Life cycle of the monarch or wanderer butterfly, Danaus plexippus. (After photographs by P.J. Gullan.) Chapter 6 INSECT DEVELOPMENT AND LIFE HISTORIES TIC06 5/20/04 4:45 PM Page 141 142 Insect development and life histories In this chapter we discuss the pattern of growth from egg to adult – the ontogeny – and life histories of insects. The various growth phases from the egg, through immature development, to the emergence of the adult are dealt with. Also, we consider the significance of different kinds of metamorphosis and suggest that com- plete metamorphosis reduces competition between conspecific juveniles and adults, by providing a clear differentiation between immature and adult stages. Amongst the different aspects of life histories covered are voltinism, resting stages, the coexistence of differ- ent forms within a single species, migration, age deter- mination, allometry, and genetic and environmental effects on development. The influence of environmen- tal factors, namely temperature, photoperiod, humid- ity, toxins, and biotic interactions, upon life-history traits is vital to any applied entomological research. Likewise, knowledge of the process and hormonal regu- lation of molting is fundamental to insect control. Insect life-history characteristics are very diverse, and the variability and range of strategies seen in many higher taxa imply that these traits are highly adaptive. For example, diverse environmental factors trigger ter- mination of egg dormancy in different species of Aedes although the species in this genus are closely related. However, phylogenetic constraint, such as the restrained instar number of Nepoidea (Box 5.5), undoubtedly plays a role in life-history evolution in insects. We conclude the chapter by considering how the potential distributions of insects can be modeled, using data on insect growth and development to answer questions in pest entomology, and bioclimatic data associated with current-day distributions to predict past and future patterns. 6.1 GROWTH Insect growth is discontinuous, at least for the sclerot- ized cuticular parts of the body, because the rigid cuticle limits expansion. Size increase is by molting – periodic formation of new cuticle of greater surface area and shedding of the old cuticle. Thus, for sclerite-bearing body segments and appendages, increases in body dimensions are confined to the postmolt period imme- diately after molting, before the cuticle stiffens and hardens (section 2.1). Hence, the sclerotized head cap- sule of a beetle or moth larva increases in dimensions in a saltatory manner (in major increments) during development, whereas the membranous nature of body cuticle allows the larval body to grow more or less continuously. Studies concerning insect development involve two components of growth. The first, the molt increment, is the increment in size occurring between one instar (growth stage, or the form of the insect between two successive molts) and the next. Generally, increase in size is measured as the increase in a single dimension (length or width) of some sclerotized body part, rather than a weight increment, which may be misleading because of variability in food or water intake. The second component of growth is the intermolt period or interval, better known as the stadium or instar duration, which is defined as the time between two successive molts, or more precisely between successive ecdyses (Fig. 6.1 and section 6.3). The magnitude of both molt increments and intermolt periods may be affected by food supply, temperature, larval density, and physical damage (such as loss of appendages) (section 6.10), and may differ between the sexes of a species. In collembolans, diplurans, and apterygote insects, growth is indeterminate – the animals continue to molt until they die. There is no definitive terminal molt in such animals, but they do not continue to increase in size throughout their adult life. In the vast majority of insects, growth is determinate, as there is a distinctive instar that marks the cessation of growth and molting. All insects with determinate growth become reproduct- ively mature in this final instar, called the adult or imaginal instar. This reproductively mature individual is called an adult or imago (plural: imagines or ima- gos). In most insect orders it is fully winged, although secondary wing loss has occurred independently in the adults of a number of groups, such as lice, fleas, and certain parasitic flies, and in the adult females of all scale insects (Hemiptera: Coccoidea). In just one order of insects, the Ephemeroptera or mayflies, a subimagi- nal instar immediately precedes the final or imaginal instar. This subimago, although capable of flight, only rarely is reproductive; in the few mayfly groups in which the female mates as a subimago she dies without molting to an imago, so that the subimaginal instar actually is the final growth stage. In some pterygote taxa the total number of pre-adult growth stages or instars may vary within a species depending on environmental conditions, such as developmental temperature, diet, and larval density. In many other species, the total number of instars (although not necessarily final adult size) is genetically TIC06 5/20/04 4:45 PM Page 142 determined and constant regardless of environmental conditions. 6.2 LIFE-HISTORY PATTERNS AND PHASES Growth is an important part of an individual’s onto- geny, the developmental history of that organism from egg to adult. Equally significant are the changes, both subtle and dramatic, that take place in body form as insects molt and grow larger. Changes in form (morphology) during ontogeny affect both external structures and internal organs, but only the external changes are apparent at each molt. We recognize three broad patterns of developmental morphological change during ontogeny, based on the degree of external altera- tion that occurs in the postembryonic phases of development. The primitive developmental pattern, ametaboly, is for the hatchling to emerge from the egg in a form essentially resembling a miniature adult, lacking only genitalia. This pattern is retained by the primitively wingless orders, the silverfish (Zygentoma) and bristle- tails (Archaeognatha) (Box 9.3), whose adults con- tinue to molt after sexual maturity. In contrast, all pterygote insects undergo a more or less marked change in form, a metamorphosis, between the immature phase of development and the winged or secondarily wingless (apterous) adult or imaginal phase. These insects can be subdivided according to two broad patterns of development, hemimetaboly (partial or incomplete metamorphosis; Fig. 6.2) and holomet- aboly (complete metamorphosis; Fig. 6.3 and the vignette for this chapter, which shows the life cycle of the monarch butterfly). Developing wings are visible in external sheaths on the dorsal surface of nymphs of hemimetabolous insects except in the youngest immature instars. The term exopterygote has been applied to this type of “external” wing growth. In the past, insect orders with hemimetabolous and exopterygote development were grouped into “Hemimetabola” (also called Exoptery- gota), but this group is recognized now as applying to a grade of organization rather than to a monophyletic phylogenetic unit (Chapter 7). In contrast, pterygote orders displaying holometabolous development share the unique evolutionary innovation of a resting stage or pupal instar in which development of the major structural differences between immature (larval) and adult stages is concentrated. The orders that share this unique, derived pattern of development represent a clade called the Endopterygota or Holometabola. In the early branching Holometabola, expression of all adult features is retarded until the pupal stage; however, in more derived taxa including Drosophila, uniquely adult structures including wings may be pre- sent internally in larvae as groups of undifferentiated Life-history patterns and phases 143 Fig. 6.1 Schematic drawing of the life cycle of a non-biting midge (Diptera: Chironomidae, Chironomus) showing the various events and stages of insect development. TIC06 5/20/04 4:45 PM Page 143 144 Insect development and life histories cells called imaginal discs (or buds) (Fig. 6.4), although they are scarcely visible until the pupal instar. Such wing development is called endopterygote because the wings develop from primordia in invaginated pockets of the integument and are everted only at the larval–pupal molt. The evolution of holometaboly allows the immature and adult stages of an insect to specialize in different Fig. 6.2 The life cycle of a hemimetabolous insect, the southern green stink bug or green vegetable bug, Nezara viridula (Hemiptera: Pentatomidae), showing the eggs, nymphs of the five instars, and the adult bug on a tomato plant. This cosmopolitan and polyphagous bug is an important world pest of food and fiber crops. (After Hely et al. 1982.) TIC06 5/20/04 4:45 PM Page 144 resources, contributing to the successful radiation of the group (see section 8.5). 6.2.1 Embryonic phase The egg stage begins as soon as the female deposits the mature egg. For practical reasons, the age of an egg is estimated from the time of its deposition even though the egg existed before oviposition. The beginning of the egg stage, however, need not mark the commencement of an individual insect’s ontogeny, which actually begins when embryonic development within the egg is triggered by activation. This trigger usually results from fertilization in sexually reproducing insects, but in parthenogenetic species appears to be induced by various events at oviposition, including the entry of oxygen to the egg or mechanical distortion. Following activation of the insect egg cell, the zygote nucleus subdivides by mitotic division to produce many daughter nuclei, giving rise to a syncytium. These nuclei and their surrounding cytoplasm, called cleavage energids, migrate to the egg periphery where the membrane infolds leading to cellularization of the superficial layer to form the one-cell thick blastoderm. This distinctive superficial cleavage during early em- bryogenesis in insects is the result of the large amount of yolk in the egg. The blastoderm usually gives rise to all the cells of the larval body, whereas the central yolky part of the egg provides the nutrition for the developing embryo and will be used up by the time of eclosion, or emergence from the egg. Regional differentiation of the blastoderm leads to the formation of the germ anlage or germ disc (Fig. 6.5a), which is the first sign of the developing embryo, whereas the remainder of the blastoderm becomes a thin membrane, the serosa, or embryonic cover. Next, the germ anlage develops an infolding in a process called gastrulation (Fig. 6.5b) and sinks into the yolk, forming a two-layered embryo containing the amniotic cavity (Fig. 6.5c). After gastrulation, the germ anlage becomes the germ band, which externally is charac- terized by segmental organization (commencing in Fig. 6.5d with the formation of the protocephalon). The germ band essentially forms the ventral regions of the future body, which progressively differentiates with the head, body segments, and appendages becoming increasingly well defined (Fig. 6.5e–g). At this time the embryo undergoes movement called katatrepsis which brings it into its final position in the egg. Later, near the end of embryogenesis (Fig. 6.5h,i), the edges of the germ band grow over the remaining yolk and fuse mid-dorsally to form the lateral and dorsal parts of the insect – a process called dorsal closure. In the well-studied Drosophila, the complete embryo is large, and becomes segmented at the cellularization stage, termed “long germ” (as in all studied Diptera, Coleoptera, and Hymenoptera). In contrast, in “short- germ” insects (phylogenetically earlier branching taxa, including locusts) the embryo derives from only a small region of the blastoderm and the posterior segments are added post-cellularization, during subsequent growth. In the developing long-germ embryo, the syncytial phase is followed by cell membrane intrusion to form the blastoderm phase. Functional specialization of cells and tissues occurs during the latter period of embryonic development, so that by the time of hatching (Fig. 6.5j) the embryo is a tiny proto-insect crammed into an eggshell. In ametabolous and hemimetabolous insects, this stage may be recognized as a pronymph – a special hatching stage (section 8.5). Molecular developmental processes involved in organizing the polarity and differentiation of areas of the body, including segmentation, are reviewed in Box 6.1. 6.2.2 Larval or nymphal phase Hatching from the egg may be by a pronymph, nymph, Life-history patterns and phases 145 Fig. 6.3 Life cycle of a holometabolous insect, a bark beetle, Ips grandicollis, showing the egg, the three larval instars, the pupa, and the adult beetle. (After Johnson & Lyon 1991.) TIC06 5/20/04 4:45 PM Page 145 146 Insect development and life histories or larva: eclosion conventionally marks the beginning of the first stadium, when the young insect is said to be in its first instar (Fig. 6.1). This stage ends at the first ecdysis when the old cuticle is cast to reveal the insect in its second instar. Third and often subsequent instars generally follow. Thus, the development of the immat- ure insect is characterized by repeated molts separated by periods of feeding, with hemimetabolous insects generally undergoing more molts to reach adulthood than holometabolous insects. All immature holometabolous insects are called larvae. Immature terrestrial insects with hemimeta- bolous development such as cockroaches (Blattodea), grasshoppers (Orthoptera), mantids (Mantodea), and bugs (Hemiptera) always are called nymphs. How- ever, immature individuals of aquatic hemimetabolous insects (Odonata, Ephemeroptera, and Plecoptera), although possessing external wing pads at least in later instars, also are frequently, but incorrectly, referred to as larvae (or sometimes naiads). True larvae look very different from the final adult form in every instar, whereas nymphs more closely approach the adult appearance at each successive molt. Larval diets and lifestyles are very different from those of their adults. In Fig. 6.4 Stages in the development of the wings of the cabbage white or cabbage butterfly, Pieris rapae (Lepidoptera: Pieridae). A wing imaginal disc in an (a) first-instar larva, (b) second-instar larva, (c) third-instar larva, and (d) fourth-instar larva; (e) the wing bud as it appears if dissected out of the wing pocket or (f ) cut in cross-section in a fifth-instar larva. ((a–e) After Mercer 1900.) TIC06 5/20/04 4:45 PM Page 146 contrast, nymphs often eat the same food and coexist with the adults of their species. Competition thus is rare between larvae and their adults but is likely to be preval- ent between nymphs and their adults. The great variety of endopterygote larvae can be classified into a few functional rather than phylogen- etic types. Often the same larval type occurs conver- gently in unrelated orders. The three commonest forms are the polypod, oligopod, and apod larvae (Fig. 6.6). Lepidopteran caterpillars (Fig. 6.6a,b) are character- istic polypod larvae with cylindrical bodies with short thoracic legs and abdominal prolegs (pseudopods). Symphytan Hymenoptera (sawflies; Fig. 6.6c) and most Mecoptera also have polypod larvae. Such larvae are rather inactive and are mostly phytophagous. Oligopod larvae (Fig. 6.6d–f ) lack abdominal prolegs but have functional thoracic legs and frequently pro- gnathous mouthparts. Many are active predators but others are slow-moving detritivores living in soil or are phytophages. This larval type occurs in at least some members of most orders of insects but not in the Lepidoptera, Mecoptera, Diptera, Siphonaptera, or Life-history patterns and phases 147 Fig. 6.5 Embryonic development of the scorpionfly Panorpodes paradoxa (Mecoptera: Panorpodidae): (a–c) schematic drawings of egg halves from which yolk has been removed to show position of embryo; (d–j) gross morphology of developing embryos at various ages. Age from oviposition: (a) 32 h; (b) 2 days; (c) 7 days; (d) 12 days; (e) 16 days; (f ) 19 days; (g) 23 days; (h) 25 days; (i) 25–26 days; (j) full grown at 32 days. (After Suzuki 1985.) TIC06 5/20/04 4:45 PM Page 147 148 Insect development and life histories Box 6.1 Molecular insights into insect development The formation of segments in the early embryo of Drosophila is understood better than almost any other complex developmental process. Segmentation is controlled by a hierarchy of proteins known as trans- cription factors, which bind to DNA and act to enhance or repress the production of specific messages. In the absence of a message, the protein for which it codes is not produced; thus ultimately transcription factors act as molecular switches, turning on and off the pro- duction of specific proteins. In addition to controlling genes below them in the hierarchy, many transcription factors also act on other genes at the same level, as well as regulating their own concentrations. Mechanisms and processes observed in Drosophila have much wider relevance, including to vertebrate development, and information obtained from Drosophila has provided the key to cloning many human genes. However, we know Drosophila to be a highly derived fly, and it may not be a suitable model from which to derive gen- eralities about insect development. During oogenesis (section 6.2.1) in Drosophila, the anterior–posterior and dorsal–ventral axes are estab- lished by localization of maternal messenger RNAs (mRNAs) or proteins at specific positions within the egg. For example, the mRNAs from the bicoid (bcd) and nanos genes become localized at anterior and poster- ior ends of the egg, respectively. At oviposition, these messages are translated and proteins are produced that establish concentration gradients by diffusion from each end of the egg. These protein gradients differ- entially activate or inhibit zygotic genes lower in the segmentation hierarchy – as in the upper figure (after Nagy 1998), with zygotic gene hierarchy on the left and representative genes on the right – as a result of their differential thresholds of action. The first class of zygotic genes to be activated is the gap genes, for example Kruppel (Kr), which divide the embryo into broad, slightly overlapping zones from anterior to posterior. The maternal and gap proteins establish a complex of overlapping protein gradients that provide a chemical framework that controls the periodic (altern- ate segmental) expression of the pair-rule genes. For example, the pair-rule protein hairy is expressed in seven stripes along the length of the embryo while it is still in the syncytial stage. The pair-rule proteins, in addition to the proteins produced by genes higher in the hierarchy, then act to regulate the segment polarity genes, which are expressed with segmental periodicity and represent the final step in the determination of segmentation. Because there are many members of the various classes of segmentation genes, each row of cells in the anterior–posterior axis must contain a unique combination and concentration of the transcription factors that inform cells of their position along the anterior–posterior axis. Once the segmentation process is complete each developing segment is given its unique identity by the homeotic genes. Although these genes were first discovered in Drosophila it has since been established that they are very ancient, and a more or less complete TIC06 5/20/04 4:45 PM Page 148 Molecular insights into insect development 149 subset of them is found in all multicellular animals. When this was realized it was agreed that this group of genes would be called the Hox genes, although both terms, homeotic and Hox, are still in use for the same group of genes. In many organisms these genes form a single cluster on one chromosome, although in Droso- phila they are organized into two clusters, an anteriorly expressed Antennapedia complex (Antp-C) and a posteriorly expressed Bithorax complex (Bx-C). The composition of these clusters in Drosophila is as follows (from anterior to posterior): (Antp-C) – labial (lab), proboscidea (pb), Deformed (Dfd), Sex combs reduced (Scr), Antennapedia (Antp); (Bx-C) – Ultrabithorax (Ubx), abdominal-A (abd-A), and Abdominal-B (Abd-B), as illustrated in the lower figure of a Drosophila embryo (after Carroll 1995; Purugganan 1998). The evolutionary conservation of the Hox genes is remarkable for not only are they conserved in their primary structure but they follow the same order on the chromosome, and their temporal order of expression and anterior border of expression along the body correspond to their chromosomal position. In the lower figure the anterior zone of expression of each gene and the zone of strongest expression is shown (for each gene there is a zone of weaker expression posteriorly); as each gene switches on, protein production from the gene anterior to it is repressed. The zone of expression of a particular Hox gene may be morphologically very different in different organisms so it is evident that Hox gene activities demarcate relative positions but not particular morphological structures. A single Hox gene may regulate directly or indirectly many targets; for example, Ultrabithorax regulates some 85–170 genes. These downstream genes may operate at different times and also have multiple effects (pleiotropy); for example, wingless in Drosophila is involved successively in segmentation (embryo), Malpighian tubule formation (larva), and leg and wing development (larva–pupa). Boundaries of transcription factor expression are important locations for the development of distinct morphological structures, such as limbs, tracheae, and salivary glands. Studies of the development of legs and wings have revealed something about the processes involved. Limbs arise at the intersection between expression of wingless, engrailed, and decapentaplegic (dpp), a protein that helps to inform cells of their posi- tion in the dorsal–ventral axis. Under the influence of the unique mosaic of gradients created by these gene products, limb primordial cells are stimulated to express the gene distal-less (Dll) required for proximodistal limb growth. As potential limb primordial cells (anlage) are present on all segments, as are limb-inducing protein gradients, prevention of limb growth on inappropriate segments (i.e. the Drosophila abdomen) must involve repression of Dll expression on such segments. In Lepidoptera, in which larval prolegs typically are found on the third to sixth abdominal segments, homeotic gene expression is fundamentally similar to that of Drosophila. In the early lepidopteran embryo Dll and Antp are expressed in the thorax, as in Drosophila, with abd-A expression dominant in abdominal segments including 3–6, which are prospective for proleg development. Then a dramatic change occurs, with abd-A protein repressed in the abdominal proleg cell anlagen, followed by activation of Dll and up-regulation of Antp expression as the anlagen enlarge. Two genes of the Bithorax complex (Bx-C), Ubx and abd-A, repress Dll expression (and hence prevent limb formation) in the abdomen of Drosophila. Therefore, expression of prolegs in the caterpillar abdomen results from repres- sion of Bx-C proteins thus derepressing Dll and Antp and thereby permitting their expression in selected target cells with the result that prolegs develop. A somewhat similar condition exists with respect to wings, in that the default condition is presence on all thoracic and abdominal segments with Hox gene repres- sion reducing the number from this default condition. In the prothorax, the homeotic gene Scr has been shown to repress wing development. Other effects of Scr expression in the posterior head, labial segment, and prothorax appear homologous across many insects, including ventral migration and fusion of the labial lobes, specification of labial palps, and development of sex combs on male prothoracic legs. Experimental mutational damage to Scr expression leads, amongst other deformities, to appearance of wing primordia from a group of cells located just dorsal to the prothoracic leg base. These mutant prothoracic wing anlagen are situated very close to the site predicted by Kukalová-Peck from paleontological evidence (section 8.4, Fig. 8.4b). Furthermore, the apparent default condition (lack of repression of wing expression) would produce an insect resembling the hypothesized “proto- pterygote”, with winglets present on all segments. Regarding the variations in wing expression seen in the pterygotes, Ubx activity differs in Drosophila between the meso- and metathoracic imaginal discs; the anterior produces a wing, the posterior a haltere. Ubx is unexpressed in the wing (mesothoracic) imaginal disc but is strongly expressed in the metathoracic disc, where its activity suppresses wing and enhances haltere formation. However, in some studied non- dipterans Ubx is expressed as in Drosophila – not in the fore-wing but strongly in the hind-wing imaginal disc – despite the elaboration of a complete hind wing as in butterflies or beetles. Thus, very different wing morphologies seem to result from variation in “down- stream” response to wing-pattern genes regulated by Ubx rather than from homeotic control. TIC06 5/20/04 4:45 PM Page 149 150 Insect development and life histories Strepsiptera. Apod larvae (Fig. 6.6g–i) lack true legs and are usually worm-like or maggot-like, living in soil, mud, dung, decaying plant or animal matter, or within the bodies of other organisms as parasitoids (Chapter 13). The Siphonaptera, aculeate Hymenoptera, nema- toceran Diptera, and many Coleoptera typically have apod larvae with a well-developed head, whereas in the maggots of higher Diptera the mouth hooks may be the only obvious evidence of the cephalic region. The grub-like apod larvae of some parasitic and gall- inducing wasps and flies are greatly reduced in external structure and are difficult to identify to order level even by a specialist entomologist. Furthermore, the early- instar larvae of some parasitic wasps resemble a naked embryo but change into typical apod larvae in later instars. A major change in form during the larval phase, such as different larval types in different instars, is called larval heteromorphosis (or hypermetamor- phosis). In the Strepsiptera and certain beetles this involves an active first-instar larva, or triungulin, fol- lowed by several grub-like, inactive, sometimes legless, later-instar larvae. This developmental phenomenon occurs most commonly in parasitic insects in which a Clearly, much is yet to be learnt concerning the multiplicity of morphological outcomes from the interaction between Hox genes and their downstream interactions with a wide range of genes. It is tempting to relate major variation in Hox pathways with morpholo- gical disparities associated with high-level taxonomic rank (e.g. animal classes), more subtle changes in Hox regulation with intermediate taxonomic levels (e.g. orders/suborders), and changes in downstream regulatory/functional genes perhaps with suborder/ family rank. Notwithstanding some progress in the case of the Strepsiptera (q.v.), such simplistic relationships between a few well-understood major developmental features and taxonomic radiations may not lead to great insight into insect macroevolution in the immediate future. Estimated phylogenies from other sources of data will be necessary to help interpret the evolutionary significance of homeotic changes for some time to come. Fig. 6.6 Examples of larval types. Polypod larvae: (a) Lepidoptera: Sphingidae; (b) Lepidoptera: Geometridae; (c) Hymenoptera: Diprionidae. Oligopod larvae: (d) Neuroptera: Osmylidae; (e) Coleoptera: Carabidae; (f ) Coleoptera: Scarabaeidae. Apod larvae: (g) Coleoptera: Scolytidae; (h) Diptera: Calliphoridae; (i) Hymenoptera: Vespidae. ((a,e–g) After Chu 1949; (b,c) after Borror et al. 1989; (h) after Ferrar 1987; (i) after CSIRO 1970.) TIC06 5/20/04 4:45 PM Page 150 [...]... allometric, i.e the parts grow at rates peculiar to themselves, and often very different from the growth rate of the body as a whole The horned adornments on the head and thorax of Onthophagus dung beetles discussed in section 5.3 exemplify the trade-offs associated with allometric growth TIC 06 5/20/04 4:45 PM Page 166 166 Insect development and life histories 6. 9.2 Age-grading of adult insects The age of an... aegypti in the equation given above, the value of K can be calculated for each of the experimental temperatures from 14 to 36 C: 24 2921 Thus, the K-value for Ae aegypti is approximately independent of temperature, except at extremes (14 and 28 2 866 30 2755 32 2 861 34 3415 36 3882 34– 36 C), and averages about 2740 hour-degrees or 114 day-degrees between 16 and 32°C TIC 06 5/20/04 4:45 PM Page 169 Environmental... aestivation In some species the life-history stage in which photoperiod is assessed is in advance of the stage that reacts, as is the case when the photoperiodic response of the maternal generation of silkworms affects the eggs of the next generation The ability of insects to recognize seasonal photoperiod and other environmental cues requires some means of measuring time between the cue and the subsequent onset... initiates the next molt, the epidermal cells produce pupal cuticle as a result of the activation of many new genes The decline in ecdysteroid level towards the end of each molt seems to be essential for, and may be the physiological trigger causing, ecdysis to occur It renders the tissues sensitive to EH and permits the release of EH into the hemolymph (see section 6. 2.4 for further discussion of the actions... alone The options available for avoidance of the extremes are behavioral avoidance, such as by burrowing into soil of a more equable temperature, migration (section 6. 7), diapause (section 6. 5), and in situ tolerance/ survival in a very altered physiological condition, the topic of the following sections 6. 6.1 Cold Biologists have long been interested in the occurrence of insects at the extremes of the. .. occurs after each molt (the molt increment) Thus, it should be possible to determine the actual number of instars in the life history of a species from a frequency histogram of measurements of a sclerotized body part (Fig 6. 11) Entomologists have sought to quantify this size progression for a range of insects One of the earliest TIC 06 5/20/04 4:45 PM Page 165 Age-grading 165 Fig 6. 11 Growth and development... independence from the influence of the brain, especially during the pupal instar In most insects, a reduction in the amount of circulating juvenile hormone (as a result of reduction of corpora allata activity) is essential to the initiation of metamorphosis (The physiological events are described in section 6. 3.) The molt into the pupal instar is called pupation, or the larval–pupal molt Many insects survive... division of the epidermal cells leading to increases in the volume and surface area of the epidermis The subcuticular or apolysial space formed after apolysis becomes filled with the secreted but inactive molting fluid The chitinolytic and proteolytic enzymes of the molting fluid are not activated until the epidermal cells have laid down the protective outer layer of a new cuticle Then the inner part of the. .. between the ecdysteroid and JH titers and the cuticular changes that occur in the last two larval instars and in prepupal development During the molt at the end of the fourth larval instar, the epidermis responds to the surge of ecdysteroid by halting synthesis of endocuticle and the blue pigment insecticyanin A new epicuticle is synthesized, much of the old cuticle is digested, and resumption of endocuticle... Fortunately, the limiting effects of climate on a species usually can be estimated reliably from observations on the geographical distribution The climatic tolerances of the species are inferred from the climate of the sites where the species is known to occur and are described by the stress indices of the CLIMEX model The values of the stress indices are progressively adjusted until the CLIMEX predictions . differentiation of the blastoderm leads to the formation of the germ anlage or germ disc (Fig. 6. 5a), which is the first sign of the developing embryo, whereas the remainder of the blastoderm. hemolymph (see section 6. 2.4 for further discus- sion of the actions of eclosion hormone). Apolysis at the end of the fifth larval instar marks the beginning of a prepupal period when the developing. considerable independence from the influence of the brain, espe- cially during the pupal instar. In most insects, a reduc- tion in the amount of circulating juvenile hormone (as a result of reduction of corpora