Julio S. Bernal

Evaluation of lectin-expressing transgenic sugarcane against stalkborers (Lepidoptera: Pyralidae) : Effects on life history parameters

Abstract The impact of snowdrop lectin (Galanthus nivalis agglutinin, GNA) expressed in transgenic sugarcane on life history parameters of Mexican rice borer [Eoreuma loftini (Dyar)] and sugarcane borer [Diatraea saccharalis (F.)] (both Lepidoptera: Pyralidae) was evaluated. In the laboratory, lyophilized sugarcane leaf sheath tissue was incorporated in a meridic diet resulting in a GNA concentration of 0.47% of total protein, and used for insect bioassays over two successive generations. Deleterious effects of GNA were not observed on survival, weight, and developmental periods of larvae and pupae, nor on adult fecundity and egg viability of D. saccharalis. Moreover, in the first generation, addition of transgenic sugarcane tissue to the diet enhanced larval growth in D. saccharalis resulting in higher larval and pupal weight compared with diet with nontransgenic sugarcane, but this effect was not observed in the second generation. In contrast, larval survival, percent adult emergence, and female fecundity of E. loftini were significantly reduced when fed transgenic sugarcane diet compared with nontransgenic sugarcane diet. In addition, a substantial reduction of female pupal weight of E. loftini was observed in the second generation. For both species, the only consistent effect of GNA in both generations was a reduction in adult female longevity. Life table parameters showed that GNA at the level found in the transgenic diet negatively affected development and reproduction of E. loftini, whereas it had a nil to positive effect on development and reproduction of D. saccharalis.

Host influences on sex ratio, longevity, and egg load of two Metaphycus species parasitic on soft scales : implications for insectary rearing

Metaphycus flavus (Howard) and M. stanleyi Compere (Hymenoptera: Encyrtidae) are currently being screened for use as augmentative biological control agents of citrus‐infesting soft scales (Homoptera: Coccidae). Two factors were investigated, host quality‐dependent sex allocation and local mate competition, which likely influence these parasitoids sex allocation strategies and are therefore of interest for their mass‐rearing. The results of these studies suggested that, under the mass‐rearing protocol that is envisioned for these parasitoids, offspring sex ratios in both M. flavus and M. stanleyi are dominated by host quality (= size) influences, but not by interactions with other females. These results indicated that host size strongly influences offspring sex ratios and brood sizes; larger hosts led to more female offspring and larger broods. In contrast, increasing the number of parental females did not lead to fewer female offspring as expected under local mate competition. Additionally, within‐brood sex ratios did not vary with brood size; this result is inconsistent with expected sex ratios due to local mate competition. Other results also indicated that host quality was a dominant influence on M. flavus‘ and M. stanleyis sex ratios. Larger hosts led to a larger size in the emerging wasps, and larger wasps had greater egg loads and lived longer than smaller wasps. However, wasp longevity, and the influence of wasp size on longevity were mediated by a wasps diet. Metaphycus flavus females lived the longest when they had access to hosts, honey, and water, followed by honey and water, and shortest when they had access to water alone; M. stanleyi females lived longest with honey and water, followed by hosts, honey, and water, and shortest with water alone. Greater wasp size led to greater longevity in females only when they had access to food (honey, or hosts and honey). Finally, other results suggested that both M. flavus and M. stanleyi are facultatively gregarious. Wasp size did not decrease with brood size as expected under superparasitism. Overall, the results of these studies suggested that holding newly emerged females of both M. flavus and M. stanleyi for several days in the presence of an appropriate food source before field release could enhance a females performance as an augmentative biological control agent. It increases their initial life expectancy following release, and maximizes the females‘ egg load (both Metaphycus species) and resources for replacing oviposited eggs (M. flavus only).

Effects of Snowdrop Lectin (Galanthus nivalis Agglutinin) Expressed in Transgenic Sugarcane on Fitness of Cotesia flavipes (Hymenoptera: Braconidae), a Parasitoid of the Nontarget Pest Diatraea saccharalis (Lepidoptera: Crambidae)

Abstract Cotesia flavipes (Cameron) is a parasitoid responsible for maintaining populations of sugarcane borer, Diatraea saccharalis (F.), below economic levels in south Texas sugarcane fields. Transgenic sugarcane expressing the snowdrop lectin (Galanthus nivalis agglutinin, GNA) was developed against the Mexican rice borer, Eoreuma loftini (Dyar), the primary pest of south Texas sugarcane. The potential impact of GNA-expressing sugarcane on various biological and fitness parameters of Cotesia flavipes (Cameron) was studied in the laboratory to gain insight on likely effects of the transgenic sugarcane on biological control of sugarcane borer by C. flavipes. Females of C. flavipes were offered sugarcane borer larvae fed one of two diet treatments for oviposition for two successive generations: (1) artificial diet containing transgenic sugarcane tissue or (2) artificial diet containing nontransgenic sugarcane tissue. Small to marginal negative effects of artificial diet containing transgenic sugarcane tissue were evident in the rate of host suitability, number of cocoons and adult parasitoids emerging per host, percentage cocoons yielding parasitoids, and sex ratio and adult lifespan of parasitoids. These effects were variable between the two parasitoid generations examined. In contrast, differences were not detected between diet treatments in rates of host acceptance, egg load of females, and egg to adult developmental periods. The negative effects of transgenic sugarcane on C. flavipes detected in this study are important because GNA levels in the diet (≈0.49% of total protein content) containing transgenic sugarcane tissue were ≈50% of the level expressed in transgenic sugarcane plants. Results are discussed in relation to potential impacts of the transgenic cultivar on biological control of sugarcane borer by C. flavipes.

Facilitated by nature and agriculture: performance of a specialist herbivore improves with host‑plant life history evolution, domestication, and breeding

Plant defenses against herbivores are predicted to change as plant lineages diversify, and with domestication and subsequent selection and breeding in the case of crop plants. We addressed whether defense against a specialist herbivore declined coincidently with life history evolution, domestication, and breeding within the grass genus Zea (Poaceae). For this, we assessed performance of corn leafhopper (Dalbulus maidis) following colonization of one of four Zea species containing three successive transitions: the evolutionary transition from perennial to annual life cycle, the agricultural transition from wild annual grass to primitive crop cultivar, and the agronomic transition from primitive to modern crop cultivar. Performance of corn leafhopper was measured through seven variables relevant to development speed, survivorship, fecundity, and body size. The plants included in our study were perennial teosinte (Zea diploperennis), Balsas teosinte (Zea mays parviglumis), a landrace maize (Zea mays mays), and a hybrid maize. Perennial teosinte is a perennial, iteroparous species, and is basal in Zea; Balsas teosinte is an annual species, and the progenitor of maize; the landrace maize is a primitive, genetically diverse cultivar, and is ancestral to the hybrid maize; and, the hybrid maize is a highly inbred, modern cultivar. Performance of corn leafhopper was poorest on perennial teosinte, intermediate on Balsas teosinte and landrace maize, and best on hybrid maize, consistent with our expectation of declining defense from perennial teosinte to hybrid maize. Overall, our results indicated that corn leafhopper performance increased most with the agronomic transition, followed by the life history transition, and least with the domestication transition.

Sex ratios in field populations of two parasitoids (Hymenoptera: Chalcidoidea) of Coccus hesperidum L. (Homoptera: Coccidae)

Abstract We tested several assumptions and predictions of host-quality-dependent sex allocation theory (Charnov et al. 1981) with data obtained for the parasitoid Metaphycus stanleyi Compere on its host, brown soft scale (Coccus hesperidum L.), in a California citrus grove and in the laboratory. Scales ceased growing after parasitization by M. stanleyi. Thus, M. stanleyi may gauge host quality (=size) at oviposition. Host size positively influenced adult parasitoid size, and parasitoid size in turn influenced adult longevity of M. stanleyi. However, parasitoid fitness gains with host size and adult size were similar in males versus females. Sex allocation to individual hosts by M. stanleyi depended on host size; females consistently emerged from larger hosts than males. Host size was important in a relative sense; the mean host sizes of females versus males, and of solitary versus gregarious parasitoids varied with the available host size distribution. The offspring sex ratio of M. stanleyi reflected the available host size distribution; the sex ratio of emerging parasitoids varied with the available host size distribution. We did not detect a “critical host size” below which males emerged, and above which females emerged; rather, only females emerged from hosts in the upper size range, and a variable ratio of males and females emerged from hosts in the lower size range. We conclude that the sex ratio of field populations of M. stanleyi is driven largely by the available size distribution of C. hesperidum. In addition, we tested predictions resulting from theoretical analyses of sex allocation in autoparasitoids with data obtained on Coccophagus semicircularis (Förster) parasitizing brown soft scale in the field. The sex ratio of C. semicircularis was consistently and strongly female biased (ca. 90% females). Based on available theoretical analyses, we suggest that this sex ratio pattern may have resulted from a very low encounter rate of secondary hosts coupled with a strong time limitation in C. semicircularis females. This explanation was the most plausible given constraints stemming from the detection of secondary hosts, their variable location within primary hosts, and their handling times. Finally, the size of hosts which yielded single versus multiple parasitoids, and the sizes of these parasitoids, were compared. These comparisons suggested that: (1) M. stanleyi females gauge host sizes precisely, and in terms of female offspring; thus a fitness penalty is not incurred by females which share a host, while males benefit from sharing a host, and; (2) instances where multiple C. semicircularis emerged from a single host were probably the result of parasitism by different females, or during different encounters by a single female.

Substrate influences mating success and transmission of courtship vibrations for the parasitoid Cotesia marginiventris

The influences of artificial and natural rearing substrates on mating success were investigated for the parasitoid wasp Cotesia marginiventris (Cresson) (Hymenoptera: Braconidae), a candidate for augmentative biological control of various lepidopteran pests. Five rearing substrates were tested: plastic, glass, chiffon fabric, and leaves of two host plants, bean [Vigna unguiculata (L.) (Fabaceae)] and maize [Zea mays L. (Poaceae)]. Mating success was highest on chiffon, lowest on plastic and glass, and intermediate on maize and bean. The transmission characteristics of one component (buzz 1) of the courtship vibrations produced by male wing fanning were investigated using laser vibrometry. The duration of buzz 1 was longer on maize, bean, and chiffon than on plastic and glass. The fundamental frequency of buzz 1 (~300 Hz) was lowest on bean and highest on glass, and intermediate among other substrates. The relative amplitude of buzz 1 was higher on chiffon than on plastic, glass, or bean, and intermediate on maize. The relative importance of airborne sound and substrate vibration as courtship signals was also investigated with experiments that manipulated the production of courtship vibrations and the mating substrates. The amplitude of courtship vibrations on chiffon was significantly higher for winged males than for dealated males. The mating success of males was impacted by both the presence of wings and the mating substrate. These findings suggest that mating success and transmission of courtship vibrations are influenced by the rearing substrate, and that courtship vibrations are critical to mating success in C. marginiventris. Future efforts to mass rear this parasitoid and other insects should consider the potential influences of rearing substrates on mating.

Genetically modified crops deserve greater ecotoxicological scrutiny

Historians are keen to remind us that history tends to rhyme, even if it does not repeat itself. In a historical context, the story of today’s genetically modified (GM) crops resembles that of the synthetic organic insecticides beginning circa the second half of the last century. In practice, GM crops include crop cultivars that have been modified by incorporating one or more genes (using genetic engineering techniques) originating from species other than the crop species itself. Presently available GM crops include crop cultivars expressing tolerance to particular herbicides, or resistance to insect pests or diseases (or combinations). Beginning in the late 1940s, synthetic organic insecticides became widely available for agricultural use, were received by farmers and agricultural scientists with excitement and little scepticism, and soon became indispensable if farmers were to remain competitive (Perkins 1982). Synthetic insecticides were rapidly used indiscriminately and the environmental consequences of such indiscriminate use became clearly evident within three decades, including impacts on communities of nontarget organisms (NTO) (Perkins 1982). The human-health consequences were evident sooner, but were largely ignored. Such consequences remain easily seen today, for example in the public’s attitude toward pesticide use in agriculture (due in good measure to Rachel Carson’s Silent Spring), in the ways we attempt to manage agricultural pests (e.g., Integrated Pest Management) and in some of our environmental and agricultural policies (e.g., creation

Olfactory Responses of Anaphes iole (Hymenoptera: Mymaridae) to Volatile Signals Derived from Host Habitats

Anaphes iole Girault is a frequent parasitoid of Lygus spp. eggs in the United States, and has potential as a biological control agent against Lygus hesperus Knight in different crops. Feeding and oviposition by L. hesperus induce emission of plant volatiles, but studies to date do not address the role of plant volatiles in the host-searching behavior of A. iole. In this study, a four-arm olfactometer was used to test the responses of female parasitoids to odors emanating from cotton (Gossypium hirsutum L., Malvaceae) plants damaged by L. hesperus females, L. hesperus males, larvae of the nonhost Spodoptera exigua Hubner, or mechanically, or to odors from L. hesperus females alone. In addition, various plants damaged by L. hesperus females were evaluated in the olfactometer: cotton, alfalfa (Medicago sativa L., Fabaceae), common groundsel (Senecio vulgaris L., Asteraceae), annual ragweed (Ambrosia artemisifolia L., Asteraceae), and redroot pigweed (Amaranthus retroflexus L., Amaranthaceae). In all olfactometry bioassays, treatment odors were compared against three controls (humidified air). Results showed that A. iole females were consistently attracted to odors derived from different plant–L. hesperus complexes, while odors from plants subjected to nonhost (S. exigua) or mechanical damage and L. hesperus females alone were not attractive or only variably attractive. These findings suggest that while searching for hosts A. iole females use specific volatiles induced by L. hesperus feeding and oviposition to locate hosts inhabiting a wide variety of plants, including annual and perennial species from four plant families. It was suggested that future research should seek to identify the chemical elicitors involved in the release of plant volatiles attractive to A. iole females.

Effect of continuous rearing on courtship acoustics of five braconid parasitoids, candidates for augmentative biological control of Anastrepha species

The courtship acoustics of five species of parasitoid wasps (Hymenoptera: Braconidae), potential candidates for augmentative biological control of Anastrepha (Schiner) species (Diptera: Tephritidae), were compared between recently colonized individuals and those continuously reared 70–148 generations. During courtship, males of these parasitoid species fan their wings and produce a series of low amplitude pulses. The first series of 15 or more continuous courtship pulses was used to measure the pulse duration, frequency, and interpulse interval (IPI) from the beginning, middle, and end of the pulse series. Each parameter was compared between young and old colonies, and among species. Several differences in courtship acoustics were detected in colonies that had been continuously reared. The pulse duration at the end of the pulse series was longer in old colonies for Doryctobracon crawfordi (Viereck) (Hymenoptera: Braconidae), but shorter for old colonies of Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae). The IPI of the middle pulse was shorter in old colonies of Opius hirtus (Fischer) (Hymenoptera: Braconidae), and was also shorter at the last pulse for old colonies of both Utetes anastrephae (Viereck) (Hymenoptera: Braconidae) and D. longicaudata. The duration of the middle pulse distinguished the three native species, and separated the two introduced species from each other. We discuss our findings in light of their biological and applied implications, particularly those dealing with quality control of mass-reared parasitoids.

Population genetic structure of a specialist leafhopper on Zea: likely anthropogenic and ecological determinants of gene flow

Corn leafhopper, Dalbulus maidis DeLong & Wolcott (Hemiptera: Cicadellidae), is a specialist herbivore on the genus Zea (Poaceae). The genera Dalbulus and Zea evolved in central Mexico. We sought to determine whether population genetic structuring is prevalent in corn leafhoppers inhabiting three of its host plants: (1) the highland species perennial teosinte (Zea diploperennis Iltis, Doebley & Guzman), (2) the mid‐ to lowland‐species Balsas teosinte (Zea mays ssp. parviglumis Iltis & Doebley), and (3) the ubiquitous domesticated maize (Zea mays ssp. mays L.). We used amplified fragment length polymorphisms to detect population structuring and genetic differentiation among corn leafhoppers on the three host plants in western‐central and ‐northern Mexico. Our results showed that corn leafhopper in Mexico is composed of at least two genetically discrete populations: an ‘Itinerant’ population associated with the annual hosts maize and Balsas teosinte, which appears to be widely distributed in Mexico, and a ‘Las Joyas’ population restricted to perennial teosinte and confined to a small mountain range (Sierra de Manantlán) in western‐central Mexico. Our results further suggested that population structuring is not due to isolation by distance or landscape features: Las Joyas and Itinerant corn leafhopper populations are genetically distinct despite their geographic proximity (ca. 4 km), whereas Itinerant corn leafhoppers separated by hundreds of kilometers (>800 km), mountain ranges, and a maritime corridor (Sea of Cortez) are not genetically distinct. Based on our results and on published ethnohistorical and archaeological data, we propose pre‐Columbian and modern scenarios, including likely ecological and anthropogenic influences, in which the observed genetic population structuring of corn leafhopper could have originated and could be maintained. Also, we hypothesize that after evolving on the lowland Balsas teosinte, corn leafhopper expanded its host range to include maize and then the highland perennial teosinte, following the domestication and spread of maize within the last 9 000 years.