Acclimatisation in the green spruce aphid, Elatobium abietinum

Thin layer chromatography separation of 80% ethanol extracts of adult Elatobium abietinum revealed the presence of the polyhydric alcohol mannitol in aphids overwintering outdoors but not in aphids kept permanently indoors at 15°C. After 3 days at 15°C no traces of mannitol were left in overwintering aphids. Mean freezing temperatures of outdoor, unfed instar I nymphs were about 4°C lower than those of unfed instar I nymphs produced at 15°C. Mean freezing temperatures of overwintering adults were considerably higher than those of unfed instar I nymphs and showed no changes associated with time at 15°C following transference indoors. Similarly, mean freezing temperatures of Sitka spruce needles transferred to 15°C did not change. It was concluded that, although freezing was mainly avoided by supercooling, the presence of mannitol lowered the true freezing temperature of aphid haemolymph and, consequently, the actual freezing temperatures of nymphs produced under cold conditions. However, the considerable increase in freezing point temperatures caused by imbibition of plant sap masked these acclimatisation changes in feeding aphids.     

A comparison of Elatobium abietinum populations on Sitka spruce trees differing in needle retention during aphid outbreaks

Summary No difference in the rate of symptom development and needle loss was evident between resistant and susceptible trees. Population growth rates, which were higher on current year needles in the upper crown, adult weights, reproductive state and instar ratios were similar for aphids from both resistant and susceptible trees. Overwintering survival rates were higher on susceptible trees and were the main cause of the higher number of aphids found on susceptible trees and on the lower crown in comparison with the resistant trees and the upper crown. The supercooling points of needles from resistant trees were higher than those from susceptible trees during the three coldest months of the year-December, January and February. As the aphid freezes at the same temperature as the needle on which it feeds differences in needle freezing temperatures can result in greater aphid mortality on the resistant trees at the temperatures prevailing in NE Scotland during the period of the study. No correlation was found between provenance latitude and supercooling ability of the needles although a very great variability was evident between individual trees. Changes in the form of the super-cooling temperature frequency curves indicated a gradual increase in the quantity of nucleators present in the spruce needles during the winter, the change being more pronounced in the resistant needles.     

Damage caused by the Green Spruce Aphid to Norway and Sitka spruce needles

Three-month-old needles of Sitka spruce were less susceptible to Elatobium abietinutn than 15-month-old needles. Symptoms appeared after longer aphid feeding times but only a proportion of damaged needles fell. After short feeding periods symptoms appeared in more Norway spruce needles than in Sitka spruce, whereas longer feeding periods resulted in more needles producing symptoms in the Sitka spruce. The symptoms took 4—6 days longer to appear in Norway spruce, and needle fall followed a longer feeding period than on Sitka spruce. Following 72 h feeding, needle fall occurred more quickly on Sitka spruce than on Norway spruce. The time taken for needle fall to occur was inversely related to the feeding time in Sitka spruce but such a response was not evident in Norway spruce. The results are discussed in relation to the differences exhibited in the probing behaviour of the aphid on the two spruces.     

Seasonal changes in the distribution of green spruce aphid Elatobium abietinum (Walker) (Homoptera: Aphididae) in the canopy of Sitka spruce

1 Seasonal changes in the distribution of green spruce aphid Elatobium abietinum (Walker) within the canopy of 20–25‐year‐old Sitka spruce are described based on data from two low‐altitude sites (310–420 m above sea‐level), two mid‐altitude sites (500–550 m a.s.l) and one high‐altitude site (610 m a.s.l). 2 Aphids were counted throughout the canopy on shoots representative of all needle age‐classes present at each whorl of branches. Counts were made during the middle week of each month from September to July for 4 years (1999–2003), and mean E. abietinum densities at each canopy position were calculated separately for each month and for the low‐, mid‐ and high‐altitude sites. 3 During September to November, the highest densities of E. abietinum occurred on 3–4‐year‐old needles on branches low in the canopy. Over the winter and spring, the centre of the aphid’s distribution shifted outward and upward, so that by June of the next year the highest aphid densities occurred on current and 1‐year‐old needles on branches near the top of the tree. 4 The aphid distribution was re‐set each year during July, at the time when the nutrient quality of the host was in decline and E. abietinum populations were decreasing. Aphid densities decreased less on 3–4‐year‐old needles than on current and 1‐year‐old needles, suggesting that older needles were a superior food resource at this time of year and in the autumn. However, other factors, such as higher temperatures in the upper canopy during the summer or differential mortality caused by natural enemies, could also have contributed to the change in distribution. 5 The outward and upward shift in the aphid distribution over the winter period provided no evidence that aphids at positions lower and deeper in the canopy were better insulated from freezing temperatures and had higher over‐winter survival rates. Mean air temperatures at the top and bottom of the canopy during the winter were also found to differ by only 0.1–0.2 °C. 6 The percentage of the total aphids per tree that occurred on current or 1‐year‐old needles varied widely between seasons and between sites. Consequently, sampling programmes designed to estimate total population numbers of E. abietinum have little option but to sample needles throughout the canopy, and at regular intervals during the period when the aphid is abundant.     

Factors affecting the cold hardiness of the peach-potato aphid Myzus persicae

Supercooling point studies were used to investigate the factors influencing the cold hardiness of the peach-potato aphid Myzus persicae, a freezing-susceptible insect. Overwintering adults lost cold hardiness as winter progressed, with a variable proportion showing a marked reduction in supercooling ability. Cold hardiness increased in spring so that all individuals demonstrated extensive supercooling ability typical of aphids reared in the laboratory at 20°C with a long photoperiod; these levels of cold hardiness were maintained in the field during summer and early autumn. First instar nymphs demonstrated considerable cold hardiness all year. Surface moisture caused inoculative freezing in some first instar nymphs and adults when supercooled, but the majority were unaffected. In the laboratory, adults starved for 7 days at 5°C showed distinct losses of supercooling potential equivalent to those observed in the field during mid to late winter. No loss of cold hardiness was found in first instar nymphs starved under the same conditions. The results demonstrate that the cold hardiness characteristics of M. persicae are atypical of those observed in other freezing-susceptible insects and it is suggested that continued feeding during mild winter conditions allows maintenance of cold hardiness particularly in adult aphids, and provides a possible explanation for the successful anholocyclic overwintering of M. persicae during such winters.     

Cold hardiness and overwintering of the grain aphid Sitobion avenae

ABSTRACT.

• 1 Cold hardiness as measured by supercooling ability in the active stages of the grain aphid Sitobion avenae (F.) decreased progressively with maturation.
• 2 Aphids showed no acclimation response when maintained at low temperatures.
• 3 Starvation did not improve supercooling ability.
• 4 In a single exposure, surface moisture caused inoculation above the inherent supercooling point in a small proportion of a population.
• 5 Field populations show a seasonal change in supercooling ability, which is at a maximum in summer and a minimum in late winter.
• 6 It is concluded that the act of feeding on healthy plant tissue may confer extensive supercooling ability.

     

Supercooling of Myzus persicae in relation to gut content

The supercooling distribution curves of Myzus persicae varied in response to feeding position, diet and age. Unfed instar I aphids supercooled to temperatures lower than –20 °C. Feeding aphids supercooled to higher temperatures, the earlier instars on subsidiary veins supercooling to lower temperatures than the late instars and adults on the main veins. Starvation decreased supercooling ability in some adult aphids. The supercooling ability of aphids feeding on sucrose solutions through a Parafilm membrane decreased in response to an increasing concentration, the position being reversed on 25% sucrose. Varying the amino acid concentration had no effect. The possible effects of this variable supercooling ability on the winter survival of the aphid are discussed.     

Summer survival of the green spruce aphid,Elatobium abietinum,in North East Scotland

Summary In North East Scotland small numbers of all developmental stages of the green spruce aphid survived on Sitka spruce needles during the summer months despite the nutritional inadequacy of these needles for aphid survival following population collapse in early summer. The surviving adults lost weight and fat reserves in response to time and contained low numbers of embryos. No metabolic acclimatization of the respiration rate occurred in response to exposure to different temperatures. Aphid respiration rates during summer were significantly higher than those in winter. Therefore, it was concluded that no summer aestivation occurred. Summer survival was dependent on the ability of a few aphids of all instars to survive on the nutritionally inadequate host, these aphids possibly possessing higher than normal fat reserves or feeding on marginally nutritionally superior trees or shoots.     

Interactions between green spruce aphid (Elatobium abietinum (Walker)) and Norway and Sitka spruce under high and low nutrient conditions

1 Green spruce aphid (Elatobium abietinum) is a serious pest of spruce (Picea spp.) in north‐west Europe, causing defoliation of one‐year‐old and older needles. 2 Relationships between population development of E. abietinum, needle loss and tree growth were compared for five pure genotypes of Sitka spruce and mixed‐genotype material of Sitka and Norway spruce, grown under high and low nutrient conditions. 3 Despite wide differences in flushing date between spruce genotypes, E. abietinum populations peaked on the same date on each genotype and on the mixed‐genotype material, irrespective of nutrient supply. 4 Larger aphid populations developed on trees grown under high nutrient conditions than under low nutrients. However, more needles were lost per aphid in the low nutrient treatment and overall defoliation rates in the two nutrient treatments were similar. 5 Total aphid numbers differed significantly between Sitka spruce genotypes within nutrient treatments, but not in relation to bud‐burst or needle terpene content. Reductions in height growth caused by infestation were greater (15–44%), and related to mean aphid densities and defoliation, in the low nutrient treatment, but were smaller (11–27%) and not related to aphid density and defoliation in the high nutrient treatment. 6 Development of E. abietinum populations was similar on Norway and Sitka spruce, but Norway spruce lost fewer needles. However, the effects of infestation on tree growth were more closely related to aphid density and were similar for Norway and Sitka spruce. 7 Infestation caused a decrease in total root dry weight of Norway and Sitka spruce in proportion to the reductions observed in above‐ground growth.     

Aphid honeydew and its effect on the phyllosphere microflora of Picea abies (L.) Karst

Aphids of the genus Cinara, feeding on Norway spruce, excrete copious amounts of honeydew, a carbon-rich waste product, which accumulates locally on needles and twigs. We investigated the role of honeydew as a potential source of energy which might promote the growth of micro-organisms in the phyllosphere of conifer trees. To approach this question, we followed the population dynamics of Cinara spp. in a natural forest stand over two seasons. We also studied the amounts of honeydew produced by individual aphids and identified potential parameters which might influence honeydew production. Finally, we determined the growth of micro-organisms on infested and uninfested needles of Norway spruce during the growing season. Confined to Picea abies, the investigated Cinara species only became abundant in midsummer, when needles and shoots were expanding. The populations showed only a single peak in abundance, the timing and magnitude of which may vary from year to year due to weather conditions, changes in plant quality in a yearly cycle or the impact of natural enemies. The amount of honeydew produced by individual aphids was dependent on the developmental stage of the aphid, the nutritional supply of its host plant and on the developmental state of the Norway spruce (e.g. bud burst, end of shoot extension). The presence of honeydew significantly increased the growth of bacteria, yeast and filamentous fungi on the surface of needles and there was a pronounced seasonal trend, with the highest abundance in midsummer correlating with the period of peak aphid abundance. Taken together, these findings indicate that aphids have an influence on microbial ecology in the phyllosphere of trees. The implication of our study, from interactions at the population level to effects and potential consequences for C and N fluxes at the level of forest ecosystems, is discussed.     

Variation in summer cold-hardiness of the Antarctic oribatid mite Alaskozetes antarcticus from contrasting habitats on King George Island

Summary The Antarctic oribatid mite Alaskozetes antarcticus was collected from several field habitats near Great Wall Station (62°13S, 58°58W) on King George Island during January and February 1990. The tritonymphs and adults were examined for their supercooling ability and survival at subzero temperatures in relation to inoculative freezing. The active tritonymphs and adults showed a wide range of supercooling points probably due to their polyphagous feeding activity and humid habitat conditions, with means ranging from -3.8° to -22.4°C. Detrivores were inferior to algivores in their supercooling ability. The former seemed to be transiently exposed to the hazard of freezing during the cool Antarctic summer. The resting (premoulting) tritonymphs exhibited the lowest mean supercooling point of -28.3°C. Inoculative freezing reduced the survival of A. antarcticus. Its effect became conspicuous at temperatures below -20°C and was serious in the deeply supercooled individuals, such as resting tritonymphs and algivorous adults. During the active season, spontaneous freezing probably started from the gut contents seemed to be more fatal than inoculative freezing for this freeze intolerant species.     

Freezing stress and membrane injury of Norway spruce (Picea abies) tissues

Effects of sub-zero temperatures (−5 to −35°C) on the tissues of needles, buds and shoots of Norway spruce [ Picea abies (L.) Karst.] were studied. The freezing caused increased efflux of cellular electrolytes. Freezing injury of the primordial shoots and 1-year-old shoots was the result of the spontaneous freezing of a deep supercooled cellular water. The crystallization injures the cellular membranes leading to the loss of semipermeability and to the drastic efflux of K⁺. In the needles there was no deep supercooling of water and two patterns of changes in the membranes, depending upon the range of the applied temperatures, could be distinguished. At 0 to – 25°C, which do not kill the cells, we observed a disturbance in the membrane semipermeability as monitored by electrolytes efflux within a few hours after thawing of the needles. At lower temperatures (−35°C) we observed irreversible loss of the membrane semipermeability, and death of the tissue. Those changes occurred 10 h after thawing and were probably caused by the released lytic enzymes and some toxic compounds, which acted on the cellular membranes.     

Colony orientation and successful defence behaviour in the conifer aphid, Schizolachnus pineti

We studied colony structure and body orientation behaviour of Schizolachnus pineti, a specialised aphid feeding on needles of Scots pine. Aphids feeding at the margin of a colony always face towards the centre of the colony, bounding the colony with their legs. This behaviour increased the defence success aginst the specialised parasitoid (Pauesia unilachni) and the specialised predator (Scymnus nigrinus). Both enemies had less success when attacking S. pineti from the abdominal site when compared to the front. We assume that the defence orientation displayed by marginally feeding aphids has an adaptive nature and is particularly effective when feeding in a row on linear plant structures such as needles.     

The effect of needle age on the acceptability of Sitka spruce needles to the aphid,Elatobium abietinum (Walker)

Summary A comparison of the feeding behaviour of E. abietinum on current year needles and previous year needles of P. sitchensis revealed that during the summer months the aphids preferentially settled on previous year needles, this response not being evident in winter. Aphids on current year needles in summer took a much longer period of time to commence sap uptake than in winter, intake ceasing following a very short feeding period. On previous year needles sap uptake in December commenced after a shorter feeding period than in June. Analysis of total and soluble nitrogen levels in Sitka spruce needles showed that current year needles had initially higher levels during shoot elongation in May and early June, but that previous year needles had higher levels for most of the remainder of the year. Quantitative analyses of amino acids revealed that in current year needles the levels were generally lower than in previous year needles. Less marked proportional differences were observed between previous year needles in May and in July/August when the needles were unsuitable. Addition of amino acids in solution into cut current year shoots resulted in increased longevity on shoots containing introduced iso-leucine, histidine and methionine and revealed a general imbalance of the amino acids.     

Effects of temperature on overwintering populations of the green spruce aphid Elatobium abietinum

Recorded minimum temperatures of – 7 ^oC or lower noticeably reduced overwintering populations of the green spruce aphid, Elatobium abietinum, in north-east Scotland. It is suggested that, at these temperatures, ice formation in the needles of the host Sitka spruce, caused attached aphids to freeze. Aphid mortality also occurred when maximum temperatures did not rise above + 6 ^oC for prolonged periods, possibly as a result of starvation following an extended chill coma. A diagrammatic representation of the main factors affecting anholocyclic populations of E. abietinum during the winter is presented to emphasize the governing role played by temperature. The balance between mortality and recruitment determines the size of the population at the end of the winter, and this in turn determines the subsequent summer infestation. It should be possible, therefore, to predict aphid outbreaks either from winter temperatures or from the number of aphids present at the end of the winter. Temperature records obtained from integrating thermometers indicated that the inside of the lower crown tended to be the warmest part of the tree during the winter, resulting in greater aphid survival in the lower branches.     

Seasonal variation in the cold-hardiness of a free-living predatory antarctic nematode,Coomansus gerlachei (mononchidae)

Summary Coomansus gerlachei shows a strong seasonal variation in its supercooling ability, with most individuals freezing between -5° and -7°C, in a discrete high group, or between -15° and -26°C in a more diffuse low group. Increased cold-hardiness in winter is achieved by a strategy of freeze avoidance with an increase in the proportion of the population showing extensive supercooling ability. Smaller life-stages also exhibit lower supercooling points. Ice-nucleation at high sub-zero temperatures increases the probability of surviving freezing, a capacity which is enhanced in larger life-stages; adults and J4 stage juveniles show 50% survival at a supercooling point of -8.65° C. Laboratory incubations of field-fresh worms suggest that recent feeding is responsible for the movement between low and high groups. The cold-hardiness data for C. gerlachei provide interesting comparisons to the available data for microarthropods and create a precedent for seasonal changes between strategies of freezing tolerance and freezing intolerance in a field invertebrate population.     

Deep supercooling enabled by surface impregnation with lipophilic substances explains the survival of overwintering buds at extreme freezing

The frost survival mechanism of vegetative buds of angiosperms was suggested to be extracellular freezing causing dehydration, elevated osmotic potential to prevent freezing. However, extreme dehydration would be needed to avoid freezing at the temperatures down to ?45°C encountered by many trees. Buds of Alnus alnobetula, in common with other frost hardy angiosperms, excrete a lipophilic substance, whose functional role remains unclear. Freezing of buds was studied by infrared thermography, psychrometry, and cryomicroscopy. Buds of A. alnobetula did not survive by extracellular ice tolerance but by deep supercooling, down to ?45°C. An internal ice barrier prevented ice penetration from the frozen stem into the bud. Cryomicroscopy revealed a new freezing mechanism. Until now, supercooled buds lost water towards ice masses that form in the subtending stem and/or bud scales. In A. alnobetula, ice forms harmlessly inside the bud between the supercooled leaves. This would immediately trigger intracellular freezing and kill the supercooled bud in other species. In A. alnobetula, lipophilic substances (triterpenoids and flavonoid aglycones) impregnate the surface of bud leaves. These prevent extrinsic ice nucleation so allowing supercooling. This suggests a means to protect forestry and agricultural crops from extrinsic ice nucleation allowing transient supercooling during night frosts.     

Quantifying aphid behavioral responses to environmental change

Aphids are the most common vector of plant viruses, and their feeding behavior is an important determinant of virus transmission. Positive effects of global change on aphid performance have been documented, but effects on aphid behavior are not known. We assessed the plant‐mediated behavioral responses of a generalist aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae), to increased CO₂ and nitrogen when feeding on each of three host species: Amaranthus viridis L. (Amaranthaceae), Polygonum persicaria L. (= Persicaria maculosa Gray) (Polygonaceae), and Solanum dulcamara L. (Solanaceae). Via a family of constrained Markov models, we tested the degree to which aphid movements demonstrate preference among host species or plants grown under varying environmental conditions. Entropy rates of the estimated Markov chains were used to further quantify aphid behavior. Our statistical methods provide a general tool for assessing choice and quantitatively comparing animal behavior under different conditions. Aphids displayed strong preferences for the same host species under all growth conditions, indicating that CO₂‐ and N‐induced changes in plant chemistry have minimal effects on host preference. However, entropy rates increased in the presence of non‐preferred hosts, even when preferred hosts were available. We conclude that the presence of a non‐preferred host species affected aphid‐feeding behavior more than changes in plant leaf chemistry when plants were grown under elevated CO₂ and increased N availability.     

Freezing avoidance by supercooling in Olea europaea cultivars: the role of apoplastic water,solute content and cell wall rigidity

Plants can avoid freezing damage by preventing extracellular ice formation below the equilibrium freezing temperature (supercooling). We used Olea europaea cultivars to assess which traits contribute to avoid ice nucleation at sub‐zero temperatures. Seasonal leaf water relations, non‐structural carbohydrates, nitrogen and tissue damage and ice nucleation temperatures in different plant parts were determined in five cultivars growing in the Patagonian cold desert. Ice seeding in roots occurred at higher temperatures than in stems and leaves. Leaves of cold acclimated cultivars supercooled down to ?13 °C, substantially lower than the minimum air temperatures observed in the study site. During winter, leaf ice nucleation and leaf freezing damage (LT₅₀) occurred at similar temperatures, typical of plant tissues that supercool. Higher leaf density and cell wall rigidity were observed during winter, consistent with a substantial acclimation to sub‐zero temperatures. Larger supercooling capacity and lower LT₅₀ were observed in cold‐acclimated cultivars with higher osmotically active solute content, higher tissue elastic adjustments and lower apoplastic water. Irreversible leaf damage was only observed in laboratory experiments at very low temperatures, but not in the field. A comparative analysis of closely related plants avoids phylogenetic independence bias in a comparative study of adaptations to survive low temperatures.     

Comparative analysis of overwintering physiology in nine species of semi‐aquatic bugs (Heteroptera: Gerromorpha)

In semi‐aquatic bugs (Heteroptera: Gerromorpha), the strategies of overwintering in a cryothermic state (i.e. at body temperatures below the equilibrium freezing point) remain largely unexplored. The present study provides an analysis of the ecophysiological aspects of overwintering in nine gerromorphan species. All nine species avoid ice formation by means of a more or less extensive supercooling of their body fluids. There is a tight correlation between the supercooling point (SCP) and the lower lethal temperature. Different species use different physiological adjustments to increase the likelihood of survival in a supercooled state. These include stabilization of the supercooled state by active antifreeze factors that cause thermal hysteresis between equilibrium melting and freezing points, the accumulation of low‐molecular weight sugars and polyols with putative cryoprotective functions, or by having a relatively high body fluid osmolality, combined with a low level of hydration. The majority of species under study overwinter only as adults, whereas Velia caprai Tamanini can overwinter either as an adult or in the egg stage. The supercooling capacity of V. caprai adults is insufficient to prevent the risk of lethal freezing. The adults therefore survive only opportunistically in suitable microhabitats, and/or during mild winters. The survival of V. caprai in winter is assured by extensive supercooling and having overwintering eggs that are highly cold tolerant.