The genetic basis of host range inHeliothis virescens: larval survival and growth

Larval feeding was assayed in a generalist caterpillar (Heliothis virescens (F.)), a specialist caterpillar (Heliothis subflexa (Gn.)), their F₁ hybrids and a backcross withH. s. to gain a preliminary understanding of the genetic basis of host use inH. v. Plants used in these experiments were tobacco (Nicotiana tabaccum), cotton (Gossypium hirsutum), soybean (Glycine max) (hosts ofH. v.) and ground cherry (Physalis pubescens) (host ofH.s.). A feeding study ofH.v., H.s. and their reciprocal hybrids showed that, after the first eight days of feeding,H.v. had its highest survival and weight gain on soybean and had lowest survival and weight gain on its non-host,Physalis. H.s. had its highest survival and weight gain on its host plant,Physalis, and performed very poorly on the three non-host plants. The two F₁ hybrids were SV₀ (offspring ofH.s. female andH.v. male) and VS₀ (offspring ofH.v. female andH.s. male). The hybrids did not differ from each other, indicating no sex linkage or maternal effects, except that VS₀ had greater weight gain on tobacco than did SV₀. The hybrids, unlike their parents, survived well on all four host plants and their weight gain was intermediate on all four host plants. In a separate experiment the VS₀ hybrid was mated to the specialist to produce the VS₁ backcross. In contrast to the F₁, the backcross had significantly lower survival than the generalist on soybean, cotton, tobacco and weight gain was lower on soybean, cotton and tobacco but higher onPhysalis. Survival and weight gain on cotton and tobacco were inherited as partially dominant traits; onPhysalis there was overdominance for survival and complete dominance for weight gain; on soybean both survival and weight gain were additive. Survival on cotton,Physalis, soybean and tobacco and weight gain onPhysalis could not be completely explained by a model that included only dominance and additive effects. These traits may be influenced by epistatic and/or environmental effects.     

Host plant direct defence against eggs of its specialist herbivore,Heliothis subflexa

1. The insect Heliothis subflexa Guenée is a specialist on plants in the genus Physalis. In the present study, the physical response of Physalis leaves to egg deposition by H. subflexa is described. 2. It was observed that the leaves of Physalis plants respond to the eggs of H. subflexa, while co‐occurring non‐host plants do not. Leaves of Physalis angulata L. and Physalis pubescens L. respond to H. subflexa eggs by the formation of (i) necrotic tissue, (ii) undifferentiated cells that form a bump (neoplasm) under the eggs of this herbivore, or (iii) both types of responses. 3. Greenhouse experiments showed that 64% of eggs laid on P. angulata elicited a response, and that a response to an egg decreased the probability of hatching. Further experiments in the field with P. angulata showed that the mean response to eggs by leaves was 31%, and that this response increased as temperature increased. Field experiments also confirmed that a plant response to an egg decreased the probability of hatching and increased the probability of removal from the plant by physical dislodgement or predation. 4. Eggs that elicited a response had a 25% lower probability of hatching and a 28% lower probability of remaining on the plant, resulting in an average fitness cost of 19.3% for H. subflexa. This is the first study to show an induced direct physical defence of a plant against eggs of a noctuid moth.     

THE GENETIC BASIS OF DIFFERENCES IN GROWTH AND BEHAVIOR OF SPECIALIST AND GENERALIST HERBIVORE SPECIES: SELECTION ON HYBRIDS OF HELIOTHIS VIRESCENS AND HELIOTHIS SUBFLEXA (LEPIDOPTERA)

Two species of moths and their hybrids were studied to determine the genetic basis of host range. One species, Heliothis virescens (HV), has a broad host range and is an agricultural pest on cotton (Malvaceae), soybean (Fabaceae), and tobacco (Solanaceae). The other species, Heliothis subflexa (HS), has a narrow host range, feeding on plants in the genus Physalis (Solanaceae). Experiments were done to determine whether the abilities to feed on cotton, soybean, and tobacco were each under separate genetic control in HV (genetically uncorrelated) or whether feeding on all three hosts was genetically correlated. By repeatedly backcrossing hybrids to HS while selecting for high larval survival and weight on soybean, loci conferring the ability to feed on soybean were moved from HV into the genetic background of the specialist, HS. After six generations of selection on soybean we tested the selected line on soybean, cotton, Physalis, and tobacco to determine if ability to feed on soybean was correlated with feeding ability on these other hosts. We found no evidence to support the hypothesis that feeding on all three hosts was correlated and controlled by a single locus. Compared to HS, the survival and weight of the selected line were significantly higher on soybean, similar on tobacco and Physalis, and slightly, but not significantly, higher on cotton. We also conducted a behavioral choice test to determine if larval preference for soybean was correlated with preference for other hosts. The selected line larvae had higher preference for soybean and cotton than HS. Preference for tobacco was equal in the selected line and in HS. These results support the trends seen in survival and growth tests where performances on soybean and cotton appeared partially correlated and performances on soybean and tobacco were uncorrelated. In conclusion, the generalist, HV, did not have a single set of loci that governed feeding on soybean, cotton and tobacco. It is likely that some of the loci governing performance and preference for soybean also contributed to performance and preference for cotton and that the soybean loci examined did not interact negatively to lower performance and preference for Physalis, the specialist's host. This approach can be applied more widely to address evolutionary questions about host range and other ecological traits.     

The role of alcohols in pheromone biosynthesis by two noctuid moths that use acetate pheromone components

Primary alcohols varying in chain length from C₁₃ to C₁₆, and in number, position, and geometric configuration of double bonds, were applied in dimethyl sulfoxide to the surface of the female sex pheromone glands of Heliothis subflexa (Gn.) and Hydraecia micacea (Esper). Capillary gas chromatographic analysis of extracts of the treated glands indicated that the alcohols were converted to the corresponding aldehydes by H. subflexa females and to the acetates by H. micacea females. Conversions of the alcohols showed no preferences for molecular weight, number, position, or geometry of the double bonds in either species. Application of the acetates of the primary alcohols to the gland surface of H. subflexa females resulted in the production of both the corresponding alcohols and aldehydes, while neither alcohols nor aldheydes were produced when acetates were applied to the glands of H. micacea. In addition, application of the acetates to the gland surface of Heliothis virescens (F.) resulted in the production of both the corresponding alcohols and aldehydes. However, no evidence was found to indicate that acetates are ever produced by the pheromone gland of females of H. virescens.     

Alkanes of four related moth species,Helicoverpa and Heliothis

Comparison of the presence and quantities of cuticular hydrocarbons has been used successfully for identifying sibling species and races of several groups of insects. This approach has been extended to four species of moths previously regarded as belonging to the same genus, Heliothis. Gas chromatography was used to quantify the numerous high-molecular weight alkanes found on the cuticle of two pairs of closely related species: Helicoverpa zea and Helicoverpa armigera, and Heliothis virescens and Heliothis subflexa. Both sexes of H. zea and H. armigera contained different quantities of several alkanes that could be used for unambiguous identification. Similar comparisons of H. subflexa and H. virescens showed four peak ratios that were different for each species. Sexual dimorphism was minor in H. subflexa and H. virescens.     

Association of a sperm-specific protein with the mitochondrial F1F0-ATPase in Heliothis: Implications for sterility of H. virescens × H. subflexa backcross hybrids

Evidence is presented that a mitochondrial protein that displays a species-specific net charge in the Heliothis spp complex is associated with subunits of the F₁-ATPase. The expression of this 63 kd polypeptide, p63, was restricted to sperm and its developmental pattern of synthesis and accumulation paralleled that of the putative β-subunit of the F₁-ATPase complex. Comparisons of the enzyme from fertile Heliothis virescens and sterile (H. virescens × H. subflexa) backcross hybrid males revealed two differences. First, the specific activity of the F₁-ATPase isolated from sterile males was half that of preparations from fertile males; and second, the p63 protein was bound less tightly to the complex in backcross sperm. The implications of these findings in relation to both the identification of the cause of backcross male sterility and prospects for future research are discussed.     

A comparison of responses from olfactory receptor neurons of<Emphasis Type="Italic"> Heliothis subflexa</Emphasis> and<Emphasis Type="Italic"> Heliothis virescens</Emphasis> to components of their sex pheromone

Single-cell electrophysiological recordings were obtained from olfactory receptor neurons in sensilla trichodea on male antennae of the heliothine species Heliothis subflexa and the closely related congener H. virescens. A large percentage of sensilla (72% and 81%, respectively, of all sensilla sampled) contained a single odor-responsive receptor neuron tuned to the major pheromone component of both species, Z-11-hexadecenal. A second population of sensilla on H. subflexa antennae (18%) housed receptor neurons that were tuned to Z-9-hexadecenal but also responded with less sensitivity to Z-9-tetradecenal. A similar population of sensilla (4%) on H. virescens male antennae housed receptor neurons that were shown to be tuned specifically only to Z-9-tetradecenal, with no response to even high dosages of Z-9-hexadecenal. A third population of sensilla (comprising 8% and 16% of the sensilla sampled in H. subflexa and H. virescens, respectively) housed two olfactory receptor neurons, one of which was tuned to Z-11-hexadecenyl acetate and the other tuned to Z-11-hexadecenol. In H. subflexa the Z-11-hexadecenyl acetate-tuned neuron also responded to Z-9-tetradecenal with nearly equivalent sensitivity. The behavioral requirements of males of these two species for distinct pheromonal blends was, therefore, reflected by the subtle differences in the tuning properties of antennal olfactory receptor neurons.Abbreviations MGC macroglomerular complex - ORN olfactory receptor neuron - Z9–14:Ald (Z)-9-tetradecenal - Z9–16:Ald (Z)-9-hexadecenal - Z11–16:Ac (Z)-11-hexadecenyl acetate - Z11–16:Ald (Z)-11-hexadecenal - Z11–16:OH (Z)-11-hexadecenol     

Effect of Heliothis subflexa herbivory on fruit abscission by Physalis species: the roles of mechanical damage and chemical factors

1. Insect oral secretions are important for the induction of a number of plant responses, but the relative role of mechanical damage in the induction of these responses is often not well understood. Damage from the frugivore Heliothis subflexa, a specialist on Physalis species, causes herbivore‐induced fruit abscission. In this field study, we examined the separate and combined effects of mechanical damage and H. subflexa oral secretions on Physalis fruit abscission. 2. To determine the relative role of mechanical and chemical factors, the following treatments were administered to fruit: (1) three levels of mechanical damage, (2) natural herbivore damage by control larvae and by larvae surgically treated to inhibit saliva secretion, and (3) injection of H.subflexa oral secretions and a water control. Abscission of mechanically damaged fruit with and without the addition of oral secretions was also compared. 3. Mechanical damage was sufficient to cause fruit abscission, and the addition of oral secretions to mechanically damaged fruit did not cause an increase in fruit abscission. Normal caterpillars and those treated to inhibit saliva secretion caused similar abscission rates. 4. Though most studies examining the effects of insect oral secretions on induced plant responses find these chemical stimuli to be important or essential, the results of the present study showed that oral secretions are not necessary for fruit abscission. Future work is needed to determine the relative importance of mechanical damage in herbivore‐induced plant responses in other systems.     

Role of the exoprotease InA in the pathogenicity of Bacillus thuringiensis in pupae of Hyalophora cecropia

Two geographical biotypes of Nomuraea rileyi (from Ecuador and the United States) were topically bioassayed against seven lepidopteran species, i.e., Anticarsia gemmatalis, Heliothis zea, Heliothis virescens, Heliothis subflexa, Pseudoplusia includens, Spodoptera exigua, and Trichoplusia ni. There was an average difference of 1.7-fold in mortality in how cultures of the same insect species from different sources responded to topical applications of either biotype of N. rileyi. Regression equations and LC₅₀ values were obtained for each insect species and fungal biotype combination. Larvae of S. exigua were equally susceptible to both biotypes of N. rileyi. Although larvae of A. gemmatalis were moderately susceptible to the Ecuadoran biotype, they were relatively nonsusceptible to the Mississippian biotype. Species of Heliothis (H. zea, H. virescens, and H. subflexa) were about equally susceptible to the Mississippian biotype. Larvae of H. subflexa and H. virescens, however, were significantly less susceptible than H. zea to the Ecuadoran biotype. When the integumental barrier was breached via intrahemocoelic injections, larvae of H. virescens were as susceptible as H. zea larvae to blastospores of either biotype of N. rileyi.     

Model for genetic control of Heliothis virescens

Summary A model was developed which corrects and extends an earlier one proposed for the control of the tobacco budworm, Heliothis virescens (F.), through hybrid male sterility. Population suppression is effected through the release into natural populations of the backcross progeny of a hybrid between H. virescens and a related species. Thereafter, the system perpetuates itself in nature through continual backcrossing of the fertile backcross females to native H. virescens males. When the proportion of backcross hybrid females in the total population is large enough to draw off the insemination potential of the native males, the native females fail to replace themselves. The present model demonstrated that the ratio of released backcross hybrids to natural H. virescens remains constant in a closed population. Furthermore it was shown that the release ratio necessary to achieve extinction of a closed population is related to the number of females that a male can inseminate and to the population growth rate. Release ratios required to slow natural population growth and to lessen the impact damage of releases on crop plants were also examined. Effects of selection against the backcross females on the predictions of the model were explored.     

The role of herbivore‐ and plant‐related experiences in intraspecific host preference of a relatively specialized parasitoid

Parasitoids use odor cues from infested plants and herbivore hosts to locate their hosts. Specialist parasitoids of generalist herbivores are predicted to rely more on herbivorederived cues than plant-derived cues. Microplitis croceipes (Cresson)(Hymenoptera: Braconidae) is a relatively specialized larval endoparasitoid of Heliothis virescens (F.)(Lepidoptera: Noctuidae), which is a generalist herbivore on several crops including cotton and soybean. Using M. croceipes/H. virescens as a model system, we tested the following predictions about specialist parasitoids of generalist herbivores:(i) naive parasitoids will show innate responses to herbivore-emitted kairomones, regardless of host plant identity and (ii) herbivore-related experience will have a greater influence on intraspecific oviposition preference than plant-related experience. Inexperienced (naive) female M. croceipes did not discriminate between cotton-fed and soybean-fed H. virescens in oviposition choice tests, supporting our first prediction. Oviposition experience alone with either host group influenced subsequent oviposition preference while experience with infested plants alone did not elicit preference in M. croceipes, supporting our second prediction. Furthermore, associative learning of oviposition with host-damaged plants facilitated host location. I terestingly, naive parasitoids attacked more soybeathan cotton-fed host larvae in two-choice tests when a background of host-infested cotton odor was supplied, and vice versa. This suggests that plant volatiles may have created an olfactory contrast effect. We discussed ecological significance of the results and concluded that both plant- and herbivore-related experiences play important role in parasitoid host foraging.     

Mapping quantitative trait loci controlling seed dormancy and heading date in rice, Oryza sativa L., using backcross inbred lines

 To detect quantitative trait loci (QTLs) controlling seed dormancy, 98 BC₁F₅ lines (backcross inbred lines) derived from a backcross of Nipponbare (japonica)/Kasalath (indica)//Nipponbare were analyzed genetically. We used 245 RFLP markers to construct a framework linkage map. Five putative QTLs affecting seed dormancy were detected on chromosomes 3, 5, 7 (two regions) and 8, respectively. Phenotypic variations explained by each QTL ranged from 6.7% to 22.5% and the five putative QTLs explained about 48% of the total phenotypic variation in the BC₁F₅ lines. Except for those of the QTLs on chromosome 8, the Nipponbare alleles increased the germination rate. Five putative QTLs controlling heading date were detected on chromosomes 2, 3, 4, 6 and 7, respectively. The phenotypic variation explained by each QTL for heading date ranged from 5.7% to 23.4% and the five putative QTLs explained about 52% of the total phenotypic variation. The Nipponbare alleles increased the number of days to heading, except for those of two QTLs on chromosomes 2 and 3. The map location of a putative QTL for heading date coincided with that of a major QTL for seed dormancy on chromosome 3, although two major heading-date QTLs did not coincide with any seed dormancy QTLs detected in this study. Received: 10 October 1997 / Accepted: 12 January 1998     

Validation of reference-assisted assembly using existing and novel Heliothine genomes

Chloridea subflexa and Chloridea virescens are a pair of closely related noctuid species exhibiting pheromone-based sexual isolation and divergent host plant preferences. We produced a novel Illumina short read C. subflexa genome assembly and an improved C. virescens genome assembly, which offer opportunities to study the genomic basis for evolutionarily important traits in this lepidopteran family with few genomic resources. We then examined the feasibility of reference-assisted assembly, an approach that leverages existing high quality genomic resources for genome improvement in closely related taxa and applied it to our Heliothine genomes. Our work demonstrates that reference-assisted assembly has the potential to enhance contiguity and completeness of existing insect genomic resources with minimal additional laboratory costs. We conclude by discussing both the potential and pitfalls of reference-assisted assembly according to the intended downstream assembly application.     

Estimates of marker-associated QTL effects in Monte Carlo backcross generations using multiple regression

Summary The decision of whether or not to use QTLassociated markers in breeding programs needs further information about the magnitude of the additive and dominance effects that can be estimated. The objectives of this paper are (1) to apply some of the Moreno-Gonzalez (1993) genetic models to backcross simulation data generated by the Monte Carlo method, and (2) to get simulation information about the number of testing progenies and mapping density in relation to the magnitude of gene effect estimates. Results of the Monte Carlo study show that the stepwise regression analysis was able to detect relatively small additive and dominance effects when the QTL are independently segregating. When testing selfed families derived from backcross individuals, dominance effects had a larger error standard deviation and were estimated at a lower frequency. Linked QTL require a higher marker mapping density on the genome and a larger number of progenies to detect small genetic effects. Reduction of the environmental error variance by evaluating selfed backcross families in replicate experiments increased the power of the test. Expressions of the number of progenies for detecting significant additive effects were developed for some genetic situations. The ratio of the within-backcross genetic variance to the square of a gene effect estimate is a function of the number of progenies, the heritability of the trait, the marker map density and the portion of the genetic variance explained by the model. Different values (from 0 to 1) assigned to (relative position of the QTL in the marker segment) did not cause a large shift in the residual mean square of the model.     

Field evaluation of insect resistance in a wild tomato and its effects on insect parasitoids

Populations ofHelicoverpa (=Heliothis) zea (Boddie),Heliothis virescens (F.),Manduca sexta (L.) andM. quinquemaculata (Haw.) and their egg and larval parasitoids were sampled in field plots of the: insect-resistant wild tomato,Lycopersicon hirsutum f. glabratum C. H. Mull, accession PI 134417; susceptible commercial tomato cultivar ‘Better Boy’; F₁ hybrid; and selected, moderately resistant backcross genotype. Densities ofH. zea andH. virescens eggs and small larvae were higher on resistant genotypes than on susceptible genotypes, but densities of large larvae were similar on all genotypes. Densities ofManduca spp. larvae were too low to permit similar analyses of the effects of plant genotype. Rates of egg parasitism byTrichogramma spp. andTelenomus sphingis (Ashmead) were reduced on insect-resistant genotypes. Rates of parasitism by the larval parasitoidsCampoletis sonorensis (Cameron) andCotesia congregata (Say) were reduced on resistant genotypes. No consistent effects on parasitism rates byCotesia marginiventris (Cresson) were observed and parasitism rates byCardiochiles nigriceps Viereck were unaffected.     

QTL analysis of potato tuber dormancy

The potential loss of chemical sprout inhibitors because of public concern over the use of pesticides underscores the desirability of breeding for long dormancy of potato (Solanum tuberosum L.) tubers. Quantitative trait locus (QTL) analyses were performed in reciprocal backcrosses between S. tuberosum and S. berthaultii toward defining the complexity of dormancy. S. berthaultii is a wild Bolivian species characterized by a short-day requirement for tuberization, long tuber dormancy, and resistance to several insect pests. RFLP alleles segregating from the recurrent parents as well as from the interspecific hybrid were monitored in two segregating progenies. We detected QTLs on nine chromosomes that affected tuber dormancy, either alone or through epistatic interactions. Alleles from the wild parent promoted dormancy, with the largest effect at a QTL on chromosome 2. Long dormancy appeared to be recessive in the backcross to S. berthaultii (BCB). In BCB the additive effects of dormancy QTLs accounted for 48% of the measured phenotypic variance, and adding epistatic effects to the model explained only 4% more. In contrast, additive effects explained only 16% of the variance in the backcross to S. tuberosum (BCT), and an additional 24% was explained by the inclusion of epistatic effects. In BCB variation at all QTLs detected was associated with RFLP alleles segregating from the hybrid parent; in BCT all QTLs except for two found through epistasis were detected through RFLP alleles segregating from the recurrent parent. At least three dormancy QTLs mapped to markers previously found to be associated with tuberization in these crosses.Paper number 55 of the Department of Fruit and Vegetable Science, Cornell University     

Detection and verification of malting quality QTLs using wild barley introgression lines

A malting quality quantitative trait locus (QTL) study was conducted using a set of 39 wild barley introgression lines (hereafter abbreviated with S42ILs). Each S42IL harbors a single marker-defined chromosomal segment from the wild barley accession ‘ISR 42-8’ (Hordeum vulgare ssp. spontaneum) within the genetic background of the elite spring barley cultivar ‘Scarlett’ (Hordeum vulgare ssp. vulgare). The aim of the study was (1) to verify genetic effects previously identified in the advanced backcross population S42, (2) to detect new QTLs, and (3) to identify S42ILs exhibiting multiple QTL effects. For this, grain samples from field tests in three different environments were subjected to micro malting. Subsequently, a line × phenotype association study was performed with the S42ILs in order to localize putative QTL effects. A QTL was accepted if the trait value of a particular S42IL was significantly (P < 0.05) different from the recurrent parent as a control, either across all tested environments or in a particular environment. For eight malting quality traits, altogether 40 QTLs were localized, among which 35 QTLs (87.5%) were stable across all environments. Six QTLs (15.0%) revealed a trait improving wild barley effect. Out of 36 QTLs detected in a previous advanced backcross QTL study with the parent BC₂DH population S42, 18 QTLs (50.0%) could be verified with the S42IL set. For the quality parameters α-amylase activity and Hartong 45°C, all QTLs assessed in population S42 were verified by S42ILs. In addition, eight new QTL effects and 17 QTLs affecting two newly investigated traits were localized. Two QTL clusters harboring simultaneous effects on eight and six traits, respectively, were mapped to chromosomes 1H and 4H. In future, fine-mapping of these QTL regions will be conducted in order to shed further light on the genetic basis of the most interesting QTLs.