Estimation of hymenopteran parasitism in cereal aphids by using molecular markers

Polymerase chain reaction (PCR) primers were designed and tested for identification of immature parasitoids in small grain cereal aphids and for estimation of parasitism rates. PCR technique was evaluated for 1) greenhouse-reared greenbugs, Schizaphis graminum (Rondani), parasitized by Lysiphlebus testaceipes Cresson and 2) aphids collected from winter wheat fields in Caddo County, Oklahoma. For greenhouse samples, parasitism frequencies for greenbugs examined by PCR at 0, 24, and 48 h after removal of L. testaceipes parasitoids were compared with parasitism frequencies as determined by greenbug dissection. PCR was unable to detect parasitism in greenbugs at 0 and 24 h postparasitism, but it was able to detect parasitoids 48 h after parasitoid removal at frequencies that were not significantly different from dissected samples. Field-collected samples were analyzed by rearing 25 aphids from each sample and by comparing parasitoid frequencies of mummies developed and PCR performed on another 50 aphids. Aphid samples included corn leaf aphids, Rhopalosiphum maidis (Fitch); bird cherry-oat aphids, Rhopalosiphum padi (L.); English grain aphids, Sitobion avenae (F.); and greenbugs. Mummies were isolated until adult emergence, whereupon each parasitoid was identified to species (L. testaceipes was the only parasitoid species found). Parasitism detection frequencies for PCR also were not statistically different from parasitism frequencies of reared aphids. These results indicate that PCR is a useful tool for providing accurate estimates of parasitism rates and especially for identification of immature parasitoids to species.     

Parasitism of Greenbug, Schizaphis graminum, by the parasitoid Lysiphlebus testaceipes at winter temperatures

Functional responses by Lysiphlebus testaceipes (Cresson), a common parasitoid of small grain aphids, on greenbug, Schizaphis graminum (Rondani), were measured at seven temperatures (14, 12, 10, 8, 6, 4, and 2 degrees C) during a 24-h period (12-h light: 12-h dark). Oviposition by L. testaceipes ceased at temperatures <4 degrees C. At all experimental temperatures, a type I, rather than a type II or type III, functional response was determined to be the best fit based on coefficient of determination (r2) values. L. testaceipes was observed to oviposit in greenbugs at temperatures below the developmental temperature of both the greenbug host (5.8 degrees C) and the parasitoid itself (6.6 degrees C). This ability to oviposit at subdevelopmental temperatures enables the parasitoid to increase the percentage of greenbugs that are parasitized while the greenbugs are unable to reproduce. The implications of these findings regarding population suppression of greenbugs are discussed.     

Phenotypic mechanisms of host resistance against greenbug (Homoptera: Aphididae) revealed by near isogenic lines of wheat

Interactions between biotype E greenbug, Schizaphis graminum (Rondani), and wheat, Triticum aestivum L., were investigated using resistant and susceptible near isogenic lines of the greenbug resistance gene Gb3. In an antixenosis test, the greenbugs preferred susceptible plants to resistant ones when free choice of hosts was allowed. Aphid feeding resulted in quick and severe damage to susceptible plants, which seemed to follow a general pattern spatially and was affected by the position where the greenbugs were initially placed. Symptom of damage in resistant plants resembled senescence. Within-plant distribution of aphids after infestation was clearly different between the two genotypes. Significantly more greenbugs fed on the first (oldest) leaf than on the stem in resistant plants, but this preference was reversed in the susceptible one. After reaching its peak, aphid population on the susceptible plants dropped quickly. All susceptible plants were dead in 10-14 d after infestation due to greenbug feeding. Aphid population dynamics on resistant plants exhibited a multipeak curve. After the first peak, the greenbug population declined slowly. More than 70% of resistant plants were killed 47 d after infestation. Performance of both biotype E and I greenbugs on several Gb3-related wheat germplasm lines were also examined. It seems that the preference-on-stem that was characteristic of biotype E greenbugs on the susceptible plants was aphid biotype- and host genotype-dependent. Results from this study suggested that antixenosis, antibiosis, and tolerance in the resistant plants of wheat might all contribute to resistance against greenbug feeding.     

Life history study of multiple clones of insecticide resistant and susceptible greenbug Schizaphis graminum (Homoptera: Aphididae)

Comparative differences and similarities in prereproductive time (d), progeny production in a time equal to d (Md), and intrinsic rate of increase (rm) were established for one susceptible (S) and three resistant (R) strains of the greenbug, Schizaphis graminum (Rondani), reared on sorghum hybrids Dekalb G550E and Cargill 607E. The R strains showed three patterns of elevated esterase activity. Four R1 clones, four R2 clones, one R3 clone, and four S clones were evaluated. The interaction of sorghum hybrid and greenbug strain did not significantly influence any of the parameters measured. However, R1 greenbugs exhibited a significantly longer prereproductive period than the other strains. In addition, the R1 strain had a significantly slower intrinsic rate of increase than the R2 or S greenbug strains, but did not differ significantly from the R3 strain. These results suggest that R1 greenbugs may be less fit than the other strains studied.     

Plant resistance components of two greenbug (Homoptera: Aphididae) resistant wheats

Several biotypes of the greenbug, Schizaphis graminum (Rondani), attack winter wheat, Triticum aestivum L., on the Southern Plains every year. Two wheat germplasm sources of resistance ('Largo' and 'GRS 1201') have been developed that provide protection against the three predominant greenbug biotypes (E, I, and K). Each source has agronomic and end-use quality advantages and disadvantages for the breeder to consider in choosing a greenbug-resistant breeding line. We compared these two germplasms to determine their levels of resistance against biotype E. Components of resistance (i.e., antibiosis, antixenosis, and tolerance) were measured on seedlings of GRS 1201, Largo, and 'TAM W-101' (a susceptible control). Several aphid and plant measurements (e.g., total number of aphids produced per plant, aphid selection preferences, and plant damage ratings) were recorded for each plant entry. Select data recorded for each resistance component were normalized and combined to derive a plant resistance index for each wheat entry. Results indicated that GRS 1201 had a higher level of combined resistance components than did Largo, followed by TAM W-101, the susceptible control. These data provide additional information for the breeder to consider in selecting a greenbug-resistant breeding line.     

Effect of greenbugs (Homoptera: Aphididae) on yield loss of winter wheat

The effect of greenbug, Schizaphis graminum (Rondani), feeding on the yield of four winter wheat cultivars commonly grown in Oklahoma was studied. Cultivars tested were 'Karl', a recent derivative 'Karl-92', and '2163', all greenbug-susceptible cultivars; and 'TAM-110', a cultivar with resistance to biotype E greenbugs. The objectives were to determine the effect of different greenbug densities during fall and spring on yield of winter wheat, and to develop mathematical models to quantify the effect of greenbugs on yield loss. The intensity of greenbug infestations achieved in plots by artificial infestation varied among years and growing seasons within a year, but was generally sufficient to cause a reduction in yield. Among yield components, the number of heads per square meter and the number of seeds per head were frequently negatively correlated with the accumulated number of greenbug-days per tiller. Seed weight was rarely affected by greenbug infestation. A regression model estimated yield loss for greenbug-susceptible cultivars at 0.51 kg/ha loss of yield per greenbug-day in years with near normal precipitation, and a loss of 1.17 kg/ha under severe drought conditions. The susceptible winter wheat cultivars exhibited similar yield loss in relation to the intensity of greenbug infestation, as indicated by a common slope parameter in the regression model. Results suggest that the model is robust for predicting yield loss for susceptible cultivars.     

Categories of resistance to greenbug (Homoptera: Aphididae) biotype I in Aegilops tauschii germplasm

Categories of resistance to greenbug, Schizaphisgraminum (Rondani), biotype I, were determined in goatgrass, Aegilops tauschii (Coss.) Schmal., accession 1675 (resistant donor parent), 'Wichita' wheat, Triticum aestivum L., (susceptible parent), and an Ae. tauschii-derived resistant line, '97-85-3'. Antibiosis was assessed using the intrinsic rate of increase (rm) of greenbugs confined to each of the three genotypes. Neither parent nor the resistant progeny expressed antibiosis. Mean rm values for greenbug I on Wichita (0.0956), and Ae. tauschii (0.10543) were not significantly different. Mean rm values for Wichita and 97-85-3 were also not significantly different. Antixenosis was determined by allowing aphids a choice to feed on plants of each of the three genotypes. Ae. tauschii 1675 exhibited antixenosis, but this resistance was not inherited and expressed in '97-85-3'. In experiments comparing Wichita and Ae. tauschii 1675, greenbug I population distributions were not significantly different on Wichita at 24 h, but were shifted toward Wichita at 48 h. In the second antixenosis experiment, there were no significant differences in greenbug I population distributions on 97-85-3 or Wichita at 24 or 48 h. When all three lines were compared, there were no significant differences in greenbug biotype I populations at 24 or 48 h after infestation. Comparisons of proportional dry plant weight loss (DWT) and SPAD meter readings were used to determine tolerance to greenbug I feeding. Ae. tauschii 1675 and 97-85-3 were highly tolerant compared with Wichita. Infested and uninfested Ae. tauschii 1675 DWT was nonsignificant, and infested Wichita plants weighed significantly less than uninfested plants. When Wichita and 97-85-3 were contrasted, DWT of infested and uninfested Wichita plants were significantly different, but those of 97-85-3 were not. Mean percent leaf chlorophyll losses for the three genotypes, as measured by the SPAD chlorophyll meter, were as follows: Wichita = 65%; Ae. tauschii 1675 = 25%; and 97-85-3 = 39%. Percent leaf chlorophyll losses caused by greenbug feeding was significantly different in comparisons between Wichita and Ae. tauschii 1675, and comparisons between Wichita and 97-85-3, although feeding damage was not significantly different in comparisons between Ae. tauschii 1675 and 97-85-3. These data provided further evidence of the expression of tolerance to greenbug feeding in Ae. tauschii 1675 and 97-85-3.     

Biology and biotype determination of greenbug, Schizaphis graminum (Hemiptera: Aphididae), on seashore paspalum turfgrass (Paspalum vaginatum)

Greenbug, Schizaphis graminum (Rondani) (Hemiptera: Aphididae), was first discovered damaging seashore paspalum (Paspalum vaginatum Swartz) turfgrass in November 2003 at Belle Glade, FL. Inquiries to several golf courses with seashore paspalum turf across southern Florida indicated infestation was wide spread by April 2004. Damage symptoms progress from water soaked lesions surrounding feeding sites within 24 h to chlorosis and necrosis of leaf tips within 96 h. Problems caused by greenbug feeding were initially misdiagnosed as fertilizer, disease, other insects, or water management problems because aphids were not previously found on warm season turfgrasses. Greenbug development and fecundity studies were conducted on six seashore paspalum varieties: 'Aloha,' 'SeaDwarf,' 'SeaGreen,' 'SeaIsle,' 'SeaWay,' and 'SeaWolf.' Greenbug did not survive on 'SeaWolf.' Development rates (mean +/- SEM) ranged from 7.6 +/- 0.2 to 8.2 +/- 0.2 d on the remaining varieties. Greenbug longevity and fecundity on 'Aloha' were significantly less than on the other varieties. The estimated intrinsic rate of natural increase (r(m)) for greenbug ranged from 0.24 to 0.26 across tested varieties. Values for net reproductive rate (R(o)) ranged from 12.3 on 'Aloha' to 40.4 on 'SeaWay.' In feeding trials on indicator plants, the Florida isolate of greenbug exhibited a unique biotypic profile most commonly found on noncultivated grass hosts. It was virulent on the wheat variety GRS1201 that is resistant to the principal agricultural biotypes attacking small grains and to all currently available resistant sorghum varieties.     

Molecular mapping of sorghum genes expressing tolerance to damage by greenbug (Homoptera: Aphididae)

Genetic linkage maps are fundamental for the localization of genes conferring tolerance to greenbug, Schizaphis graminum (Rondani), feeding damage in sorghum, Sorghum bicolor (L.) Moench. Thirteen linkage groups (LGs) containing 60 simple sequence repeat (SSR) loci were mapped by using a set of sorghum recombinant inbred lines (RILs) obtained from the cross '96-4121' (greenbug-tolerant parent) x Redlan (greenbug-susceptible parent). The LG spanned a distance of 603.5 cM, with the number of loci per LG varying from 2 to 14. Seventeen additional SSR loci were unlinked at a log of odds value of 3.0. Based on chlorophyll loss occurring after greenbug feeding, visual damage ratings, and soil plant analysis development (SPAD), chlorophyll-loss indices were recorded for each RIL and for the parents used in the cross. Composite-interval mapping identified three quantitative trait loci (QTLs) associated with biotype I and five QTLs associated with biotype K. The amount of phenotypic variation explained by these QTLs ranged from 9 to 19.6%. The identification of QTLs that influence greenbug tolerance will not only facilitate the use of marker-assisted selection in sorghum breeding programs but also will provide a solid foundation for detailed characterization of individual loci implicated in greenbug tolerance in sorghum.     

Spatial and temporal distribution of induced resistance to greenbug (Homoptera: Aphididae) herbivory in preconditioned resistant and susceptible near isogenic plants of wheat

Interactions between biotype E greenbugs, Schizaphis graminum (Rodani), and two near isogenic lines of the greenbug resistance gene Gb3 of wheat, Triticum aestivum L., were examined for 62 d after infestation. By comparing aphid performance and host responses on control and greenbug-preconditioned plants, we demonstrated that systemic resistance to greenbug herbivory was inducible in the resistant genotype with varying intensities and effectiveness in different parts of the plants. Preconditioning of susceptible plants resulted in modification of within-plant aphid distribution and reduction of cumulative greenbug densities, but it showed no effect on reducing greenbug feeding damage to host plant. Preconditioning of resistant plants altered greenbug population dynamics by reducing the size and buffering the fluctuation of the aphid population. Preconditioning in the first (oldest) leaf of the resistant plant had no phenotypically detectable effect in the stem and induced susceptibility locally in the first leaf within the first 2 d after infestation. The preconditioning-induced resistance reduced greenbug density, delayed aphid density peaks and extended the life of younger leaves in resistant plants. Expression of induced resistance was spatially and temporally dynamic within the plant, which occurred more rapidly, was longer in duration, and stronger in intensity in younger leaves. Host resistance gene-mediated induced resistance was effective in lowering greenbug performance and reducing damage from greenbug herbivory in host plants. Results from this study supported the optimal defense theory regarding within-plant defense allocation.     

Chemical Basis for Greenbug Resistance in Small Grains: II. Identification of the Major Neutral Metabolite of Benzyl Alcohol in Barley

((14)C)Benzyl alcohol was administered either by uptake through the roots or by injection directly into the stems of wheat (Triticum aestivum L. em Thell), sorghum (Sorghum bicolor. L Moench) and two strains of barley (Hordeum vulgare L.). One strain of barley was susceptible to the greenbug (Schizaphis granium Rondani), and the other was greenbug-resistant. In all four plants, several radioactive metabolites were formed. The major neutral metabolite has been identified as benzyl-beta-d-glucopyranoside. This glucoside was found to have no biological activity against the greenbug under conditions in which the parent compound, benzyl alcohol, inhibits the reproduction of this insect pest.     

Characterization of greenbug (Homoptera: Aphididae) resistance in synthetic hexaploid wheats

Twelve greenbug (Schizaphis graminum (Rondani)) biotype E-resistant synthetic hexaploid wheats synthesized by crossing Triticum dicoccum Schrank. and Aegilops tauschii (Coss.) Schmal. were evaluated for the three known insect resistance categories, including antibiosis, anti-xenosis, and tolerance. Different methods were evaluated for calculating antibiosis and tolerance. Calculating intrinsic rate of population increase and measuring leaf chlorophyll content with a SPAD chlorophyll meter proved to be time- and labor-efficient for antibiosis and tolerance determination, respectively. The resistance in all synthetic hexaploids proved to be the result of a combination of antibiosis, antixenosis, and tolerance, which makes them valuable sources of greenbug resistance. To assist plant breeders in selecting the best germplasm for greenbug resistance, a plant resistance index was created that revealed differences among the synthetic hexaploid wheats.     

Parasitoids use herbivore-induced information to adapt patch exploitation behaviour

Abstract.  1. Optimal foraging models ultimately predict that female parasitoids should exploit rich host patches for longer than poorer ones. At the proximate level, mechanistic models and experimental studies show that parasitoids use both chemicals produced by their hosts and direct encounters with their hosts to estimate patch quality. Although it has been extensively studied in the context of host location, the use of herbivore-induced plant response by insect parasitoids has never been considered in the context of patch time allocation.
2. In this study, the respective roles of herbivore-induced plant response and direct contact with hosts on the foraging behaviour of Lysiphlebus testaceipes females on an aphid patch were quantified. For this, the level of herbivore-induced plant response and the number of aphids on the leaf bearing the patch were manipulated independently. Different levels of plant response were obtained by varying the duration of infestation on another leaf.
3. Parasitoid residence time and number of attacks increased with both the level of plant response and the number of aphids.
4. These results suggest that L. testaceipes females use the combination of herbivore-induced response of plants and direct encounters with hosts to assess patch quality and adjust their patch use behaviour.     

Reducing plant abiotic and biotic stress: Drought and attacks of greenbugs, corn leaf aphids and virus disease in dryland sorghum

Multi-year spatial overlay patterns of plants, insects and soil water may yield insights for management for reducing biotic and abiotic stresses in dryland crops. A study of non-irrigated grain sorghum (Sorghum bicolor (L.) Moench) was conducted in a Pullman clay loam on the semi-arid High Plain of Texas during 2002–2005. The objectives of the 4-year study were to understand the mechanisms of plant spatial and temporal responses to stress from drought, infestations of greenbug, corn leaf aphid (CLA) and maize dwarf mosaic virus (MDMV) disease and soil water content (SWC) heterogeneity, and to reduce plant biotic and abiotic stress using their underlying relationships in space and time. Infrared IRt/c sensed-canopy temperature was measured at 18 or 54 sites along transects in a 6 m × 6 m grid across the years. Greenbugs, CLA, MDMV, SWC and hyperstectral reflectance were determined at each IRt/c site. Natural infestations of greenbugs and CLA on sorghum occurred in early July and insect populations peaked in late July or early August. Insect attacks resulted in plant water stress and sorghum yield loss except a late replanting in early July in 2004. Sorghum grain yield was negatively correlated with canopy temperature, greenbug and CLA (−0.38 < r < −0.75, P < 0.05), and positively correlated with SWC and plant near infrared reflectance (0.25 < r < 0.67, P < 0.05). The IRt/c temperature decreased with SWC but increased with greenbugs and CLA (0.26 < R² < 0.64). Crosscorrelation analysis showed that these insect, crop, and soil variables were correlated in space within 48–54 m. Late planting in July or spray control in late July or early August would be options to reduce dryland sorghum water stress and yield loss from drought and insect attacks.     

Genetic control of different types of the aphid resistance in cereal crops

Inheritance of the two main types of the plant resistance to insects was investigated in the sorghum-greenbug (Schizaphis graminum Rond.) and wheat-bird cherry-oat aphid (Rhopalosiphon padi L.) interaction systems. The data obtained support the hypothesis that antixenosis (avoiding of the plant by the insect, given a choice) and antibiosis (adverse effect of the plant on the insect feeding on it) are pleiotropic manifestations of the same genes. This is confirmed by the following facts. (1) Identical patterns of segregation for antixenosis and antibiosis in different cases of sorghum resistance to the greenbug: monogenic control (gene Sgr4), digenic control (Sgr1, Sgr2 and Sgr7, Sgr8), and complementary action of the genes (Sgr9 and Sgr10). (2) Correlated changes in the levels of antibiosis and antixenosis during long-term reproduction of a greenbug clone on the resistant sorghum variety k-1206 (resistance controlled by one gene). (3) Simultaneous expression of antixenosis and antibiosis in F₃ wheat hybrid families to the bird cherry-oat aphid.     

Resistance to greenbug (Heteroptera: Aphididae) biotype I in Aegilops tauschii synthetic wheats

The greenbug, Schizaphis graminum (Rondani), is a major pest of wheat in North America, reducing U.S. wheat production by 60 to 100 million dollars each year. In this research, 149 wheat lines containing genes from Aegilops tauschii (Coss.) Schmal. were evaluated for resistance to greenbug biotype I. More than 50% of the lines sustained moderate foliar chlorosis from greenbug feeding, and approximately one third of all the lines were highly resistant. All lines with chlorosis scores similar to the resistant control 'Largo' expressed high levels of antibiosis, producing greenbug populations with mean weights ranging from 0.05 to 11.8 mg. There was no significant difference between greenbug weights on these lines and those reared on 'Largo', but the mean weight of individuals reared on the susceptible control 'Thunderbird' was significantly greater than those reared on 'Largo' or any of the test lines. The mean population size of greenbugs produced on plants of each line was significantly correlated with mean greenbug weight. Tolerance was not evident in any of the lines examined, but was unexpectedly apparent in 'Thunderbird' at a level similar to that in the tolerant control cultivar 'Largo'.     

Categories of resistance to greenbug (Homoptera: Aphididae) biotype K in wheat lines containing Aegilops tauschii genes

The wheat lines (cultivars) 'Largo', 'TAM110', 'KS89WGRC4', and 'KSU97-85-3' conferring resistance to greenbug, Schizaphis graminum (Rondani), biotypes E, I, and K were evaluated to determine the categories of resistance in each line to greenbug biotype K. Our results indicated that Largo, TAM110, KS89WGRC4, and KSU97-85-3 expressed both antibiosis and tolerance to biotype K. Largo, KS89WGRC4, and KSU97-85-3, which express antixenosis to biotype I, did not demonstrate antixenosis to biotype K. The results indicate that the same wheat lines may possess different categories of resistance to different greenbug biotypes. A new cage procedure for measuring greenbug intrinsic rate of increase (r(m)) was developed, by using both drinking straw and petri dish cages, to improve the efficiency and accuracy of r(m)-based antibiosis measurements.     

番茄感染双生病毒对叶毛密度和海氏桨角蚜小蜂搜寻行为及适合性的影响

植物病毒可通过影响植物形态和生理特性从而对媒介昆虫和寄生性天敌产生作用。然而, 在植物 媒介昆虫 寄生蜂三营养级关系研究中有关植物病毒的影响很少被考虑。本研究测定和分析了番茄植株感染番茄黄化曲叶病毒（tomato yellow leaf curl virus, TYLCV)后叶毛密度的变化及对烟粉虱Bemisia tabaci (Gennadius)重要寄生性天敌海氏桨角蚜小蜂Eretmocerus hayati Zolnerowich and Rose行为与适合性的影响。结果表明： 携带TYLCV病毒番茄植株叶毛密度显著增加, 为健康植株叶毛密度的1.8倍。海氏桨角蚜小蜂在带毒植株叶片上的寄主处置时间和寄主块停留时间显著长于其在健康植株叶片上的时间, 分别为其2倍和1.5倍, 但寄生蜂的寄生率、 羽化率及发育历期差异不显著(P>0.05)。本文首次报道了双生病毒侵染可引起叶毛密度的增加, 对理解植物-双生病毒-烟粉虱-寄生蜂四方关系提供了新的数据。