Delimiting Strategic Zones for the Development of Fall Armyworm (Lepidoptera: Noctuidae) on Corn in the State of Florida.

The fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), cannot survive prolonged periods of freezing temperatures, thereby limiting where it can overwinter in North America. Climate change is anticipated to reduce the frequency of freeze days in Florida over the decades, with the potential consequence of a significant expansion of the overwintering range, whose northern limit in North America was assessed between 27 and 28°N in the last century. To assess this possibility, the development of the fall armyworm on corn leaves, one of the main host plants in the United States, was determined at five constant temperatures ranging from 14 to 30°C. Based on the development time, the thermal constant and the lower threshold temperature were used to estimate the number of generations of fall armyworm at 42 locations in the state of Florida, from 2006 to 2016. Maps were constructed to provide a visual description of the interpolated data, using GIS (Geographic Information System). The highest number of generations was observed in the counties farther south, an area that showed the highest temperatures during the years and plays a strategic role in maintaining fall armyworm populations in corn fields. Additionally, we conclude that in the absence of freeze periods, the northern limit for fall armyworm overwintering should be between 28 and 29°N.

Using intron sequence comparisons in the triose-phosphate isomerase gene to study the divergence of the fall armyworm host strains.

The noctuid moth Spodoptera frugiperda (the fall armyworm) is endemic to the Western Hemisphere and appears to be undergoing sympatric speciation to produce two subpopulations that differ in their choice of host plants. The 'rice strain' and 'corn strain' are morphologically indistinguishable, requiring the use of genetic markers for identification. Because fall armyworm is a major pest of corn and several other agricultural crops, characterizing the strains has important economic consequences. In this study, comparisons were made of the intron sequences from the triose-phosphate isomerase (Tpi) gene isolated from 85 fall armyworm specimens collected from two host plants. Sixteen new strain-specific haplotypes based on intron polymorphisms are described that can facilitate the characterization of fall armyworm populations associated with different host plants. Comparisons of genetic diversity within and between the strains provides evidence that the corn strain is undergoing active selection and supports the proposal of directional interstrain mating occurring in the wild. Comparisons of the polymorphisms indicate that each intron undergoes different patterns of mutation that in some cases corresponds to host plant preferences. The results confirm that intron sequence comparisons are an effective approach to study fall armyworm population genetics.

Modeling seasonal migration of fall armyworm moths.

Fall armyworm, Spodoptera frugiperda (J.E. Smith), is a highly mobile insect pest of a wide range of host crops. However, this pest of tropical origin cannot survive extended periods of freezing temperature but must migrate northward each spring if it is to re-infest cropping areas in temperate regions. The northward limit of the winter-breeding region for North America extends to southern regions of Texas and Florida, but infestations are regularly reported as far north as Québec and Ontario provinces in Canada by the end of summer. Recent genetic analyses have characterized migratory pathways from these winter-breeding regions, but knowledge is lacking on the atmosphere's role in influencing the timing, distance, and direction of migratory flights. The Hybrid Single-Particle Lagrangian Integrated Trajectory (HYSPLIT) model was used to simulate migratory flight of fall armyworm moths from distinct winter-breeding source areas. Model simulations identified regions of dominant immigration from the Florida and Texas source areas and overlapping immigrant populations in the Alabama-Georgia and Pennsylvania-Mid-Atlantic regions. This simulated migratory pattern corroborates a previous migratory map based on the distribution of fall armyworm haplotype profiles. We found a significant regression between the simulated first week of moth immigration and first week of moth capture (for locations which captured ≥ 10 moths), which on average indicated that the model simulated first immigration 2 weeks before first captures in pheromone traps. The results contribute to knowledge of fall armyworm population ecology on a continental scale and will aid in the prediction and interpretation of inter-annual variability of insect migration patterns including those in response to climatic change and adoption rates of transgenic cultivars.

Development and feeding of fall armyworm on Miscanthus x giganteus and switchgrass.

Observations of fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), larvae infesting plots of Miscanthus x giganteus Greef and Deuter ex Hodkinson and Renvoize prompted laboratory-based tests of survival, development, and feeding preferences on leaf tissue from M. x giganteus and switchgrass, Panicum virgatum L. Survival from hatch to pupation was >70 and 50% for fall armyworms reared on switchgrass and M. x giganteus, respectively, although survival of the S. frugiperda rice strain was significantly greater than the corn strain on both crops. Developmental times from hatch to pupation or adult emergence showed effects of crop and S. frugiperda host strain, but analysis of an interaction revealed developmental times for the rice strain were similar on both crops, whereas corn strain larvae showed delayed development on M. x giganteus relative to switchgrass. Analysis of larval (10 d) and pupal masses showed a similar pattern, with effects of crop and an interaction (at 10 d), but only the mass of corn strain larvae feeding on M. x giganteus was reduced relative to the other crop and strain combinations. In choice tests, neonates of both corn and rice strains showed a strong preference for feeding on young tissues rather than mature leaves of M. x giganteus or switchgrass, but they also clearly favored corn, Zea mays L., leaves over either of the perennial grasses. Results indicate both plants are potential hosts for S. frugiperda, but additional information is needed to understand under which scenarios and to what degree fall armyworms may damage perennial grasses grown for biofuel production.

Fall armyworm FR sequences map to sex chromosomes and their distribution in the wild indicate limitations in interstrain mating.

The fall armyworm, Spodoptera frugiperda, consists of two host strains (rice and corn) that differ in developmental, physiological and behavioural characteristics. However, because the strains are morphologically indistinguishable the investigation of strain-specific behaviour, particularly in the wild, is very difficult. This has spurred the isolation of diagnostic molecular markers. FR sequences are tandem-repeat genetic elements found in large clusters only in the rice strain. To facilitate their use as a strain marker we genetically mapped FR clusters and found they localized to the sex chromosomes. This represents one of the first examples of chromosome mapping in fall armyworm. The FR sequence and a strain-specific mitochondrial marker were then used to examine the distribution of different marker combinations in field specimens. These studies identified significant barriers to interstrain mating in the wild, specifically that corn strain females rarely, if ever, mate with rice strain males. The data also suggest that only a genetically distinct subset of the overwintering rice strain population in Florida annually migrate to Georgia. These studies demonstrate that the availability of genetically characterized molecular markers for strain identity makes possible studies on fall armyworm biology in the wild previously considered unfeasible.

The involvement of ovarian tumour in the intracellular localization of Sex-lethal protein.

The Drosophila ovarian tumour gene is required at multiple times in the germline for oogenesis. A second gene, Sex-lethal, controls sex determination in the soma and also has a separate germline function affecting similar oogenic stages as ovarian tumour. We demonstrate that ovarian tumour is not required for early Sex-lethal gene expression in the female germline, as had been previously reported. Instead, we provide evidence that ovarian tumour has a specific role in the developmentally regulated accumulation of SEX-LETHAL protein within the cytoplasm and nucleus. Furthermore, the examination of nurse cell polytene chromosomes produced by certain ovarian tumour mutations showed that SEX-LETHAL protein can associate with discrete chromosomal sites in the germline and that this pattern appears to change as the egg chamber matures. This is the first indication that SEX-LETHAL is capable of direct physical interactions with chromosomes (albeit in a mutant background) and is consistent with the developmentally regulated nuclear localization of SEX-LETHAL being important for oogenesis.

Regulatory and functional interactions between ovarian tumor and ovo during Drosophila oogenesis.

The ovo and ovarian tumor genes are required during early and late stages of Drosophila oogenesis. The ovo product, a zinc-finger transcription factor, can bind to sites and influence the level of expression of the ovarian tumor promoter. Our examination of ovo null mutant organelles demonstrate that it is required for the differentiation of XX germ cells during larval gonial stages, in addition to its known role in maintaining germ cell numbers. In contrast, ovarian tumor is required during pupal and adult stages for the cystocyte divisions that give rise to the egg chamber. Studies on sexually transformed flies indicate that both the ovo and ovarian tumor null mutant phenotypes are distinctive from and more severe than the germline defects produced when male germ cells develop in female soma. This suggests that ovo and ovarian tumor have oogenic functions other than their putative role in germline sex determination. We also demonstrate that the regulation of ovarian tumor by ovo is stage-specific, as ovarian tumor promoter activity does not require ovo during larval stages but becomes ovo-dependent in the adult ovary. This coincides with when the ovarian tumor promoter becomes responsive to sex-specific signals from the soma suggesting a convergence of somatic and germline regulatory pathways on ovarian tumor during oogenesis.

Induction of apoptosis in the germline and follicle layer of Drosophila egg chambers.

The reaper and head involution defective genes can induce apoptotic death in several Drosophila cell types, including portions of the embryo and eye. By a combination of FLP recombinase and the yeast Gal4/UAS transcription activation system, we expressed both cell death genes in discrete clones in the adult ovarian follicle cell layer. The expression of either reaper or head involution defective induced follicle cell apoptosis during all oogenic stages. Unexpectedly, the disruption of the follicle layer led to the induced degeneration of the nurse cells in an apoptotic manner, demonstrating a germline-somatic interaction required for germ cell viability. The germline apoptosis initiates at a specific time in oogenesis, coinciding with the beginning of vitellogenesis. This observation is intriguing given previous suggestions of a process to eliminate defective egg chambers at these same oogenic stages. The induce germline degeneration initiates with the transient formation of a network of filamentous actin around the nurse cell nucleus, in close association with a product of the adducin-related hu-li tai shao gene. This was immediately followed by nuclear condensation and DNA fragmentation, both characteristics diagnostic of apoptosis. Occurring concomitantly with the nuclear phenotypes were the disorganization of ring canals, and the degradation of Armadillo protein (a beta-catenin homolog) and filamentous actin. Germ cells degenerating as a normal consequence of oogenesis displayed a similar set of phenotypes, suggesting that a common apoptotic mechanism may underlie these different germline death phenomena.

Regulatory and functional interactions between the somatic sex regulatory gene transformer and the germline genes ovo and ovarian tumor.

In Drosophila, compatibility between the sexually differentiated state of the soma and the sex chromosome constitution of the germline is required for normal gametogenesis. In this study, we defined important aspects of the soma-germline interactions controlling early oogenesis. In particular, the sex-specific germline activity of the ovarian tumor promoter was found to be dependent upon somatic factors controlled by the somatic sex differentiation gene transformer. This regulation defines whether there is sufficient ovarian tumor expression in adult XX germ cells to support oogenesis. In addition, the ovarian tumor function required for female germline differentiation is dependent on the activity of another germline gene, ovo, whose regulation is transformer-independent. These and other data indicate that ovarian tumor plays a central role in coordinating regulatory inputs from the soma (as regulated by transformer) with those from the germline (involving ovo). We also demonstrate that transformer-dependent interactions influence whether XX germ cells require ovarian tumor or ovo functions to undergo early gametogenic differentiation. These results are incorporated into a model hypothesizing that the functions of ovarian tumor and ovo are dependent on an early sex determination decision in the XX germline that is at least partially controlled by somatic transformer activity.

The Drosophila ovarian tumor gene is required for the organization of actin filaments during multiple stages in oogenesis.

The ovarian tumor gene is required during both early and late stages of oogenesis. Mutations produce a range of phenotypes, including agametic ovarioles, tumorous egg chambers, and late stage oogenic arrest. We demonstrate that each of these phenotypes is associated with specific aberrations in actin distribution. In the earliest case, ovarian tumor mutations cause actin filaments to accumulate ectopically in the fusome. This correlates with abnormal fusome morphology and arrested germ cell development in the germaria. Similarly, ovarian tumor function is required for the localization of actin that is essential for the maturation of ring canals. This defect gives rise to tumorous egg chambers in which germ cell numbers and morphology are profoundly aberrant. We also confirm that ovarian tumor is required for the formation of the nurse cell cytoplasmic actin array that is essential for the nonspecific transport of cytoplasmic contents to the oocyte during late oogenesis. Our data suggest that at this stage ovarian tumor controls the site where actin filaments initiate. Taken together, these studies suggest that the diverse ovarian tumor mutant phenotypes derive from the mislocalization of actin filaments, indicating a role for this gene in organizing the female germline cytoskeleton, and that the misregulation of actin can have profound effects on germ cell division and differentiation.

A P element containing suppressor of hairy-wing binding regions has novel properties for mutagenesis in Drosophila melanogaster.

P elements are widely used as insertional mutagens to tag genes, facilitating molecular cloning and analyses. We modified a P element so that it carried two copies of the suppressor of Hairy-wing [su(Hw)] binding regions isolated from the gypsy transposable element. This transposon was mobilized, and the genetic consequences of its insertion were analyzed. Gene expression can be altered by the su(Hw) protein as a result of blocking the interaction between enhancer/silencer elements and their promoter. These effects can occur over long distances and are general. Therefore, a composite transposon (SUPor-P for suppressor-P element) combines the mutagenic efficacy of the gypsy element with the controllable transposition of P elements. We show that, compared to standard P elements, this composite transposon causes an expanded repertoire of mutations and produces alleles that are suppressed by su(Hw) mutations. The large number of heterochromatic insertions obtained is unusual compared to other insertional mutagenesis procedures, indicating that the SUPor-P transposon may be useful for studying the structural and functional properties of heterochromatin.

Developmental analysis of the ovarian tumor gene during Drosophila oogenesis.

Severe alleles of the ovarian tumor (otu) and ovo genes result in female sterility in Drosophila melanogaster, producing adult ovaries that completely lack egg chambers. We examined the developmental stage in which the agametic phenotype first becomes apparent. Germ cell development in embryos was studied using a strategy that allowed simultaneous labeling of pole cells with the determination of embryonic genotype. We found that ovo- or otu- XX embryonic germ cells were indistinguishable in number and morphology from those present in wild-type siblings. The effects of the mutations were not consistently manifested in the female germline until pupariation, and there was no evidence that either gene was required for germ cell viability at earlier stages of development. The requirement for otu function in the pupal and adult ovary is supported by temperature-shift experiments using a heat-inducible otu gene construct. We demonstrate that otu activity limited to prepupal stages was not sufficient to support oogenesis, while induction during the pupal and adult periods caused suppression of the otu mutant phenotype.

Genetic characterization of small ovaries, a gene required in the soma for the development of the Drosophila ovary and the female germline.

The small ovary gene (sov) is required for the development of the Drosophila ovary. Six EMS-induced recessive alleles have been identified. Hypomorphic alleles are female sterile and have no effect on male fertility, whereas more severe mutations result in lethality. The female-sterile alleles produce a range of mutant phenotypes that affect the differentiation of both somatic and germline tissues. These mutations generally produce small ovaries that contain few egg cysts and disorganized ovarioles, and in the most extreme case no ovarian tissue is present. The mutant egg cysts that develop have aberrant morphology, including abnormal numbers of nurse cells and patches of necrotic cells. We demonstrate that sov gene expression is not required in the germline for the development of functional egg cysts. This indicates that the sov function is somatic dependent. We present evidence using loss-of-function and constitutive forms of the somatic sex regulatory genes that sov activity is essential for the development of the somatic ovary regardless of the chromosomal sex of the fly. In addition, the genetic mapping of the sov locus is presented, including the characterization of two lethal sov alleles and complementation mapping with existing rearrangements.

The somatic sex determines the requirement for ovarian tumor gene activity in the proliferation of the Drosophila germline.

Gametogenesis in Drosophila requires sex-specific interactions between the soma and germline to control germ cell viability, proliferation, and differentiation. To determine what genetic components are involved in this interaction, we examined whether changes in the sexual identity of the soma affected the function of the ovarian tumor (otu) and ovo genes. These genes are required cell autonomously in the female germline for germ cell proliferation and differentiation. Mutations in otu and ovo cause a range of ovarian defects, including agametic ovaries and tumorous egg cysts, but do not affect spermatogenesis. We demonstrate that XY germ cells do not require otu when developing in testes, but become dependent on otu function for proliferation when placed in an ovary. This soma-induced requirement can be satisfied by the induced expression of the 98 x 10(3) M(r) OTU product, one of two isoforms produced by differential RNA splicing. These results indicate that the female somatic gonad can induce XY germ cells to become 'female-like' because they require an oogenesis-specific gene. In contrast, the requirement for ovo is dependent on a cell autonomous signal derived from the X:A ratio. We propose that differential regulation of the otu and ovo genes provides a mechanism for the female germline to incorporate both somatic and cell autonomous inputs required for oogenesis.

Molecular characterization of ovarian tumors in Drosophila.

Certain female-sterile mutations in Drosophila result in the uncontrolled proliferation of X/X germ cells. It has been proposed that this ovarian tumor phenotype results from the sexual transformation of X/X germ cells to a male identity. We present findings inconsistent with this model. We demonstrate that the tumorous cells produced by mutations in the ovarian tumor (otu), Sex-lethal (Sxl) and sans fille (snf) genes are capable of female-specific transcription and RNA processing. This indicates that these ovarian tumor cells still retain some female identity. Therefore, we propose that mutations in these genes do not cause a male transformation of the X/X germ line but instead either cause an ambiguous sexual identity or block specific stages of oogenesis. Our findings indicate that while Sxl is the master sex determination gene in somatic cells, it appears to play a more subsidiary role in the germ line. Finally, we demonstrate that the germ line function of Sxl depends on the activity of a specific OTU isoform.

Genetic and molecular characterization of P element-induced mutations reveals that the Drosophila ovarian tumor gene has maternal activity and a variable null phenotype.

The mutations in the ovarian tumor (otu) gene arrest oogenesis at several stages in development. A series of deletion mutations in the otu region were characterized, each of which causes the absence or reduction of the otu transcript. These alleles range from the most severe class, which results in ovaries lacking egg cysts, to relatively mild mutations that allow the development of late stage oocytes. Heteroallelic combinations of these mutations demonstrate that the phenotypic complexity of otu mutant ovaries is due to a dosage dependent requirement for otu activity. Reciprocal cross and developmental Northern blot studies suggest a maternal requirement for otu in the development of the female germline. In addition we demonstrate that the otu zygotic null phenotype is variable, ranging from the absence of cysts in the most extreme cases, to the presence of tumorous egg chambers.

Regulation of sex-specific RNA splicing at the Drosophila doublesex gene: cis-acting mutations in exon sequences alter sex-specific RNA splicing patterns.

Sex-specific alternative RNA splicing of the doublesex (dsx) pre-mRNA results in sex-specific polypeptides that regulate both male and female somatic sexual differentiation in Drosophila melanogaster. We have molecularly characterized a class of dsx mutations that act in cis to disrupt the regulation of dsx RNA processing, causing the dsx pre-mRNA to be spliced in the male-specific pattern regardless of the chromosomal sex of the fly. These dsx mutations are associated with rearrangements in the female-specific exon just 3' to the female-specific splice acceptor. The mutations do not affect the female-specific splice sites or intron that are identical to wild-type sequences. These results indicate that sequences in the female-specific exon are important for the regulation of sex-specific RNA splicing, perhaps by acting as sites of interaction with trans-acting regulators. Furthermore, the data suggest that female-specific regulation of dsx RNA processing occurs by promoting the usage of the female splice acceptor site, rather than by repressing the usage of the alternative male-specific splice acceptor.

The control of alternative splicing at genes regulating sexual differentiation in D. melanogaster.

The transformer (tra) and doublesex (dsx) genes produce sex-specific transcripts that are generated by differential RNA processing. We have examined the effects of mutants in other regulatory genes controlling sexual differentiation on the patterns of processing of the tra and dsx RNA transcripts. Our results demonstrate that the genes suggested by genetic studies to act upstream of tra or dsx in the sex determination hierarchy regulate these two loci at the level of RNA processing. Our data suggest that the order of interaction of the factors controlling sex is X:A greater than Sxl greater than tra greater than tra-2 greater than dsx greater than or equal to ix greater than terminal differentiation. While these results cannot preclude regulatory interactions at other levels, the regulation of RNA splicing revealed by these experiments is sufficient to account for all of the known functional interactions between the regulatory genes in this hierarchy.

Molecular and recombinational mapping of mutations in the Ace locus of Drosophila melanogaster.

The Ace locus in Drosophila melanogaster is known to be the structural gene for acetylcholinesterase. Ace is located in a region of chromosome arm 3R which has been subjected to intensive genetic and molecular analysis. Previous deletion mapping studies have identified a 40-kb region within which the Ace gene resides. This report focuses on the further localization of Ace within this 40-kb interval. Within this region, selective fine structure recombinational analysis was employed to localize three recessive Ace lethals relative to unselected restriction site variations. These three mutations fall into a segment of 7 kb within the Ace interval. Fine structure recombinational analysis was also used to confirm that the Ace phenotype of one deletion, Df(3R)AceHD1, co-segregated with the molecular deletion. This deletion does not fully remove Ace activity, but it behaves as a recessive Ace lethal. Df(3R)AceHD1 is the most distal Ace lesion identified and indicates that the Ace locus must extend at least 16 kb. Several poly(A)transcripts are detectable in the region defined by the Ace lesions. The position and extent of the Ace locus, as well as the types of transcripts found, is consistent with the recent findings which identified Torpedo-AChE homologous cDNA sequences in this region.