Tiffany Heng-Moss

Effects of temperature on development of Phormia regina (Diptera: Calliphoridae) and use of developmental data in determining time intervals in forensic entomology.

Abstract Precise developmental data for forensic indicator blow fly species are essential for accuracy in the estimate of the post-mortem interval (PMI). Why, then, does the determination of the PMI result in conflicting time frames when published conspecific developmental data are used? To answer this question, we conducted constant temperature trials between the developmental minimum temperature and upper threshold temperatures (8–32°C) on the forensically important blow fly species Phormia regina (Meigen) (Diptera: Calliphoridae). Flies were reared using two designs to quantify sources of variation. We measured rearing container temperatures and internal growth chamber temperatures by using thermocouples to accurately record temperatures experienced by larvae and to construct a degree-day model. Differences in experimental design, as seen across temperature studies for this fly species, did not significantly impact larval development. We also found that using set chamber temperatures rather than rearing container temperatures altered the final degree-day model. Using any minimum threshold (including an empirically determined true minimum) other than that from linear interpolation (x-intercept) violated degree-day assumptions and invalidated estimates of the PMI. We observed the minimum developmental temperature to be higher (14°C) than that generated under the x-intercept method (5.46°C) by using data from oviposition to adult emergence. This difference along with the noted difference in accumulated degree-days (using different base temperatures) suggests a need for additional experimentation on other forensically important fly species at low temperature thresholds to help with development of curvilinear models. Former and current estimates of the PMI may be inaccurate if the process to determine the time frame ignored degree-day model assumptions or was based upon questionable data sets.

Characterization of oxidative enzyme changes in buffalograsses challenged by Blissus occiduus

Abstract This research investigated the role of oxidative enzymes in the defense response of buffalograss, Buchloë dactyloides (Nuttall) Engelmann, to Blissus occiduus Barber. Changes in catalase and peroxidase activity were observed in both resistant and susceptible buffalograsses in response to chinch bug feeding. Susceptible plants were shown to have a lower level of catalase activity compared with their respective control plants. By contrast, catalase activities of resistant plants were similar between infested and control buffalograsses throughout the study. Resistant plants had higher levels of peroxidase activity compared with their control plants, whereas peroxidase activities for control and infested susceptible plants remained at similar levels or were slightly lower for infested plants. These findings suggest that chinch bug feeding leads to a loss in catalase activity in susceptible buffalograsses. In contrast, resistant buffalograsses may be able to tolerate chinch bug feeding by increasing their peroxidase activity. Polyphenol oxidase activities were similar between control and infested plants for the buffalograsses evaluated. Among the enzymes examined, no differences in isozyme profiles for peroxidase and polyphenol oxidase were detected between control and infested 378, NE91-118, Cody, and Tatanka plants. Gels stained for catalase identified differences in the isozyme profiles of infested and uninfested 378 plants; however, infested and control NE91-118, Tatanka, and Cody plants has similar isozyme profiles. No differences in protein profiles were observed between chinch bug-infested 378, NE91-118, Cody, and Tatanka plants and their respective uninfested controls.

Gene expression profiling of tolerant barley in response to Diuraphis noxia (Hemiptera: Aphididae) feeding

Aphids are, arguably, the single most damaging group of agricultural insect pests throughout the world. Plant tolerance, which is a plant response to an insect pest, is viewed as an excellent management strategy. Developing testable hypotheses based on genome-wide and more focused methods will help in understanding the molecular underpinnings of plant tolerance to aphid herbivory. As a first step in this process, we undertook transcript profiling with Affymetrix GeneChip Barley Genome arrays using RNA extracted from tissues of tolerant and susceptible genotypes collected at three hours, three days and six days after Diuraphis noxia introduction. Acquired data were compared to identify changes unique to the tolerant barley at each harvest date. Transcript abundance of 4086 genes was differentially changed over the three harvest dates in tolerant and susceptible barley in response to D. noxia feeding. Across the three harvest dates, the greatest number of genes was differentially expressed in both barleys at three days after aphid introduction. A total of 909 genes showed significant levels of change in the tolerant barley in response to D. noxia feeding as compared to susceptible plants infested with aphids. Many of these genes could be assigned to specific metabolic categories, including several associated with plant defense and scavenging of reactive oxygen species (ROS). Interestingly, two peroxidase genes, designated HvPRXA1 and HvPRXA2, were up-regulated to a greater degree in response to D. noxia feeding on tolerant barley plants, indicating that specific peroxidases could be important for the tolerance process. These findings suggest that the ability to elevate and sustain levels of ROS-scavenging enzymes could play an important role in the tolerant response.

Evaluation of Buffalograss Germplasm for Resistance to Blissus occiduus (Hemiptera: Lygaeidae)

Abstract Blissus occiduus Barber has emerged as an important insect pest of buffalograss, Buchloë dactyloides (Nuttall) Engelmann, in Nebraska. This research evaluated selected buffalograss germplasm for resistance to B. occiduus. Eleven buffalograss selections were screened for chinch bug resistance in three greenhouse studies and two field evaluations. Based on chinch bug damage, NE91–118, ‘Tatanka’, ‘Bonnie Brae’, and ‘Cody’ were rated highly to moderately resistant. These four buffalograsses exhibited minimal damage, even though all were heavily infested with chinch bugs. NE84-45-3 and ‘378’ were highly susceptible to B. occiduus. Field evaluations confirmed chinch bug resistance ratings under field conditions. NE91–118 displayed high levels of resistance in the field screening evaluations, whereas Cody and Tatanka showed moderate levels of resistance, and 378 was highly susceptible.

Categorizing the Resistance of Soybean Genotypes to the Soybean Aphid (Hemiptera: Aphididae)

ABSTRACT We evaluated selected soybean, Glycine max (L.) Merr., genotypes during their reproductive stages for resistance to the soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), under greenhouse conditions and documented the categories of aphid-resistant soybean. Two screening studies were performed to assess the level of resistance to the soybean aphid on six soybean genotypes during the reproductive stages of development. Significant differences in aphid damage ratings were detected among the soybean evaluated in the screening studies. Three genotypes (KS4202, K-1639-2, and K1621) were considered moderately resistant based on the assessed damage ratings. Two of these genotypes (K-1639-2 and KS4202), along with a commercial variety (‘Asgrow 2703’) were used in a follow-up greenhouse study to test for antibiosis and tolerance. For the antibiosis evaluation, KS4202 had significantly more nymphs than Asgrow 2703 and K-1639-2. In fact, KS4202 had a threefold difference in the number of nymphs compared with Asgrow 2703 (81.8 ± 14.7 and 26.2 ± 13.9 nymphs, respectively) and a fivefold difference compared with K-1639-2 (15.6 ± 13.9). Although not significant, Asgrow 2703 had more nymphs than K-1639-2. The lower aphid numbers on infested K-1639-2 plants compared with aphid numbers on Asgrow 2703 and KS4202 plants indicates antibiosis for this genotype. No significant differences in average seed weight, number of seeds per pod, or plant damage were observed between infested and control KS4202 plants; however, significant differences in biomass, total seed weight, number of pods per plant, and number of seeds per plant were detected.

Switchgrass (Panicum virgatum L) flag leaf transcriptomes reveal molecular signatures of leaf development, senescence, and mineral dynamics

Switchgrass flag leaves can be expected to be a source of carbon to the plant, and its senescence is likely to impact the remobilization of nutrients from the shoots to the rhizomes. However, many genes have not been assigned a function in specific stages of leaf development. Here, we characterized gene expression in flag leaves over their development. By merging changes in leaf chlorophyll and the expression of genes for chlorophyll biosynthesis and degradation, a four-phase molecular roadmap for switchgrass flag leaf ontogeny was developed. Genes associated with early leaf development were up-regulated in phase 1. Phase 2 leaves had increased expression of genes for chlorophyll biosynthesis and those needed for full leaf function. Phase 3 coincided with the most active phase for leaf C and N assimilation. Phase 4 was associated with the onset of senescence, as observed by declining leaf chlorophyll content, a significant up-regulation in transcripts coding for enzymes involved with chlorophyll degradation, and in a large number of senescence-associated genes. Of considerable interest were switchgrass NAC transcription factors with significantly higher expression in senescing flag leaves. Two of these transcription factors were closely related to a wheat NAC gene that impacts mineral remobilization. The third switchgrass NAC factor was orthologous to an Arabidopsis gene with a known role in leaf senescence. Other genes coding for nitrogen and mineral utilization, including ureide, ammonium, nitrate, and molybdenum transporters, shared expression profiles that were significantly co-regulated with the expression profiles of the three NAC transcription factors. These data provide a good starting point to link shoot senescence to the onset of dormancy in field-grown switchgrass.

Physiological and Biochemical Responses of Resistant and Susceptible Wheat to Injury by Russian Wheat Aphid

Abstract We examined the physiological and biochemical responses of resistant (‘Halt’ and ‘Prairie Red’) and susceptible (‘TAM 107′) wheat, Triticum aestivum L., to injury by the Russian wheat aphid, Diuraphis noxia (Mordvilko). Photosynthetic capacity was evaluated by measuring assimilation/internal CO2 (A/Ci) curves, chlorophyll fluorescence, chlorophyll, and nonstructural carbohydrate content. Total protein and peroxidase specific activity also were determined. No significant differences were detected in chlorophyll concentration between aphid-infested and control TAM 107 plants. The aphid-infested resistant cultivars had similar or significantly higher chlorophyll concentrations compared with their respective control plants. Measurements over time showed that infested Halt plants had delays in photosynthetic senescence, Prairie Red plants had photosynthetic rate changes that were similar to control plants, and TAM 107 plants displayed accelerated photosynthetic senescence patterns. The photochemical and nonphotochemical quenching coefficients were significantly higher in infested Halt plants compared with their respective control plants on day 3. Infested TAM 107 plants had significantly higher photochemical quenching compared with control plants at all times evaluated, and they had significantly higher nonphotochemical quenching on day 3. Throughout the experiment, infested Prairie Red plants exhibited photochemical and nonphotochemical quenching coefficient values that were not significantly different from control plants. Total protein content was not significantly different between aphid-infested and control plants for all cultivars. Differences between physiological responses of infested susceptible and resistant cultivars, particularly temporal changes in photosynthetic activity, imply that resistant Halt and Prairie Red wheat tolerate some impacts of aphid injury on photosynthetic integrity.

In vitro enzymatic chlorophyll catabolism in wheat elicited by cereal aphid feeding

Chlorophyll degradation is a complex phenomenon that often accompanies insect feeding damage to plants. Loss of chlorophyll can be initiated by several reactions, including oxidative bleaching, chlorophyllase activity, and Mg‐dechelatase activity. Extracts from the Russian wheat aphid [Diuraphis noxia (Mordvilko)], the bird cherry‐oat aphid [Rhopalosiphum padi (L.)], and aphid‐infested and uninfested wheat plants were assayed in vitro for activities involved in chlorophyll degradation. Although the initial infestation was the same (10 apterous adults) for both aphid species, D. noxia weight was significantly higher than R. padi after feeding for 12 days. Consequently, D. noxia feeding caused greater fresh leaf weight reduction than R. padi feeding. Chlorophyll degradation assays showed no activity from either D. noxia or R. padi extracts. Plant extract assays showed a significant difference in Mg‐dechelatase activity, while no difference was detected in either the chlorophyllase or oxidative bleaching pathways among the aphid‐infested or uninfested plant extracts. Diuraphis noxia‐infested leaf extracts showed a greater increase of Mg‐dechelatase activity than either R. padi‐infested or the uninfested plants. The findings suggest that leaf chlorosis elicited by D. noxia feeding is different from the chlorophyll degradation that occurs in natural plant senescence. Aphid‐elicited chlorosis might be the result of a Mg‐dechelatase‐driven catabolism of chlorophyll in challenged wheat seedlings, however, the factor(s) from D. noxia that elicited the increase of Mg‐dechelatase activity still remain to be determined.

Light-Induced Variability in Development of Forensically Important Blow Fly Phormia regina (Diptera: Calliphoridae)

Abstract The use of the postmortem interval (PMI) in practical applications of forensic entomology is based upon developmental data of blow flies (Diptera: Calliphoridae) generated under controlled environmental conditions. Careful review of the published forensic entomology data sets showed that experimental (environmental) parameters differed between studies. Despite the differences in study design, there are no empirical data on the effect of photoperiod on blow fly development; yet, photoperiod has been shown to alter some insect development and behavior among noncalliphorids. Consequently, will differences in photoperiod alter the developmental times of calliphorids, and thereby alter PMI estimates? To answer this question, we used a replicated design with precise temperature measurement to examine the effects of photoperiod on the forensically important blow fly Phormia regina (Meigen). We concluded that inaccurate temperature recordings by using set-chamber temperatures over rearing-container temperatures would have overshadowed any affect light had on development. Second, constant light increased variation in overall adult developmental time and significantly delayed development compared with cyclic light. Finally, not accounting for delayed development induced by photoperiod underestimated the initial empirical estimate of the PMI. These sources of variation need to be included in forensic estimates because this variation can compromise predictions of PMI based upon current data sets. Without pinpointing optimal photoperiods with which to test development, we must assume that potentially large sources of variability exist within current estimates of the PMI.

Physiological Responses of Resistant and Susceptible Buffalograsses to Blissus Occiduus (Hemiptera: Blissidae) Feeding

Abstract The impact of Blissus occiduus Barber feeding on resistant (‘Prestige’) and susceptible (‘378’) buffalograsses, Buchloë dactyloides (Nuttall) Engelmann, was evaluated through measurement of carbon exchange rate, light and carbon assimilation (A-Ci) curves, chlorophyll a fluorescence, and nonstructural carbohydrates. No significant differences in carbon exchange rates were observed between infested and control plants for 378 at 5 and 10 d after infestation; however, at 20 d after chinch bug introduction, significant differences in carbon exchange rates between infested and control 378 plants were detected. Carbon exchange rates were similar between infested and control Prestige plants at 5, 10, and 20 d after infestation, suggesting that resistant plants can allocate energy for recovery from chinch bug injury. Significant differences in the photochemical efficiency of photosystem II (PSII) and the apparent photosynthetic electron transport ratio were observed between infested and control 378 plants, whereas, no significant differences in the photochemical efficiency of PSII and the electron transport ratio were detected between control and infested Prestige plants. Blissus occiduus-infested 378 and Prestige plants consistently had similar or higher levels of nonstructural carbohydrates compared with their respective control plants. These data suggest that both resistant and susceptible buffalograsses increase levels of nonstructural carbohydrates in response to B. occiduus feeding. This research also suggests compensatory photosynthesis takes place in Prestige but not in 378.