotinylated probe of the satellite DNA showed the pericentromeric localization of these sequences in all meiotic bivalents. The presence of this repetitive DNA in other species of the genus was also tested by Southern analysis. The results showed that this satellite DNA sequence is specific to the C. americana genome and has not been found in three other species of Chrysolina with a different choice of host plants than in the former. This revised version was published online in July 2006 with corrections to the Cover Date.     

大猿叶虫夏滞育的诱导：基于定量的光周期反应

为了探明大猿叶虫Colaphellus bowringi Baly夏滞育诱导的光周期时间测量特性, 我们通过室内实验系统比较了该虫在25℃、 不同长光照条件下,夏滞育的发生以及诱导50%个体进入夏滞育所需求的光 暗循环数。结果表明：不同长光照诱导的夏滞育比率有显著差异, 其中15 h或16 h光照诱导的滞育比率最高, 短于或长于这两个光照其滞育率均明显下降。在不同光 暗循环实验中, 14 h诱导的滞育比率均低于50%, 诱导50%个体进入夏滞育所需求的光 暗循环数在L15∶D9, L16∶D8, L17∶ D7和L18∶D6分别为2.61, 3.72, 4.64和5.92 d, 处理间存在显著差异。这些结果提示该虫夏滞育的诱导是基于定量的光周期反应。     

On the laboratory rearing of green dock leaf beetles Gastrophysa viridula (Coleoptera: Chrysomelidae)

Abstract Leaf beetles Gastrophysa viridula have attracted recently increased research interest from various points of view, since they are: (i) pest insects in rhubarb crops; (ii) potential biocontrol agents of dock plants Rumex spp. in grasslands; and (iii) a model species in ecological studies on insect population dynamics, biochemistry, behavior, biomechanics and biomimetics. The continuous rearing of beetles at standardized conditions may help to unify the fitness state of different individuals, allowing a better comparison of experimental results. The present communication suggests a modular space‐ and time‐saving rearing method of G. viridula in stackable faunariums under laboratory conditions, which has been successfully established and continuously used over the last 5 years. Several developmental stages were kept in separate boxes, and multiple generations were kept simultaneously, depending on the required number of beetles.     

Cold tolerance and supercooling capacity in overwintering adults of elm leaf beetle Xanthogaleruca luteola (Coleoptera: Chrysomelidae)

Elm leaf beetle, Xanthogaleruca luteola (Muller) is one of the key pests of elm trees all over the world, and survives winter in reproductive diapause in sheltered locations. Seasonal variation of whole body supercooling points (SCPs), LT50 (temperature at which 50% of the test individuals die) and survival rate after exposure to subzero temperatures were determined in field collected adults during October 2008 to May 2009 and October 2009 to May 2010. The SCP of adults decreased significantly from October (median=-13.8°C) to January (median=-20.7°C) in first year, relatively similar results was observed in the second year. The lowest LT50 was observed in overwintering adults collected in January (-16.81°C) in the first year and December (-15.59°C) in the second year. Mortality at -15°C for 24 h was >70% in early autumn in both years whereas it decreased to lower than 45% in early winter, the highest mortality (100%) was observed in adults collected in May in both years. Cold acclimated adults (30 d, 5°C) in November 2008 exhibited significantly higher SCP (-12.21±0.64°C) than nonacclimated adults (-15.57±1.35°C). A 30-d exposure to 5°C caused >20% mortality in November, while <9% mortality was observed in adults collected in December and January 2008. Overwintering adults died upon freezing and the lower lethal temperatures were within the range of SCP, indicating that X. luteola is a freeze intolerant insect.     

Culturable bacterial microbiota of Plagiodera versicolora (L.) (Coleoptera: Chrysomelidae) and virulence of the isolated strains

Plagiodera versicolora (Laicharting, 1781) (Coleoptera: Chrysomelidae) is an important forest pest which damages many trees such as willow, poplar, and hazelnut. In order to find new microbes that can be utilized as a possible microbial control agent against this pest, we investigated the culturable bacterial flora of it and tested the isolated bacteria against P. versicolora larvae and adults. We were able to isolate nine bacteria from larvae and adults. The isolates were characterized using a combination of morphological, biochemical, and physiological methods. Additionally, we sequenced the partial sequence of the 16S rRNA gene to verify conventional identification results. Based on characterization studies, the isolates were identified as Staphylococcus sp. Pv1, Rahnella sp. Pv2, Rahnella sp. Pv3, Rahnella sp. Pv4, Rahnella sp. Pv5, Pantoea agglomerans Pv6, Staphylococcus sp. Pv7, Micrococcus luteus Pv8, and Rahnella sp. Pv9. The highest insecticidal activity against larvae and adults was obtained from M. luteus Pv8 with 50 and 40 % mortalities within 10 days after treatment, respectively. Extracellular enzyme activity of the bacterial isolates such as amylase, proteinase, lipase, cellulose, and chitinase was also determined. Consequently, our results show that M. luteus Pv8 might be a good candidate as a possible microbial control agent against P. versicolora and were discussed with respect to biocontrol potential of the bacterial isolates.     

Effects of thermoperiods on diapause induction in the cabbage beetle, Colaphellus bowringi (Coleoptera: Chrysomelidae)

Abstract. The effects of thermoperiods on diapause induction in continuous darkness or under a 12 : 12 h LD photoperiod were investigated in the cabbage beetle, Colaphellus bowringi Baly, a typical short‐day species. The diapause response curves both at different constant temperatures and at the thermocycle of format CT x: (24 ? x) h (16 : 28 °C) under continuously dark rearing conditions showed that the incidence of diapause depended mainly on whether or not the mean temperature was ≤20 °C or >20 °C. If the mean temperature was ≤20 °C, all individuals entered diapause; if >20 °C, the incidence of diapause declined gradually with increasing mean temperatures. The thermocycle (CT 12 : 12 h) with a series of different cryophases (8–22 °C) and thermophases (24–32 °C) under continuous darkness demonstrated a cryophase response threshold temperature of approximately 19 °C and a thermophase response threshold temperature of approximately 31 °C. Thermoperiodic amplitude (temperature difference between cryophase and thermophase) was shown to have a significant influence on diapause induction at the mean temperatures of 22, 23 and 24 °C, but not at ≥25 °C. Thermoperiodic responses under LD 12 : 12 h clearly showed that the incidence of diapause was influenced strongly by the photophase temperature. The thermoperiod under LD 12 : 12 h induced a much lower incidence of diapause than the thermoperiod with the same temperature in continuous darkness. The ecological significance of thermoperiodic induction of diapause in this species is discussed.     

Proteolytic activity in Plagiodera versicolora Laicharting (Coleoptera: Chrysomelidae): Characterization of digestive proteases and effect of host plants

Proteolytic profiles in the midgut of Plagiodera versicolora were studied using biochemical approaches, and the effects of host plants on possible changes in their activity were determined. Morphology of the alimentary canal revealed several areas of sections, namely bucca, pharynx, esophagus, crop, midgut, ileum, rectum and anus. A pH of 6 and 11 was found to be optimal for soluble and membrane-bound fractions, by using azocasein 2% as a substrate. Determination of specific proteases demonstrates the presence of trypsin-like, chymotrypsin-like, elastase, cathepsin B, cathepsin L and cathepsin D, as well as two exopeptidases. Regarding site of activity for each specific protease, it was found that the major activity of cathepsin B and cathepsin L was in the soluble fraction, chymotrypsin, cathepsin D and two exopeptidases in membrane-bound fraction. Additionally, trypsin-like and elastase activities had no significant differences between fractions. The presence of the above mentioned specific proteases was verified using the specific inhibitors PMSF, TLCK, TPCK, cystatin, phenanthroline and DTT. Feeding of the beetle on four host plants: including Salix aegyptica, S. alba, Populus alba and P. caspica, from the 1st larval instar to adult, revealed the highest trypsin-, chymotrypsin-like and elastase activities in the individuals fed on S. aegyptica and S. alba, respectively. Regarding cathepsins B and L, the highest activities were observed on S. alba and S. aegypticum but cathepsin D was higher in S. Alba and P. alba. Feeding on S. alba and S. aegypticum showed the highest activities of amino- and carboxy-peptidases, respectively.