John D. Burd

Mitochondrial DNA sequence divergence among greenbug (Homoptera: Aphididae) biotypes: evidence for host-adapted races

The full complement of known greenbug, Schizaphis graminum (Rondani), biotypes found in the USA were subjected to a molecular phylogenetic analysis based on a 1.2‐kb portion of the cytochrome oxidase I mitochondrial gene. In addition to these nine biotypes (B, C, E, F, G, H, I, J and K), a probable isolate of the enigmatic biotype A (NY), a ‘new biotype’ collected from Elymus canadensis (L.) (CWR), and an isolate from Germany (EUR) were included. Schizaphis rotundiventris (Signoret) was included as an outgroup. Genetic distances among S. graminum biotypes ranged from 0.08% to 6.17% difference in nucleotide substitutions. Neighbour‐joining, maximum parsimony and maximum likelihood analyses all produced dendrograms revealing three clades within S. graminum. Clade 1 contained the ‘agricultural’ biotypes commonly found on sorghum and wheat (C, E, K, I, plus J) and there were few substitutions among these biotypes. Clade 2 contained F, G and NY, and Clade 3 contained B, CWR and EUR, all of which are rarely found on crops. The rarest biotype, H, fell outside the above clades and may represent another Schizaphis species. S. graminum biotypes are a mixture of genotypes belonging to three clades and may have diverged as host‐adapted races on wild grasses.

Mitochondrial DNA sequence divergence among Schizaphis graminum (Hemiptera: Aphididae) clones from cultivated and non-cultivated hosts: haplotype and host associations.

A 1.0 kb region of the mitochondrial cytochrome oxidase subunit I gene from the greenbug aphid, Schizaphis graminum (Rondani), was sequenced for 24 field collected clones from non-cultivated and cultivated hosts. Maximum likelihood, maximum parsimony and neighbour-joining phylogenies were estimated for these clones, plus 12 previously sequenced clones. All three tests produced trees with identical topologies and confirmed the presence of three clades within S. graminum. Clones showed no relationship between biotype and mtDNA haplotype. At least one biotype was found in all three clades, suggesting exchange among clades of genetic material conditioning for crop virulence, or the sharing of a common ancestor. However, there was a relationship between host and haplotype. Clade 1 was the most homogeneous and contained 12 of 16 clones collected from cultivated hosts and five of the six collected from johnsongrass, Sorghum halepense, a congener of cultivated sorghum, S. bicolor. Four of the six clones collected from Agropyron spp. were found in clade 2. Clade 3 contained two clones from wheat, Triticum aestivum, and four from non-cultivated hosts other than Agropyron spp. A partitioning of populations by mtDNA haplotype and host suggests the occurrence of host adapted races in Schizaphis graminum.

Efficacy of Pyramiding Greenbug (Homoptera: Aphididae) Resistance Genes in Wheat

Abstract Durable resistance to greenbug, Schizaphis graminum (Rondani), in wheat is a goal of wheat improvement teams, and one that has been complicated by the regular occurrence of damaging biotypes. Simulation modeling studies suggest that pyramiding resistance genes, i.e., combining more than one resistance gene in a single cultivar or hybrid, may provide more durable resistance than sequential releases of single genes. We examined this theory by pyramiding resistance genes in wheat and testing a series of greenbug biotypes. Resistance genes Gb2, Gb3, and Gb6, and pyramided genes Gb2/Gb3, Gb2/Gb6, and Gb3/Gb6 were tested for effectiveness against biotypes E, F, G, H, and I. By comparing reactions of plants with pyramided genes to those with single resistance genes, we found that pyramiding provided no additional protection over that conferred by the single resistance genes. Based on the results of this test, we concluded that the sequential release of single resistance genes, combined with careful monitoring of greenbug population biotypes, is the most effective gene deployment strategy for greenbug resistance in wheat.

Tritrophic relationships of Russian wheat aphid (Homoptera: Aphididae), a hymenopterous parasitoid (Diaeretiella rapae McIntosh), and resistant and susceptible small grains

Abstract The tritrophic relationships between resistant small grain entries, Russian wheat aphid, Diuraphis noxia (Mordvilko), and Diaeretiella rapae McIntosh were studied. Resistant triticale entries with high levels of antibiosis affected growth and reproduction of both the aphid and the parasitoid. Parasitoid mummies from antibiotic triticale were smaller, took longer to develop, and were fewer in number compared with those from susceptible entries. The sex ratio of parasitoids emerging from the smaller mummies of aphids restricted to the resistant triticale entries was skewed toward females. The protective influence of parasitoids on plants was demonstrated in the experiment. Plants exposed to aphids without parasitoids sustained greater damage and had significantly less biomass than those so protected. A resistant wheat entry showed a reduction in aphid populations and enhanced parasitoid activity due to the fact that its leaves did not roll like those in the susceptible wheat entry.

Distribution and diversity of russian wheat aphid (Hemiptera: Aphididae) biotypes in North America.

Abstract Wheat, Triticum aestivum L., with Russian wheat aphid, Diuraphis noxia (Kurdjumov) (Hemiptera: Aphididae) resistance based on the Dn4 gene has been important in managing Russian wheat aphid since 1994. Recently, five biotypes (RWA1–RWA5) of this aphid have been described based on their ability to differentially damage RWA resistance genes in wheat. RWA2, RWA4, and RWA5 are of great concern because they can kill wheat with Dn4 resistance. In 2005, 365 Russian wheat aphid clone colonies were made from collections taken from 98 fields of wheat or barley, Hordeum vulgare L., in Oklahoma, Texas, New Mexico, Colorado, Kansas, Nebraska, and Wyoming to determine their biotypic status. The biotype of each clone was determined through its ability to differentially damage two resistant and two susceptible wheat entries in two phases of screening. The first phase determined the damage responses of Russian wheat aphid wheat entries with resistance genes Dn4, Dn7, and susceptible ‘Custer’ to infestations by each clone to identify RWA1 to RWA4. The second phase used the responses of Custer and ‘Yuma’ wheat to identify RWA1 and RWA5. Only two biotypes, RWA1 and RWA2, were identified in this study. The biotype composition across all collection sites was 27.2% RWA1 and 72.8% RWA2. RWA biotype frequency by state indicated that RWA2 was the predominant biotype and composed 73–95% of the biotype complex in Texas, Oklahoma, Colorado, and Wyoming. Our study indicated that RWA2 is widely distributed and that it has rapidly dominated the biotype complex in wheat and barley within its primary range from Texas to Wyoming. Wheat with the Dn4 resistance gene will have little value in managing RWA in the United States, based on the predominance of RWA2.

Effects of Riparian and Grassland Habitats on Ground Beetle (Coleoptera: Carabidae) Assemblages in Adjacent Wheat Fields

Abstract Natural habitats surrounding agricultural fields provide a source of natural enemies to assist in pest control. The boundaries among landscape elements filter some organisms attempting to cross them, resulting in differing communities within the landscape elements. Ground beetles are numerous and generally disperse by walking. These qualities make them excellent organisms for the study of boundary dynamics. Our goal was to determine if natural habitats adjacent to wheat fields affected the species composition of ground beetles within the wheat fields. We captured ground beetles from autumn through spring 1996–1997 at two sites using directional pitfall traps placed in wheat fields and adjacent grasslands and riparian zones. Ground beetle abundance reached two peaks, one in autumn and the other in spring. Species composition was most strongly related to these seasons. Axis 1 of a canonical correspondence analysis separated spring active beetles from autumn active beetles. Axis 2 separated winter active beetles. With the effects of season and sites removed, axes 1 and 2 of a partial canonical correspondence analysis separated beetles with respect to habitat. Axis 1 separated beetles into wheat and natural habitat assemblages. Axis 2 further distinguished assemblages in wheat fields as those adjacent to grasslands and those adjacent to riparian habitats. Axis 2 also separated grassland, grassland edge, and riparian edge assemblages from riparian assemblages. Net dispersal of beetles across the boundaries showed no consistent pattern during autumn, winter, or spring. However, mark–recapture studies showed that several species routinely cross boundaries, which resulted in different community structures and an increase in abundance of beetles in the wheat interiors during spring.

Virus Transmission Phenotype Is Correlated with Host Adaptation Among Genetically Diverse Populations of the Aphid Schizaphis graminum

ABSTRACT Schizaphis graminum is an important insect pest of several grain crops and an efficient vector of cereal-infecting luteoviruses and poleroviruses. We examined the virus transmission characteristics of several distinct populations and various developmental stages of the aphid. Seven well-characterized S. graminum biotypes maintained at the USDA-ARS laboratory in Stillwater, OK, and two biotypes maintained in New York (one collected in Wisconsin and the other collected in South Carolina) were tested for their ability to transmit five viruses that cause barley yellow dwarf disease (BYD). Four of the Oklahoma biotypes, which do not commonly colonize agronomic crops, and the Wisconsin biotype, were efficient vectors of several viruses. The three other Oklahoma biotypes, which do colonize agronomic crops, and the South Carolina biotype, were poor vectors of all five viruses. Thus, the vector specificity long associated with viruses causing BYD is not limited to the level of aphid species; it clearly extends to populations within a single species. S. graminum nymphs are reported to be more efficient vectors of Barley yellow dwarf virus (BYDV-SGV) than are adults. This was confirmed only for the Wisconsin biotype, but not for the other eight S. graminum biotypes. Thus, there does not appear to be a generalized developmentally regulated barrier to the transmission of BYDV-SGV in S. graminum. Furthermore, the developmentally regulated vector competency observed in the Wisconsin biotype did not extend to other viruses. BYDV-PAV and Cereal yellow dwarf virus-RPV were transmitted with similar efficiency by all S. graminum biotypes when acquired by nymphs or adults.

Over-Summering and Biotypic Diversity of Schizaphis graminum (Homoptera: Aphididae) Populations on Noncultivated Grass Hosts

Abstract Greenbug, Schizaphis graminum (Rondani), populations over-summering on noncultivated grass hosts may be implicated in early fall infestations in wheat. The purpose of this study was to examine the relationship between over-summering greenbugs on noncrop hosts and fall infestations on wheat. Since greenbug populations on noncultivated hosts may also act as reservoirs of virulence genes, the biotypes of collected aphids were also determined. The grass species present at three sites (two in Oklahoma and one in Kansas) were identified and a species richness curve was generated. Greenbugs were collected at these sites and their hosts and biotypes determined. At Hays, KS, a persistent over-summering greenbug population lead to an early fall infestation in wheat. At the sites in Oklahoma, where over-summering greenbugs were not detected, the fall infestation occurred 3 months later. Biotypes G, I, K, and a new biotype (i.e., previously undescribed) were found on noncultivated hosts at Hays, but only biotypes I and K were found on the cultivated wheat. Finding a new biotype supports the hypothesis that biotypic diversity (new combinations of virulence genes) is generated and maintained on noncultivated grasses, which may then act as reservoirs of virulence genes found in populations on crops.

Physiological Modification of the Host Feeding Site by Cereal Aphids (Homoptera: Aphididae)

Abstract Indole-3-acetic acid-1-14C and 14C-sucrose labels were used to study the effects of greenbugs, Schizaphis graminum (Rondani), and Russian wheat aphids, Diuraphis noxia (Mordvilko), on phloem function of wheat (Triticum aestivum L.). Greenbug feeding significantly reduced translocation from the immediate feeding site; however, phloem integrity was not impeded. In contrast, Russian wheat aphids had little effect on vein loading or phloem translocation at the feeding site. Similar results were obtained when resistant and susceptible wheats were infested with three different greenbug biotypes. Greenbugs fed artificial diets containing 14C-sucrose injected salivary material that was translocated to both root and shoot systems. The accumulation of salivary constituents in the roots of wheat seedlings fed upon by greenbugs may account for the significant reductions in root biomass that have previously been reported.

Biotypic Diversity in Greenbug (Hemiptera: Aphididae): Microsatellite-Based Regional Divergence and Host-Adapted Differentiation

ABSTRACT Nineteen isolates of the cereal aphid pest greenbug, Schizaphis graminum (Rondani) (Hemiptera: Aphididae), were collected from wheat, Triticum aestivum L.; barley, Hordeum vulgare L.; or noncultivated grass hosts in five locations from Colorado and Wyoming. Parthenogenetic colonies were established. Biotypic profiles of the 19 isolates were determined based on their abilities to damage a set of host plant differentials, and 13 new biotypes were identified. Genetic diversity among the 19 isolates and five previously designated greenbug biotypes (E, G, H, I, and K) was examined with 31 cross-species transferable microsatellite (simple sequence repeat) markers. Neighbor-joining clustering analysis of marker data revealed host-adapted genetic divergence as well as regional differentiation of greenbug populations. Host associated biotypic variation seems to be more obvious in “agricultural biotypes,” whereas isolates collected from noncultivated grasses tend to show more geographic divergence. It seems that the biotype sharing the most similar biotypic profiles and the same geographic region with current prevailing one may have the greatest potential to become the new prevailing biotype. Close monitoring of greenbug population dynamics especially biotypic variation on both crop plants and noncultivated grasses in small grain production areas may be a useful strategy for detecting potentially new prevailing virulent biotypes of the greenbug.