1995; Tanaka et al., 1996; Aguillar et al., 1999), including insects (Bennet at al., 1997; Kostal & Simek,. 1998; Ohtsu et al., 1998). Nevertheless, no change was.
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TABLE 4. Parameters of the sigmoid survival curves constructed for non-diapausing adults of Pyrrhocoris apterus exposed to –5°C prior to, and after a gradual acclimation 0°C. Treatment, temperature
Control, 25°C Acclimated t-test, P
n1
L mean ± SE
Lt50-5 mean ± SE
788 537
0.89 ± 0.12 0.58 ± 0.08 < 0.001***
7.8 ± 0.9 14.7 ± 1.8 0.0003***
Proportion of variance explained by sigmoid curve [%] 71 77
n shows the total number of adults in the experiment, degrees of freedom for calculation of the two-tailed t-test were derived from the number of true replications (i.e. groups of 10–30 adults exposed to –5°C for various periods; n = 35, control; n = 23, acclimated). 1
16.3 ± 1.2 mM after gradual acclimation to 0°C; K+: 16.3 ± 1.0 mM before and 11.1 ± 3.4 mM after (means ± SE, n = 10)].
in PCs in both tissues, the change in fat body PC being insignificant, and (d) the proportion of 16:0 FA did not change significantly.
Polyols The mean haemolymph concentrations of the four “winter” polyols (ribitol, sorbitol, arabinitol and mannitol) were relatively low, ranging between 0.15 and 0.42 mM in all samples and no statistically significant changes were observed after a gradual acclimation to 0°C (data not shown).
DISCUSSION
Phospholipids Phosphatidylethanolamines (PEs) together with phosphatidylcholines (PCs) made up more than 80% (molar) of all phospholipids in the muscle and fat body tissues. Seven molecular species dominated in the PEs and PCs from all samples. These were [fatty acid (FA) residues in positions sn1/sn2 shown]: 16:0/18:2n-6 16:0/18:1n-9 18:2n-6/18:2n-6 18:1n-9/18:2n-6 18:0/18:2n-6 18:0/18:1n-9 18:0/18:0 where 18:2n-6 is linoleic acid; 18:1n-9 is oleic acid; 16:0 is palmitic acid; and 18:0 is stearic acid. The proportions of the individual molecular species of phospholipids changed only moderately during a gradual acclimation to 0°C. The patterns of the changes were specific for each lipid class (PEs or PCs) and tissue. Thus, significant increases of PC 18:1n-9/18:2n-6, countered by decreases of PC 16:0/18:2n-6, were found in both muscle and fat body tissues. PE 18:0/18:1n-9 and PE 16:0/18:1n-9 decreased in muscle and fat body tissues. PE 16:0/18:2n-6 increased in muscle and fat body tissues. PE 18:1n-9/18:2n-6 and PE 18:0/18:2n-6 increased in the fat body, and PE 18:0/18:0 decreased in the fat body. Such minor changes (each occurring by a magnitude of a few percent) were reflected in changes in the values of four calculated parameters (Table 6): (a) the relative proportion of PEs increased, while the proportion of PCs decreased, in both tissues (the changes in muscle PEs were statistically insignificant), (b) the unsaturation coefficient UFA/SFA did not change significantly, (c) the ratio of linoleic/oleic acid increased in PEs and decreased 184
This paper shows that non-diapausing adult P. apterus can, to a limited extent, become cold-acclimated. Their chill tolerance increased in response to a gradual, 4-weeklong decrease in ambient temperature from 25 to 0°C. The “limited extent” of cold-acclimation means that, (1) chill tolerance increased in approximately 50% of the population and (2) the level of chill tolerance achieved by non-diapausing adults (Lt50-5 = 14.7 days) was lower than that achieved by comparable diapausing adults. Diapausing P. apterus had Lt50-5 an equal to 28.6 days prior to, and Lt50-5 greater than 50 days, after cold-acclimation using a similar temperature-protocol (KošĢál et al., 2001). Two aspects that play important roles in the coldacclimation of diapausing adults of P. apterus were found to be completely missing in the non-diapausing adults. These were: (a) a down-regulation of ice nucleators resulting in SCP depression, and (b) an accumulation of
Fig. 1. Chill tolerance of non-diapausing adults of Pyrrhocoris apterus measured as their survival after exposure to –5°C for different time periods. Acclimated adults (n = 537, 23 groups) were exposed to a 4-week-long gradual decrease of temperatures from 25 to 0°C prior to the test of chill tolerance. Reproducing 14-day-old adults (n = 788, 35 groups) reared at 25°C served as a control. The experimental data were fitted with sigmoid curves as described in the Materials and Methods. The parameters of the curves are summarized in Table 4.
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