Influence of simulated snow cover on the cold tolerance and freezing injury of yellow-cedar seedlings

It has been hypothesized that yellow‐cedar [Chamaecyparis nootkatensis (D. Don) Spach] decline may result from root freezing injury following climate change‐induced reductions in protective snow cover. To test this hypothesis, we measured the freezing tolerance and injury expression of yellow‐cedar seedlings in three treatments that differed in the insulative protection they provided to soils during winter and spring: (1) full exposure to ambient temperatures (exposed treatment), (2) continuous protection from ambient temperatures via addition of perlite over pots (full protection), and (3) perlite protection only during winter and exposure to ambient temperatures during spring (partial protection). Foliage from all treatments was cold tolerant enough to prevent foliar freezing injury throughout the study period. However, on all sample dates, roots of seedlings from all treatments were only tolerant to about ?5 °C – a level considerably warmer than the reported maximum cold tolerance for the species and well above the soil temperature recorded in the exposed treatment. As a result of this limited root cold tolerance, visibly uninjured roots of seedlings from the exposed treatment had significantly higher relative electrolyte leakage (REL) throughout the winter and early spring than seedlings in soil protection treatments. Seedlings from the exposed treatment also had significantly higher foliar REL values and greater visual foliar injury than seedlings from the other treatments starting in early spring. For both roots and foliage, REL measurements consistently detected tissue damage before visual injury was evident. Patterns of injury from both REL and visual injury assessments showed the same pattern: damage began with freezing injury to roots and subsequently became evident as foliar browning after spring temperatures increased. All seedlings in the exposed treatment eventually had 100% fine root damage and died. This progression of initial root damage followed by foliar browning and mortality after the onset of warming conditions is consistent with reports of yellow‐cedar decline symptom development in the field.     

Optimal sampling and spatial distribution of Ixodes pacificus, Dermacentor occidentalis and Dermacentor variabilis ticks (Acari: Ixodidae)

A common method for sampling tick populations is flagging, which is a method of dragging a white cloth over a plant substrate for a fixed distance along a transect. Flagging over rough physical surfaces or using long subtransect lengths could lead to the underestimation of tick densities. Using estimates of the drop-off rates of adult Ixodes pacificus in flag sampling, optimal sampling schemes (the length and number of subtransects) were examined using the relationships between the tick drop-off rates (c), the tick density and distribution patterns and the roughness of the sampled plant substrate. It was found that the optimal number of subtransects and Lmax, the longest subtransect length which did not significantly underestimate the tick density from c, were affected by the tick density, substrate roughness and tick distribution pattern. This study also showed that the density and distribution of I. pacificus and Dermacentor occidentalis varied greatly over time in the populations sampled, while the Dermacentor variabilis densities were low and showed no significant changes over time. Both I. pacificus and D. occidentalis had clumped distributions along trails and these clumps were aggregated. However, the clump sizes (or individual clump areas) changed significantly over time because of density fluctuation or the movement of ticks. Finally, a positive association between the number of I. pacificus and D. occidentalis adults was observed from 2 m subtransect flag collections in March 1995; no relationship was found in 1994. © Rapid Science Ltd. 1998     

Role of the male claw sensilla in perception of female mounting sex pheromone in Dermacentor variabilis,Dermacentor andersoni and Amblyomma americanum

The foreleg claw sensilla of male D. variabilis, D. andersoni and A. americanum ticks indlude the receptors that perceive the female contact mounting sex pheromone (MSP). In all three tick species, the foreleg claw sensilla comprise six anteriorly-directed setae arranged in three symmetrical pairs, two each on the opposite sides of the apotele of the claw and one on the ventral side. Morphological study and behavioral bioassays of these setae revealed that only the dorsal and middle (=lateral) pairs of claw sensilla are mechanogustatory. While the ventral pair are strictly mechanoreceptors. The dorsal and middle sensory setae exhibit a single pore-like structure located at or near their tip, a feature characteristic of mechanogustatory sensilla. These setae are similar to those found on the palps that are believed to function as pheromone receptors. In all three tick species, male mounting and postmounting behaviors were suppressed only when the dorsal and middle pairs of claw sensilla were ablated or covered with gelatin; normal behavior was restored when the gelatin was removed. Doscresponse bioassays were conducted with D. variabilis males to authenticate the results of the gelatin tests. The results of these bioassays demonstrated that the gelatin coat was impervious to the pheromone. The characteristics of the ixodid tick mating system that distinguish it from mating processes in other arthropods are discussed.     

Concurrent effects of cold and hyperkalaemia cause insect chilling injury

Chilling injury and death are the ultimate consequence of low temperature exposure for chill susceptible insects, and low temperature tolerance is considered one of the most important factors determining insect distribution patterns. The physiological mechanisms that cause chilling injury are unknown, but chronic cold exposure that causes injury is consistently associated with elevated extracellular [K⁺], and cold tolerant insects possess a greater capacity to maintain ion balance at low temperatures. Here, we use the muscle tissue of the migratory locust (Locusta migratoria) to examine whether chill injury occurs during cold exposure or following return to benign temperature and we specifically examine if elevated extracellular [K⁺], low temperature, or a combination thereof causes cell death. We find that in vivo chill injury occurs during the cold exposure (when extracellular [K⁺] is high) and that there is limited capacity for repair immediately following the cold stress. Further, we demonstrate that that high extracellular [K⁺] causes cell death in situ, but only when experienced at low temperatures. These findings strongly suggest that that the ability to maintain ion (particularly K⁺) balance is critical to insect low temperature survival, and highlight novel routes of study in the mechanisms regulating cell death in insects in the cold.     

Kinetics of ingested host immunoglobulin G in hemolymph and whole body homogenates during nymphal development of Dermacentor variabilis and Ixodes scapularis ticks (Acari: Ixodidae)

Patterns in the utilization of host immunoglobulin G (IgG) during nymphal development differed between Dermacentor varibilis (Say) and Ixodes scapularis Say ticks. In unfed nymphs of D. variabilis, host IgG was readily detectable in both hemolymph and whole body homogenates. In unfed nymphs of I. scapularis, host IgG was absent in hemolymph and at very low concentrations in whole body homogenates. Host IgG in unfed nymphs was undoubtedly the remnants of IgG acquired during the larval bloodmeal that persisted through metamorphosis to the nymphal stage. In both tick species, host IgG crossed the midgut into the hemocoel during the latter phases of engorgement. Concentrations of host IgG in I. scapularis declined considerably after replete nymphs molted to the adult stage. In contrast, concentrations of host IgG in D. variabilis remained elevated throughout metamorphosis to the adult stage. When larval D. variabilis were fed on a rat, then 2 months later as nymphs on a rabbit, the rat IgG (“old IgG”) present in unfed nymphs was totally replaced by rabbit IgG (“new IgG”) within 2 d of nymphs attaching to the rabbit. Presumably, the old IgG acquired from a previous bloodmeal was secreted via saliva into the new host. This revised version was published online in July 2006 with corrections to the Cover Date.     

Accumulation of glycinebetaine during cold acclimation and freezing tolerance in leaves of winter and spring barley plants

A study was performed to examine whether or not betaine (glycinebetaine), a compatible solute, is accumulated in response to cold stress and is involved in mechanisms that protect plants from freezing injury. For this purpose, we used near-isogenic lines of barley, with each line differing only in a single gene for the spring type of growth habit; the various lines were produced by back-crosses to a recurrent cultivar of the winter type. The winter type of growth habit requires a low temperature for triggering of flower development (vernalization), whereas the spring type does not. Betaine was accumulated to five times the basal level over the course of 3 weeks at low temperature (5 °C) in the winter-type cultivar and in a spring-sh line having the sh gene for the spring-type growth habit, but the level was only doubled in the spring-Sh3 line, which carried the Sh₃ gene for the spring-type growth habit. Among near-isogenic lines of the same cultivar, the levels of betaine accumulated in leaves at low temperature were well correlated with the percentages (on a dry weight basis) of green leaves that survived freezing injury (-5 °C). This observation indicates the possibility, separate from the recognized role of betaine in the response to salinity and/or drought, that betaine accumulates in response to cold stress and that the accumulation of betaine during cold acclimation is associated to some extent with freezing tolerance in leaves of barley plants.     

Environmental regulation and physiological basis of freezing tolerance in woody plants

Cold acclimation of plants is a complex process involving a number of biochemical and physiological changes. The ability to cold acclimate is under genetic control. The development of freezing tolerance in woody plants is generally triggered by non-freezing low temperatures but can also be induced by mild drought or exogenous abscisic acid, as well as by short photoperiod. In nature, the extreme freezing tolerance of woody plants is achieved during sequential stages of cold acclimation the first of which is initiated by short photoperiods and non-freezing low temperatures, and the second by freezing temperatures. Although recent breakthroughs have increased our knowledge on the physiological molecular basis of freezing tolerance in herbaceous species, which acclimate primarily in response to non-freezing low temperatures, very little is known about cold acclimation of woody plants. This article attempts to review our current understanding of the physiological aspects that underline cold acclimation in woody plants.     

Acquisition of freezing tolerance in early autumn and seasonal changes in gall water content influence inoculative freezing of gall fly larvae,Eurosta solidaginis (Diptera,Tephritidae)

We examined seasonal changes in freeze tolerance and the susceptibility of larvae of the gall fly, Eurosta solidaginis to inoculative freezing within the goldenrod gall (Solidago sp.). In late September, when the water content of the galls was high (approximately 55%), more than half of the larvae froze within their galls when held at -2.5 degrees C for 24 h, and nearly all larvae froze at -4 or -6 degrees C. At this time, most larvae survived freezing at > or = -4 degrees C. By October plants had senesced, and their water content had decreased to 33%. Correspondingly, the number of larvae that froze by inoculation at -4 and -6 degrees C also decreased, however the proportion of larvae that survived freezing increased markedly. Gall water content reached its lowest value (10%) in November, when few larvae froze during exposure to subzero temperatures > or = -6 degrees C. In winter, rain and melting snow transiently increased gall water content to values as high as 64% causing many larvae to freeze when exposed to temperatures as high as -4 degrees C. However, in the absence of precipitation, gall tissues dried and, as before, larvae were not likely to freeze by inoculation. Consequently, in nature larvae freeze earlier in the autumn and/or at higher temperatures than would be predicted based on the temperature of crystallization (T(c)) of isolated larvae. However, even in early September when environmental temperatures are relatively high, larvae exhibited limited levels of freezing tolerance sufficient to protect them if they did freeze.     

Nymphal survival and habitat distribution of Ixodes scapularis and Amblyomma americanum ticks (Acari: Ixodidae) on Fire Island, New York, USA

The distribution and survival of Ixodes scapularis and Amblyomma americanum were studied in deciduous and coniferous wooded habitats and in open habitats on Fire Island, New York, USA. The survival of nymphal I. scapularis in field enclosures was greater in forests than in open habitats, suggesting that greater survival contributes to the higher tick population in the woods. The nymphs of each species were more common in deciduous thickets (predominantly Aronia arbutifolia and Vaccinium corynbosum) than in coniferous woods (mostly Pinus rigida) in most but not all years. Larval I. scapularis were more common in coniferous sites in 1994, while the same ticks, as nymphs, were more common in deciduous sites in 1995. The survival of the nymphs was not consistently greater in either the deciduous or coniferous woods. Therefore, factors other than nymphal survival (e.g. larval overwintering survival and tick movement on hosts) probably influenced the relative nymph abundance in different forest types. Overall, the survival of A. americanum was far higher than that of I. scapularis.     

Magnetic resonance imaging (MRI) of water during cold acclimation and freezing in winter wheat

Nuclear magnetic resonance (NMR) spectroscopy and magnetic resonance imaging (MRI) were used to analyse changes in the physical state of water in wheat crowns during cold acclimation and during the freezing/thawing cycle. Spectroscopically measured average spin-spin relaxation times (T₂) decreased during cold acclimation and increased when plants were grown at normal temperature. Spin-spin relaxation images whose contrast is proportional to T₂, times were calculated allowing association of water relaxation with regions of tissue in spin-echo images during acclimation and freezing. Images taken during freezing revealed nonuniform freezing of tissue in crowns and roots. Acclimated and non-acclimated wheat crowns were imaged during freezing and after thawing. Spin-echo image signal intensity and T₂ times decreased dramatically between -4°C and -8°C as a result of a decrease in water mobility during freezing. Images collected during thawing were diffuse with less structure and relaxation times were longer, consistent with water redistribution in tissue after membrane damage.     

Redox changes during cold acclimation affect freezing tolerance but not the vegetative/reproductive transition of the shoot apex in wheat

Cold acclimation is necessary for winter wheat (Triticum aestivum L.) to achieve its genetically determined maximum freezing tolerance, and cold also fulfils the vernalisation requirement. Chromosome 5A is a major regulator of these traits. The aim of the present study was to discover whether changes in the half‐cell redox potential of the glutathione/glutathione disulphide (GSH/GSSG) and ascorbate/dehydroascorbate (AA/DHA) couples induced by cold acclimation are related to freezing tolerance and vernalisation requirement in a specific genetic system including chromosome 5A substitution lines. The amounts of H₂O₂ and AA, and the AA/DHA ratio showed a rapid and transient increase in the crown of all genotypes during the first week of acclimation, followed by a gradual increase during the subsequent 2 weeks. The amount of GSH and its ratio compared to GSSG quickly decreased during the first day, while later these parameters showed a continuous slow increase. The H₂O₂, AA and GSH concentrations, AA/DHA and GSH/GSSG ratios and the half‐cell reduction potential of the GSH/GSSG couple were correlated with the level of freezing tolerance after 22 days at 2 °C; hence these parameters may have an important role in the acclimation process. In contrast to H₂O₂ and the non‐enzymatic antioxidants, the lipid peroxide concentration and activity of the four antioxidant enzymes exhibited a transient increase during the first week, with no significant difference between genotypes. None of the parameters studied showed any relationship with the vegetative/generative transition state monitored as apex morphology and vernalisation gene expression.     

The effect of female body size on male mounting behaviour in Dermacentor variabilis and D. andersoni

Abstract. This study was undertaken to determine if body size of female ticks was an important factor in eliciting the mating behaviour of Dermacentor variabilis and D. andersoni males. Dummy female ticks (DFTs) representing varying sizes of feeding females were prepared from plastic beads. Size of DFT was shown to be important in determining both the time spent by males in contact with DFTs and the extent of male mating response released. Dermacentor varlabilis males preferred the smaller-sized DFT, whereas D. andersoni males preferred the larger sizes. Males of both species were able to discriminate between size of DFT only when mounting sex pheromone (MSP) was present.
Size of the DFT was more important than the composition of the MSP extract in determining the time males spent in contact with DFTs.
Males of both species were sensitive to variations in concentration of extract, and spent 2–6 times longer in contact with DFTs when the concentration was optimal than when it was not. No significant difference in the male's mating response was seen in response to variation in concentration of heterospecific extract.
Male mating response, i.e. the behaviour of the male as it progresses through the initial contact, climbing onto the dorsal surface and turning onto the venter, was shown not to be dependent upon how long the male spent in contact with the DFT. Male D. andersoni had a much lower mating response to DFTs than D. variabilis males, suggesting that some further stimulus may be required.     

热激处理对冷藏枇杷果实冷害的生理作用

枇杷(Eriobotrya japonica Lindl.)果实采后经48～52℃、10 min的热激处理,然后2～5℃贮藏,通过对贮藏期间果实冷害级别、呼吸速率、过氧化物酶、过氧化氢酶、苯丙氨酸解氨酶活性和质膜相对透性变化的分析,研究贮前热激处理对冷藏枇杷果实冷害的生理作用.结果表明,2～5℃低温可诱导枇杷果实呼吸速率和苯丙氨酸解氨酶活性异常升高,果实冷害程度与苯丙氨酸解氨酶活性之间呈正相关,相关系数r=0.926.热激处理能降低冷藏条件下(2～5℃)枇杷果实呼吸速率的异常升高,减轻由于低温胁迫造成的果肉细胞膜损伤,提高枇杷果实的过氧化物酶和过氧化氢酶活性,降低苯丙氨酸解氨酶的活性.贮前热激处理有推迟和减轻枇杷果实冷害症状发生、降低果肉低温劣变的作用.贮前热激处理结合低温冷藏是延长枇杷果实贮藏寿命的有效措施之一.     

Evolution of freezing susceptibility and freezing tolerance in terrestrial arthropods

Arthropods have evolved various adaptations to survive adverse seasons and it has long been discussed why some arthropods are freezing-susceptible and some are freezing-tolerant. However, which mode of frost resistance came first during the course of evolution? A commonly held opinion is that no choice of strategy has been offered in evolution, because each species of arthropod may have its own evolutionary and natural history, leading to cold-hardiness. Freezing tolerance is more frequent in holometabolous insect orders and partially used by certain vertebrates, like some terrestrially hibernating amphibians and reptiles. Supported by phylogenetic, ontogenetic and ecological arguments, we suggest here that freezing tolerance is more recent than freezing susceptibility in the course of arthropods evolution. In addition, we observe that three basic modes of freezing resistance in insect species exist in the field: (i) permanent or year-round freezing-susceptible species, (ii) alternative or seasonal freezing-susceptible/freezing-tolerant species, (iii) permanent or year-round freezing tolerant species.     

Effect of thawing time, cooling rate and boron nutrition on freezing point of the primordial shoot in norway spruce buds

BACKGROUND: Effects of cooling rates on bud frost hardiness have been studied but there is little information on bud responses to thawing. Since the cell wall pore size has been found to increase with boron (B) deficiency, B deficiency may affect the supercooling ability of buds in winter. METHODS: The effects of duration of thawing time and rate of cooling on bud frost hardiness of Norway spruce (Picea abies) were studied in a B fertilization trial in February 2003 and March 2005. Frost hardiness of apical buds was determined by differential thermal analysis (DTA) and visual scoring of damage. KEY RESULTS: In 2003, the freezing point of primordial shoots of buds (T(f)), i.e. the low-temperature exotherm (LTE), was, on average, -39 degrees C when buds were thawed for less than 3 h and the T(f) increased to -21 degrees C after 18 h of thawing. During the first 4 h of thawing, the rate of dehardening was 6 degrees C h(-1). In 2005, buds dehardened linearly from -39 degrees C to -35 degrees C at a rate of 0.7 degrees C h(-1). In 2003, different cooling rates of 1-5 degrees C h(-1) had a minor effect on T(f) but in 2005 with slow cooling rates T(f) decreased. In both samplings, at cooling rates of 2 and 1 degrees C h(-1), T(f) was slightly higher in B-fertilized than in non-fertilized trees. By contrast, at very short thawing times in 2003, T(f) was somewhat lower in B-fertilized trees. CONCLUSIONS: There was little evidence of reduced frost hardiness in trees with low B status. This study showed that buds deharden rapidly when exposed to above-freezing temperatures in winter, but if cooled again they reharden more slowly. According to this study, rapid dehardening of buds has to be taken into account in assessments of frost hardiness.     

Heterosis in the freezing tolerance, and sugar and flavonoid contents of crosses between Arabidopsis thaliana accessions of widely varying freezing tolerance

Heterosis is defined as the increased vigour of hybrids in comparison to their parents. We investigated 24 F(1) hybrid lines of Arabidopsis thaliana generated by reciprocally crossing either C24 or Col with six other parental accessions (Can, Co, Cvi, Ler, Rsch, Te) that differ widely in their freezing tolerance. The crosses differed in the degree of heterosis for freezing tolerance, both in the non-acclimated state and after a 14 d cold acclimation period. Crosses with C24 showed more heterosis than crosses with Col, and heterosis was stronger in acclimated than in non-acclimated plants. Leaf content of soluble sugars and proline showed more deviation from mid-parent values in crosses involving C24 than in those involving Col, and deviations were larger in acclimated than in non-acclimated plants. There were significant correlations between the content of different sugars and leaf freezing tolerance, as well as between heterosis effects in freezing tolerance and sugar content. Flavonoid content and composition varied between accessions, and between non-acclimated and acclimated plants. In the crosses, large deviations from the mid-parent values in the contents of different flavonols occurred, and there were strikingly strong correlations between both flavonol content and freezing tolerance, and between heterosis effects in freezing tolerance and flavonol content.     

Effects of salicylic acid and cold on freezing tolerance in winter wheat leaves

The effects of salicylic acid (SA) (0.01, 0.1 and 1 mM) and cold on freezing tolerance (freezing injury and ice nucleation activity) were investigated in winter wheat (Triticum aestivum cv. Dogu-88) grown under control (20/18 °C for 15, 30 and 45-day) and cold (15/10 °C for 15-day, 10/5 °C for 30-day and 5/3 °C for 45-day) conditions. Cold acclimatisation caused a decrease of injury to leaf segments removed from the plants and subjected to freezing conditions. Exogenous SA also decreased freezing injury in the leaves grown under cold (15/10 °C) and control (15 and 30-day) conditions. Cold conditions (10/5 and 5/3 °C) caused an increase in ice nucleation activity by apoplastic proteins, which were isolated from the leaves. For the first time, it was shown that exogenous SA caused an increase in ice nucleation activity under cold (15/10 and 10/5 °C) and control conditions. These results show that salicylic acid can increase freezing tolerance in winter wheat leaves by affecting apoplastic proteins.