Genomic and Proteomic Analysis of Schizaphis graminum Reveals Cyclophilin Proteins Are Involved in the Transmission of Cereal Yellow Dwarf Virus

Yellow dwarf viruses cause the most economically important virus diseases of cereal crops worldwide and are transmitted by aphid vectors. The identification of aphid genes and proteins mediating virus transmission is critical to develop agriculturally sustainable virus management practices and to understand viral strategies for circulative movement in all insect vectors. Two cyclophilin B proteins, S28 and S29, were identified previously in populations of Schizaphisgraminum that differed in their ability to transmit the RPV strain of Cereal yellow dwarf virus (CYDV-RPV). The presence of S29 was correlated with F2 genotypes that were efficient virus transmitters. The present study revealed the two proteins were isoforms, and a single amino acid change distinguished S28 and S29. The distribution of the two alleles was determined in 12 F2 genotypes segregating for CYDV-RPV transmission capacity and in 11 genetically independent, field-collected S . graminum biotypes. Transmission efficiency for CYDV-RPV was determined in all genotypes and biotypes. The S29 isoform was present in all genotypes or biotypes that efficiently transmit CYDV-RPV and more specifically in genotypes that efficiently transport virus across the hindgut. We confirmed a direct interaction between CYDV-RPV and both S28 and S29 using purified virus and bacterially expressed, his-tagged S28 and S29 proteins. Importantly, S29 failed to interact with a closely related virus that is transported across the aphid midgut. We tested for in vivo interactions using an aphid-virus co-immunoprecipitation strategy coupled with a bottom-up LC-MS/MS analysis using a Q Exactive mass spectrometer. This analysis enabled us to identify a third cyclophilin protein, cyclophilin A, interacting directly or in complex with purified CYDV-RPV. Taken together, these data provide evidence that both cyclophilin A and B interact with CYDV-RPV, and these interactions may be important but not sufficient to mediate virus transport from the hindgut lumen into the hemocoel.     

Genetics coupled to quantitative intact proteomics links heritable aphid and endosymbiont protein expression to circulative polerovirus transmission

Yellow dwarf viruses in the family Luteoviridae, which are the causal agents of yellow dwarf disease in cereal crops, are each transmitted most efficiently by different species of aphids in a circulative manner that requires the virus to interact with a multitude of aphid proteins. Aphid proteins differentially expressed in F2 Schizaphis graminum genotypes segregating for the ability to transmit Cereal yellow dwarf virus-RPV (CYDV-RPV) were identified using two-dimensional difference gel electrophoresis (DIGE) coupled to either matrix-assisted laser desorption ionization-tandem mass spectrometry or online nanoscale liquid chromatography coupled to electrospray tandem mass spectrometry. A total of 50 protein spots, containing aphid proteins and proteins from the aphid's obligate and maternally inherited bacterial endosymbiont, Buchnera, were identified as differentially expressed between transmission-competent and refractive aphids. Surprisingly, in virus transmission-competent F2 genotypes, the isoelectric points of the Buchnera proteins did not match those in the maternal Buchnera proteome as expected, but instead they aligned with the Buchnera proteome of the transmission-competent paternal parent. Among the aphid proteins identified, many were involved in energy metabolism, membrane trafficking, lipid signaling, and the cytoskeleton. At least eight aphid proteins were expressed as heritable, isoelectric point isoform pairs, one derived from each parental lineage. In the F2 genotypes, the expression of aphid protein isoforms derived from the competent parental lineage aligned with the virus transmission phenotype with high precision. Thus, these isoforms are candidate biomarkers for CYDV-RPV transmission in S. graminum. Our combined genetic and DIGE approach also made it possible to predict where several of the proteins may be expressed in refractive aphids with different barriers to transmission. Twelve proteins were predicted to act in the hindgut of the aphid, while six proteins were predicted to be associated with the accessory salivary glands or hemolymph. Knowledge of the proteins that regulate virus transmission and their predicted locations will aid in understanding the biochemical mechanisms regulating circulative virus transmission in aphids, as well as in identifying new targets to block transmission.     

Resistance to multiple cereal aphids in wheat–alien substitution and translocation lines

Rhopalosiphum padi, Schizaphis graminum, and Sitobion avenae are three of the most destructive aphid species of wheat (Triticum aestivum L.). They can significantly reduce wheat yields directly by feeding and indirectly by transmitting viruses. This study aimed to search for resistance to these aphid species among lines derived from different rye (Secale cereale) origins and from Aegilops speltoides, all in the genetic background of the wheat cultivar Pavon F76. Resistance was quantified as aphid weight (R. padi, S. avenae, and S. graminum) and the number of aphids and percentage of infested leaf area exhibiting chlorosis (S. graminum). The most resistant genotypes reduced R. padi and S. avenae weight by 24.2 and 34.3 %, respectively, at the seedling stage, compared with Pavon F76 control plants. Strong S. graminum resistance was found only in A. speltoides-derived lines, the most resistant of which (7A.7S-L5) sustained just 3 % chlorosis and reduced S. graminum colony weight by 67.7 %. One line carrying the 1AL.1RS_am wheat–rye translocation from Amigo wheat (originally from Insave rye) reduced S. avenae weight by 23.2 and 21.8 % in seedling and adult plants, respectively. Single genotypes carrying the complete 1R chromosome or the 1RS chromosome arm derived from E12165 wheat and Presto triticale proved to be resistant to both R. padi and S. avenae at the seedling stage. Further research should be conducted to unravel the genetic basis of resistance to these aphids in 1RS genotypes. The sources of resistance identified here may be useful for incorporating multiple aphid species resistance in wheat breeding programs, particularly for R. padi and S. avenae, to which no resistant wheats have been bred.     

Glycine-betaine accumulation influences susceptibility of water-stressed barley to the aphid Schizaphis graminum

Water deficit increased susceptibility of barley to the aphid Schizaphis graminum. Proline and glycine-betaine accumulated in the stressed plants. These compounds were incorporated into artificial diets to test their effects on aphids. Survival of S. graminum was not affected by proline and glycine-betaine. In addition, glycine-betaine increased reproduction of the greenbug at concentrations similar to those found in stressed barley plants. When glycine-betaine was added to detached shoots of barley, population growth rate of S. graminum increased in that plant material kept in the betaine solutions. It is suggested that glycine-betaine accumulation may be responsible for the increased susceptibility of water-stressed barley to the greenbug.     

Cryptic Virulence and Avirulence Alleles Revealed by Controlled Sexual Recombination in Pea Aphids

Although aphids are worldwide crop pests, little is known about aphid effector genes underlying virulence and avirulence. Here we show that controlling the genetics of both aphid and host can reveal novel recombinant genotypes with previously undetected allelic variation in both virulence and avirulence functions. Clonal F₁ progeny populations were derived from reciprocal crosses and self-matings between two parental genotypes of pea aphid (Acyrthosiphon pisum) differing in virulence on a Medicago truncatula host carrying the RAP1 and RAP2 resistance genes. These populations showed Mendelian segregation consistent with aphid performance being controlled largely by a dominant virulence allele derived from only one parent. Altered segregation ratios on near-isogenic host genotypes differing in the region carrying RAP1 were indicative of additional heritable functions likely related to avirulence genes originating from both parents. Unexpectedly, some virulent F₁ progeny were recovered from selfing of an avirulent parent, suggesting a reservoir of cryptic alleles. Host chlorosis was associated with virulence, whereas necrotic hypersensitive-like response was not. No maternal inheritance was found for any of these characteristics, ruling out sex-linked, cytoplasmic, and endosymbiotic factors. Our results demonstrate the tractability of dissecting the genetic basis of pest-host resistance mechanisms and indicate that the annual sexual cycle in aphids may lead to frequent novel genotypes with both increased and decreased virulence. Availability of genomes for both pest and host can facilitate definition of cognate gene-for-gene relationships, potentially leading to selection of crop genotypes with multiple resistance traits.     

Tangible benefits of the aphid Acyrthosiphon pisum genome sequencing for aphid proteomics: Enhancements in protein identification and data validation for homology-based proteomics

Homology-driven proteomics promises to reveal functional biology in insects with sparse genome sequence information. A proteomics study comparing plant virus transmission competent and refractive genotypes of the aphid Schizaphis graminum isolated numerous candidate proteins involved in virus transmission, but limited genome sequence information hampered their identification. The complete genome of the pea aphid, Acyrthosiphon pisum, released in 2008, enabled us to double the number of protein identifications beyond what was possible using available EST libraries and other insect sequences. This was concomitant with a dramatic increase of the number of MS and MS/MS peptide spectra matching the genome-derived protein sequence. LC-MS/MS proved to be the most robust method of peptide detection. Cross-matching spectral data to multiple EST sequences and error tolerant searching to identify amino acid substitutions enhanced the percent coverage of the Schizaphis graminum proteins. 2-D electrophoresis provided the protein pI and MW which enabled the refinement of the candidate protein selection and provided a measure of protein abundance when coupled to the spectral data. Thus, the homology-based proteomics pipeline for insects should include efforts to maximize the number of peptide matches to the protein to increase certainty in protein identification and relative protein abundance.     

Virus-Vector Cell Interactions Regulating Transmission Specificity of Soybean Dwarf Luteoviruses

Transmission of soybean dwarf viruses (SbDV) indigenous to Japan (SbDV‐D) and to the eastern United States (SbDV‐Va19) were compared in vector and nonvector aphid species. Absolute vector‐specificity was maintained when Aulacorthum solani, Acyrthosiphon pisum, and Myzus persicae were allowed to feed on solutions of either virus (100 μg/ml) through Parafil© membranes. SbDVD was transmitted only by A. solani, and SbDV‐Va19 was transmitted only by A. pisum and M. persicae. Similar results were obtained when individual aphids were micro‐injected with 2 ng virus and subsequently allowed to feed on healthy plants. Ultrastructural studies of A. solani and M. persicae indicated that both SbDV‐D and SbDV‐Va20 were acquired specifically through the aphid hindgut. No difference in hindgut acquisition specificity was observed, and both A. solani and M. persicae were able to transport SbDV‐D and SbDV‐Va20 into the haemocoel by endocytotic/exocytotic pathways. When injected, SbDV was shown to be associated with only the accessory salivary glands (ASG) in aphids, indicating a high level of tissue specificity. Two different interactions with the ASG were observed for SbDV‐D and SbDV‐Va20 in A. solani and M. persicae. SbDV‐D penetrated the ASG basal lamina of A. solani, but was never observed in the basal lamina of M. persicae. The ASG basal lamina was a barrier to SbDV‐D transmission by M. persicae. SbDV‐Va19 penetrated the ASG basal lamina of both A. solani and M. persicae. However, SbDV‐Va20 was not observed in the ASG cytoplasm in A. solani, indicating that the basal plasmalemma functioned as the transmission barrier. Observations indicated that capsid protein structure, aphid basal lamina composition and cell membrane components influenced virus‐aphid interactions regulating SbDV transmission.     

Proteins Identified from Saliva and Salivary Glands of the Chinese Gall Aphid Schlechtendalia chinensis

Aphid saliva plays an essential role in the interaction between aphids and their host plants. Several aphid salivary proteins have been identified but none from galling aphids. Here the salivary proteins from the Chinese gall aphid are analyzed, Schlechtendalia chinensis, via an LC‐MS/MS analysis. A total of 31 proteins are identified directly from saliva collected via an artificial diet, and 141 proteins are identified from extracts derived from dissected salivary glands. Among these identified proteins, 17 are found in both collected saliva and dissected salivary glands. In comparison with salivary proteins from ten other free‐living Hemipterans, the most striking feature of the salivary protein from S. chinensis is the existence of high proportion of proteins with binding activity, including DNA‐, protein‐, ATP‐, and iron‐binding proteins. These proteins maybe involved in gall formation. These results provide a framework for future research to elucidate the molecular basis for gall induction by galling aphids.     

Comparative transmission of pea mosaic virus by one strain ofMyzus persicae Sulz. and two strains ofAcyrthosiphon pisum Harris

Experiments were conducted to compare the efficiency of transmission of a strain of pea mosaic virus (PMV) isolated in Czechoslovakia by two strains (clones) of the pea aphidAcyrthosiphon pisum Harris (green and red) and one strain ofMyzus persicae Sulz. PMV is a nonpersistent virus and the preliminary fasting of aphids before acquisition feeding increases the efficiency of aphids in transmission of this virus. In our experiments two hour fasted individuals were used and two periods of acquisition feeding on the source (1 and 5 minutes). On the healthy test plants the aphids were left over night. As the source and test plants pea (Pisum sativum L.) of the cv. Raman were used. During the one minute acquisition period on the source of infection the aphids were observed under the stereoscopic microscope. They usually made two to three probes. During the five minute acquisition feeding time the aphids were not observed and they were taken from the source of infection after a lapse of five minutes. To compare the efficiency in transmission of this virus by these aphids only one aphid per tested plant was used and in all trials only two to four day old nymphs were taken. Differences in transmission efficiency between two strains ofAcyrthosiphon pisum Harris were highly significant. The green strain of pea aphid was the less efficient vector in comparison with the green peach aphid and the red strain of the pea aphid, the latter being the most efficient vector of this virus.     

Effects of indole alkaloids from gramineae on aphids

The indole bases N_∞-methyltryptamine, 5-methoxy-N, N-dimethyltryptamine, 3-N,N-dimethylaminomethylindole (gramine) and 5-methoxytryptamine decreased survival of nymphs of Rhopalosiphum maidis with LD₅₀ of 3.8, 3.5, 2.9 and 2.3 mM, respectively, after 48 hr of feeding with a synthetic diet. Gramine also decreased survival of Schizaphis graminum with an LD₅₀ of 0.7 mM after 48 hr of feeding. The four compounds tested showed feeding deterrent activity on the aphids at concentrations as low as 0.5 mM. In addition, gramine showed toxic effects on S. graminum. Since the deleterious effects of gramine and related compounds in synthetic diets are observed at concentrations similar to those found in plants, it is possible that these molecules may have a role in protecting the plant against aphids     

Insight into watery saliva proteomes of the grain aphid,Sitobion avenae

The grain aphid, Sitobion avenae, is an economically important cereal pest worldwide. Aphid saliva plays an essential role in the interaction between aphids and their host plants. However, limited information is available regarding the proteins found in the saliva of S. avenae. Here, the watery saliva proteins from S. avenae were collected in an artificial diet and identified using a liquid chromatography–mass spectrometry/mass spectrometry analysis. A total of 114 proteins were identified in S. avenae saliva, including several enzymes, binding proteins, and putative effectors, as well as other proteins with unknown functions. In comparison with salivary proteins from nine other aphid species, the most striking feature of the salivary protein from S. avenae was the different patterns of protein functions. Several orthologous proteins secreted by other aphid species such as glucose dehydrogenase, elongation factors, and effector C002 were also detected in S. avenae saliva and speculated to play a significant role in aphid–plant interactions. These results provide further insight into the molecular basis between aphids and cereal plant interactions.     

Biometrical genetic analysis of luteovirus transmission in the aphid Schizaphis graminum

The aphid Schizaphis graminum is an important vector of the viruses that cause barley yellow dwarf disease. We studied the genetic architecture of virus transmission by crossing a vector and a non-vector genotype of S. graminum. F1 and F2 hybrids were generated, and a modified line-cross biometrical analysis was performed on transmission phenotype of two of the viruses that cause barley yellow dwarf: Cereal yellow dwarf virus (CYDV)-RPV and Barley yellow dwarf virus (BYDV)-SGV. Our aims were to (1) determine to what extent differences in transmission ability between vectors and non-vectors is due to net additive or non-additive gene action, (2) estimate the number of loci that determine transmission ability and (3) examine the nature of genetic correlations between transmission of CYDV-RPV and BYDV-SGV. Only additive effects contributed significantly to divergence in transmission of both CYDV-RPV and BYDV-SGV. For each luteovirus, Castle-Wright's estimator for the number of effective factors segregating for transmission phenotype was less than one. Transmission of CYDV-RPV and BYDV-SGV was significantly correlated in the F2 generation, suggesting that there is a partial genetic overlap for transmission of these luteoviruses. Yet, 63% of the F2 genotypes transmitted CYDV-RPV and BYDV-SGV at significantly different rates. Our data suggest that in S. graminum, the transmission efficiency of both CYDV-RPV and BYDV-SGV is regulated by a major gene or set of tightly linked genes, and the transmission efficiency of each virus is influenced by a unique set of minor genes.     

Detection of fungal metabolites of various Trichoderma species by the aphid Schizaphis graminum

The feeding preferences of alate and apterous morphs of the aphid Schizaphis graminum (Rondani) (Homoptera: Aphididae) were evaluated using leaves treated with powdered rice cultures of four fungal isolates belonging to different species of the genus Trichoderma (Deuteromycotina: Hyphomycetes). All of the fungal isolates restrained alate morphs of S. graminum from visiting treated leaves, but only Trichoderma citrinoviride Bisset also influenced the preference of apterous morphs. Trials carried out with supernatants obtained by centrifuging aqueous suspensions of the fungal cultures showed that the feeding preference of aphids was maintained in the absence of fungal spores and mycelia, supporting the hypothesis that at least part of the fungal metabolites responsible for this effect were water‐soluble compounds. Electrophysiological studies showed that the structures involved in the perception of the fungal metabolites are located on the aphid tarsomeres.     

Chromosome‐level genome assembly for the horned‐gall aphid provides insights into interactions between gall‐making insect and its host plant

The aphid Schlechtendalia chinensis is an economically important insect that can induce horned galls, which are valuable for the medicinal and chemical industries. Up to now, more than twenty aphid genomes have been reported. Most of the sequenced genomes are derived from free‐living aphids. Here, we generated a high‐quality genome assembly from a galling aphid. The final genome assembly is 271.52 Mb, representing one of the smallest sequenced genomes of aphids. The genome assembly is based on contig and scaffold N50 values of the genome sequence are 3.77 Mb and 20.41 Mb, respectively. Nine‐seven percent of the assembled sequences was anchored onto 13 chromosomes. Based on BUSCO analysis, the assembly involved 96.9% of conserved arthropod and 98.5% of the conserved Hemiptera single‐copy orthologous genes. A total of 14,089 protein‐coding genes were predicted. Phylogenetic analysis revealed that S. chinensis diverged from the common ancestor of Eriosoma lanigerum approximately 57 million years ago (MYA). In addition, 35 genes encoding salivary gland proteins showed differentially when S. chinensis forms a gall, suggesting they have potential roles in gall formation and plant defense suppression. Taken together, this high‐quality S. chinensis genome assembly and annotation provide a solid genetic foundation for future research to reveal the mechanism of gall formation and to explore the interaction between aphids and their host plants.     

Genotypic interactions in an aphid-host plant relationship: Uroleucon rudbeckiae and Rudbeckia laciniata

Summary Four clones of the aphid Uroleucon rudbeckiae were grown on four clones of the host plant Rudbeckia laciniata. Age-specific fecundities were used to determine the fitness, F _i , of each individual aphid. The analysis of variance for F _i revealed that (1) plant genotype has a significant effect on aphid fitness; (2) there is an interaction between aphid and host plant genotypes with respect to aphid fitness; and (3) aphid fitness is affected by phenotypic differences among individual host plants. Result (2) supports the hypothesis that genotypic interactions between aphid and host may maintain genetic diversity in aphid populations. The results of a preference test, while not significant at customary probability levels, suggested that aphids will choose to feed on plants which confer greater fitness.     

Vector specificity of barley yellow dwarf virus serotypes and variants in southwestern Idaho

Plants with symptoms of barley yellow dwarf virus (BYDV) obtained in infection feeding assays of aphids collected in the field in Idaho between 1986 and 1988 were tested for virus transmissibility by possible aphid vectors. Isolates obtained during 1987–1988 were also tested with a range of polyclonal antisera which distinguished PAV, MAV, SGV, RPV and RMV serotypes. In 1989 some Idaho (ID) BYDV isolates, maintained as standards for comparison, were serotyped and tested for aphid transmissibility, using 11 species of aphids. There was not always the expected correspondence between serotype and vector specificity for ID isolates. For isolates obtained from field-collected Rhopalosiphum padi, vector transmissibility and serotype corresponded with previous reports; however, 44% of isolates which were serotyped as RMV were also transmissible by species other than Rhopalosiphum maidis. Similarly, the transmissibility of the ID laboratory standards did not always conform to the reported vector specificity of serotypes. The laboratory ID-MAV culture was transmitted by Metopolophium dirhodum and Myzus persicae as well as by Sitobion avenae. The laboratory ID-SGV culture was transmitted by R. padi and 5. avenae as well as by Schizaphis graminum. The ID-RPV culture was transmitted by S. graminum and Rhopalosiphum insertum as well as R. padi. Both of two laboratory ID-RMV cultures were transmissible by R. insertum and R. padi transmitted one of them. The results indicate that, for isolates collected in Idaho, vector specificity cannot be assumed from their serotypes.     

Tripartite Interactions of Barley Yellow Dwarf Virus,Sitobion avenae and Wheat Varieties

The tripartite interactions in a pathosystem involving wheat (Triticum aestivum L.), the Barley yellow dwarf virus (BYDV), and the BYDV vector aphid Sitobion avenae were studied under field conditions to determine the impact of these interactions on aphid populations, virus pathology and grain yield. Wheat varietal resistance to BYDV and aphids varied among the three wheat varieties studied over two consecutive years. The results demonstrated that (1) aphid peak number (APN) in the aphid + BYDV (viruliferous aphid) treatment was greater and occurred earlier than that in the non-viruliferous aphid treatment. The APN and the area under the curve of population dynamics (AUC) on a S. avenae-resistant variety 98-10-30 was significantly lower than on two aphid-susceptible varieties Tam200(13)G and Xiaoyan6. (2) The production of alatae (PA) was greater on the variety 98-10-30 than on the other varieties, and PA was greater in the aphid + BYDV treatment on 98-10-30 than in the non-viruliferous aphid treatment, but this trend was reversed on Tam200(13)G and Xiaoyan6. (3) The BYDV disease incidence (DIC) on the variety 98-10-30 was greater than that on the other two varieties in 2012, and the disease index (DID) on Tam200(13)G was lower than on the other varieties in the aphid + BYDV and BYDV treatments in 2012, but not in 2011 when aphid vector numbers were generally lower. (4) Yield loss in the aphid + BYDV treatment tended to be greater than that in the aphid or BYDV alone treatments across varieties and years. We suggested that aphid population development and BYDV transmission tend to promote each other under field conditions. The aphids + BYDV treatment caused greater yield reductions than non-viruliferous aphids or virus treatment. Wheat varietal resistance in 98-10-30 affects the aphid dispersal, virus transmission and wheat yield loss though inhibits aphid populations from increasing.     

两种麦蚜取食诱导小麦抗性品种后对后来取食蚜生物学特性的影响

刺吸式昆虫在刺吸作物韧皮部取食后会影响作物的正常生长发育,随着作物抗性及昆虫种类的不同,作物反过来也会对昆虫造成或正或反的影响,然而一种刺吸式昆虫取食后对后来者有什么样的影响目前尚不确定。本研究通过严谨的实验方法,即选定3个不同国家的小麦抗性品种98-10-30、Amigo和Batis,在所有条件严格统一的人工智能控制温室内,通过各处理在同一植株上先后不同的接蚜方式,分别测定了麦二叉蚜Schizaphis graminum (Rondani)和麦长管蚜Sitobion avenae Fab.在前期被蚜虫危害的抗虫品种98-10-30、Amigo和Batis上的发育历期、体重差及相对日均体重增长率(mean relative growth rate MRGR)等生物学参数。结果表明：品种不同,蚜虫在不同处理条件下取食作物时受到的影响也不同,即在品种98-10-30上,通过与前期不接虫的对照及前期接不同麦蚜的相关处理比较,麦长管蚜对后期取食的麦二叉蚜(MRGR=0.0974±0.0071)具有抑制作用,而麦二叉蚜对后期取食的麦长管蚜(MRGR=0.1614±0.0048)却有促进作用;在品种Amigo上,前期麦蚜的危害对麦长管蚜的取食具有促进作用,而在品种Batis上前期危害对麦二叉蚜的取食具有促进作用。同时明确了3个品种对两种蚜虫的抗性状况,即在前期无蚜虫危害时,品种98-10-30和Batis对两种麦蚜的抗性相当;在前期有麦蚜危害时,品种98-10-30对麦二叉蚜的抗性较好,而品种Batis对麦长管蚜的抗性较好;品种Amigo无论在任何处理下均对麦二叉蚜的抗性较好。     

Categories of Resistance to Greenbug and Yellow Sugarcane Aphid (Hemiptera: Aphididae) in Three Tetraploid Switchgrass Populations

Switchgrass, Panicum virgatum L., has been targeted as a bioenergy feedstock. However, little is currently known of the mechanisms of insect resistance in this species. Here, two no-choice studies were performed to determine the categories (antibiosis and tolerance) and relative levels of resistance of three switchgrass populations (Kanlow–lowland ecotype, Summer–upland ecotype, and third generation derivatives between Kanlow?×?Summer plants, K×S) previously identified with differential levels of resistance to the greenbug, Schizaphis graminum (Rondani), and yellow sugarcane aphid, Sipha flava (Forbes). No-choice studies indicated that Kanlow possessed multi-species resistance, with high levels of antibiosis to both aphid species, based on aphid survival at 7 and 14 days after aphid introduction and cumulative aphid days, while K×S possessed low-to-moderate levels of antibiosis to S. flava. Further, functional plant loss indices based on plant height and biomass indicated that tolerance is an important category of resistance for Summer plants to S. graminum. These studies also indicated that Summer lacks both tolerance and antibiosis to S. flava, relative to the other switchgrasses tested, whereas K×S lack tolerance and antibiosis to S. graminum. These studies are the first attempt to analyze the categories of resistance in switchgrass and provide critical information for characterizing the biological mechanisms of resistance and improving our knowledge of the plant–insect interactions within this system.