Tritrophic interaction of parasitoid Lysiphlebus testaceipes (Hymenoptera: Aphidiidae), greenbug, Schizaphis graminum (Homoptera: Aphididae), and greenbug-resistant sorghum hybrids

Interactions of the parasitoid Lysiphlebus testaceipes (Cresson) and the greenbug, Schizaphis graminum (Rondani), on greenbug-resistant 'Cargill 607E' (antibiosis), 'Cargill 797' (primarily tolerance), and -susceptible 'Golden Harvest 510B' sorghum, Sorghum bicolor (L.) Moench, were tested using three levels of biotype I greenbug infestation. The parasitoid infestation rate was 0.5 female and 1.0 male L. testaceipes per plant. For all three greenbug infestation levels, the parasitoid brought the greenbug under control (i.e., prevented the greenbugs from killing the plants) on both resistant hybrids, but it did not prevent heavy leaf damage at the higher greenbug infestation rates. At the low greenbug infestation rate (50 greenbugs per resistant plant when parasitoids were introduced), greenbugs damaged 5 and 18% of the total leaf area on 'Cargill 797' and 'Cargill 607E', respectively, before greenbugs were eliminated. Leaf damage was higher for the intermediate infestation study (120 greenbugs per plant), 21% and 30% leaf area were damaged on the resistant sorghum hybrids 'Cargill 797' and 'Cargill 607E', respectively. At the high greenbug infestation rate (300 greenbugs per plant), heavy damage occurred: 61% on 'Cargill 607E' and 75% on 'Cargill 797'. The parasitoids did not control greenbugs on the susceptible sorghum hybrid 'Golden Harvest 510B'. L. testaceipes provided comparable control on both greenbug-resistant hybrids. This study supports previous studies indicating that L. testaceipes is effective in controlling greenbugs on sorghum with antibiosis resistance to greenbugs. Furthermore, new information is provided indicating that L. testaceipes is also effective in controlling greenbugs on a greenbug-tolerant hybrid.     

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.     

Mitochondrial DNA sequences of greenbug (Homoptera: Aphididae) biotypes

Sequence comparisons were made for 738-bp of mtDNA cloned from seven greenbug, Schizaphis graminum, biotypes (B, C, E, F, G, H and I) obtained from laboratory colonies maintained by USDA-ARS, Stillwater, OK. These sequences include parts of the genes for 16S ribosomal subunit (16S rRNA), tRNAleu, tRNAser, cytochrome b (cytb) and NADH dehydrogenase (ND) subunits one and four. Sequence data revealed considerable variation in 86 (12%) nucleotide sites over the 738-bp sequenced among the seven greenbug biotypes. Nucleotide invariance was observed within the seven greenbug biotypes from both the laboratory colonies and field collected biotype E greenbugs from Kansas, Nebraska, Oklahoma, and Texas.     

Molecular analysis of sorghum resistance to the greenbug (Homoptera: Aphididae)

Chromosomal regions of sorghum, Sorghum bicolor (L.) Moench, conferring resistance to greenbug, Schizaphis graminum (Rondani), biotypes C, E, I, and K from four resistance sources were evaluated by restriction fragment-length polymorphism (RFLP) analysis. At least nine loci, dispersed on eight linkage groups, were implicated in affecting sorghum resistance to greenbug. The nine loci were named according to the genus of the host plant (Sorghum) and greenbug (Schizaphis graminum). Most resistance loci were additive or incompletely dominant. Several digenic interactions were identified, and in each case, these nonadditive interactions accounted for a greater portion of the resistance phenotype than did independently acting loci. One locus in three of the four sorghum crosses appeared responsible for a large portion of resistance to greenbug biotypes C and E. None of the loci identified were effective against all biotypes studied. Correspondingly, the RFLP results indicated resistance from disparate sorghums may be a consequence of allelic variation at particular loci. To prove this, it will be necessary to fine map and clone genes for resistance to greenbug from various sorghum sources.     

Mitochondrial DNA sequences of greenbug (Homoptera: Aphididae) biotypes

Sequence comparisons were made for 738-bp of mtDNA cloned from seven greenbug, Schizaphis graminum, biotypes (B, C, E, F, G, H and I) obtained from laboratory colonies maintained by USDA-ARS, Stillwater, OK. These sequences include parts of the genes for 16S ribosomal subunit (16S rRNA), tRNA^leu, tRNA^ser, cytochrome b (cytb) and NADH dehydrogenase (ND) subunits one and four. Sequence data revealed considerable variation in 86 (12%) nucleotide sites over the 738-bp sequenced among the seven greenbug biotypes. Nucleotide invariance was observed within the seven greenbug biotypes from both the laboratory colonies and field collected biotype E greenbugs from Kansas, Nebraska, Oklahoma, and Texas.     

Identifying insecticide-resistant greenbugs (Homoptera: Aphididae) with diagnostic assay tests

Laboratory bioassays were used to develop a diagnostic assay test for identifying greenburg, Schizaphis graminum (Rondani), populations that are insecticide-resistant. Petri dish assays with chlorpyrifos showed greenbug mortality should be monitored after 2 h of exposure. One-hour exposure did not kill a high percentage of susceptible greenbugs, and a 3-h exposure killed too many resistant greenbugs. Ethanol and methanol were both good solvents for mixing with chlorpyrifos in the petri dish assay. From the laboratory bioassays, four diagnostic concentrations of chlorpyrifos (3, 10, 30, and 100 ppm) were evaluated in the field by Texas A&M University agricultural research and extension entomologists across the Texas High Plains. Results from the diagnostic assay tests were compared with gel-electrophoresis resistance tests to validate resistance detection. The diagnostic assay tests gave the same greenbug resistance identification as the gel-electrophoresis analysis in 21 of 22 field bioassays in 1994 and 35 of 39 field bioassays in 1995. Diagnostic concentrations of 30 and 100 ppm chlorpyrifos killed > or = 85 and > or = 90%, respectively, of greenbugs identified by gel-electrophoresis as susceptible and < 40% and < 55%, respectively, of resistant greenbugs. The diagnostic assay technique is a quick, reliable, and inexpensive method for detecting insecticide resistance in greenbug populations.     

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.     

Inheritance of greenbug resistance in several forms of grain sorghum and sudangrass

The inheritance of resistance against the Krasnodar population of common greenbug Schizaphis graminum Rond. was analyzed in nine accessions of grain sorghum and sudangrass. The dominant gene of cultivar Capbam (k-455, United States) was effective against some greenbug clones and differed from the Sgr1-Sgr11 resistance genes. The gene was designated as Sgr12. The cultivar Capbam was proposed for use as a differentiator in population genetic studies in S. graminum. The cultivar Sarvasi (k-3852, Hungary) contains not only the dominant Sgr1 gene, but also a recessive gene (most likely Sgr2), which is effective against some greenbug clones. Grain sorghum accessions k-928 and k-929 (Gugara Belaya, western China) each carry two highly effective dominant resistance genes, which differ from Sgr1-Sgr4, Sgr6, Sgr9, and Sgr10. In addition, the resistance genes of accession k-929 differ from the Sgr5 gene. Accession k-928 proved to contain an additional dominant resistance gene, which is expressed in response to some greenbug clones. The gene was designated as Sgr13. Sudangrass accessions k-100 and k-122 (Ukraine) each carry two dominant resistance genes. Accessions k-62, k-99 (Ukraine), and k-96 (Russia) each carry one dominant and one recessive resistance gene. The dominant resistance genes that are expressed in the cultivar Odesskaya 25 (k-122) in response to infestation with some clones from the natural greenbug population were designated as Sgr14 and Sgr15.     

Biotypic diversity in greenbug (Hemiptera: Aphididae): characterizing new virulence and host associations

Biotypic diversity of the greenbug, Schizaphis graminum (Rondani) (Hemiptera: Aphididae), was assessed among populations collected from cultivated wheat, Triticum aestivum L., and sorghum, Sorghum bicolor (L.) Moench, and their associated noncultivated grass hosts. Greenbugs were collected during May through August 2002 from 30 counties of Kansas, Nebraska, Oklahoma, and Texas. Discounting the presumptive biotype A, five of the remaining nine letter-designated greenbug biotypes were collected; however, biotypes C, F, J, and K were not detected. Biotypes E and I exhibited the greatest host range and were the only biotypes collected in all four states. Sixteen greenbug clones, collected from eight plant species, exhibited unique biotype profiles. Eleven were collected from noncultivated grasses, three from wheat, and two from sorghum. The most virulent biotypes were collected from noncultivated hosts. The great degree of biotypic diversity among noncultivated grasses supports the contention that the greenbug species complex is composed of host-adapted races that diverged on grass species independently of, and well before, the advent of modern agriculture.     

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.     

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'.     

Identification of a major quantitative trait locus conditioning resistance to greenbug biotype E in sorghum PI 550610 using simple sequence repeat markers

Greenbug, Schizaphis graminum (Rondani), represents the most important pest insect of sorghum, Sorghum bicolor (L.) Moench, in the Great Plains of the United States. Biotype E is the most widespread and dominant type not only in sorghum and wheat, Triticum aestivum L., fields, but also on many noncultivated grass species. This study was designed to determine sorghum accession PI 550610 resistance to greenbug biotype E, to map the resistance quantitative trait loci (QTLs) by using an established simple sequence repeat (SSR) linkage map and to identify SSR markers closely linked to the major resistance QTLs. In greenhouse screening tests, seedlings of PI 550610 showed strong resistance to the greenbug at a level similar to resistant accession PI550607. For QTL mapping, one F2 population containing 277 progeny and one population containing 233 F2:3 families derived from Westland A line x PI 550610 were used to genotype 132 polymorphic SSR markers and to phenotype seedling resistance to greenbug feeding. Phenotypic evaluation of sorghum seedling damage at 7, 12, 17, and 21 d postinfestation in the F2:3 families revealed that resistance variation was normally distributed. Single marker analysis indicated 16 SSRs spread over five chromosomes were significant for greenbug resistance. Composite interval and multiple interval mapping procedures indicated that a major QTL resided in the interval of 6.8 cM between SSR markers Xtxp358 and Xtxp289 on SBI-09. The results will be valuable in the development of new greenbug biotype E resistant sorghum cultivars and for the further characterization of major genes by map-based cloning.     

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.     

Development and validation of a binomial sequential sampling plan for the greenbug (Homoptera: Aphididae) infesting winter wheat in the southern plains

From 1997 to 1999, Schizaphis graminum (Rondani), intensity (number per tiller) was estimated on 115 occasions from hard red winter wheat fields located throughout the major wheat growing regions of Oklahoma. A total of 32 and 83 fields was sampled during the fall and spring, respectively. The parameters of linear regressions relating the mean number of greenbugs per tiller (m) and the proportion of infested tillers (PT) differed significantly between fall and spring infestations. The PT-m linear model provided a good fit for data on S. graminum for fall and spring infestations at tally thresholds of 0, 1, 2, and 3. A tally threshold (T) represents the number of greenbugs present on a tiller before the tiller is classified as infested by >T greenbugs. A regression model with a tally threshold of 2 was the most precise for classifying S. graminum populations during fall growth of winter wheat because it explained a greater amount of the variation in the PT-m relationship (97%) than models with other tally thresholds. A separate spring model with a tally threshold of 1 was the most precise for classifying S. graminum populations during spring growth of winter wheat. Sequential sampling stop lines based on sequential probability ratio tests were calculated for economic thresholds of 3 or 6 greenbugs per tiller for fall infestations and 6 or 9 greenbugs per tiller for spring infestations. With the newly developed parameters, the average sample number required to classify greenbug populations near economic thresholds (as above or below the economic threshold) varied from 69 to 207. We expect that the sampling plans for greenbugs in winter wheat developed during this study will be efficient and useful tools for consultants and producers in the southern plains.     

Using high-throughput amplified fragment length polymorphism to distinguish sorghum greenbug (Homoptera: Aphididae) biotypes

Abstract 1 The greenbug Schizaphis graminum (Rondani) is a serious pest of Sorghum bicolor L. and small grains in the Southern Plains of the U.S.A. Use of resistant cultivars, the major greenbug management strategy, has been challenged by the rapid development of new greenbug biotypes that overcome plant resistance. 2 We used a high‐throughput amplified fragment length polymorphism (AFLP) fingerprinting method to examine genetic divergence among eight greenbug biotypes (B, C, E, G, I and K, New York and South Carolina). Clustering analysis based on 1775 scored AFLP markers clearly showed that biotypes (C, E, I and K), which are able to infest sorghum fields, share more common polymorphisms among themselves than with other biotypes. 3 This result suggests that common genetic factors exist among these biotypes, enabling them to predominate and thrive in monoculture crops. Our study demonstrated the sensitivity of AFLP in obtaining large quantities of biotype‐associated polymorphic information across the entire greenbug genome.     

Inheritance of greenbug resistance in several forms of grain sorghum and sudangrass

The inheritance of resistance against the Krasnodar population of common greenbug Schizaphis graminum Rond. was analyzed in nine accessions of grain sorghum and sudangrass. The dominant gene of cultivar Capbam (k-455, United States) was effective against some greenbug clones and differed from the Sgr1–Sgr11 resistance genes. The gene was designated as Sgr12. The cultivar Capbam was proposed for use as a differentiator in population genetic studies in S. graminum. The cultivar Sarvasi (k-3852, Hungary) contains not only the dominant Sgr1 gene, but also a recessive gene (most likely Sgr2), which is effective against some greenbug clones. Grain sorghum accessions k-928 and k-929 (Gugara Belaya, western China) each carry two highly effective dominant resistance genes, which differ from Sgr1–Sgr4, Sgr6, Sgr9, and Sgr10. In addition, the resistance genes of accession k-929 differ from the Sgr5 gene. Accession k-928 proved to contain an additional dominant resistance gene, which is expressed in response to some greenbug clones. The gene was designated as Sgr13. Sudangrass accessions k-100 and k-122 (Ukraine) each carry two dominant resistance genes. Accessions k-62, k-99 (Ukraine), and k-96 (Russia) each carry one dominant and one recessive resistance gene. The dominant resistance genes that are expressed in the cultivar Odesskaya 25 (k-122) in response to infestation with some clones from the natural greenbug population were designated as Sgr14 and Sgr15.     

Effect of insecticides used in corn, sorghum, and alfalfa on the predator Orius insidiosus (Hemiptera: Anthocoridae).

Orius insidoisus (Say) is an important predator in corn, sorghum, and alfalfa. Foliar insecticides commonly used on corn (permethrin, bifenthrin, and fipronil); sorghum (chlorpyrifos, carbofuran, dimethoate, and cyfluthrin); and both crops (A-cyhalothrin and ethyl parathion) were evaluated in 1998 and 1999 for their residual effects on O. insidiosus by caging adults on treated plants at several time intervals: at application (day 0) and 2, 3, and 6 d after application. In addition, imidacloprid, fipronil, and thiamethoxam used as seed treatments on corn and sorghum were tested for their effects on O. insidiosus by caging adults on plants in the presence and absence of greenbugs, Schizaphis graminum (Rondani). Finally, six of the same insecticides that also are used on alfalfa were evaluated in the field for their effects on O. insidiosus and other insects. On day 0, ethyl parathion. bifenthrin, and [lambda]-cyhalothrin on corn caused significantly higher mortality to O. insidiosus than permethrin and fipronil. Ethyl parathion and carbofuran on sorghum caused significantly higher mortality than chlorpyrifos, dimethoate, and A-cyhalothrin, which differed significantly from the control. Mortality with cyfluthrin did not differ significantly from that in the control. Insecticides had no significant effects on O. insidiosus 3 and 6 d after application in 1998 with the exception of permethrin on day 3. Similar patterns of mortality were observed in 1999 experiments. No significant differences in mortality of adults occurred with fipronil and thiamethoxam in the presence and absence of greenbugs. Imidachloprid caused significantly higher mortality to O. insidiosus adults than thiamethoxam or fipronil in some instances when greenbugs were not supplied as food. In alfalfa, the insecticides caused significant mortality to most of the insects evaluated. Ethyl parathion, permethrin, chlorpyrifos, and cyfluthrin caused significantly higher mortality to O. insidiosus than carbofuran and A-cyhalothrin, which differed significantly from the control in 1998. In 1999, all treatments significantly reduced O. insidiosus numbers and did not differ significantly from each other.     

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.