Impact of Lepidoptera (Crambidae, Noctuidae, and Pyralidae) Pests on Corn Containing Pyramided Bt Traits and a Blended Refuge in the Southern United States.

Blended refuge for transgenic plants expressing Bacillus thuringiensis (Bt) toxins has been approved in the northern United States as a resistance management strategy alternative to a structured refuge. A three-year study (2012-2014) was conducted with 54 trials across nine states in the southern United States to evaluate plant injury from lepidopteran pests of corn and yield in a corn hybrid expressing Cry1F × Cry1Ab × Vip3Aa20 (Pioneer Brand Optimum Leptra) planted as a pure stand and in refuge blends of 5, 10, and 20% in both early and late plantings. Injury by corn earworm, Helicoverpa zea Boddie (Lepidoptera: Noctuidae), and fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), was generally proportional to the percentage of non-Bt corn within each refuge blend. Across locations, ear injury in plots with 100% Cry1F × Cry1Ab × Vip3Aa20 (Optimum Leptra) corn ranged from no injury to a maximum of 0.42 cm(2) per ear in Mississippi in 2013. Leaf injury ratings in 100% non-Bt plots in early and late planted trials in 2014 were 86- and 70-fold greater than in 100% Cry1F × Cry1Ab × Vip3Aa20 (Optimum Leptra) plots. Plants in plots with blended refuges had significantly greater leaf injury in 2012 (5, 10, and 20% refuge blends), in the early-planted corn in 2013 (10 and 20% only), and in both early- and late-planted corn in 2014 (20% only) as compared with leaf injury in a pure stand of Cry1F × Cry1Ab × Vip3Aa20 (Optimum Leptra) seen during these years. Corn ears in plots with blended refuges also had significantly greater area of kernels injured in 2012 (5, 10, and 20%), in early- and late-planted corn in 2013 (5, 10, and 20%), and in early (10 and 20% only)- and late-planted corn (5, 10, and 20%) in 2014 as compared with ear injury in a pure stand of Cry1F × Cry1Ab × Vip3Aa20 (Optimum Leptra) seen during these years. Infestations of southwestern corn borer, Diatraea grandiosella Dyar (Lepidoptera: Crambidae), were also significantly reduced by Cry1F × Cry1Ab × Vip3Aa20 (Optimum Leptra). Despite these differences in injury, yield averaged across locations varied among refuge blends only in the late-planted trials in 2013, with greater yields in the 0% refuge blend than in the 20% blend; however, when examining yield separately by location, only two of nine locations had higher yields in the 100% Bt plots than in any of the blended refuge plots. As a complement to studying the contribution of blended refuge to delaying resistance, quantifying injury and yield in a range of refuge blends is a necessary step to provide management information on the range of lepidopteran pests that occur in the southern United States.

Mammary epithelial cell phagocytosis downstream of TGF-ß3 is characterized by adherens junction reorganization.

After weaning, during mammary gland involution, milk-producing mammary epithelial cells undergo apoptosis. Effective clearance of these dying cells is essential, as persistent apoptotic cells have a negative impact on gland homeostasis, future lactation and cancer susceptibility. In mice, apoptotic cells are cleared by the neighboring epithelium, yet little is known about how mammary epithelial cells become phagocytic or whether this function is conserved between species. Here we use a rat model of weaning-induced involution and involuting breast tissue from women, to demonstrate apoptotic cells within luminal epithelial cells and epithelial expression of the scavenger mannose receptor, suggesting conservation of phagocytosis by epithelial cells. In the rat, epithelial transforming growth factor-ß (TGF-ß) signaling is increased during involution, a pathway known to promote phagocytic capability. To test whether TGF-ß enhances the phagocytic ability of mammary epithelial cells, non-transformed murine mammary epithelial EpH4 cells were cultured to achieve tight junction impermeability, such as occurs during lactation. TGF-ß3 treatment promoted loss of tight junction impermeability, reorganization and cleavage of the adherens junction protein E-cadherin (E-cad), and phagocytosis. Phagocytosis correlated with junction disruption, suggesting junction reorganization is necessary for phagocytosis by epithelial cells. Supporting this hypothesis, epithelial cell E-cad reorganization and cleavage were observed in rat and human involuting mammary glands. Further, in the rat, E-cad cleavage correlated with increased γ-secretase activity and ß-catenin nuclear localization. In vitro, pharmacologic inhibitors of γ-secretase or ß-catenin reduced the effect of TGF-ß3 on phagocytosis to near baseline levels. However, ß-catenin signaling through LiCl treatment did not enhance phagocytic capacity, suggesting a model in which both reorganization of cell junctions and ß-catenin signaling contribute to phagocytosis downstream of TGF-ß3. Our data provide insight into how mammary epithelial cells contribute to apoptotic cell clearance, and in light of the negative consequences of impaired apoptotic cell clearance during involution, may shed light on involution-associated breast pathologies.

Lepidoptera (Crambidae, Noctuidae, and Pyralidae) Injury to Corn Containing Single and Pyramided Bt Traits, and Blended or Block Refuge, in the Southern United States.

Fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae); corn earworm, Helicoverpa zea Boddie (Lepidoptera: Noctuidae); southwestern corn borer, Diatraea grandiosella Dyar (Lepidoptera: Crambidae); sugarcane borer, Diatraea saccharalis F. (Lepidoptera: Crambidae); and lesser cornstalk borer, Elasmopalpus lignosellus Zeller (Lepidoptera: Pyralidae), are lepidopteran pests of corn, Zea mays L., in the southern United States. Blended refuge for transgenic plants expressing the insecticidal protein derivative from Bacillus thuringiensis (Bt) has recently been approved as an alternative resistance management strategy in the northern United States. We conducted a two-year study with 39 experiments across 12 states in the southern United States to evaluate plant injury from these five species of Lepidoptera to corn expressing Cry1F and Cry1Ab, as both single and pyramided traits, a pyramid of Cry1Ab×Vip3Aa20, and a pyramid of Cry1F×Cry1Ab plus non-Bt in a blended refuge. Leaf injury and kernel damage from corn earworm and fall armyworm, and stalking tunneling by southwestern corn borer, were similar in Cry1F×Cry1Ab plants compared with the Cry1F×Cry1Ab plus non-Bt blended refuge averaged across five-plant clusters. When measured on an individual plant basis, leaf injury, kernel damage, stalk tunneling (southwestern corn borer), and dead or injured plants (lesser cornstalk borer) were greater in the blended non-Bt refuge plants compared to Cry1F×Cry1Ab plants in the non-Bt and pyramided Cry1F×Cry1Ab blended refuge treatment. When non-Bt blended refuge plants were compared to a structured refuge of non-Bt plants, no significant difference was detected in leaf injury, kernel damage, or stalk tunneling (southwestern corn borer). Plant stands in the non-Bt and pyramided Cry1F×Cry1Ab blended refuge treatment had more stalk tunneling from sugarcane borer and plant death from lesser cornstalk borer compared to a pyramided Cry1F×Cry1Ab structured refuge treatment. Hybrid plants containing Cry1F×Cry1Ab within the pyramided Cry1F×Cry1Ab blended refuge treatment had significantly less kernel damage than non-Bt structured refuge treatments. Both single and pyramided Bt traits were effective against southwestern corn borer, sugarcane borer, and lesser cornstalk borer.

Differential expression of candidate salivary effector proteins in field collections of Hessian fly, Mayetiola destructor.

Evidence is emerging that some proteins secreted by gall-forming parasites of plants act as effectors responsible for systemic changes in the host plant, such as galling and nutrient tissue formation. A large number of secreted salivary gland proteins (SSGPs) that are the putative effectors responsible for the physiological changes elicited in susceptible seedling wheat by Hessian fly, Mayetiola destructor (Say), larvae have been documented. However, how the genes encoding these candidate effectors might respond under field conditions is unknown. The goal of this study was to use microarray analysis to investigate variation in SSGP transcript abundance amongst field collections from different geographical regions (southeastern USA, central USA, and the Middle East). Results revealed significant variation in SSGP transcript abundance amongst the field collections studied. The field collections separated into three distinct groups that corresponded to the wheat classes grown in the different geographical regions as well as to recently described Hessian fly populations. These data support previous reports correlating Hessian fly population structure with micropopulation differences owing to agro-ecosystem parameters such as cultivation of regionally adapted wheat varieties, deployment of resistance genes and variation in climatic conditions.

El Niño-Southern Oscillation (ENSO) effects on Hessian fly (Diptera: Cecidomyiidae) infestation in the southeastern United States.

Climate variability is expected to have an influence on the population of Hessian fly, Mayetiola destructor Say (Diptera: Cecidomyiidae), a serious insect pest of winter wheat in the southeastern United States. This study had two objectives: 1) to examine the effects of El Niño-Southern Oscillation (ENSO) on Hessian fly infestation and 2) to develop a weather-based Hessian fly infestation model for wheat yield loss prediction. At least 20 years of Hessian fly infestation and wheat yield records from two locations in South Georgia were used for this study. The yearly values of infestation were separated by ENSO phase and tested to assess the infestation differences across ENSO phases. Each year, yield losses from infestation were calculated by subtracting the yields of resistant varieties from those of susceptible ones. The yield losses were then separated by ENSO phase and tested. Multiple regression analyses were conducted to identify the contribution of monthly weather variables and changes in wheat acreage to Hessian fly infestation. Results showed that Hessian fly infestation and yield losses were greatest during the La Niña and least during the El Niño phase. The weather conditions that significantly increased the risk for infestation were those of the August-February period. The risk of infestation was higher during August-September under wetter, cooler conditions and during October-February under drier, warmer conditions. These findings could help wheat growers reduce the risk of infestation in the years that are expected to have more infestation through the adoption of necessary mitigation measures before the crop season.

Insect Damage, Aflatoxin Content, and Yield of Bt Corn in Alabama.

Isoline pairs of hybrid corn, similar except for presence or absence of a Bt trait, were planted at eight sites across Alabama over three years. This study evaluated insect damage, yield, and aflatoxin levels as affected by the Bt traits, YieldGard Corn Borer (expressing Cry1Ab), Herculex I (expressing Cry1F), Genuity VT Triple PRO (expressing Cry1A.105 and Cry2Ab2), Agrisure Viptera 3111 (expressing Vip3Aa20 and Cry1Ab), and Genuity SmartStax (expressing Cry1A.105, Cry2Ab2, and Cry1F). When examined over all sites and years, hybrids with any of the included Bt traits had lower insect damage and higher yields. However, insect damage was not consistently correlated to yield. Bt traits expressing multiple proteins provided greater protection from corn earworm feeding than did traits for single proteins. Yields and aflatoxin levels were highly variable among sites although irrigated sites had higher yields than nonirrigated sites. Aflatoxins commonly accumulate in corn in the southeastern United States because of prevailing high temperatures and frequent dry conditions. Aflatoxin levels were not consistently associated with any factors that were evaluated, including Bt traits.

GATA3 inhibits lysyl oxidase-mediated metastases of human basal triple-negative breast cancer cells.

Discovery of mechanisms that impede the aggressive and metastatic phenotype of human basal triple-negative-type breast cancers (BTNBCs) could provide novel targets for therapy for this form of breast cancer that has a relatively poor prognosis. Previous studies have demonstrated that expression of GATA3, the master transcriptional regulator of mammary luminal differentiation, can reduce the tumorigenicity and metastatic propensity of the human BTNBC MDA-MB-231 cell line (MB231), although the mechanism for reduced metastases was not elucidated. We demonstrate through gene expression profiling that GATA3 expression in 231 cells resulted in the dramatic reduction in the expression of lysyl oxidase (LOX), a metastasis-promoting, matrix-remodeling protein, in part, through methylation of the LOX promoter. Suppression of LOX expression by GATA3 was further confirmed in the BTNBC Hs578T cell line. Conversely, reduction of GATA3 expression by small interfering RNA in luminal BT474 cells increased LOX expression. Reconstitution of LOX expression in 231-GATA3 cells restored metastatic propensity. A strong inverse association between LOX and GATA3 expression was confirmed in a panel of 51 human breast cancer cell lines. Similarly, human breast cancer microarray data demonstrated that high LOX/low GATA3 expression is associated with the BTNBC subtype of breast cancer and poor patient prognosis. Expression of GATA3 reprograms BTNBCs to a less aggressive phenotype and inhibits a major mechanism of metastasis through inhibition of LOX. Induction of GATA3 in BTNBC cells or novel approaches that inhibit LOX expression or activity could be important strategies for treating BTNBCs.

Smad3 loss confers resistance to the development of trinitrobenzene sulfonic acid-induced colorectal fibrosis.

BACKGROUND: Transforming growth factor-beta (TGF-beta)/Smad3 signalling plays a central role in tissue fibrogenesis, acting as a potent stimulus of extracellular matrix (ECM) protein accumulation. The aim of this study was to evaluate the potential role of Smad3 in the pathogenesis of colonic fibrosis induced by trinitrobenzene sulfonic acid (TNBS) in Smad3 null mice. MATERIALS AND METHODS: Chronic colitis-associated fibrosis was induced in 15 Smad3 null and 13 wild-type mice by intra-rectal administration of TNBS. Each mouse received an incremental dose of TNBS (0.5-1.0 mg per week) over a 6-week period. The colon was excised for macroscopic examination and histological, morphometric and immunohistochemical analyses. For immunohistochemistry, alpha-smooth muscle actin (alpha-SMA), collagen types I-III, TGF-beta1, connective tissue growth factor (CTGF), Smad3, Smad7, and CD3 antibodies were used. RESULTS: At macroscopic examination, the colon of Smad3 wild-type mice appeared significantly harder, thicker and shorter than that of the Smad3 null mice. Of the wild-type mice, 50% presented colonic adhesions and strictures. Histological and morphometric evaluation revealed a significantly higher degree of colonic fibrosis and accumulation of collagen in the Smad3 wild-type compared to null mice, whereas the degree of colonic inflammation did not differ between the two groups of mice. Immunohistochemical evaluation showed a marked increase in CTGF, collagen I-III, TGF-beta and Smad3 staining in the colon of Smad3 wild-type compared to null mice, whereas Smad7 was increased only in null mice. CONCLUSIONS: These results indicate that Smad3 loss confers resistance to the development of TNBS-induced colonic fibrosis. The reduced fibrotic response appears to be due to a reduction in fibrogenic mesenchymal cell activation and ECM production and accumulation. Smad3 could be a novel target for potential treatment of intestinal fibrosis, especially in inflammatory bowel disease.

Acoustic indicators for mapping infestation probabilities of soil invertebrates.

Acoustic and traditional excavation methods were used in consecutive summers to conduct two geospatial surveys of distributions of white grubs and other soil invertebrates in two forage fields. Indicator variables were constructed from listener- and computer-based assessments of sounds detected at each recording site and then applied in geostatistical analysis, contingency analysis, and spatial analysis of distance indices (SADIE) of soil invertebrate distributions. Significant relationships were identified between the acoustic indicators and the counts of sound-producing soil invertebrates in a majority of the geostatistical and contingency analyses. Significant clusterings and overall spatial associations were identified also in most of the SADIE analyses. In addition, significant local spatial associations were identified between acoustic indicators and counts of sound-producing soil invertebrates that could be of potential value in selection of specific sites as targets for treatment or for untreated reserves in integrated pest management programs. An example is presented of the relative efficiency of acoustic surveys for targeting of white grub treatments.

SNIP1 inhibits NF-kappa B signaling by competing for its binding to the C/H1 domain of CBP/p300 transcriptional co-activators.

SNIP1 is a 396-amino acid nuclear protein shown to be an inhibitor of the TGF-beta signal transduction pathway and to be important in suppressing transcriptional activation dependent on the co-activators CBP and p300. In this report we show that SNIP1 potently inhibits the activity of NF-kappa B, which binds the C/H1 domain of CBP/p300, but does not interfere with the activity of transcription factors such as p53, which bind to other domains of p300, or factors such as VP16, which are independent of these co-activators. Inhibition of NF-kappa B activity is a function of the N-terminal domain of SNIP1 and involves competition of SNIP1 and the NF-kappa B subunit, RelA/p65, for binding to p300, similar to the mechanism of inhibition of Smad signaling by SNIP1. Immunohistochemical staining shows that expression of SNIP1 is strictly regulated in development and that it colocalizes, in certain tissues, with nuclear staining for RelA/p65 and for p300, suggesting that they may regulate NF-kappa B activity in vivo in a spatially and temporally controlled manner. These data led us to suggest that SNIP1 may be an inhibitor of multiple transcriptional pathways that require the C/H1 domain of CBP/p300.

Heterozygous mice for the transforming growth factor-beta type II receptor gene have increased susceptibility to hepatocellular carcinogenesis.

The transforming growth factor-beta (TGF-beta) receptor complex and its downstream signaling intermediates constitute a tumor suppressor pathway. In many cancers, expression of TGF-beta type II receptor (TbetaR-II) is markedly decreased. In the present study, we show that the hepatocytes isolated from 15-day-old, but not 9-month-old, mice heterozygous for the deletion of the TbetaR-II gene are slightly less sensitive to the growth-inhibitory effect of TGF-beta when compared with wild-type littermates of same age. In addition, the proliferation index of hepatocytes as indicated by bromodeoxyuridine incorporation is mildly increased in the heterozygous mice. These subtle changes in cellular phenotype did not result in either gross or microscopic abnormality of the liver. The treatment of these mice with the chemical carcinogen, diethylnitrosamine, results in a significantly enhanced tumorigenesis in the liver when compared with the wild-type littermates. Our results demonstrate the gene-dosage effect of TbetaR-II and indicate that the reduced expression of TbetaR-II in mice increases susceptibility to tumorigenesis in the liver.

Is Smad3 a major player in signal transduction pathways leading to fibrogenesis?

Transforming growth factor (TGF)-beta plays a central role in fibrosis, contributing both to the influx and activation of inflammatory cells, as well as to activation of fibroblasts to elaborate extracellular matrix. In the past few years, new insight has been gained into signal transduction pathways downstream of the TGF-beta receptor serine-threonine kinases with the identification of a family of evolutionarily conserved Smad proteins. Two receptor-activated Smad proteins, Smad2 and Smad3, are phosphorylated by the activated TGF-beta type I receptor kinase, after which they partner with the common mediator, Smad4, and are translocated to the nucleus to where they participate in transcriptional complexes to control expression of target genes. We have shown in wound healing studies of mice null for Smad3, that loss of this key signaling intermediate interferes with the chemotaxis of inflammatory cells to TGF-beta as well as with their ability to autoinduce TGF-beta. Moreover, studies with mouse embryo fibroblasts null for Smad3 show that TGF-beta-dependent induction of c-Jun and c-Fos, important in induction of collagen as well as in autoinduction of TGF-beta, is mediated by Smad3. Based on these observations, we hypothesize that loss of Smad3 will confer resistance to fibrosis and result in reduced inflammatory cell infiltrates, reduced autoinduction of TGF-beta, important to sustain the process, and reduced elaboration of collagen. Preliminary observations in a model of radiation-induced fibrosis confirm this hypothesis and suggest that inhibitors of Smad3 might have clinical application both to improve wound healing and to reduce fibrosis.

Transforming growth factor-beta1 expression in early biopsy specimen predicts long-term graft function following pediatric renal transplantation.

The main cause of late graft loss or declining long-term graft function is chronic allograft nephropathy (CAN), characterized by progressive interstitial fibrosis. Transforming growth factor (TGF)-beta1 plays a key role in fibrogenesis. We immunohistochemically investigated whether the degree of TGF-beta1 expression in early biopsy specimens routinely obtained from stable allografts at 100 d could predict fibrosis and graft dysfunction in the late phase. Patients were children with grafts from related donors. We immunohistochemically determined intracellular and extracellular expression of TGF-beta1 in the graft using LC antibody (LC) for intracellular TGF-beta1 and CC antibody (CC) for extracellular TGF-beta1. The change in creatinine clearance between 100 d and 3 yr after transplantation (DeltaCcr) was used as an index of long-term graft function. We also used image analysis to calculate the relative area involved by interstitial fibrosis in the trichrome-stained section of graft biopsy specimens at 100 d and 3 yr, designating the change as DeltaFI. DeltaCcr was -4.2+/-9.4 mL/min in subjects with minimal early immunoreactivity for CC and -20.5+/-15.9 mL/min in subjects with strong reactivity (p<0.05). DeltaCcr was -14.5+/-18.6 mL/min in subjects with minimal early immunoreactivity for LC and -11.7+/-12.8 mL/min in those with strong reactivity. DeltaFI in subjects with minimal CC reactivity (1.28+/-4.11%) tended to be lower than that in subjects with strong reactivity (8.45+/-15.47%). Neither fibrosis at 100 d nor DeltaFI differed between subjects with minimal and strong LC reactivity. Thus, strong extracellular TGF-beta1 expression in grafts at 100 d after transplantation is associated with a long-term decline in graft function and tends to be associated with increased graft fibrosis at 3 yr.

Oviposition site and food preference of the green June beetle (Coleoptera: Scarabaeidae).

Relative preferences of green June beetle, Cotinis nitida (L.), adults and grubs for different organic fertilizers were determined in field and laboratory choice experiments. Six organic fertilizer treatments (low rate of broiler litter, high rate of broiler litter, cow manure, hay, Milorganite, or no fertilizer [the control]) were applied to sandy-loam soil and exposed to adults in 2.7 by 3.7 by 2.4-m screen cages. More eggs and larvae were found in pots treated with broiler litter (43%), cow manure (23%), and hay (30%) than in pots treated with Milorganite (4%) or no fertilizer (0%). Orientation preferences of third-instar grubs were tested in Y-tube and satellite olfactometers. Of the five treatments (broiler litter, cow manure, hay, Milorganite, and a blank control), preference was greatest for broiler litter and cow manure, but all organic fertilizer treatments were generally preferred over the blank control. These experiments suggest that use of organic fertilizers may result in higher densities of green June beetle grubs both by attracting the ovipositing females, and by acting as a food attractant for the mobile larvae.

Transforming growth factors-beta are not good biomarkers of chemopreventive efficacy in a preclinical breast cancer model system.

Using a carcinogen-initiated rat model of mammary tumorigenesis, we tested the hypothesis that transforming growth factor (TGF)-betas are useful biomarkers of chemopreventive efficacy in the breast. The chemopreventive agents tested were tamoxifen and the retinoids 9-cis-retinoic acid (9cRA) and N-(4-hydroxyphenyl)retinamide (4-HPR), because both antiestrogens and retinoids have previously been shown to upregulate TGF-betas in vitro. Despite demonstrable chemopreventive efficacy in this model, none of these agents, alone or in combination, had any significant impact on the expression of TGF-betas in the mammary ductal epithelium or periductal stroma as determined by immunohistochemistry. These data suggest that TGF-betas are not likely to be useful biomarkers of chemopreventive efficacy in a clinical setting.

Immunohistochemical expression of Smads 1-6 in the 15-day gestation mouse embryo: signaling by BMPs and TGF-betas.

The eight mammalian Smad proteins mediate cellular signaling from members of the transforming growth factor-beta (TGF-beta), bone morphogenetic protein (BMP), and activin families. Smads 1, 5, and 8 transmit signals from BMPs, while Smads 2 and 3 transmit signals from TGF-betas and activin. Smad 4 is a common mediator of both pathways, while Smads 6 and 7 inhibit signaling. Signal transduction involves translocation of Smad complexes to the nucleus and subsequent gene activation. Little is known about the expression of endogenous Smad proteins during development. We identified commercially available Smad antibodies that specifically recognize a unique Smad protein and are suitable for immunohistochemistry. Here we compare the localization of Smads 1, 2, 3, 4, 5, and 6 in tissues of the 15-day gestation mouse embryo. Immunoreactive Smad proteins are seen in many tissues with differences in the localization being dependent upon the cell type. All tissues express Smad 4 and at least one each of the BMP-specific and TGF-beta-specific Smads, while expression of Smad 6 is more restricted. Differences are observed in the nuclear versus cytoplasmic localization among the Smads in different cell types or tissues, suggesting selective activation of Smads during this stage of development.

Mapping of soil insect infestations sampled by excavation and acoustic methods.

Geostatistical analysis was used to map traditionally and acoustically sampled populations of soil invertebrates at two different times in a hayfield at Grove Hill, AL, and once in an Auburn, AL, hayfield. The distributions of nearly all the soil invertebrates and their sounds were nonrandom in all three mapping studies. The maps constructed by excavation and acoustic sampling methods were compared by correlating the estimated (kriged) soil invertebrate counts with the estimated (kriged) counts of sounds per minute (pulse rate). Acoustic and traditional estimates were positively correlated in the Auburn study. Kriged estimates for green June beetle grub counts overlapped significantly with kriged estimates of sound pulse rate (R2 = 0.47). Overlap with sound pulse rates increased slightly when other soil organisms were counted along with green June beetle grubs: estimates of sound pulse rates were significantly correlated with counts of all white grubs (R2 = 0.50), all white grubs with earthworms (R2 = 0.52), all white grubs with earthworms and earwigs (R2 = 0.59), and total invertebrates (R2 = 0.59). The correlation between acoustic and traditional estimates was not significant at Grove Hill in either year, possibly because of a lack of experience in signal analysis or because the soil invertebrates may not have generated enough sounds to be detected above the background noise levels. These results suggest that acoustic technology is a promising tool for detecting insect pests in soil, but that further study and additional analysis are needed to improve interpretation of acoustic data obtained in the field.

Characterization of the mouse Smad1 gene and its expression pattern in adult mouse tissues.

Smad1 belongs to a family of receptor-activated proteins which mediate signals from TGF-beta superfamily ligands, including TGF-beta and BMPs. Although much is known about the biochemistry of Smad1 signal transduction, the role of Smad1 in vivo is still unclear. Here we present the first description of the genomic structure of the mouse Smad1 gene and the characterization of its expression pattern in adult mouse tissues by immunohistochemistry. The Smad1 gene contains 7 exons and spans >42 kb of genomic DNA. Its coding region is contained within 6 exons and all introns, except intron 1, follow the GT/AG rule. Immunohistochemical analysis shows that Smad1 is widely expressed in adult mouse tissues, with a varying degree of nuclear localization in different cell types, suggesting a regulated function for this protein. This study assigns all of the exon-intron boundaries of the mouse Smad1 gene and provides the basis for assessing the functional significance of this gene using targeted gene manipulation in the mouse.

Eavesdropping on insects hidden in soil and interior structures of plants.

Accelerometer, electret microphone, and piezoelectric disk acoustic systems were evaluated for their potential to detect hidden insect infestations in soil and interior structures of plants. Coleopteran grubs (the scarabaeids Phyllophaga spp. and Cyclocephala spp.) and the curculionids Diaprepes abbreviatus (L.) and Otiorhynchus sulcatus (F.) weighing 50-300 mg were detected easily in the laboratory and in the field except under extremely windy or noisy conditions. Cephus cinctus Norton (Hymenoptera: Cephidae) larvae weighing 1-12 mg could be detected in small pots of wheat in the laboratory by taking moderate precautions to eliminate background noise. Insect sounds could be distinguished from background noises by differences in frequency and temporal patterns, but insects of similarly sized species could not be distinguished easily from each other. Insect activity was highly variable among individuals and species, although D. abbreviatus grubs tended to be more active than those of O. sulcatus. Tests were done to compare acoustically predicted infestations with the contents of soil samples taken at recording sites. Under laboratory or ideal field conditions, active insects within approximately 30 cm were identified with nearly 100% reliability. In field tests under adverse conditions, the reliability decreased to approximately 75%. These results indicate that acoustic systems with vibration sensors have considerable potential as activity monitors in the laboratory and as field tools for rapid, nondestructive scouting and mapping of soil insect populations.

Biotype composition of Hessian fly (Diptera: Cecidomyiidae) populations from the southeastern, midwestern, and northwestern United States and virulence to resistance genes in wheat.

Twenty-three Hessian fly, Mayetiola destructor (Say), populations collected in the southeastern (Alabama and Mississippi), midwestern (Indiana), and northwestern (Idaho and Washington) United States from 1995 to 1999 were evaluated for biotype composition based on response to Hessian fly resistance genes H3, H5, H6, and H7H8 in wheat, Triticum aestivum L. Biotypes L and O, combined, made up at least 60% of all Alabama populations. Biotype L was predominant in the northern third of Alabama and biotype O in the southern two-thirds of the state. Based on biotype data, wheat cultivars with H7H8 resistance should be highly effective in central and southern Alabama. Fifty-four percent of the Mississippi population consisted of biotype L, and the remaining virulent biotypes (B, D, E, G, J, and O) ranged in frequency from 1 to 17%. The Mississippi population also contained 4% of the avirulent biotype GP. Only biotypes D and L were found in Indiana populations, but biotype L was predominant. Hessian fly populations from Idaho and Washington contained one or more of the virulent biotypes D-H, J, and L-O; however, only biotypes E, F, and G occurred at frequencies > 12%. The avirulent biotype GP made up 25-57% of Idaho and Washington populations, a much higher percentage than found in populations from the eastern United States. Although the highest level of virulence in Idaho and Washington populations was found to resistance genes H3 and H6, the frequency of biotype GP would indicate that the currently deployed gene H3 would provide a moderate to high level of resistance, depending on location. Nine of the populations, plus populations collected from the mid-Atlantic state area in 1989 and 1996, also were tested against the wheat cultivar 'INW9811' that carries H13 resistance to Hessian fly biotype L and two Purdue wheat lines with unidentified genes for resistance. The H13 resistance in INW9811 was highly effective against all populations tested from the eastern and northwestern U.S. wheat production areas, except Maryland and Virginia. Population studies also indicated that wheat line CI 17960-1-1-2-4-2-10 likely carries the H13 resistance gene, based on the similarity of its response and that of INW9811 to eight fly populations. Continued monitoring of biotype frequency in Hessian fly populations is required for optimal deployment and management of resistance genes in all wheat production areas.