Comparative rearing parameters for bisexual and genetic sexing strains of Zeugodacus cucurbitae and Bactrocera dorsalis (Diptera: Tephritidae) on an artificial diet

The Sterile Insect Technique (SIT) is an important component of area wide programs to control invading or established populations of pestiferous tephritids. The SIT involves the production, sterilization, and release of large numbers of the target species, with the goal of obtaining sterile male x wild female matings, which yield infertile eggs. A major advance in SIT involved sex-linked, genetic manipulations that allowed the production and release of male-only strains (also termed genetic sexing strains, GSS). The use of GSS avoids matings between sterile males and females, which may divert males from seeking and mating with wild females, and studies show that male-only releases result in greater suppression of wild populations than standard bisexual releases (i.e., those including both males and females). GSS based on sex-linked pupal color exist for Zeugodacus cucurbitae (Coquillett) and Bactrocera dorsalis (Hendel), two important agricultural pest species, but their rearing characteristics have not been documented in detail. The goal of the present study was to compare the pupal color sexing and bisexual strains for each of these species with respect to important rearing parameters, including egg production and eclosion of larvae from eggs (egg hatch), pupal recovery, and weight, emergence rate, and flight ability. In both species, most of these parameters were significantly greater for the bisexual strain than the GSS, and, for a given number of eggs, the production of flight-capable adults was approximately 2 times greater in the bisexual strains of both species. The potential usefulness of GSS in SIT against Z. cucurbitae and B. dorsalis is assessed based on these findings.     

Repair of a genetically-caused defect in oogenesis in Drosophila melanogaster by transplantation of cytoplasm from wild-type eggs and by injection of pyrimidine nucleosides

Eggs produced by homozygous mutant rudimentary (r⁹, 154.5) females of Drosophila melanogaster die during embryogenesis, apparently because the mutant female fails to incorporate certain substances into the egg during oogenesis. These eggs can be rescued by injecting them at the preblastoderm stage with wild-type nuclei and cytoplasm or wild-type cytoplasm alone from unfertilized eggs. Some preblastoderm eggs injected with 1.5% of egg volume of cytoplasm from unfertilized wild-type eggs were able to complete both embryonic and postembryonic development and emerged as adults, whereas not a single uninjected control egg was able to complete embryonic development. The eggs of rudimentary mothers can also be rescued by injecting each egg at the blastoderm stage with 0.01 μg of pyrimidine nucleosides. The results demonstrate that a pyrimidine deficiency is the cause of abortive embryogenesis, and confirm the previous finding that the rudimentary mutation leads to pyrimidine auxotrophy.     

Biological studies onTrichogrammatoidea armigera Nagaraja a new dimorphic egg parasite ofHeliothis armigera (Hübner) in India

A new species ofTrichogrammatoidea which is being described byH. Nagaraja asT. armigera, has been reared from eggs ofHeliothis armigera onPolianthes tuberosa and from those of an unidentified Lepidopteron onCajanus cajan. In the laboratory the parasite was successfully bred onCorcyra cephalonica, Achaea janata, Gnorimoschema operculella andPlutella xylostella; it did not show any perceptible preference for any of these hosts. It also parasitised eggs ofSpodoptera litura, but although development proceeded to the adult stage, adults failed to emerge, suggesting unsuitability of this host. The males ofT. armigera are found to exhibit dimorphism — one form being alate and the other typically apterous. The apterous males were almost exclusively produced only by fertilised females and in the progeny of virgin females these forms were extremely rare (1 apterous male: 1,500 winged males). Among the progeny of a single mated female, an apterous male developed invariably in association with a female, but this rule did not apply when a singleCorcyra egg was parasitised by more than one parental female. However, in no case did an apterous male alone emerge from a single host egg. The biology ofT. armigera has been studied at 25°C±1°C. and R.H. 75%, usingC. cephalonica eggs. The parasite completed its life-cycle in 7–9 days — the egg, larval and pupal periods occupying, 1, 2–3 and 4–5 days, respectively. When fed honey, the average longevity of females was 7 (max. 11) days, of alate males 6 (max. 10) days and of apterous males 1 (max. 2) day. The maximum fecundity was 118 while the average was about 63. From 2–26 (average 9) eggs were parasitised per day. The sex-ratio was 62% females: 38% males (35% alate and 3% apterous). An alate male during its lifetime inseminated upto 10 (Av. 9) females while an apterous one inseminated up to 4 (Av. 3) females.     

A Sex-Specific, Temperature-Sensitive Maternal Effect of the Daughterless Mutation of DROSOPHILA MELANOGASTER

Every aspect of the phenotype of the daughterless (2–41.5) maternal effect mutation was found to be strongly temperature sensitive. Above 22°, da/da mothers produced no daughters; however female progeny did survive the da maternal effect if the last 60 hours of oogenesis and the first three hours of embryonic development took place at 18°. The females which survived under these conditions displayed morphological abnormalities in a variety of adult cuticular structures, characteristic of cell death during development. In contrast, their male siblings were morphologically normal. Upon prolonged exposure to 29°, da/da females became sterile but continued to lay eggs. Some sexually mosaic progeny from da/da mothers survived even at 25°, but the distribution and development of the female tissue in these mosaics were abnormal. It is suggested that there are multiple functions of the da⁺ gene during oogenesis, one of which may be required specifically for the subsequent survival of female cells throughout the embryo. In addition to and distinct from its effect during oogenesis, the da mutation acted in both sexes of progeny as a recessive temperature-sensitive lethal mutation with a TSP during the first half of embryonic development.     

Effect of hosts and parasite density on the egg parasiteTrichogramma pretiosum [Hym.: Trichogrammatidae]

When females ofTrichogramma pretiosum Riley were confined with host eggs at a density of 2/150 eggs, they produced 12 times more female progeny on eggs of potato tuber moth than on eggs ofHeliothis armigera (Hübner) and 13,6 times more on eggs ofSitotroga cerealella (Olivier) than on eggs ofHeliothis. At a density of 4/150 eggs, the correspondent figures were 13 and 8 times. The percentage emergence fromHeliothis eggs was from 0,29 to 0,14 times as great as from tuber moth orSitotroga. From 15 to 140 times more runts were observed amongTrichogramma fromHeliothis eggs than among those from tuber moth eggs and 8 times more thant among those fromSitotroga eggs. This may explain the low recoveries in South Africa ofT. pretiosum in eggs ofH. armigera collected in cotton fields after mass liberation of the parasite. An increase in parasite density from 1/300 eggs to 16/300 eggs resulted in a decrease from 29 to 14 in the hosts parasitised per female, a decrease in the proportion of female progeny from 72 to 39%, a decrease in the female progeny per female from 18 to 4,8, and an increase in the proportion of runts from 2,4 to 12,4%. It is suggested that in mass culture ofTrichogramma unduly high parasite densities should be avoided in order to reduce the effect of mutual interference and raise the output of female progeny.     

Biology ofTrichogramma ostriniae (Hym.: trichogrammatidae) reared onOstrinia nubilalis (Lep.: pyralidae) and survey for additional hosts

Trichogramma ostriniae has been identified as a candidate for biological control ofOstrinia nubilalis. As little was known about the biology of this parasitoid when reared onO. nubilalis, we undertook experiments to quantify biological parameters important to mass-rearing and ase ofT. ostriniae. When reared continuously onO. nubilalis, femaleT. ostriniae on average lived 2.7 days and produced 24 progeny. Continuous access to honey resulted in a four-fold increase in longevity and fecundity and a significant increase in the percentage of females parasitizing eggs. Rates of fecundity and parasitism decreased with age of female. Likewise, emergence rates and percentage of female progeny decreased with age of parental female.T. ostriniae successfully parasitizedO. nubilalis eggs until the blackhead stage. Most parasitism of eggs and eclosion of adults occurred during the first half of photophase. Eggs of 13 Lepidopterans were parasitized byT. ostriniae. Eggs of the Noctuidae, Pyralidae, and Plutellidae experienced higher levels of parasitism than others tested.T. ostriniae appears to be similar to other species ofTrichogramma in several respects and does not possess any characteristics that limit its potential for mass rearing and use for augmentative biological control ofO. nubilalis.     

Factors affecting sex ratio in rearing ofCoeloides sordidator (Hym.: Braconidae)

Five factors known to affect the sex ratio (% of males) in parasitic Hymenoptera were investigated forCoeloides sordidator, a parasitoid ofPissodes weevils. The host age, the age of ovipositing females, and the host of origin had a significant impact on the sex ratio of offspring. In contrast, the number of ovipositing females had an insignificant effect on sex ratio whereas the effect of host density could not be clearly defined. The sex ratio decreased with host age, probably because, like many other hymenopteran parasitoids, females tend to lay male eggs on small hosts and female eggs on larger hosts in order to maximize the size and fitness of their female offspring. The sex ratio also varied with the age of the mother, younger females laying more male eggs and older females more female eggs. The host of origin also had an influence on sex ratio. The strain fromPissodes castaneus was significantly more male-biased than the strain fromP. validirostris, which corroborates previous observations made on field populations     

Thiotepa, a reliable marker for sperm precedence measurement in a polyandrous moth

The determination of paternity of offspring produced by polyandrous females is essential for the understanding of sperm competition mechanisms. The sterile male technique using radiation is one of the most commonly employed methods for this purpose. However, sterilization using radiation is likely to be restricted by the equipment availability and cost. Chemosterilization may thus be a cheaper and easier alternative for sterilizing male insects in sperm competition studies. Here we report a reliable chemomarker, thiotepa (N,N′,N″-triethylenethiophosphoramide), for the study of sperm competition and precedence in a polyandrous moth, Ephestia kuehniella (Lepidoptera: Pyralidae). Dipping heads of male moths in 1% thiotepa aqueous solution for 10 s resulted in complete sterilization, i.e. their sperm still fertilized eggs but those eggs did not hatch. The sterilization treatment did not significantly affect male copulation ability, female fecundity, and sperm transfer, motility and fertilization. Our results indicate that 86% offspring of the twice-copulated females were fathered by the second males and 14% by the first males. Males treated with 5% thiotepa aqueous solution died within 24 h while those treated with 0.5% thiotepa were not fully sterilized.     

Isolation and Characterization of Sex-Linked Female-Sterile Mutants in DROSOPHILA MELANOGASTER

The purpose of the experiments described was to identify X chromosome genes functioning mainly or exclusively during oogenesis. Two mutagenesis experiments were carried out with ethyl methane sulfonate. Following treatment inducing 60% lethals, 9% of the treated X chromosomes carried a female sterility mutation which did not otherwise seriously affect viability. Among —95 isolated mutants, 19 were heat-sensitive and 5 cold-sensitive. The mutants have been classified as follows: I (16 mutants; 12 complementation groups): the females laid few or no eggs; the defect concerned either ovulation or oogenesis. II (37 mutants; 18 complementation groups): the female laid morphologically abnormal eggs, often with increased membrane permeability. III A (13 mutants; at least 8 complementation groups): the homozygous females were sterile if mated to mutant males; their progeny (homo- and hemizygous) died at a late embryonic stage (11 mutants), at the larval stage (1 mutant) or at the pupal stage (1 mutant). However fertility was partly restored by breeding to wild-type males as shown by survival of some heterozygous descendants. III B (29 mutants; 22 complementation groups): the fertility of the females was not restored by breeding to a wild-type male. Most of the eggs of 13 of the mutants died at a late stage of embryogenesis. The eggs of the others ceased development earlier or, perhaps, remained unfertilized. The distribution of the number of mutants per complementation group led to an estimation of a total of about 150 X-linked genes involved in female fertility. The females of three mutants, heat-sensitive and totally sterile at 29°, produced at a lower temperature descendants morphologically abnormal or deprived of germ cells. Three other mutants not described in detail showed a reduction in female fertility with many descendants lacking germ cells. A desirable mutant which was not recovered was one with normal fertile females producing descendants which, regardless of their genotype, bore specific morphological abnormalities. The value of the mutants isolated for analysis of the complex processes leading to egg formation and initiation of development is discussed.     

Requirement offlex (femalelethal onX) in the development of the female germ line ofDrosophila melanogaster

Drosophila melanogaster females homozygous forflex, an X-linked recessive mutation, do not survive. Hemizygous males are unaffected. Homozygous embryos appear to lack SXL, the product of theSex-lethal (Sxl) gene, apparently as a result of disruption ofSxl splicing. It is known that bothSxl and its somatic splicing regulators [snf andfl(2)d] also function in the development of the female germ line. For this reason, we investigated the role offlex in the germ line by generatingflex/flex clones inflex/+ females. Females carrying such clones in their germ lines do not lay eggs whereas females carryingflex+ eggs lay viable eggs. Additionally, DAPI staining of ovarioles showed that diploid germ cells that are homozygous mutant forflex do not complete oogenesis. These results indicate that theflex+ gene product may be required for the development of the female germ line.     

Glomus fasciculatum,a Weak Pathogen of Heterodera glycines

The occurrence ofchlamydospores of Glomus fasciculatum (Gf) within cysts of the soybean cyst nematode, Heterodera glycines, and the effects of vesicular-arbuscular mycorrhizae on nematode population dynamics and soybean (Glycine max) plant growth were investigated. Chlamydospores occupied 1-24% of cysts recovered from field soil samples. Hyphae of Missouri isolate Gfl penetrated the female nematode cuticle shortly after she ruptured the root epidermis. Convoluted hyphae filled infected eggs, and sporogenesis occurred within infected eggs. G. microcarpum, G. mosseae, and two isolates of Gf were inoculated with H. glycines on plants of ''Essex'' soybeans. Each of the two Gf isolates infected about 1% of the nematode eggs in experimental pot cuhures. The Gfl isolate decreased the number of first-generation adult females 26%, compared with the nonmycorrhizal control. The total numbers of first-generation plus second-generation adult females were similar for both Gf isolates and 29-41% greater than the nonmycorrhizal control. Soybean plants with Gf and H. glycines produced more biomass than did nonmycorrhizal plants with nematodes, but only Gfl delayed leaf senescence.     

The egg size determining gene,Esd, is a unique morphological marker on the W chromosome ofBombyx mori

Triploid females of the silkworm,Bombyx mori, were produced by crossing a tetraploid female with a male carrying asch (sex-linked chocolate) gene on its Z chromosomes. Since thesch gene expresses the color of reddish brown (chocolate) in newly hatched larvae, the females with ZWW chromosomes become chocolate, while those with ZZW have normal color (black). The effects of the W chromosome on oogenesis were studied by comparing these two types of 3n females. Despite the fact that the theoretical value of the ratio of chocolate larvae to black larvae should be 1:5, the actual ratio was found to be 1:80. The chocolate larvae revealed very low hatchability. Thirteen adult ZWW female moths were obtained, all laying normal-shaped eggs and even larger than those of 4n females which also have two W chromosomes. Eighty percent of the eggs of ZZW 3n females were irregular-shaped, and the size of the normal-shaped eggs was as large as that of 2n females which possess one W chromosome. It is hypothesized that egg size is determined by a gene on the W chromosome. The higher frequency of normal-shaped eggs in the ZWW female group than the ZZW female group suggests the presence of another kind of gene on the W chromosome, which leads to normal oogenesis.     

Observations onCheyletus cacahuamilpensis,a predator of the tenuipalpid miteDolichotetranychus floridanus

Aspects of the biology ofCheyletus cacahuamilpensis Baker a predatory mite of the associated lawn grass pestDolichotetranychus floridanus (Banks) were studied. Apart from a larval stage immature females ofC. cacahuamilpensis pass through 2 nymphal stages, while the male has only 1 before reaching adulthood. At 27°C the life cycle and adult longevity respectively averaged 30.1 & 34.4 days for the female and 22.8 & 16.4 days for the male. In a 20.4 days oviposition period 28 eggs were laid. Immature and adult females devoured an average of 80.4 & 143.2D. floridanus, while those of the males consumed 40.8 & 63.6 individuals respectively. Thus the total number ofD. floridanus attacked during the predator's life span averaged 223.6 per female and 104.4. per male.     

Reduction in fitness of female Asian longhorned beetle (Anoplophora glabripennis) infected with Metarhizium anisopliae

Bioassays were conducted to document the effects of Metarhizium anisopliae infection on adult female Asian longhorned beetle (Anoplophora glabripennis) reproduction before death and subsequent survival of offspring. The effect of infection on fecundity was evaluated for females already laying eggs and for newly eclosed females using M. anisopliae isolates ARSEF 7234 and 7711, respectively. Decreased longevity and oviposition compared with controls were observed in females that were already laying eggs when exposed to M. anisopliae ARSEF 7234. Newly eclosed females exposed to M. anisopliae ARSEF 7711 displayed shortened longevity (10.0 ± 0.7 days vs 74.3 ± 6.8 days for controls) and decreased oviposition (1.3 ± 0.7 eggs per ARSEF 7711-exposed female vs 97.2 ± 13.7 eggs per female for controls) compared with controls. Percentages of eggs that did not hatch were greater for both groups of fungal-treated females compared with controls and 60.0% of unhatched eggs contained signs of fungal infection. The percentage of larvae dying within 9 weeks of oviposition was higher for sexually mature females exposed to ARSEF 7234 compared with controls and >40% of dead larvae displayed signs of fungal infection. Thus, for both stages of females and both fungal isolates, fewer surviving larvae were produced after female fungal infection compared with controls. M. anisopliae infection affects female fitness by decreasing female longevity, by decreasing female oviposition before death and through horizontal transmission of M. anisopliae to offspring.     

Egg laying and host stage preference at constant temperatures inEncarsia tricolor [Hym.: Aphelinidae]

Encarsia tricolor Foërster is a facultative autoparasitoid that develops on the important pestTrialeurodes vaporariorum (Westwood) in outdoor crop conditions, which makes this aphelinid species promising for biological control programs in regions where field and protected crops coexist. In this paper we report the results obtained in the study of daily and totalE. tricolor egg laying and of adult female preference for different host stages in which to lay eggs at constant temperatures in the range 10 to 32 °C. Only whitefly nymphs were present in the searching arena (tomato leaflets). The mean number of eggs laid per female in one day ranged from 4.0 (10 °C and 32 °C) to 15.2 (24 °C). The mean total number of eggs increased with temperature from 10 to 28 °C, reaching a maximum of 123 eggs per female at 28 °C, and decreased sharply from 28 to 32 °C. The relation between the intrinsic rate of increase (r_m) and temperature in the range 10 to 28 °C followed a straight line whose equation was r_m=?0.076+0.011*T (R²=0.99). The r_m ofE. tricolor was greater than the r_m ofT. vaporariorum when temperature was higher than 9.2 °C. The preference for any particular host instar in which to lay eggs was not always significant. However, N4 was the host instar preferred whenever preference was statistically significant.     

Functional response and reproductive attributes of the aphidophagous ladybird beetle,Harmonia dimidiata (Fabricius) in oak trees of sericultural importance

Harmonia dimidiata (Fabricius) (Coleoptera: Coccinellidae) is the dominant predator of the aphid species Cervaphis quercus Takahashi. This aphid is a serious pest of oak trees in several parts of north-east India. Young leaves of oak trees are used in sericulture by rural people and by industry in several parts of north-east India. The effect of different aphid densities on food consumption and fecundity of H. dimidiata was studied in the laboratory. Female beetles were maintained from the time of eclosion till death at a fixed density of 25, 50, 75,100 or 125 aphids. Both the functional response and the reproductive numerical response showed the upper asymptote at 100 adult aphids/female. At this density, females matured earlier and produced more eggs over a longer reproductive period. At lower prey densities, females matured late and they either did not produce eggs or produced fewer eggs. At the higher prey densities, females did not produce more eggs than the asymptote. Results suggested that H. dimidiata are an effective predator of C. quercus aphids on oak trees and could be exploited as a biological control agent in the rising phase of aphid population growth.     

Sperm precedence in the dragonfly Erythemis simplicicollis

Females of the dragonfly Erythemis simplicicollis (Say) (Odonata, Libellulidae) store enough sperm to fertilize 6–13 clutches of eggs laid on consecutive days. Nonetheless, they usually mate one or more times per day. Males wait for females at ponds containing surface vegetation on which the females lay eggs. Some males defend vegetation while other act as satellites. After mating, both types of males attempt to guard females against takeover by other males. Sperm precedence by male E. simplicicollis was studied using sterility produced by gamma irradiation to label sperm. After a dose-response analysis, males receiving a dose of 25 kiloroentgens (>99.9% sterile) were returned to their home pond as territory residents and satellites. Both types of males fertilized an average of 99.5% (range 97.3–100%) of the female's remaining clutch. After mating with a sterile male, females were isolated in a large cage, and eggs collected for several consecutive days. These clutches revealed that sperm mixing in the bursa of the females is essentially complete after 24 to 48 h and that the last male to mate had replaced an average of more than 57–75% of the sperm stored by female from previous matings. Thus, the last sperm in is the first sperm out fertilizing essentially all of the eggs laid soon (5–6 min) after the mating. Sperm from the most recent mating competes for fertilizations with sperm stored from previous matings only if the female oviposits on the following day without remating.     

Proportion of mated females, female mating experience, and sex ratio of the osmund sawfly, Strongylogaster osmundae (Hymenoptera, Tenthredinidae)

The proportion of mated females (M _f) of the osmund sawfly, Strongylogaster osmundae, and the sex ratio of the eggs they deposited (r, proportion of males) were estimated in the wild by collecting egg masses. The proportion of mated females at oviposition varied from 0 to 1.0. M _f was high (often 1.0) among the females that emerged after hibernation, and lower in the subsequent generations. Mated females of the hibernated generation deposited equal numbers of eggs of both sexes. Mated females of the first and subsequent generations produced more female than male eggs. These results qualitatively agreed with the prediction provided by an evolutionarily stable strategy (ESS) model (if M _f < 1 then r < 0.5). However, the quantitative prediction provided by the model [M _f (1 − r) = 0.5] was not always observed in the wild, especially where the population density and M _f were high. The value of r was often lower than the predicted one. The following simple hypothesis was tested by experimentation: “Females that encounter males frequently estimate the proportion of mated females to be high and deposit eggs with a 1:1 sex ratio.” However, results did not support this hypothesis. Females that copulated soon after emergence and were courted by males two or more times did not show a higher offspring sex ratio than those which mated 1 or 2 days after emergence and experienced no other sexual encounter. Another mechanism for determination of r is suggested, and the reason why the population sex ratio of sawflies is often female-biased (r < 0.5) is discussed.     

Effect of hexythiazox and spiromesifen resistance on the life cycle of the predatory mite Neoseiulus californicus (Acari: Phytoseiidae)

Neoseiulus californicus (McGregor) (Acari: Phytoseiidae) is one of the most important and effective predators of Tetranychus spp. (Acari: Tetranychidae). This study analyzed the effects of hexythiazox and spiromesifen resistance on biological characteristics of N. californicus. Pre-oviposition, oviposition, post-oviposition times, adult lifespan, total number of eggs laid per female, net reproductive rate (R₀), intrinsic rate of increase (r_m), doubling time (DT), mean generation time (T) and finite rate of increase (λ) were compared among three populations: 63.9-fold hexythiazox resistant (HEX14), 53.6-fold spiromesifen resistant (SPR13) and a susceptible base population. Pre-oviposition and oviposition times, mean number of eggs/female, adult lifespan, R₀ and r_m values were all significantly higher for the two resistant populations than for the susceptible population. Life tables of HEX14, SPR13 and the susceptible population showed that R₀ was 35.0, 26.5 and 19.4 (females/female generation), r_m was 0.35, 0.32 and 0.30 (females/female/day), DT was 1.92, 2.07 and 2.27 days, T was 9.8, 9.8 and 9.7 days, and λ was 1.43, 1.39 and 1.35 (individuals/female/day), respectively.