Search for biocontrol agents among endophytic lipopeptide-synthesizing bacteria Bacillus spp. to protect wheat plants against Greenbug aphid (Schizaphis graminum).

Beneficial endophytic bacteria can suppress the development of insect pests through direct antagonism, with the help of metabolites, or indirectly by the induction of systemic resistance through the regulation of hormonal signaling pathways. Lipopeptides are bacterial metabolites that exhibit direct antagonistic activity against many organisms, including insects. Also, lipopeptides are able to trigger induced systemic resistance (ISR) in plants against harmful organisms, but the physiological mechanisms of their action are just beginning to be studied. In this work, we studied ten strains of bacteria isolated from the tissues of wheat and potatoes. Sequencing of the 16S rRNA gene showed that all isolates belong to the genus Bacillus and to two species, B. subtilis and B. velezensis. The genes for lipopeptide synthetase - surfactin synthetase (Bs_srf ), iturin synthetase (Bs_ituA, Bs_ituB) and fengycin synthetase (Bs_fenD) - were identified in all bacterial isolates using PCR. All strains had high aphicidal activity against the Greenbug aphid (Schizaphis graminum Rond.) due to the synthesis of lipopeptides, which was proven using lipopeptide-rich fractions (LRFs) isolated from the strains. Endophytic lipopeptide-synthesizing strains of Bacillus spp. indirectly affected the viability of aphids, the endurance of plants against aphids and triggered ISR in plants, which manifested itself in the regulation of oxidative metabolism and the accumulation of transcripts of the Pr1, Pr2, Pr3, Pr6 and Pr9 genes due to the synthesis of lipopeptides, which was proven using LRF isolated from three strains: B. subtilis 26D, B. subtilis 11VM, and B. thuringiensis B-6066. We have for the first time demonstrated the aphicidal effect of fengycin and the ability of the fengycin-synthesizing strains and isolates, B. subtilis Ttl2, Bacillus sp. Stl7 and B. thuringiensis B-6066, to regulate components of the pro-/antioxidant system of aphid-infested plants. In addition, this work is the first to demonstrate an elicitor role of fengycin in triggering a systemic resistance to S. graminum in wheat plants. We have discovered new promising strains and isolates of endophytes of the genus Bacillus, which may be included in the composition of new biocontrol agents against aphids. One of the criteria for searching for new bacteria active against phloem-feeding insects can be the presence of lipopeptide synthetase genes in the bacterial genome.

Examining Innovative Technologies: Nano-Chelated Fertilizers for Management of Wheat Aphid (Schizaphis graminum Rondani).

The use of nanofertilizers has both advantages and concerns. One benefit is that nano-fertilizers can enhance plant resistance against insect pests, making them a valuable strategy in integrated pest management (IPM). This study focused on the effect of wheat leaves treated with nano-chelated fertilizers and nitrogen (N) fertilizer on the wheat aphid (Schizaphis graminum Rondani), a harmful pest of wheat plants that transmits dangerous viruses. The nano-Cu treatment showed the longest pre-adult longevity. Additionally, the nano-Cu treatment resulted in the lowest adult longevity, fecundity, nymphoposition day number, intrinsic rate of population growth (r), finite rate of population increase (λ), and net reproductive rate (R0) and gross reproductive rate (GRR). Also, nano-Cu treatment led to the highest amount of (T). The N treatment led to the highest levels of fecundity, nymphoposition days, r, λ, and R0. Nano-Fe and nano-Zn demonstrated fewer negative effects on S. graminum life table parameters than nano-Cu. Our results indicate that N treatment yielded numerous advantageous effects on the wheat aphid while simultaneously impeding the efficacy of the aphid control program. Conversely, nano-Cu treatment exhibited a detrimental influence on various parameters of the aphid's life table, resulting in a reduction in the pest's fitness. Consequently, the integration of nano-Cu should be seriously considered as a viable option in the IPM of the wheat aphid.

Newly Developed Restorer Lines of Sorghum [Sorghum bicolor (L.) Moench] Resistant to Greenbug.

Eight lines of grain sorghum [Sorghum bicolor (L.) Moench], which can be used as a promising source material in heterotic hybrid breeding as pollen fertility restorers and donors of resistance to the greenbug (Schizaphis graminum Rondani), are characterized. The new restorer lines (R-lines) were developed by crossing the maternal sterile line Nizkorosloe 81s (CMS A1) with two lines selected from the grain sorghum collection accessions VIR-928 and VIR-929 as the paternal forms. The R-lines were genotyped using PCR markers, and also characterized by height, duration of the seedling-flowering period, and some of the technological properties of flour. With the use of microsatellite markers linked to the Rf genes and by hybridological analysis, it was shown that the new lines carry the dominant allele of the gene Rf2. The PCoA analysis demonstrated clear differences of each R-line from the parents. The genotypes of the new lines and their parental forms for the Rf2 locus were confirmed by applying three allele-specific codominant CAPS markers which detected SNPs in the candidate Rf2 gene. All new lines were highly fertile, as demonstrated by cytological analysis of acetocarmine-stained pollen preparations. A high resistance to the greenbug was demonstrated for each new R-line both in the laboratory and field conditions against a severe aphid infestation. Grain quality parameters such as protein content and dough rheological properties varied widely and were quite satisfactory in some R-lines. Characteristics common to all eight sorghum lines studied, such as the ability to restore pollen fertility in the F1 generation, good pollen quality, greenbug resistance, early ripening, spreading panicle, and low stature, allow us to recommend them for producing commercial F1 hybrids with satisfactory grain quality for both fodder and food purposes.

Genetic Diversity of Barley Accessions from East Asia for Greenbug Resistance.

The greenbug, Schizaphis graminum, is a dangerous pest of barley and other grain crops in the south of Russia. An effective and environmentally friendly way to control this insect is to cultivate resistant varieties. The differential interaction between the phytophage and host plants necessitates the search for new donors of resistance. Seven hundred and seventy-eight accessions of barley from East Asian countries (313 from China, 450 from Japan, and 15 from Nepal) were evaluated for greenbug resistance. The Krasnodar population of the insect and clones isolated from it were used in the experiments. Forty heterogeneous accessions were identified, in which plants with a high level of resistance to the aphid were found. As a result of damage assessment by the 108 S. graminum clones of 11 lines selected from heterogeneous accessions, 52 insect virulence phenotypes were identified. Experiments with aphid test clones showed that all 11 lines possess diverse greenbug resistance alleles, which differ from the previously identified Rsg1, but their efficiency is low. The frequency of clones virulent to ten lines and the cultivar Post (a carrier of the Rsg1 gene) varies from 60.4% to 98.0%. The exception is line 15903, which is resistant to the aphid population and protected by one dominant gene. The high resistance of other lines against a part of the natural population of S. graminum is also under oligogenic control. Lines 15600 and 16190 each have one dominant resistance gene, and line 28129 is protected by two genes, the dominant and recessive ones. A recessive resistance gene is presumably present in line 15600. Lines 16237/1 and 16237/2, isolated from the same collection accession, each have one dominant gene effective against individual aphid clones. The loss of effectiveness of distinctly manifested resistance genes causes the expression of previously masked genes with a weak phenotypic manifestation, which differentially interact with insect genotypes.

Untapped Sources of Dual Resistance to Hessian Fly and Greenbug in Synthetic Hexaploid Wheats.

The Hessian fly (Hf) and greenbugs (Gb) are major pests of wheat, causing severe economic losses globally. Deploying resistant wheat is the most effective strategy for managing these destructive insects. However, the resistance is not effective against all Hf or Gb biotypes and can impose selection pressure on insects, resulting in the development of virulent biotypes. These challenges must be met through the discovery of new and novel sources of resistance to these pests. Synthetic Hexaploid Wheat (SHW)-developed cultivars are a rich source of resistance against a diverse array of pathogens and pests. In this study, 80 SHW lines were evaluated for their resistance to Hf and Gb under controlled environmental conditions. Of these, a total of 36 SHW lines showed resistance independently to Hf biotype L and Gb biotype E, while 27 lines showed combined resistance to both Hf and Gb. Further, a subset of 10 SHW lines showed resistance to additional Hf biotypes, Great Plains and vH13. The identification of SHW lines resistant to multiple insects and biotypes offers an invaluable resource to breeders who are looking to stack resistance traits to develop elite cultivars as a strategy to alleviate economic impacts upon global wheat production.

Some Physiological Effects of Nanofertilizers on Wheat-Aphid Interactions.

The increasing use of nanofertilizers in modern agriculture and their impact on crop yield and pest management require further research. In this study, the effects of nano-Fe, -Zn, and -Cu (which are synthesized based on nanochelating technology), and urea (N) fertilizers on the antioxidant activities of wheat plants (cv. Chamran), and the wheat green aphid Schizaphis graminum (Rondani) are investigated. The authors observed the highest levels of phenolics in non-infested nano-Zn-treated plants (26% higher compared with control). The highest H2O2 levels are in the infested and non-infested nano-Zn-treated and infested nano-Fe-treated plants (in infested nano-Zn and nano-Fe treated plants, 18% and non-infested nano-Zn-treated plants, 28% higher compared with control). The highest peroxidase (POX) activity is observed in the infested and non-infested N-treated and non-infested water-treated plants (almost 14%, 37%, and 46% higher than control, respectively). The lowest activity is in the infested plants' nano-Zn and -Fe treatments (almost 7 and 5 folds lower compared to the control, respectively). The highest and lowest catalase (CAT) activity are in the infested N-treated plants (almost 42% higher than control) and water-treated plants, respectively. The infested nano-Zn, -Fe, -Cu and Hoagland-treated plants showed the highest superoxide dismutase (SOD) activity. Regarding the antioxidant enzyme activities of S. graminum, the highest POX activity is in the nano-Cu treatment (more than two folds higher compared with control); the highest CAT and SOD activities are in the nano-Cu and -Zn treatments. It can be concluded that the application of nanofertilizers caused increasing effects on the wheat plant's antioxidant system and its resistance to S. graminum.

Heat-avoidance behavior associates with thermal sensitivity rather than tolerance in aphid assemblages.

How to predict animals' heat-avoidance behaviors is critical since behavior stands the first line for animals dealing with frequent heat events under ongoing climate warming. However, the discrepancy between the scarcity of research on heat-avoidance behaviors and the commonness of eco-physiological data for thermal tolerance and for thermal sensitivity such as the temperature-dependent survival time makes it difficult to link physiological thermal traits to heat-avoidance behavior. Aphids usually suck plant sap on a fixed site on the host plants at moderate temperatures, but they will leave and seek cooler feeding sites under stressful temperatures. Here we take the cereal aphid assemblages comprising different species with various development stages as a model system. We tested the hypotheses that heat tolerance (critical thermal maximum, CTmax) or heat sensitivity (temperature-dependent declining rate of survival time, similarly hereinafter) would associate with the temperature at which aphid activate heat-avoidance behavior. Specifically, we hypothesized the aphids with less heat tolerance or greater heat sensitivity would take a lower heat risk by leaving the host plant earlier. By mimicking the linear increase in ambient temperature during the daytime, we measured the CTmax and the heat-avoidance temperature (HAT, at which aphids leave the host plant to find cooler places) to understand their heat tolerance and heat-avoidance behavior. Then, we tested the survival time of aphids at different temperatures and calculated the slope of survival time declining with temperature to assess their heat sensitivity (HS). Finally, we examined the relationships between CTmax and HAT and between HS and HAT to understand if the heat-avoidance behavior associates with heat tolerance or with heat sensitivity. The results showed that HS and HAT had a strong correlation, with more heat sensitive individuals displayed lower HAT. By contrast, CTmax and HAT had a weak correlation. Our results thus provide evidence that heat sensitivity is a more reliable indicator than thermal tolerance linking with the heat-avoidance behavior in the aphid assemblages. Most existing studies use the indexes related to thermal tolerance to predict warming impacts. Our findings highlight the urgency to incorporate thermal sensitivity when predicting animal responses to climate change.

Population dynamics of migrant wheat aphids in China's main wheat production region and their interactions with bacterial symbionts.

Sitobion miscanthi, Rhopalosiphum padi, and Schizaphis graminum are the three main pests in Chinese wheat-producing regions. In 2020, they are classified into the Chinese Class I list of agricultural diseases and pests, due to their severe harm to wheat plantings. S. miscanthi, R. padi, and S. graminum are migrant pests, and understanding their migration patterns and simulating their migration trajectories would improve forecasting and controlling them. Furthermore, the bacterial community of the migrant wheat aphid is also less known. In this study, we employed a suction trap to uncover the migration patterns of the three wheat aphid species in Yuanyang county, Henan province, during 2018 to 2020. And then the migration trajectories of S. miscanthi and R. padi were simulated using the NOAA HYSPLIT model. The interactions between wheat aphids and bacteria were further revealed by specific PCR and 16S rRNA amplicon sequencing. The results showed that the population dynamics of migrant wheat aphids was varied. Most of the trapped samples were identified to be R. padi, and S. graminum was the least collected sample. Typically, R. padi had two migration peaks in the 3 years, whereas S. miscanthi and S. graminum only exhibited one migration peak in 2018 and 2019. Moreover, the aphid migration trajectories varied over the years. Generally, the aphids originated from the south and migrated to the north. Herein, the infections of three main aphid facultative bacterial symbionts, Serratia symbiotica, Hamiltonella defensa, and Regiella insercticola, were detected in S. miscanthi and R. padi with specific PCR. Rickettsiella, Arsenophonus, Rickettsia, and Wolbachia were further identified with 16S rRNA amplicon sequencing. Biomarker searching indicated that Arsenophonus was significantly enriched in R. padi. Furthermore, diversity analyses showed that the bacterial community of R. padi had a higher richness and evenness than that of S. miscanthi. In conclusion, this study expands our knowledge about the migration patterns of aphids in the main wheat plant region of China and reveals the interactions between bacterial symbionts and migrant aphids.

Efficacy of Imidacloprid Seed Treatments against Four Wheat Aphids under Laboratory and Field Conditions.

Imidacloprid seed treatments are effective at reducing the cohorts of many insect pests on crops such as cotton, corn, and cereals. The effects of imidacloprid seed treatments depend on the aphid species. In China, there are four wheat aphid species-Sitobion avenae (Fabricius), Rhopalosiphum padi (Linnaeus), Schizaphis graminum (Rondani), and Metopolophium dirhodum (Walker)-and for a given region, these four aphid species differ in dominance with changes in cultivation practices and climate. Therefore, it is necessary to evaluate the effects of imidacloprid seed treatments on the four different aphid species. In experiments in the laboratory, imidacloprid seed treatments significantly reduced the survival rates of S. avenae, R. padi, and S. graminum to 57.33 ± 2.86%, 12.67 ± 1.92%, and 20.66 ± 2.33%, respectively, but for M. dirhodum, there was no significant difference between the control (96.33 ± 1.08%) and the treatment (97.00 ± 0.98%). The fecundities of the four aphid species were much reduced, especially for R. padi when feeding on treated wheat plants. For the field survey, only three aphid species were considered because the density of S. graminum was too low to be analyzed. The effects of imidacloprid seed treatment on the three aphid species in the field were consistent with the laboratory results. Imidacloprid seed treatment reduced the population sizes of S. avenae and R. padi at rates of 70.30 ± 3.15% and 87.62 ± 2.28%, respectively, for the whole wheat season in the field. For M. dirhodum, imidacloprid seed treatments were less effective, and the densities of M. dirhodum increased on four sample days. From this study, we confirmed that the effect of imidacloprid seed treatment varied with the composition of aphid species, being especially less effective for M. dirhodum.

Effects of Crop Resistance on the Tritrophic Interactions between Wheat Lines, Schizaphis graminum (Hemitera: Aphididae), and Propylaea japonica (Coleoptera: Coccinellidae).

Crop resistance and biological control are both considered efficient and environmentally friendly methods of sustainable pest control. In this study, we aimed at investigating the direct influence of four wheat lines with varying resistance level on the life-history traits of the greenbug, Schizaphis graminum, and the mediational effect on the functional response of a predatory ladybird, Propylaea japonica, under laboratory conditions. Results showed that the aphid fitness was the lowest for aphids that had been feeding on wheat line '98-10-19' for one year. These aphids had the longest development time, and least adult mass, minimal mean relative growth rate, and lowest reproductive fitness. In contrast, the aphids that fed on wheat line '98-10-30' were the fittest, with the shortest development time and highest levels of reproductive fitness. The predatory activities of the ladybeetle, especially the adult male significantly decreased following the consumption of aphids belonging to the '98-10-19'-acclimated population. However, there were no significant differences in predatory efficiency (net attack frequency) among the four aphid acclimated populations. Our results showed that the wheat line '98-10-19' has a relative higher resistance to S. graminum than the other three wheat lines, which could further decrease the amount of prey available for consumption. However, the ecological effect of the resistance of '98-10-19' to S. graminum posed no negative influence on the biocontrol potential of P. japonica to these aphids, as their predatory efficiency increases at the fourth instar larvae phase.

The salivary effector protein Sg2204 in the greenbug Schizaphis graminum suppresses wheat defence and is essential for enabling aphid feeding on host plants.

Aphids secrete diverse repertoires of salivary effectors into host plant cells to promote infestation by modulating plant defence. The greenbug Schizaphis graminum is an important cereal aphid worldwide. However, the secreted effectors of S. graminum are still uncharacterized. Here, 76 salivary proteins were identified from the watery saliva of S. graminum using transcriptome and proteome analyses. Among them, a putative salivary effector Sg2204 was significantly up-regulated during aphid feeding stages, and transient overexpression of Sg2204 in Nicotiana benthamiana inhibited cell death induced by BAX or INF1. Delivering Sg2204 into wheat via the type III secretion system of Pseudomonas fluorescens EtAnH suppressed pattern-triggered immunity (PTI)-associated callose deposition. The transcript levels of jasmonic acid (JA)- and salicylic acid (SA)-associated defence genes of wheat were significantly down-regulated, and the contents of both JA and SA were also significantly decreased after delivery of Sg2204 into wheat leaves. Additionally, feeding on wheat expressing Sg2204 significantly increased the weight and fecundity of S. graminum and promoted aphid phloem feeding. Sg2204 was efficiently silenced via spray-based application of the nanocarrier-mediated transdermal dsRNA delivery system. Moreover, Sg2204-silenced aphids induced a stronger wheat defence response and resulted in negative impacts on aphid feeding behaviour, survival and fecundity. Silencing of Sg2204 homologues from four aphid species using nanocarrier-delivered dsRNA also significantly reduced aphid performance on host plants. Thus, our study characterized the salivary effector Sg2204 of S. graminum involved in promoting host susceptibility by suppressing wheat defence, which can also be regarded as a promising RNAi target for aphid control.

Entomotoxic Activity of the Extracts from the Fungus, Alternaria tenuissima and Its Major Metabolite, Tenuazonic Acid.

The study of fungal antibiotics in their competitive interactions with arthropods may lead to the development of novel biorational insecticides. Extracts of Alternaria tenuissima MFP253011 obtained using various methods showed a wide range of biological activities, including entomotoxic properties. Analysis of their composition and bioactivity allowed us to reveal several known mycotoxins and unidentified compounds that may be involved in the entomotoxic activity of the extracts. Among them, tenuazonic acid (TeA), which was the major component of the A. tenuissima extracts, was found the most likely to have larvicidal activity against Galleria mellonella. In the intrahaemocoel injection bioassay, TeA was toxic to G. mellonella and of Zophobas morio with an LT50 of 6 and 2 days, respectively, at the level of 50 µg/larva. Administered orally, TeA inhibited the growth of G. mellonella larvae and caused mortality of Acheta domesticus adults (LT50 7 days) at a concentration of 250 µg/g of feed. TeA showed weak contact intestinal activity against the two phytophages, Tetranychus urticae and Schizaphis graminum, causing 15% and 27% mortality at a concentration of 1 mg/mL, respectively. TeA was cytotoxic to the Sf9 cell line (IC50 25 µg/mL). Thus, model insects such as G. mellonella could be used for further toxicological characterization of TeA.

Seed priming with calcium chloride enhances wheat resistance against wheat aphid Schizaphis graminum Rondani.

BACKGROUND: Calcium is an essential macronutrient for plant growth. Although it has been shown that exogenous Ca application can increase plant resistance to abiotic stress, little is known about its potential to enhance plant tolerance to biotic stress. Here, we investigated whether pretreatment of wheat (Triticum aestivum L.) seeds with calcium chloride (CaCl2 ) improves plant resistance against wheat aphid (Schizaphis graminum Rondani). The developmental time, population size, feeding behavior of aphids on plants grown from CaCl2 - and water-pretreated seeds, and plant defense responses to aphid attack were investigated. RESULTS: Seed pretreatment with CaCl2 extended aphid development time and reduced aphid population size and feeding efficiency. In addition, the pretreatment significantly increased the concentration of Ca2+ in wheat leaves, and upregulated expression levels of TaCaM genes and callose synthase genes (TaGSL2, TaGSL8, TaGSL10, TaGSL12, TaGSL19, TaGSL22 and TaGSL23). Callose concentration in the leaves of plants grown from CaCl2 -pretreated seeds increased significantly upon aphid attack. Further, callose deposition was observed mainly in the phloem. CONCLUSION: These results suggest that seed pretreatment with CaCl2 primes the plant response against wheat aphid attack, leading to modulation of callose deposition in the phloem in response to aphid attack. © 2021 Society of Chemical Industry.

Transcriptome analysis reveals rapid defence responses in wheat induced by phytotoxic aphid Schizaphis graminum feeding.

BACKGROUND: Schizaphis graminum is one of the most important and devastating cereal aphids worldwide, and its feeding can cause chlorosis and necrosis in wheat. However, little information is available on the wheat defence responses triggered by S. graminum feeding at the molecular level. RESULTS: Here, we collected and analysed transcriptome sequencing data from leaf tissues of wheat infested with S. graminum at 2, 6, 12, 24 and 48 hpi (hours post infestation). A total of 44,835 genes were either up- or downregulated and differed significantly in response to aphid feeding. The expression levels of a number of genes (9761 genes) were significantly altered within 2 hpi and continued to change during the entire 48 h experiment. Gene Ontology analysis showed that the downregulated DEGs were mainly enriched in photosynthesis and light harvesting, and the total chlorophyll content in wheat leaves was also significantly reduced after S. graminum infestation at 24 and 48 hpi. However, a number of related genes of the salicylic acid (SA)-mediated defence signalling pathway and MAPK-WRKY pathway were significantly upregulated at early feeding time points (2 and 6 hpi). In addition, the gene expression and activity of antioxidant enzymes, such as peroxidase and superoxide dismutase, were rapidly increased at 2, 6 and 12 hpi. DAB staining results showed that S. graminum feeding induced hydrogen peroxide (H2O2) accumulation at the feeding sites at 2 hpi, and increased H2O2 production was detected with the increases in aphid feeding time. Pretreatment with diphenylene iodonium, an NADPH oxidase inhibitor, repressed the H2O2 accumulation and expression levels of SA-associated defence genes in wheat. CONCLUSIONS: Our transcriptomic analysis revealed that defence-related pathways and oxidative stress in wheat were rapidly induced within hours after the initiation of aphid feeding. Additionally, NADPH oxidase plays an important role in aphid-induced defence responses and H2O2 accumulation in wheat. These results provide valuable insight into the dynamic transcriptomic responses of wheat leaves to phytotoxic aphid feeding and the molecular mechanisms of aphid-plant interactions.

Two novel alkaloids from Corydalis curviflora Maxim. and their insecticidal activity.

BACKGROUND: Botanical pesticide plays an essential role in the control of agricultural pests. Corydalis curviflora Maxim. is used as a cholagogue and larvicide in the rural areas of Northwest China. In this study, our objective was to identify the insect active ingredients of C. curviflora extract. RESULTS: Bioassay-guided isolation of the high active fraction led to the identification of two novel N-demethyl hexahydrobenzophenanthridine-type alkaloids, Curviflorain A (1) and Curviflorain B (2), together with nine known alkaloids, ambiguanine A (3), ambiguanine B (4), ambiguanine C (5), 6-acetylambinine (6), 1,1-dimethyl-6-methoxy-7-hydroxyl-1,2,3,4-tetrahydroisoquinoline (7), hendersine B (8), coryximine (9), isochotensine (10) and corysolidine (11). Compounds 1, 2, and 6 showed promising activity to the larvae of Culex pipiens pallens Coq. and Aedes albopictus Skuse. These compounds were also tested against the insect pests, Mythimna separata walker. and Schizaphis graminum Rondani. CONCLUSION: These findings provide a better understanding of the insecticidal activity of C. curviflora extract and the active compounds. This has the potential to lead to a more effective botanical insecticide.

Comparative transcriptome and histological analyses of wheat in response to phytotoxic aphid Schizaphis graminum and non-phytotoxic aphid Sitobion avenae feeding.

BACKGROUND: Infestation of the phytotoxic aphid Schizaphis graminum can rapidly induce leaf chlorosis in susceptible plants, but this effect is not observed with the nonphytotoxic aphid Sitobion avenae. However, few studies have attempted to identify the different defence responses induced in wheat by S. graminum and S. avenae feeding and the mechanisms underlying the activation of chlorosis by S. graminum feeding. RESULTS: S. graminum feeding significantly reduced the chlorophyll content of wheat leaves, and these effects were not observed with S. avenae. A transcriptomic analysis showed that the expression levels of genes involved in the salicylic acid, jasmonic acid and ethylene signalling defence pathways were significantly upregulated by both S. avenae and S. graminum feeding; however, more plant defence genes were activated by S. graminum feeding than S. avenae feeding. The transcript levels of genes encoding cell wall-modifying proteins were significantly increased after S. graminum feeding, but only a few of these genes were induced by S. avenae. Furthermore, various reactive oxygen species-scavenging genes, such as 66 peroxidase (POD) and 8 ascorbate peroxidase (APx) genes, were significantly upregulated after S. graminum feeding, whereas only 15 POD and one APx genes were induced by S. avenae feeding. The activity of four antioxidant enzymes was also significantly upregulated by S. graminum feeding. Cytological examination showed that S. graminum feeding induced substantial hydrogen peroxide (H2O2) accumulation in wheat leaves. The chlorosis symptoms and the loss of chlorophyll observed in wheat leaves after S. graminum feeding were reduced and inhibited by the scavenging of H2O2 by dimethylthiourea, which indicated that H2O2 plays important role in the induction of chlorosis by S. graminum feeding. CONCLUSIONS: S. graminum and S. avenae feeding induces the JA, SA and ET signalling pathways, but S. graminum activated stronger plant defence responses than S. avenae. S. graminum feeding triggers strong ROS-scavenging activity and massive H2O2 production in wheat leaves, and the accumulation of H2O2 induced by S. graminum feeding is involved in the activation of chlorosis in wheat leaves. These results enhance our understanding of mechanisms underlying aphid-wheat interactions and provide clues for the development of aphid-resistant wheat varieties.

Plant-Mediated Interactions between Two Cereal Aphid Species: Promotion of Aphid Performance and Attraction of More Parasitoids by Infestation of Wheat with Phytotoxic Aphid Schizaphis graminum.

Here, we investigated changes in physiological characteristics in wheat affected by phytotoxic-aphid Schizaphis graminum feeding and nonphytotoxic-aphid Sitobion avenae feeding. We also determined whether shared host-mediated interspecific interactions occur between S. graminum and S. avenae. S. graminum feeding but not S. avenae feeding induced significant chlorophyll loss and hydrogen peroxide accumulation in wheat. Gene-expression analysis and GC/MS metabonomic results indicated that S. graminum infestation induced stronger salicylic acid mediated defense responses than S. avenae did and significantly increased the contents of several amino acids in wheat leaves. Feeding on wheat preinfested with S. graminum significantly increased the reproduction of both aphids and shortened the development time of S. graminum. However, olfactometer bioassays showed that the parasitoid wasp Aphidius gifuensis was more attracted to the odors of S. graminum infested wheat than to those of control and S. avenae infested wheat. This study demonstrates that S. graminum and S. avenae feeding induced different defense responses and changes in plant nutritional quality. Additionally, plant-mediated interactions occurred between these cereal aphids.

The Discovery of Resistant Sources of Spring Barley, Hordeum vulgare ssp. spontaneum, and Unique Greenbug Biotypes.

The genetic sources for host-plant resistance to the greenbug (Schizaphis graminum Rondani) in barley (Hordeum vulgare ssp. spontaneum) are limited in that only two single dominant genes Rsg1 and Rsg2 are available for the complex of greenbug biotypes. We evaluated four new barley lines from the Wild Barley Diversity Collection (WBDC) that previously showed potential for greenbug resistance. Three of those entries, WBDC 53, WBDC 117, WBDC 336, exhibited very dominant sources of resistance to older known biotypes B, C, E, F, H, I, and TX1, which also add to the host-plant differentials used to separate these greenbug biotypes. We also re-evaluated the earlier known set of greenbug biotypes that have been in culture for several years against the known host-plant differentials, and included seven newer greenbug isolates collected from Wyoming to the full complement of small grain differentials. This resulted in the discovery of five new greenbug biotypes, WY10 MC, WY81, WY10 B, WY12 MC, and WY86. Wyoming isolates WY4 A and WY4 B were identical in their phenotypic profile, and should be combined as a single unique greenbug biotype. These barley trials resulted in finding new sources of host-plant resistance, although more research needs to be conducted on what type of resistance was found, and how it can be used. We also document that the Wheatland, Wyoming area serves as a very conducive environment for the development of new greenbug biotypes.

Comparative Life Histories of Greenbugs and Sugarcane Aphids (Hemiptera: Aphididae) Coinfesting Susceptible and Resistant Sorghums.

Host-plant resistance has been a fundamental component of aphid management in cereal crops. Over decades, various sources of resistance to greenbug, Schizaphis graminum (Rondani), were bred into cultivars of sorghum, Sorghum bicolor (L.) Moench, to counter recurring virulent greenbug biotypes. The recent invasion of sugarcane aphid, Melanaphis sacchari (Zehntner), raised questions about plant-mediated interactions between the two aphids and the possibility of using greenbug antibiosis against sugarcane aphid. The present work was undertaken to characterize the impact of PI 550610 resistance to 'biotype I' greenbug, expressed in seed parental line KS 116B, on aphid life histories and to observe plant-mediated interactions between aphid species in its presence and absence. At 23°C, sugarcane aphid nymphs matured 1.5 d faster than greenbug nymphs on susceptible hybrid P8500, but at similar rates on the resistant line, which delayed maturity by 1-1.5 d in both species and increased juvenile mortality by three- to fourfold. Sugarcane aphid reproductive rate was double that of greenbug on susceptible sorghum (4.45 vs. 2.30 nymphs per female per day), but not significantly different on the resistant one (3.09 vs. 2.27). Thus, PI 550610 expresses antibiosis, not tolerance, to these aphids. Coinfestation of P8500 had a positive effect on greenbug intrinsic rate of increase (rm), which changed to negative on KS 116B, whereas the rm of sugarcane aphid was unaffected by coinfestation with greenbug on either cultivar. The results indicate that KS 116B will be useful for producing sugarcane aphid-resistant hybrids, and that PI 550610 antibiosis changes the sugarcane aphid-greenbug interspecific relationship from commensalism to amensalism.

Indigenous Aphid Predators Show High Levels of Preadaptation to a Novel Prey, Melanaphis sacchari (Hemiptera: Aphididae).

The performance of four aphid predators, Hippodamia convergens Guerin-Meneville, Coleomegilla maculata DeGeer, Chrysoperla carnea Stephens and Orius insidiosus Say was compared on three prey species: Schizaphis graminum Rondani, Melanaphis sacchari (Zehntner), and Ephestia kuehniella Zeller eggs. Species predatory in both life stages (all except Ch. carnea) were reared on E. kuehniella eggs and switched to aphid prey for assessment of reproduction. Differences were greater between the E. kuehniella and aphid diets than between the two aphid species. Juvenile survival was high for all predators on all prey, except for O. insidiosus, which had survival on E. kuehniella > S. graminum > M. sacchari. The fastest development of Ch. carnea and O. insidiosus was obtained on E. kuehniella, whereas H. convergens developed fastest on S. graminum, and C. maculata did not differ among diets. S. graminum also yielded the largest H. convergens adults, whereas the largest adults of other predators were obtained on E. kuehniella. Female fecundity and egg viability were similarly high on both aphid diets for H. convergens and C. maculata, whereas, on E. kuehniella, 50% of the former entered reproductive diapause and the latter species had reduced fecundity. Reproductive success of Ch. carnea was S. graminum = M. sacchari > E. kuehniella, but it was similar among treatments for O. insidiosus, although female infertility ranged from 25 to 37.5%. We concluded that all the predators studied are preadapted to utilize sugarcane aphid as prey and have excellent potential to provide sustainable biological control of this newly invasive pest.