A review of JH mimics and their action on insects and other organisms Johari Jalinas School of Environmental and Natural

Resource Sciences, Faculty of Science and Technology, Universiti Kebangsaan Malaysia e-mail: johari_j@ukm.my

Juvenile Hormone Juvenile hormone (JH) is a sesquiterpenoid hormone that regulates almost aspects of an insects life such as growth, development, stimulating aspects of reproduction(Wyatt 1997; Riddiford 2008), and repressing metamorphosis in insects (Wyatt 1997; Madhavan 1972). JH was first discovered by Wigglesworth (1934) in Rhodnius prolixus as a key factor that could prevent metamorphosis of final instar nymph into the adult. Insect JH is first produced in the late embryo and been synthesized in the corpora allata (figure 1.0) (Minakuchi and Riddiford 2006; Kamita et al. 2003; Riddiford 2002).

Figure 1.0 shows the juvenile hormone (JH) system of insect larvae. The diagrams shows the location of corpora which responsible in production of JH in Manduca (left) and Drosophila (right) (Riddiford 2002) JH is considered to have status quo (Kremen and Nijhout 1989; Riddiford 2008; Minakuchi and Riddiford 2006; Aribi et al. 2006) action in insect metamorphosis by controlling ecdysone-induced molts (Madhavan 1972; Minakuchi and Riddiford 2006). JH acting by itself can suppress disc morphogenesis and growth (Palli 2009). JH is very essential for formation of the larval cuticle, diffentiation of the midgut, and normal dorsal closure. The copora allata continue to produce the hormone throughout the larval life until the final instar. JH is also a key regulator of reproductive maturation especially in egg maturation (Wyatt 1997; Riddiford 2008). The hormone could control the gene expression at the translational level (Madhavan 1972; Minakuchi and Riddiford 2006). Intermolt juvenile hormone influences maintenance of larval-specific organs and behavior, and the production of the prothoracicotropic hormone (Krishnakumaran and Schneiderman 1965). JH and Ecdysteroid along with other hormones regulate vitellogenesis, oogenesis, fertilization and choriogenesis during reproductive maturation of insects (Palli 2009). JH plays a key role in the regulation of vitellogenin genes. JH regulates other transformative processes such as diapauses (Bean, Goodman, and Beck 1983), migratory behavior (Riddiford 2008), wing length, and color polymorphism and caste determination in social insect (Cornette, Koshikawa, and Miura 2008). The important of the Juvenile Hormone in insects had lead to either in the finding of compounds that mimic to JH or in synthesizing the compounds that can inhibit the action of the JH. The implication of using hazardous chemical insecticides to control insect pests have

promoted researchers to search for new approach to control insect pest (Palli 2009). The development of synthetic or analog JH and identification of the natural JHs are the best strategy that had been explored by the scientists in developing new insecticides which less harmful to environment. Mimic Juvenile Hormone Juvenile hormone (JH) analogs or mimic juvenile hormones are substances of natural and synthethic origin which act in the same way as endogenous JH. The mimic JH interrupts the molting and metamorphosis reproduction (Kremen and Nijhout 1989; Partasarathy, Tan, and Palli 2008), both larval and adult diapauses (Madhavan 1972; Bean, Goodman, and Beck 1983), and caste determination in social insects (Wyatt 1997; Cornette, Koshikawa, and Miura 2008). The mimic (JH) is an insect growth regulator that has been developed comprehensively recently. Some JH mimics have been used as insecticides and might represent a new generation of pesticides, with selective action on different insect pest species. These compounds are toxic during the last larval, embroyonic and reproductive stages of insects (Aribi et al. 2006). JH analogs (JHA) analogs might make a contribution to insect pest management when the insect pest showing genetic resistance to the chemical synthetic pesticides (Wyatt 1997). Recent studies of the enzymes in JH biosynthetis and metabolic pathways should be helpful for the discovery of more potent analogs and also for the establishment of new means of pest management using recombinant DNA technology (Riddiford 2002). Numerous chemical compounds with juvenilizing activity, also called juvenoids or insect growth regulators, have been synthesized during the past few years for their pest control potential. The examples of the Juvenile hormone-mimicking substances are farnesol, farnesyl methyl ether, farnesyl

diethylamine, nerolidol, and dodecyl methyl ether (Krishnakumaran and Schneiderman 1965; Palli 2009). Many JH mimics had been extracted and isolated from the plants such as juvabione and juvocimenes (Minakuchi and Riddiford 2006). Juvabione is a JH mimic that had been extracted from American gymnosperm (balsam fir) and juvocimenes was isolated from the sweet basil Ocimum basilicum (Minakuchi and Riddiford 2006). Acetone extract of the wood of Cedrus deodara exhibited JH activity towards Dysdercus koenigii . The present of juvabione in very small amounts could penetrate insectss cuticles and prevent normal development and reproduction (Wyatt 1997). Juvocimenes could cause the disruption for the endocrine system of insects. JH homolog also had been isolated from Crustacea; such as Methyl farnesoate that effect the reproductive maturation of insects (Wyatt 1997). Recently insect growth regulators (IGR) that worked as mimicking JH and molting hormone, 20-hydroyecdysone had been synthesized (Oberlander and Silhacek 1998). These compounds disrupt the normal development of insects by interfering with chitin synthesis (Wyatt 1997). Several JH analogs have been synthesized and are commercially available as IGR such as methoprene, fenoxycarb, hydropene, kinoprene, JH III, pyriproxyfen, and diafenolan (figure 2.0) (Palli 2009). These JH analogs are stable and in the environment and resistant to metabolism of insect and might be effective in controlling insects pest. However, there are reports of development of resistance JH analogs (Palli 2009). Although, the mechanisms of insect resistance to JH analogs are not well understood, the molecular mode of action of JH and JHA would be the basis for future study in the developing of new pesticides (Palli 2009).

Figure 2.0 shows the chemical structures of JH III and JH analogs, pyriproxyfen, fenoxycarb, methoprene, hydropene and kinoprene (Palli 2009). JH mimics and their action on insects JH mimics are known to disrupt embryonic, molting (Barret 1974), larval insect development (Palli 2009) and induce sterility in adults. The application of natural JH or JH mimics at appropriate times and doses can result in disruption of normal development (Barret 1974). The definite of JH mimics function is for insects pest control. However, the results depend largely on type of analog, the dose and time of application and also the optimal sensitive period of the insects (Barret 1974).

The JH mimics stimulated the prothoracic glands and consequently disrupted the larval stages of insect (Krishnakumaran and Schneiderman 1965; Aribi et al. 2006). The JH analogue had caused effect in caste determination of social insects such as termites. These hormones induced soldier differentiation (Cornette, Koshikawa, and Miura 2008) by changing the size of corpora allata resulting hypertrophy of corpora allata (Cornette, Koshikawa, and Miura 2008). JH mimics such as t,t-farnesol, dichlorofarnesenic acid ethyl ester (DFAEE) and a synthetic racemic or isomeric mixture C18 JH have caused morphogenetic effects on adult development of Drosophilia (Madhavan 1972), Rhodnius (Barret 1974) and Lepidoptera (Krishnakumaran and Schneiderman 1965) . The JH mimics also caused derangement of adult stage of Drosophilia such as retardation in the synthesis of brown eye pigments, general pigmentation, and development of internal organ and cuticular structures of the abdomen (Madhavan 1972). JH analogs such as hyropene, kinoprene pyripoxyfen and methoprene have been used to control several insects pest such as whiteflies (Aribi et al. 2006) and mosquitoes (Palli 2009). Methoprene (Figure 1.0) is successful been used for controlling fleas(Wyatt 1997) and the application of precocene in low dose could temporary inhibit the protein synthesis (Wyatt 1997). Methoprene blocks and disrupts the developing of mosquitoes pupae and resulting larvae die during the pupal stage (Palli 2009). Methoprene had also caused the delayed onset of the diapauses by inducing a moult in the European Corn borer, Ostrinia nubilalis (Bean, Goodman, and Beck 1983). JH mimics such as methoprene is not only affecting the insects pest but also affecting the natural enemies of the insect such as parasitoid insects (Beckage and Riddiford 1982). The

topical application of methoprene to parasitized Manduca sexta larvae inhibited the emergence of the endoparasitic wasp, Apanteles congregatus (Beckage and Riddiford 1982). Meanwhile, hydropene blocked larval-pupal metamorphosis of T. casteaneum and induced supernumerary larval molts (Palli 2009). JH pyriproxyfen is an insect growth regulator that has been developed comparatively recently. Pyriproxyfen showed larvicidal effects on Culex pipiens (El-Shazly and Refaie 2002) and caused significant inhibition of ecdysteroid amounts resulting significant inhibition of thickness of new cuticle secretion on development of the mealworm, Tenebrio molitor (Aribi et al. 2006). Electron micrographs showed the destruction of procuticle lamellae, formation of cuticular vacuoles, deformed mitochondria, and destruction of nuclear envelopes and the epidermal layer when pyriproxyfen was applied to Culex pipiens (El-Shazly and Refaie

2002).Then the treatment with pyriproxyfen on Tenebrio molitor caused a blockage of the pupaladult development (Aribi et al. 2006). The usage of insect regulators such as pyriproxyfen that work as larvicides is a proven strategy in the control of some mosquito species (El-Shazly and Refaie 2002). Fenoxycarb, JH mimics caused significant inhibitory effect on the metamorphosis from pupae to adult in the Beet Armyworm, Spodoptera exigua (Kim, Kim, and Lee 2000) and could control the sexual behavior and reproduction of the Black Cutworm, Agrotis ipsilon (Gadenne 1993) . Fenoxycarb was very active to induce follicle cell patency. The effectiveness of using fenoxycarb can be increased if it is applied during the early pupal stage of Spodoptera exigua (Kim, Kim, and Lee 2000).

New juvenile hormone mimic (JHM), NC-170, affected the metamorphosis and diapause of the small brown planthopper, Laodelphax striatellus. NC-170 (JHM) is selectively active against leafhoppers and planthoppers had a strong metamorphosis-inhibiting activity (Miyake et al. 1992). Subsequently, JHM caused morphogenetic effects by producing supernumery larvae and larval-pupal intermediates. Thus, its strong morphogenetic and sterile activities have shown that it can be a practical new hopper-control agent. Although juvenile hormone mimics had successfully been used to control the insects, but few study also showed that the JH mimics such as JH R394 could give increased the growth and development of silk gland on Silkworm (Bombyx mori L.). The high doses of juvenile hormone (JH) mimics induced the prolongation of last instar larval duration of Bombyx mori L. resulting more silk could be produced due to the increasing cocoon weight (Gangwar 2009). JH mimics on other organisms The toxicity of JH analogs is low to non-target organisms. However, nontarget organisms may also be contaminated through the food-chain or by residues of the exogenous hormone degradation. Some of these JH mimic are toxic to aquatic insects such as the dragonfly and the back swimmer and to crustaceans (Laufer et al. 1987) such as shrimp, crabs (Payen and Costlow 1997) and lobsters (Wyatt 1997). JH analogs also caused the disruption the development of the beneficial insects and natural enemies when the pest is induced or treated with JH mimics such as methoprene (Beckage and Riddiford 1982). Previous studies showed that natural synthetic JH such as methoprene (Figure 3.0) had affected crustaceans (Payen and Costlow 1997). Injection of synthetic JH-I caused vitellogenic inhibition resulting a physiological effect on in the amphipod Orchestia gammarellus by (Payen

and Costlow 1997). The morphological effects are blocking of egg development in the isopod Armadillidium vulgare with topical application of both analogs and natural JH-I. The JH mimic could disrupt sex determination of the crustacean Dapnia magna (Wyatt 1997). The structure of Juvenile and Mimic Juvenile Hormone

Figure 3.0 shows the chemical structure of the mimic juvenile hormone, methoprene. Adopted from article Effects of a Juvenile Hormone Mimic on Male and Female Gametogenesis of the non-target organisms Mud-Crab, Rhithropanopeus harrisii (Gould) (Brachyura: Xanthidae) (Payen and Costlow 1997) JH mimics had caused a precocious metamorphosis without settlement in the acorn barnacle, Balanus galeatus, reared with the synthetic juvenile hormone analog (Laufer et al. 1987). The exposure of the JH mimics reduced the survical rate of larvae of the mud-crab Rhithropanopeus harrissii (Payen and Costlow 1997). In addition, female and male gametogenesis of the crab R.harrisii indicated a progressive inhibition of vitellogenesis and stimulation of spermatogenesis in the presence of methoprene reared in the laboratory after a short time of exposure (12-15 days) (Payen and Costlow 1997) In consequence, the JH mimics indirectly can cause other arthropod other than insects to get gametogenesis disorder and stoppage of vitellogenesis is characterized by a blockage of oocytes at terminal previtellogenesis and proliferation of the follicles cells involved in

vitelogenic oocyte lysis. Spermatogenic degeneration begins with the amalgam of spermatocyte clusters in meiotic prophase and pycnosis of gonia (Payen and Costlow 1997). Therefore, it is very important that the effects and toxicity of these JH mimics to nontarget organisms in the environment to be carefully considered when using these JH mimics as a control strategy for a particular pest insect. The beneficial organism may also affect them if the usage of JH mimics is no controlled properly and carefully in the field.

References: Aribi, N., G. Smagghe, S. Lakbar, N. Soltani-Mazouni, and N. Soltani. 2006. Effects of Pyriproxyfen, a Juvenile Hormone Analog, on Development of the Mealworm, Tenebrio molitor. Pesticide Biochemistry and Physiology 84:55-62. Barret, F. Michael. 1974. Effect of Tropical Application of a Juvenile Hormone Mimic on the Duration of the Moulting Cycle in Fifth Instar Rhodnius. Journal of Insect Physiology 20:1507-1514. Bean, Daniel W., Walter G. Goodman, and Stanley D. Beck. 1983. Regulation of Juvenile Hormone Esterase Activity in the European Corn Borer, Ostrinia nubilalis. Journal of Insect Physiology 29 (12):877-883. Beckage, Nancy E., and Lynn M. Riddiford. 1982. Effects of Methoprene and Juvenile Hormone on Larval Ecdysis, Emergence, and Metamorphosis of the Endoparasitic Wasp, Apanteles congregatus. Journal of Insect Physiology 28 (4):329-334.

Cornette, R., S. Koshikawa, and T. Miura. 2008. Histology of the Hormone-Producing Glands in the Damp-Wood Termite Hodotermopsis sjostedti (Isoptera, Termopsidae): A focus on soldier differentiation. Insect Social 55:407-416. El-Shazly, Mohamed M., and Baraka M. Refaie. 2002. Larvicidal Effect of the Juvenile Hormone Mimic Pyriproxyfen on Culex pipiens. Journal of the American Mosquito Control Association 18 (4):321-328. Gadenne, Christophe. 1993. Effects of Fenoxycarb, Juvenile Hormone Mimetic, on Female Sexual Behaviour of the Black Cutworm, Agrotic ipsilon (Lepidoptera: Noctuidae). Journal of Insect Physiology 39 (1):25-29. Gangwar, S. K. 2009. Effect of Juvenile Hormone Mimic R394 on Silkworm (Bombyx mori L.) Growth and Development of Silk Gland. ARPN Journal of Agriculture and Biological Science 4 (6):65-67. Kamita, Shizua G., Andrew C. Hinton, Craig E. Wheelock, Mark D. Wogulis, David K. Wilson, Nicola M. Wolf, Jeanette E. Stock, Bertold Hock, and Bruce D. Hammock. 2003. Juvenile Hormone (JH) Esterase: Why are you so JH specific? Insect Biochemistry and Molecular Biology 33:1261-1273. Kim, Yonggyun, Donki Kim, and Jungeon Lee. 2000. Disturbance of Adult Eclosion by Fenoxycarb, a Juvenile Hormone Mimic, in the Beet Armyworm, Spodoptera exigua. Journal of Asia-Pacific Entomology 3 (2):103-111. Kremen, Claire, and H. Frederik Nijhout. 1989. Juvenile Hormone Controls the Onset of Pupal Commitment in the Imaginal Disks and Epidermis of Precis Coenia

(Lepidoptera:Nymphalidae). Journal of Insect Physiology 35 (8):603-612.

Krishnakumaran, A., and H. A. Schneiderman. 1965. Prothoracotrophic Activity of Compounds that Mimic Juvenile Hormone. Journal of Insect Physiology 11:1517-1532. Laufer, Hans, Matthew Landau, Ellen Homola, and David W. Borst. 1987. Methyl Farnesoate: Its Site of Synthesis and Regulation of Secretion in a Juvenile Crustacean. Insect Biochemistry 17 (7):1129-1131. Madhavan, Kornath. 1972. Morphogenetic Effects of Juvenile Hormone and Juvenile Hormone Mimics on Adult Development of Drosophila. Journal of Insect Physiology 19:441-453. Minakuchi, Chieka, and Lynn M. Riddiford. 2006. Insect Juvenile Hormone Action as a Potential Target of Pest Management. Journal of Pesticide Science 31 (2):77-84. Miyake, Toshiro, Hiroshi Haruyama, Takashi Mitsui, and Akira Sakurai. 1992. Effects of a New Juvenile Hormone Mimic, NC-170, on Metamorphosis and Diapause of the Small Brown Planthopper, Laodelphax striatellus. Journal of Pesticide Science 17:75-82. Oberlander, Herbet, and Donald L. Silhacek. 1998. Extende Summaries 9th International Congress of Pesticide Chemistry. Pesticide Science 54:300-322. Palli, Subba Reddy. 2009. Recent Advances in the Mode of Action of Juvenile Hormones and Their Analog. Biorational Control of Arthropod Pests:111-129. Partasarathy, R., Anjiang Tan, and Subba R. Palli. 2008. bHLH-PAS- Family Transcription Factor Methoprene-Tolerent Plays a Key Role in JH action in Preventing the Premature Development of Adult Structures during Larval-Pupal Metamorphosis. Mechanisms of Development 125:601-616. Payen, Geneviele G., and John D. Costlow. 1997. Effects of a Juvenile Hormone Mimic on Male and Female Gametogenesis of the Mud-Crab, Rhithropanopeus harrisii (Gould) (Brachyura: Xanthidae). Biology Bulletin 152:199-208.

Riddiford, James W. Truman and Lynn M. 2002. Insect Developmental Hormones and Their Mechanism of Action. Hormones, Brain and Behavior 2:841-873. Riddiford, Lynn M. 2008. Juvenile Hormone Action: A 2007 Perpective. Journal of Insect Physiology 54:895-901. Wyatt, Gerard R. 1997. Juvenile hormone in insect reproduction- a paradox? Europe Journal of Entomology 94:323-333.