Role Of Maternal Effects Research Articles

Impact of mother genetic and resource environment on her offspring's growth features in Munjal sheep.

The present study evaluated maternal and additive influences that contribute to phenotypic variation in various growth traits in Munjal sheep. The targeted traits that pertained to 2278 records of 706 lambs were birth weight (BWT), weaning weight (WT3), 6-month body weight (WT6), 12-month body weight (WT12), average daily gain (ADG1: 0-3 months; ADG2: 3-6 months, ADG3: 6-12 months of age) and their corresponding Kleiber ratios designated as KR1, KR2 and KR3. The direct heritability estimates for BWT, WT3, WT6, WT12, ADG1, ADG2, ADG3, KR1, KR2 and KR3 under animal models were 0.20 ± 0.08, 0.28 ± 0.08, 0.17 ± 0.07, 0.47 ± 0.09, 0.33 ± 0.08, 0.09 ± 0.06, 0.36 ± 0.10, 0.33 ± 0.08, 0.09 ± 0.06 and 0.32 ± 0.10, respectively. The estimates of maternal genetic effects contributed significantly and were 8% and 7% for BWT and WT3 traits, respectively, which highlighted the considerable role of maternal effects on early growth traits. Genetic and phenotypic correlations ranged from moderate to high between weaning and post-weaning traits. It was concluded that early selection that considered additive as well as maternal effects at weaning age may be delivered to the desired genetic progress in Munjal sheep.

The role of maternal effects on offspring performance in familiar and novel environments.

Maternal effects are an important evolutionary force that may either facilitate adaptation to a new environment or buffer against unfavourable conditions. The degree of variation in traits expressed by siblings from different mothers is often sensitive to environmental conditions. This could generate a Maternal-by-Environment interaction (M × E) that inflates estimates of Genotype-by-Environment effects (G × E). We aimed to test for environment-specific maternal effects (M × E) using a paternal full-sib/half-sib breeding design in the seed beetle Callosobruchus maculatus, where we split and reared offspring from the same mother on two different bean host types-original and novel. Our quantitative genetic analysis indicated that maternal effects were very small on both host types for all the measured life-history traits. There was also little evidence that maternal oviposition preference for a particular host type predicted her offspring's performance on that host. Further, additive genetic variance for most traits was relatively high on both hosts. While there was higher heritability for offspring reared in the novel host, there was no evidence for G × Es, and most cross-host genetic correlations were positive. This suggests that offspring from the same family ranked similarly for performance on both host types. Our results point to a genetic basis of host adaptation in the seed beetle, rather than maternal effects. Even so, we encourage researchers to test for potential M × Es because, due to a lack of testing, it remains unclear how often they arise.

A Maternal Effect on Queen Production in Honeybees

Influences from the mother on offspring phenotype, known as maternal effects, are an important cause of adaptive phenotypic plasticity [1, 2]. Eusocial insects show dramatic phenotypic plasticity with morphologically distinct reproductive (queen) and worker castes [3, 4]. The dominant paradigm for honeybees (Apis mellifera) is that castes are environmentally rather than genetically determined, with the environment and diet of young larvae causing caste differentiation [5-9]. A role for maternal effects has not been considered, but here we show that egg size also influences queen development. Queens laid significantly bigger eggs in the larger queen cells than in the worker cells. Eggs laid in queen cells (QE), laid in worker cells (WE), and 2-day old larvae from worker cells (2L) were transferred to artificial queen cells to be reared as queens in a standardized environment. Newly emerged adult queens from QE were heavier than those from the other two groups and had more ovarioles, indicating a consequence of egg size for adult queen morphology. Gene expression analyses identified several significantly differentially expressed genes between newly emerged queens from QE and those from the other groups. These included a disproportionate number of genes involved in hormonal signaling, body development, and immune pathways, which are key traits differing between queens and workers. That egg size influences emerging queen morphology and physiology and that queens lay larger eggs in queen cells demonstrate both a maternal effect on the expression of the queen phenotype and a more active role for the queen in gyne production than has been realized previously.

Gene expression in the phenotypically plastic Arctic charr (Salvelinus alpinus): A focus on growth and ossification at early stages of development.

Gene expression during development shapes the phenotypes of individuals. Although embryonic gene expression can have lasting effects on developmental trajectories, few studies consider the role of maternal effects, such as egg size, on gene expression. Using qPCR, we characterize relative expression of 14 growth and/or skeletal promoting genes across embryonic development in Arctic charr (Salvelinus alpinus). We test to what extent their relative expression is correlated with egg size and size at early life‐stages within the study population. We predict smaller individuals to have higher expression of growth and skeletal promoting genes, due to less maternal resources (i.e., yolk) and prioritization of energy toward ossification. We found expression levels to vary across developmental stages and only three genes (Mmp9, Star, and Sgk1) correlated with individual size at a given developmental stage. Contrary to our hypothesis, expression of Mmp9 and Star showed a non‐linear relationship with size (at post fertilization and hatching, respectively), whilst Sgk1 was higher in larger embryos at hatching. Interestingly, these genes are also associated with craniofacial divergence of Arctic charr morphs. Our results indicate that early life‐stage variation in gene expression, concomitant to maternal effects, can influence developmental plasticity and potentially the evolution of resource polymorphism in fishes.

Seed Heteromorphism: An Important Adaptation of Halophytes for Habitat Heterogeneity.

Seed germination is a very critical and important step for seedling establishment under saline environments, as high level of salinity in the soil can prevent seed germination. However halophytes exhibit an interesting mechanism to cope with salt stress. Many halophytes produce heteromorphic seeds, which have different dormancy and germination behavior under saline conditions. This characteristic is related to the structural and physiological differences among heteromorphic seeds. It was unclear that how heteromorphic seeds differently accumulate organic and inorganic substances under saline conditions, and what are the physiological and molecular mechanisms involved in the production of heteromorphic seeds, and in the development of transgenerational plasticity in heteromorphic seeds. In the current brief review, dormancy and germination and the possible role of seed coat and storage compounds in this process of heteromorphic seeds development have been discussed. Moreover, the role of maternal effects on heteromorphic seeds production under saline environments and growth and reproduction capability of the descendants from them have been highlighted.

Telomere length and dynamics of spotless starling nestlings depend on nest-building materials used by parents

Nest materials used by animals can have profound effects on developing offspring. They can modify the bacterial and parasitic environment of the nest, and can influence parental investment through sexual signalling processes. In spotless starlings, Sturnus unicolor, green plants and feathers are known nest materials with such functions. The aim of our study was to experimentally assess their influence on nestlings' telomere length and attrition, which are good predictors of their survival prospects. In a full-factorial experiment, we explored these effects in two different populations, together with the potential effects of hatching date, ectoparasitism, bacterial environment and nestling growth. Telomere length and attrition largely depended on population identity and hatching date. After correcting for these effects, the addition of feathers resulted in higher rates of telomere attrition. The addition of plants did not affect nestling telomeres in general, but did in interaction with location: in Hueneja, the experimental addition of green plants resulted in longer telomeres. Feather pigmentation also did not affect telomere length or attrition in general, but did in interaction with location: in Hueneja, the experimental addition of unpigmented feathers resulted in nestlings with longer telomeres and lower attrition rates. Moreover, prevalence of staphylococci on the skin of 8-day-old nestlings was negatively related to telomere lengths of fledglings. Taken together, these results suggest a direct link between nest material composition and nestling telomere length and dynamics. This relationship could be partially mediated by the antimicrobial and/or antiparasitic properties of nest materials or by sexual signalling processes. We discuss possible roles of maternal effects, parasites, immunity and nestling growth in explaining these experimental effects.

Roles of maternal effects in maintaining genetic variation: Maternal storage effect.

Understanding the maintenance of genetic variation remains a central challenge in evolutionary biology. Recent empirical studies suggest the importance of temporally varying selection, as allele frequencies have been found to fluctuate substantially in the wild. However, previous theory suggests that the conditions for the maintenance of genetic variation under temporally fluctuating selection are quite restrictive. Using mathematical models, we demonstrate that maternal genetic effects, whereby maternal genotypes affect offspring phenotypes, can facilitate the maintenance of polymorphism in temporally varying environments. Maternal effects result in mismatches between genotypes and phenotypes, thereby buffering the influence of selection on allele frequency. This decreases the magnitude of allele-frequency fluctuations and creates conditions for the maintenance of variation when selection causes fluctuations. Therefore, maternal effects may result in a temporal storage effect ("maternal storage effect"). On the other hand, when selection does not cause fluctuations (e.g., linear negative frequency-dependent selection), maternal genetic effects moderate the relative importance of selection compared to genetic drift and promote stochastic allele extinction in finite populations. Thus, maternal effects can play an important role in the maintenance of polymorphism, but the direction of the effect depends on the nature of selection.

Coadaptation of offspring begging and parental provisioning: A role for prenatal maternal effects?

Recent studies on birds have shown that offspring begging and parental provisioning covary at the phenotypic level, which is thought to reflect genetic correlations. However, prenatal maternal factors, like yolk testosterone, may also facilitate parent-offspring coadaptation via their effects on offspring begging and development. In fact, maternal effects are thought to adjust offspring phenotype to the environmental conditions they will experience after birth, which are in turn strongly dependent on the levels of parental provisioning. Using cross-fostering experiments in canaries, we tested the role of maternal effects on parent-offspring coadaptation from two different approaches. First, we analyzed whether females deposit yolk testosterone in relation to their own or their partner's prospective parental provisioning, measured as the rate of parental feeding to foster nestlings. Second, we investigated whether females deposit yolk testosterone in relation to costs they incurred when raising a previous brood, as this likely impinges on their capacity to provide parental care in the near future. However, from the results of both experiments we have no evidence that canary females deposit yolk testosterone in order to match offspring begging to the levels of care they and/or their partners provide. We therefore found no evidence that yolk testosterone facilitates parent-offspring coadaptation. In addition, our results suggest that the functional consequences of yolk testosterone deposition may relate to hatching asynchrony since it primarily varied with egg laying order.

The role of maternal effects in adaptation to different diets

The environmental influences of mothers on offspring traits, or maternal effects, often arise from dietary differences experienced by mothers. However, few studies have explored if and how maternal effects facilitate adaptation to new host plants. To address this, we compared the maternal and direct effects arising from dietary differences in two populations of the large milkweed bug, Oncopeltus fasciatus that live on and feed on the seeds from different hosts. We compared a laboratory population, which has been reared for over 400 generations on sunflower seeds and is now adapted to use these as a host, to the wild population, which is adapted to the ancestral diet of toxic milkweed seeds. We first tested for changes in maternal effects, and then examined offspring performance and survivorship. We found evidence for evolution of the maternal effect facilitating the use of a novel host. However, the strongest effects were population differences and direct dietary effects for all traits. Offspring performance was more strongly influenced by diet than maternal effects. Survivorship depended on population and offspring diet, and their interaction, but was unaffected by maternal diet or other interactions. In the artificially evolved population, diet breadth was increased rather than evolving specialization. Our results suggest changes in maternal effects are likely to be weak compared to direct effects of host plants and other adaptations in adaptation to a novel host.

Melanic coloration differentially predicts transfer of immune factors to eggs with daughters or sons

Maternal allocation shapes offspring phenotype by being tailored to parental phenotype and offspring sex. Because melanogenesis and immunity have partly common genetic control, maternal effects via egg immune factors and parental melanization may covary. Here, we tested whether barn swallows Hirundo rustica allocated antibodies against a novel antigen to their eggs according to embryo sex depending on maternal melanin-based coloration while controlling for paternal tail ornamentation. Egg antibody concentration increased with maternal plumage darkness in broods with relatively more females, whereas it decreased in broods with relatively more males. Thus, darker females allocated more antibodies to their eggs when offspring sex ratio was female biased, whereas paler females allocated more antibodies to the eggs when offspring sex ratio was male biased. These effects were independent of laying order. Our results are compatible with the hypotheses that maternal allocation to sons depends on their reproductive value as predicted by heritable melanin-based coloration and with the hypothesis of differential susceptibility of pheomelanic daughters to parasitism. These effects may depend on pleiotropy of the genes that control melanogenesis and immunity, which may differentially act on investment in self-maintenance and maternal effects. This study is the first study in vertebrates of the association between maternal effects on either sex via egg quality and parental melanization. The links between the genetic control of melanogenesis and of major fitness traits like immunity and the role of maternal effects in reproductive strategies prompt for more studies of the covariation between parental coloration and maternal sex-dependent allocation via egg quality.

Roles of maternal effects and nuclear genetic composition change across the life cycle of crop-wild hybrids.

• Premise of the study: The fitness of an offspring may depend on its nuclear genetic composition (via both parental genotypes) as well as on genetic maternal effects (via only the maternal parent). Understanding the relative importance of these two genetic factors is particularly important for research on crop-wild hybridization, since traits with important genetic maternal effects (e.g., seed size) often differ among crops and their relatives. We hypothesized that the effects of these genetic factors on fitness components would change across the life cycle of hybrids.• Methods: We followed seed, plant size, and reproductive traits in field experiments with wild and four crop-wild hybrids of sunflower (Helianthus annuus), which differed in nuclear genetic composition and maternal parent (wild or F1 hybrid).• Key results: We identified strong genetic maternal effects for early life cycle characteristics, with seeds produced on an F1 mother having premature germination, negligible seed dormancy, and greater seedling size. Increased percentages of crop alleles also increased premature germination and reduced dormancy in seeds produced on a wild mother. For mature plants, nuclear genetic composition dominated: greater percentages of crop alleles reduced height, branching, and fecundity.• Conclusions: Particular backcrosses between hybrids and wilds may differentially facilitate movement of crop alleles into wild populations due to their specific features. For example, backcross seeds produced on wild mothers can persist in the seed bank, illustrating the importance of genetic maternal effects, whereas backcross individuals with either wild or F1 mothers have high fecundity, resulting from their wild-like nuclear genetic composition.

Genotype-by-Environment Interactions for Seedling Establishment Across Native and Degraded-Forest Habitats in a Long-Lived Cycad

Habitat differences might promote adaptive differentiation among populations that can be evidenced by genotype-by-environment interactions (GxE). I examined GxE in seed germination and seedling survival in demes of a rainforest cycad across their native and degraded-forest habitats, and explored the role of maternal effects and resource availability on the observed GxE. A reciprocal-transplant experiment showed a home-site advantage in terms of establishmen of the demes. Germination in a manipulative greenhouse experiment mirrored the patterns in natural environments, with GxE in response to light and water availability. Overall germination was lower in the degraded-forest habitat and under high-light and low-water conditions in the greenhouse. Several analysis suggested that maternal effects related to size on germination are weak, but maternal effects are suggested by better survival of larger seedlings in the degraded-forest habitat. With weak maternal effects, GxE in establishment of individuals suggest some adaptive differentiation across demes in this cycad, which could have implications for population persistence in its habitats.

Maternal effects alter natural selection on phytochromes through seed germination

Summary1. Phytochromes regulate seed germination in response to light and temperature, and different phytochromes contribute to germination under different environmental conditions.2. Using Arabidopsis thaliana mutants with different combinations of non‐functional phytochromes, we tested which phytochromes contribute to germination and other life‐history traits under field conditions and whether that contribution changes with seed‐maturation temperature. We also quantified natural selection on phytochrome variants through their influence on seed germination.3. We found that some phytochromes contributed to germination under field conditions and that the phytochrome that contributed most strongly depended on seed‐maturation temperature. Specifically, when seeds were matured under warm temperature, phyA and phyE null plants had the most strongly reduced germination, with phyA not able to germinate late in the season. In contrast, when seeds were matured under cool temperature, phyB nulls had the most reduced germination, and effects of the phyA mutation were apparent only on a phyB background.4. These effects on germination translated to effects on total lifetime fitness, such that selection on phytochromes that contributed to germination sometimes depended on seed‐maturation conditions.5. Synthesis. Natural selection on phytochromes occurs through their effects on seed germination, and maternal effects alter phytochrome contributions to germination. Therefore, maternal effects can alter natural selection on phytochromes. The results demonstrate a novel role of maternal effects in contributing to variable natural selection on specific genes associated with plant responses to climatic conditions.

Disentangling sources of maternal effects in the heterocarpic species Rumex bucephalophorus

Maternal effects are one of the most important influences on the offspring phenotype and their adaptive significance has been increasingly recognized. In this study we document the role of two sources of maternal effects, position and heterocarpy, on juvenile and adult traits of Rumex bucephalophorus, an annual plant that occurs in heterogeneous and unstable environments. Up to three different diaspore types can be found in populations of this species: buried diaspores (BD) at the plant base, and fixed diaspores (FD) and long dispersible diaspores (LD) both borne along aerial stems. The two sources of maternal effects affected most stages of the life cycle of R. bucephalophorus: seed size, dispersability, seedling size and survival, and adult plant fecundity. Maternal effects due to position affected mainly juvenile traits, while those due to heterocarpy were stronger and more persistent. In general, seedlings from FD showed higher survival and originated plants with higher fecundity but with lower dispersability than those derived from BD and LD. Dispersability was markedly affected by the two maternal effects at two different stages: first, there was a trend to place the slighter and more easily detachable diaspores (LD) at the optimal positions for dispersal, and second maternal effect due to heterocarpy surprisingly affected dispersability of the second generation progeny. Our study shows that maternal effects can play an important role in ecology, dynamic and evolution of populations and it adds to growing evidence about the important role of maternal effects.

Maternal effects alter progeny's response to disturbance and nutrients in two Plantago species

Environmental stress leading to a decrease in growth may be compensated for later in ontogeny by a growth plastic response. Such response could be also transmitted to the next generation, which is called transgenerational plasticity. In this study, two Plantago species were used to test whether compensation for biomass loss after disturbance is driven by maternal effects (ME) due to nutrients and disturbance, i.e. by the form of transgenerational plasticity. Additionally, we tested whether ME could contribute to a different performance of progeny having different disturbance histories. We also tested whether ME are adaptive and whether they differ between related species. The degree of (over)compensation for biomass loss was affected by ME. Maternal effects resulted in different performance of disturbed over undisturbed progeny in relation to nutrient status of the progeny environment along with disturbance and nutrients experienced by mothers. Progeny of P. lanceolata grew more leaf biomass when grown in the same nutrient conditions as experienced by their mothers suggesting that maternal effects might be adaptive. Although overall, there was a consistent role of ME in biomass compensation after disturbance among the two species, there were also some species-specific effects. We conclude that compensation for biomass loss is driven both by maternal effects and by progeny environment. This may lead to the different success of regenerative strategies in environments with contrasting nutrient levels. The different role of ME even between related species may contribute to ecological diversity among species.

Use of a mathematical model to study the role of maternal effects in population dynamics and diapause formation in Daphnia

An individual-based model of a population of Daphnia longispina, a common species of zooplankton of lakes and temporary water-bodies of the temperate zone of the Palearctic is described. The concept of this model is based upon an individual's potential for growth and reproduction and for the change from parthenogenesis to gamogenesis, which was controlled by the life conditions of three consecutive generations of Daphnia. This was achieved by simulating the effects of temperature, food conditions and photoperiod experienced by grandmothers and mothers on an individual's potential for growth and reproduction. The model was used to test hypotheses on the significance of maternal transmission of information in population dynamics of Daphnia. The results supported several important conclusions, e.g. the corroboration of the importance of maternal effects for seasonal adaptations of crustaceans. The possibility of accumulation of maternal effects in successive generations apparently increases the resistance of the population to inter-annual fluctuations of environmental parameters. The model also confirms that maternal effects along with intra-population polymorphism are a mechanism providing stability of biological systems on the species (population) level.

Maternal effects, maternal body size and offspring energetics: A study in the common woodlouse Porcellio laevis

What are the consequences of the natural variation in maternal body mass on offspring energetic performance? How are performance traits related to thermal physiology and energetics phenotypically integrated on offspring? To answer these questions, fifty breeding pairs of the common terrestrial isopod Porcellio laevis were set up in the lab. Physiological performance, thermal tolerance and thermal sensitivity were measured in F1 adults. Maternal effects were estimated as: the direct influence of maternal body mass and the variation associated with mothers. Phenotypic integration was evaluated using path analysis. Our results show that: (1) maternal body size affects positively offspring long-term metabolism, (2) maternal variation was significant in many of the physiological traits and (3) there is an intricate set of relationships among traits and importantly, that offspring exhibited compensational strategies among metabolism, thermal sensitivity and thermal tolerance traits. Even if we cannot clearly state whether these maternal influences were because of the genes or the environment that mothers provided and thus no predictions can be done regarding their evolutionary consequences, it seems clear that the role of maternal effects on physiology can no longer be ignored. In this sense, there's a lot to be gained by incorporating explicit experimental protocols to test for maternal effects.

GEOGRAPHIC VARIATION IN ACID STRESS TOLERANCE OF THE MOOR FROG, RANA ARVALIS. II. ADAPTIVE MATERNAL EFFECTS

Abstract The knowledge about the relative contributions of additive genetic and maternal effects, as well as the proximate determinants of maternal effects variation, on population differentiation remains elusive. Likewise, although embryonic performance is often an important component of fitness, it has been relatively little explored in respect to population differentiation. By conducting reciprocal crosses between an acid and a neutral origin population of moor frogs (Rana arvalis), we investigated the relative importance of additive genetic versus maternal effects in local adaptation to acidity in embryonic traits. Furthermore, by performing removal experiments of gelatinous egg capsules (jelly), we evaluated the possibility that differences in the extraembryonic membranes might explain the interpopulation variation in embryonic acid tolerance found in this and earlier studies. Embryos were raised from fertilization to hatching at three different pH levels (pH 4.0, 4.25, and 7.5) in the laboratory, and acid stress tolerance was measured in terms of embryonic survival, growth and development (i.e., size and age at hatching). The results show that the higher acid tolerance of acid population embryos (in terms of survival) was maternally determined, indicating adaptive maternal effects. The jelly removal experiment revealed that adaptation to acidity in embryonic survival may arise through variation related to structure/composition of the egg capsules. There was no evidence for a genetic basis in acid tolerance in sublethal effects, but additive and nonadditive genetic effects were found in embryonic growth and development, independently of treatment. The results indicate a role for maternal effects in local adaptation to acidity in amphibians, and genetically based differences in early life-histories among the populations.

GEOGRAPHIC VARIATION IN ACID STRESS TOLERANCE OF THE MOOR FROG, RANA ARVALIS. II. ADAPTIVE MATERNAL EFFECTS

The knowledge about the relative contributions of additive genetic and maternal effects, as well as the proximate determinants of maternal effects variation, on population differentiation remains elusive. Likewise, although embryonic performance is often an important component of fitness, it has been relatively little explored in respect to population differentiation. By conducting reciprocal crosses between an acid and a neutral origin population of moor frogs (Rana arvalis), we investigated the relative importance of additive genetic versus maternal effects in local adaptation to acidity in embryonic traits. Furthermore, by performing removal experiments of gelatinous egg capsules (jelly), we evaluated the possibility that differences in the extraembryonic membranes might explain the interpopulation variation in embryonic acid tolerance found in this and earlier studies. Embryos were raised from fertilization to hatching at three different pH levels (pH 4.0, 4.25, and 7.5) in the laboratory, and acid stress tolerance was measured in terms of embryonic survival, growth and development (i.e., size and age at hatching). The results show that the higher acid tolerance of acid population embryos (in terms of survival) was maternally determined, indicating adaptive maternal effects. The jelly removal experiment revealed that adaptation to acidity in embryonic survival may arise through variation related to structure/composition of the egg capsules. There was no evidence for a genetic basis in acid tolerance in sublethal effects, but additive and nonadditive genetic effects were found in embryonic growth and development, independently of treatment. The results indicate a role for maternal effects in local adaptation to acidity in amphibians, and genetically based differences in early life-histories among the populations.

Maternal temperature has different effects on the photoperiodic response and duration of larval diapause in blow fly (Calliphora vicina) strains collected at two latitudes

Abstract.The blow fly Calliphora vicina Robineau‐Desvoidy (Diptera: Calliphoridae) has a wide distribution across northern and temperate Europe. It has a facultative, maternally‐induced larval diapause in response to short days. The photoperiodic response, measured at 15 and 20°C, of two populations was compared. A southern population (originating at 51° N) was sensitive to temperature at all daylengths; the incidence of diapause was greatly reduced at 20°C compared with 15°C. The photoperiodic response of a northern population (from 65° N) was sensitive to temperature only in long days; in short days (< 14 h of light) the response of this strain was identical at each temperature.Variation in parental photoperiod and temperature were found to affect the duration of larval diapause, indicating a role for maternal effects in diapause intensity as well as incidence. However, the between‐strain variation was greater than that within strains, indicating qualitative differences in diapause response. These differences may arise from the ecological conditions at the points of origin of the two strains. The northern strain from the harsher climate has a more intense diapause that follows a relatively temperature‐insensitive photoperiodic response. In contrast, the southern strain has a shallow diapause and its photoperiodic response may be overridden by the experience of concurrent high temperature.