Yasmin Akhtar

Comparative growth inhibitory and antifeedant effects of plant extracts and pure allelochemicals on four phytophagous insect species

Antifeedant and growth inhibitory effects of crude plant extracts (Melia volkensii and Origanum vulgare) and pure allelochemicals (digitoxin, cymarin, xanthotoxin, toosendanin, thymol and trans‐anethole) were investigated in the cabbage looper (Trichoplusia ni), and in the armyworm (Pseudaletia unipuncta) using different bioassays. Antifeedant effects of M. volkensii, O. vulgare and thymol were investigated in larvae of the diamondback moth (Plutella xylostella), and of O. vulgare and thymol in the Mexican bean beetle (Epilachna varivestis), using leaf disc choice bioassays. M. volkensii was the most potent growth inhibitor for T. ni and P. unipuncta (dietary EC50 = 7.6 and 12.5 p.p.m., respectively) of all the test substances. Cymarin was the second most potent growth inhibitor (EC50 = 132.0 p.p.m.) for T. ni. The most effective feeding deterrents for third instar T. ni larvae were xanthotoxin andM. volkensii (DC50 = 0.9 and 8.3 μg/cm2, respectively). M. volkensii was also the most potent feeding deterrent for third instar P. unipuncta, P. xylostella and adult E. varivestis (DC50 = 10.5, 20.7 and 2.3 μg/cm2, respectively). Because of interspecific differences in response to feeding deterrents and the lack of a strong relationship between EC50 and DC50 values, we recommend testing a battery of bioassay species with candidate compounds and the use of more than one bioassay. Based on their growth inhibitory and feeding deterrent properties, some of these plant extracts and pure allelochemicals have potential for use as alternative crop protectants against a number of pest species.

Comparative bioactivity of selected extracts from Meliaceae and some commercial botanical insecticides against two noctuid caterpillars, Trichoplusia ni and Pseudaletia unipuncta

Plant-derived extracts and phytochemicals have long been a subject of research in an effort to develop alternatives to conventional insecticides but with reduced health and environmental impacts. In this review we compare the bioactivities of some plant extracts with those of commercially available botanical insecticides against two important agricultural pests, the cabbage looper, Trichoplusia ni and the armyworm, Pseudaletia unipuncta. Test materials included extracts of Azadirachta indica (neem), A. excelsa (sentang), Melia volkensii, M. azedarach (Chinaberry) and Trichilia americana, (all belonging to the family Meliaceae) along with commercial botanical insecticides ryania, pyrethrum, rotenone and essential oils of rosemary and clove leaf. Most of the extracts and botanicals tested proved to be strong growth inhibitors, contact toxins and significant feeding deterrents to both lepidopteran species. However, there were interspecific differences with T. ni generally more susceptible to the botanicals than the armyworm, P. unipuncta. All botanicals were more inhibitory to growth and toxic (through feeding) to T. ni than to P. unipuncta, except for M. azedarach which was more toxic to P. unipuncta than to T. ni. Athough, pyrethrum was the most toxic botanical to both noctuids, A. indica, A. excelsa, and M. volkensii were more toxic than ryania, rotenone, clove oil and rosemary oil for T. ni. As feeding deterrents, pyrethrum was the most potent against T. ni, whereas A. indica was the most potent against the armyworm. Based upon growth inhibition, chronic toxicity, and antifeedant activity, some of these plant extracts have levels of activity that compare favorably to botanical products currently in commercial use and have potential for development as commercial insecticides.

Comparative Toxicity of Essential Oils of Litsea pungens and Litsea cubeba and Blends of Their Major Constituents against the Cabbage Looper, Trichoplusia ni

Contact toxicity of essential oils of Litsea pungens Hemsl. and L. cubeba (Lour.) Pers. (Lauraceae) and of blends of their major constituents was assessed against third-instar Trichoplusia ni larvae via topical application. Both oils showed moderate activity against T. ni larvae with LD(50) values of 87.1 and 112.5 microg/larva, respectively. 1,8-Cineole from the essential oil of L. pungens and gamma-terpinene from the oil of L. cubeba accounted for much of the toxicity of the oils to T. ni larvae. The toxicity of blends of selected constituents indicated a synergistic effect among putatively active and inactive constituents, with the presence of all constituents necessary for full toxicity of the natural oils. The results show that essential oils of L. pungens and L. cubeba and some of their constituents have potential for development as botanical insecticides.

Feeding responses of specialist herbivores to plant extracts and pure allelochemicals: effects of prolonged exposure

We investigated the possibility of decreased feeding deterrent response following prolonged exposure in three specialist herbivores, Plutella xylostella (larvae), Pseudaletia unipuncta (larvae), and Epilachna varivestis (adults) through leaf disc choice and no‐choice tests. Neonate larvae (<24 h old) of the three species were reared on their host plants sprayed with the antifeedants until tested. Our results demonstrated that Ps. unipuncta did not show a decrease in feeding deterrent response to extracts of Melia volkensii (choice and no‐choice tests) or oregano (Origanum vulgare) (choice test) following prolonged exposure. Plutella xylostella exhibited a decrease in feeding deterrent response to M. volkensii in a choice test only, but not to oregano. Although not significant, we did find a trend toward decreasing feeding deterrent response to M. volkensii by both species in no‐choice tests. However, both species exhibited a significant decrease in feeding deterrent response to pure allelochemicals (Ps. unipuncta to thymol, and P. xylostella to thymol and toosendanin) compared with the naïve groups, following prolonged exposure in leaf disc choice tests. Epilachna varivestis showed a significant decrease in feeding deterrent responses to both oregano and thymol in leaf disc choice tests. We conclude from our present and previous studies that not only are there interspecific differences between generalist and specialist species but also among specialist species.

Decreased response to feeding deterrents following prolonged exposure in the larvae of a generalist herbivore, Trichoplusia ni (Lepidoptera: Noctuidae)

We investigated the role of experience with several antifeedants on the feeding behavior of a generalist herbivore, Trichoplusia ni. Second-, third-, and fifth-instar larvae of T. ni were examined for their feeding responses to plant extracts (Melia volkensii, Origanum vulgare) and individual plant allelochemicals (cymarin, digitoxin, xanthotoxin, toosendanin, and thymol), after being exposed to them continually beginning as neonates. All tested instars of T. ni were capable of showing a decreased antifeedant response following prolonged exposure to most of the antifeedants tested compared with their naive conspecifics. Cardenolides (digitoxin and cymarin) were the exceptions. The response to oregano was affected as a result of previous exposure to different concentrations of oregano, unlike M. volkensii, leading us to conclude that T. ni sensitivity varies between stimuli and cannot be generalized. To demonstrate that decreased deterrence following prolonged exposure to M. volkensii was the result of learned habituation, three aversive stimuli were used. A high concentration of the particular antifeedant, xanthotoxin, acted as a noxious stimulus and dishabituated (reversed) the decrement in the antifeedant response to M. volkensii. Cold shock or CO2 was marginally effective in dishabituating the response. The fact that the decrease in antifeedant response can be dishabituated has implications for pest management.

Dialkoxybenzene and Dialkoxyallylbenzene Feeding and Oviposition Deterrents against the Cabbage Looper, Trichoplusia ni: Potential Insect Behavior Control Agents

The antifeedant, oviposition deterrent, and toxic effects of individual dialkoxybenzene compounds/sets and of hydroxy- or alkoxy-substituted allylbenzenes, obtained through Claisen rearrangement of substituted allyloxybenzenes, were assessed against the cabbage looper, Trichoplusia ni , in laboratory bioassays. Most of the compounds/sets strongly deterred larval feeding, with some exhibiting mild toxic and oviposition deterrent effects as well. Some of the compounds/sets were more active than the commercial insect repellent, DEET (N,N-diethyl-m-toluamide), as both feeding and oviposition deterrents against the cabbage looper. On the basis of the obtained oviposition data a general hypothesis was proposed regarding the oviposition sites: one binding mode with the alkyl and allyl groups on the same side of the benzene ring resulted in deterrence, the other with alkyl and allyl groups on opposite sides of the benzene ring resulted in stimulation. The results suggest some structure-activity relationships useful in improving the efficacy of the compounds and designing new, nontoxic insect control agents for agriculture.

Insecticidal and feeding deterrent activities of essential oils in the cabbage looper, Trichoplusia ni (Lepidoptera: Noctuidae)

Ten essential oils were tested against the cabbage looper, Trichoplusia ni larvae for contact, residual and fumigant toxicities and feeding deterrent effects. Against third instar T. ni, Syzygium aromaticum (LD50 = 47.8 μg/larva), Thymus vulgaris (LD50 = 52.0 μg/larva) (the two positive controls) and Cinnamomum glanduliferum (LD50 = 76.0 μg/larva) were the most toxic via topical application. Litsea pungens (LD50 = 87.1 μg/larva), Ilex purpurea (LD50 = 94.0 μg/larva), Cinnamomum cassia (LD50 = 101.5 μg/larva) and Litsea cubeba (LD50 = 112.4 μg/larva) oils were equitoxic. Thymus vulgaris (LC50 = 4.8 mg/ml) and S. aromaticum (LC50 = 6.0 mg/ml) oils were the most toxic in residual bioassays. Cymbopogon citratus (LC50 = 7.7 mg/ml) and C. cassia (LC50 = 8.5 mg/ml) oils were equitoxic followed by Cymbopogon nardus (LC50 = 10.1 mg/ml) in this bioassay. The remaining five oils showed little or no residual effects. In a fumigation bioassay, L. cubeba (LC50 = 16.5 μl/l) and I. purpurea (LC50 = 22.2 μl/l) oils were the most toxic. Cinnamomum glanduliferum (LC50 = 29.7 μl/l) and Sabina vulgaris (LC50 = 31.2 μl/l) oils were equitoxic. Interestingly, S. aromaticum did not exhibit any fumigant toxicity. Cymbopogon citratus, C. nardus and C. cassia strongly deterred feeding by third instar T. ni (DC50s = 26.9, 33.8 and 39.6 μg/cm2, respectively) in a leaf disc choice bioassay. The different responses of T. ni larvae to the oils in different bioassays suggest that these essential oils exhibit different modes of action. Based on their comparable efficacy with essential oils already used as active ingredients in many commercial insecticides (i.e. clove oil and thyme oil), some of these essential oils may have potential as botanical insecticides against T. ni.

Binary mixtures of feeding deterrents mitigate the decrease in feeding deterrent response to antifeedants following prolonged exposure in the cabbage looper, Trichoplusia ni (Lepidoptera: Noctuidae)

Summary. The effect of rearing larvae of Trichoplusia ni on individual feeding deterrents or on binary mixtures of deterrents on their subsequent gustatory sensitivity was measured in paired choice leaf disc bioassays. Our working hypothesis was that mixtures of antifeedants (pure allelochemicals) would mitigate decreased feeding deterrent response following prolonged exposure in this generalist herbivore. Neonate larvae were reared on cabbage leaves treated with individual feeding deterrents (digitoxin, thymol, toosendanin or xanthotoxin), or with binary mixtures of these until the third instar. Feeding deterrent responses to each antifeedant or mixture was then determined in a leaf disc choice bioassay. All of the mixtures produced additive deterrence when presented to naïve larvae. Larvae reared on individual antifeedants showed a significantly decreased feeding deterrent response (except to digitoxin), whereas larvae reared on binary mixtures of antifeedants did not show a decreased feeding deterrent response to any of them. Such mixtures were synergistic in terms of their feeding deterrence to “experienced” larvae. Our experiment supports the hypothesis (Jermy 1986) that mixtures of deterrents can prevent decreased feeding deterrent response following prolonged exposure, and provides one explanation for the multiplicity of chemical defenses found in many plants.

Relationship between adult and larval host plant selection and larval performance in the generalist moth, Trichoplusia ni

Adult oviposition preferences are expected to correlate with host plant suitability for the development of their offspring. For most lepidopteran species, this is particularly important as the hatching neonate larvae of many species are relatively immobile. Thus, the site of oviposition chosen by a female adult can greatly influence the probability of survival for her offspring. In the present study, we investigated the oviposition preference of adult Trichoplusia ni moths for six plant species to determine whether they could accurately rank the suitability of the plants for larval development. We also compared oviposition preferences to neonate larval acceptance and preference to determine whether the adult host range matched that of larval diet breath. Our results indicate that in two-choice and no-choice tests adult T. ni were able to rank the plants accurately, with the exception of anise hyssop. However, when given a choice of all six plants together, they laid more eggs on a plant that was not suitable for larval survival. Larvae accepted and fed on all plants in no-choice tests, and accurately ranked them according to larval performance. We conclude that neonate larvae are better able than adults to rank plants according to larval performance, and that larval diet breadth is wider than the range of plants accepted by adults. We also provide a discussion of the reduced accuracy of adult oviposition preference with increased plant choices.

GENERALIZATION OF A HABITUATED FEEDING DETERRENT RESPONSE TO UNRELATED ANTIFEEDANTS FOLLOWING PROLONGED EXPOSURE IN A GENERALIST HERBIVORE, Trichoplusia ni

The possibility of generalization of habituated response to unrelated feeding deterrents following prolonged exposure was examined in third instar Trichoplusia ni (Lepidoptera: Noctuidae) larvae by rearing them on antifeedants and then testing with other unrelated antifeedants. We introduced neonate larvae (<24-hr old) onto cabbage leaves treated with crude seed extracts of Melia volkensii (Meliaceae) or oil of Origanum vulgare (“oregano”) (Lamiaceae) and allowed them to feed until early in the third instar. Naïve larvae were reared on cabbage leaves treated with carrier solvent alone. Both experienced and naïve larvae were tested for feeding deterrent response with the same and the different extracts in a leaf disc choice bioassay. Habituation was generalized to both M. volkensii and oregano following prolonged exposure to either plant extract and also to a pure allelochemical, thymol, following prolonged exposure to either digitoxin or xanthotoxin. However, there was no generalization of the habituated response to oregano following prolonged exposure to digitoxin or thymol, or to thymol or xanthotoxin following prolonged exposure to oregano or M. volkensii. Our results demonstrate that habituated response to feeding deterrents in a polyphagous insect herbivore can be generalized among and between plant extracts and pure allelochemicals, but not in all situations. The implications of such behavioral plasticity in herbivorous insects for the use of antifeedants as crop protectants or for host plant shifts is discussed.