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The cereal cyst nematode, Heterodera avenae is distributed worldwide and causes substantial damage in bread wheat, Triticum aestivum. This nematode is extremely difficult to manage because of its prolonged persistence as unhatched eggs encased in cysts. Due to its sustainable and target-specific nature, RNA interference (RNAi)-based strategy has gained unprecedented importance for pest control. To date, RNAi strategy has not been exploited to manage H. avenae in wheat. In the present study, 40 H. avenae target genes with different molecular function were rationally selected for in vitro soaking analysis in order to assess their susceptibility to RNAi. In contrast to target-specific downregulation of 18 genes, 7 genes were upregulated and 15 genes showed unaltered expression (although combinatorial soaking showed some of these genes are RNAi susceptible), suggesting that a few of the target genes were refractory or recalcitrant to RNAi. However, RNAi of 37 of these genes negatively altered nematode behavior in terms of reduced penetration, development and reproduction in wheat. Subsequently, wheat plants were transformed with seven H. avenae target genes (that showed greatest abrogation of nematode parasitic success) for host-induced gene silencing (HIGS) analysis. Transformed plants were molecularly characterized by PCR, RT-qPCR and Southern hybridization. Production of target gene-specific double- and single-stranded RNA (dsRNA/siRNA) was detected in transformed plants. Transgenic expression of galectin, cathepsin L, vap1, serpin, flp12, RanBPM and chitinase genes conferred 33.24-72.4 % reduction in H. avenae multiplication in T 1 events with single copy ones exhibiting greatest reduction. A similar degree of resistance observed in T 2 plants indicated the consistent HIGS effect in the subsequent generations. Intriguingly, cysts isolated from RNAi plants were of smaller size with translucent cuticle compared to normal size, dark brown control cysts, suggesting H. avenae developmental retardation due to HIGS. Our study reinforces the potential of HIGS to manage nematode problems in crop plant., (Copyright © 2020 Elsevier B.V. All rights reserved.)

When a guild of species exploit the same limited resources, interspecific competition induces the exclusion of inferior competitors, in which case, interspecific recognition mechanisms are needed. Here, we address resource partitioning and interspecific competition among three main solitary parasitoid species attacking the same host resource, the aphid Sitobion avenae in wheat fields. Optimal host acceptance models predict that parasitoid species should prefer attacking unparasitized hosts when they are available in order to maximize their fitness, as already parasitized hosts are less valuable for laying eggs, especially for inferior competitors. Therefore, we expected the level of competition (multiparasitism) in the field to increase at low host density. By using a combination of taxonomical (determination) and molecular (PCR-based) approaches, we assessed the species of all parasitoid adults and immature stages within aphid hosts. Our results demonstrate that, early in the season, the multiparasitism rates were low, whereas they were high in the mid-late season, corresponding to an aphid density decrease over time. Moreover, parasitoid species could not have been exploiting host resources randomly and the better competitor, Aphidius ervi, seemed to be foraging preferentially on hosts already parasitized by the inferior competitor A. rhopalosiphi, even when unparasitized hosts were still available. This could be due to differences in their host detection capability, as species with a narrow host range may be better at detecting their hosts in comparison with species with a greater host range, such as A. ervi, with a greater host range within the guild. Our study suggests differences in the host exploitation of two prevalent parasitoid species through the main period of aphid colonization, which still allowed the coexistence of a third inferior competitor (A. rhopalosiphi) within the assemblage, in spite of some negative interactions (multiparasitism) and redundancies.

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.

Cereal cyst nematodes (CCNs) are among the most important nematode pests that limit production of small grain cereals like wheat and barley. These nematodes alone are estimated to reduce production of crops by 10% globally. This necessitates a huge enhancement of nematode resistance in cereal crops against CCNs. Nematode resistance in wheat and barley in combination with higher grain yields has been a preferential research area for cereal nematologists. This usually involved the targeted genetic exploitations through natural means of classical selection breeding of resistant genotypes and finding quantitative trait luci (QTLs) associated with resistance genes. These improvements were based on available genetic diversity among the crop plants. Recently, genome-wide association studies have widely been exploited to associate nematode resistance or susceptibility with particular regions of the genome. Use of biotechnological tools through the application of various transgenic strategies for enhancement of nematode resistance in various crop plants including wheat and barley had also been an important area of research. These modern approaches primarily include the use of gene silencing, exploitation of nematode effector genes, proteinase inhibitors, chemodisruptive peptides and a combination of one or more of these approaches. Furthermore, the perspective genome editing technologies including CRISPR-Cas9 could also be helpful for improving CCN resistance in wheat and barley. The information provided in this review will be helpful to enhance resistance against CCNs and will attract the attention of the scientific community towards this neglected area., Competing Interests: The authors declare no conflict of interest.

Aphids are major pests in cereal crops that cause direct and indirect damage leading to yield reduction. Despite the fact that wheat provides 20% of the world's caloric and protein diet, its metabolic responses to aphid attack, in general, and specifically its production of benzoxazinoid defense compounds are poorly understood. The objective of this study was to compare the metabolic diversity of durum wheat seedlings (Triticum turgidum ssp. durum) under attack by three different cereal aphids: i) the English grain aphid (Sitobion avenae Fabricius), ii) the bird cherry-oat aphid (Rhopalosiphum padi L.), and iii) the greenbug aphid (Schizaphis graminum Rondani), which are some of the most destructive aphid species to wheat. Insect progeny bioassays and metabolic analyses using chromatography/Q-Exactive/mass spectrometry non-targeted metabolomics and a targeted benzoxazinoid profile were performed on infested leaves. The insect bioassays revealed that the plants were susceptible to S. graminum, resistant to S. avenae, and mildly resistant to R. padi. The metabolic analyses of benzoxazinoids suggested that the predominant metabolites DIMBOA (2,4-dihydroxy-7-methoxy-1,4-benzoxazin- 3-one) and its glycosylated form DIMBOA-glucoside (Glc) were significantly induced upon both S. avenae, and R. padi aphid feeding. However, the levels of the benzoxazinoid metabolite HDMBOA-Glc (2-hydroxy-4,7-dimethoxy-1,4-benzoxazin-3-one glucoside) were enhanced due to the feeding of S. avenae and S. graminum aphids, to which Svevo was the most resistant and the most susceptible, respectively. The results showed a partial correlation between the induction of benzoxazinoids and aphid reproduction. Overall, our observations revealed diverse metabolic responses of wheat seedlings to cereal aphid feeding., Competing Interests: The authors have declared that no competing interests exist.

Like pathogens, galling insects deliver effectors into plant tissues that induce gall formation. The gall midge Mayetiola destructor, also called Hessian fly, can convert a whole wheat seedling into a gall by inducing the formation of nutritive cells at the feeding site, inhibiting wheat growth, and reprogramming metabolic pathways of the attacked plants. Here we demonstrated the identification of a secreted Hessian fly protein, the nucleoside diphosphate kinase (NDK), in infested wheat plants through liquid chromatography-tandem mass spectrometry (LC-MS/MS) and western blots. In association with the NDK presence, enzymatic activity of NDK increased significantly in wheat tissues at the feeding site. In addition, there was a sudden increase in ATP abundance at the feeding site of infested susceptible wheat seedlings 24 h following Hessian fly larval infestation. Even though a direct link between cause and effect remains to be established, our data points to the direction that Hessian fly larvae injected NDK into wheat tissues as an effector protein, which plays a role in manipulating host plants and converting the plants into galls., (Copyright © 2018. Published by Elsevier Ltd.)

Plants respond in a similar manner to aphid feeding as to pathogen attack. Diuraphis noxia is a specialist aphid, feeding only on selected grasses that include wheat, barley, and oats. The wheat-Diuraphis noxia interaction is characterized by responses very similar to those seen in wheat-pathogen interactions with none of the underlying resistance pathways and genes characterized yet. From wheat harboring the Dn1 resistance gene, we have identified a nucleotide-binding leucine-rich repeat (NLR) gene containing two integrated domains (IDs). These are three C-terminus ankyrin repeat domains and an N-terminus WRKY domain. The NLR core of the gene can be traced through speciation events within the grass family, with a recent WRKY domain integration that is Triticum-specific. Virus-induced gene silencing of the gene in a resistant wheat line resulted in the abolishment of the resistance response and induced a highly susceptible phenotype. Silenced plants supported a higher number of aphids, similar to the susceptible near-isogenic line (NIL), and the intrinsic rate of increase of the aphids matched that of aphids feeding on the susceptible NIL. The presence of the gene is necessary for Dn1 resistance and we have named the gene Associated with Dn resistance 1 (Adnr1) to reflect this function.

Cereal cyst nematode (Heterodera avenae) is attracted to and aggregated around wheat roots to initiate infection, but this interaction between wheat and the nematode is not fully understood. The transcriptional responses of both wheat and H. avenae were examined during their early contact stage by mRNA sequencing analysis; certain numbers of the differentially expressed genes (DEGs) were validated using quantitative real-time PCR. The immobile host wheat root only had 93 DEGs (27 up-regulated and 66 down-regulated), while the mobile plant parasitic nematode reacted much more actively with 879 DEGs (867 up-regulated and 12 down-regulated). Among them, a number of wheat DEGs (mostly down-regulated) were involved in biotic stress pathways, while several putative effector genes were up-regulated in the nematode DEGs. One putative chitinase-like effector gene of H. avenae was able to suppress BAX-triggered programmed cell death in Nicotiana benthamiana. Results of these experiments demonstrated that nematode responded more actively than wheat during the contact stage of parasitism. The parasite's responses mainly involved up-regulation of genes including at least one anti-plant-defence effector gene, whereas the host responses mainly involved down-regulation of certain defence-related genes.

Parasitoids are important natural enemies of aphids in wheat fields of northern China, and interest in them has increased in recent years. However, little is known regarding parasitoids of wheat aphids, which has hindered the study and understanding of aphid-parasitoid interactions. In the present study, three primary parasitoids and 15 hyperparasitoids were collected in wheat fields during a 2-year survey in northern China (2014, 2015) and a 2-year investigation at Langfang, Hebei Province (2015, 2016). Among them, Aphidius uzbekistanicus Luzhetski was found most frequently among the primary parasitoids, while Pachyneuron aphidis (Bouché) dominated the hyperparasitoid community. Investigation of the dynamics of wheat aphids and parasitoids revealed that the primary parasitoids appeared early in the growing period and that the hyperparasitoids appeared later. Analysis of the seasonal dynamics revealed that growth of the parasitoid population followed that of the aphid population and that the parasitism rates were highest in the late growing period.

The wheat curl mite (WCM), Aceria tosichella Keifer, is a major pest of cereals worldwide that also comprises a complex of at least 16 genetic lineages with divergent physiological traits, including host associations and specificity. The goal of this study was to test the extent to which host-plant species and landscape spatial variation influence WCM presence and population density across the entire area of Poland (>311,000 km2). Three important findings arose from the results of the study. (1) The majority of WCM lineages analyzed exhibited variation in patterns of prevalence and/or population density on both spatial and host-associated scales. (2) Areas of occurrence and local abundance were delineated for specific WCM lineages and it was determined that the most pestiferous lineages are much less widespread than was expected, suggesting relatively recent introductions into Poland and the potential for further spread. (3) The 16 WCM lineages under study assorted within four discrete host assemblages, within which similar host preferences and host infestation patterns were detected. Of these four groups, one consists of lineages associated with cereals. In addition to improving basic ecological knowledge of a widespread arthropod herbivore, the results of this research identify high-risk areas for the presence of the most pestiferous WCM lineages in the study area (viz. the entirety of Poland). They also provide insight into the evolution of pest species of domesticated crops and facilitate testing of fundamental hypotheses about the ecological factors that shape this pest community., Competing Interests: The authors have declared that no competing interests exist.

During parasitoid development, the immature parasitoid is confined to the host species. As a result, any potential to modify the physiology or behaviour of the host could play an important role in parasitoid fitness. The potential for host manipulation by the aphid parasitoid Aphidius avenae to increase cold thermotolerance was investigated using the aphid host species Metopolophium dirhodum and Sitobion avenae. Aphids were parasitized at L3/L4 instar stage (5 d old) and allowed to develop into pre-reproductive adults (10 d old) containing a 5 d old parasitoid larva. A control group was created of non-parasitized pre-reproductive adults (10 d old). The inherent physiological thermotolerance (LT50) and potential behavioural thermoregulation (behaviour in a declining temperature regime) of parasitized and non-parasitized aphids were investigated. Results revealed no effect of parasitism on the physiological thermotolerance of S. avenae and M. dirhodum. Significant differences in the behaviour of parasitized and non-parasitized aphids were observed, in addition to differences between host species, and such behaviours are discussed in view of the potential for host manipulation., Competing Interests: The authors have declared that no competing interests exist.

Phenotypic plasticity has recently been considered a powerful means of adaptation, but its relationships with corresponding life-history characters and plant specialization levels of insects have been controversial. To address the issues, Sitobion avenae clones from three plants in two areas were compared. Varying amounts of life-history trait plasticity were found among S. avenae clones on barley, oat and wheat. In most cases, developmental durations and their corresponding plasticities were found to be independent, and fecundities and their plasticities were correlated characters instead. The developmental time of first instar nymphs for oat and wheat clones, but not for barley clones, was found to be independent from its plasticity, showing environment-specific effects. All correlations between environments were found to be positive, which could contribute to low plasticity in S. avenae. Negative correlations between trait plasticities and fitness of test clones suggest that lower plasticity could have higher adaptive value. Correlations between plasticity and specialization indices were identified for all clones, suggesting that plasticity might evolve as a by-product of adaptation to certain environments. The divergence patterns of life-history plasticities in S. avenae, as well as the relationships among plasticity, specialization and fitness, could have significant implications for evolutionary ecology of this aphid.

The genetic tractability of the Hessian fly (HF, Mayetiola destructor) provides an opportunity to investigate the mechanisms insects use to induce plant gall formation. Here we demonstrate that capacity using the newly sequenced HF genome by identifying the gene (vH24) that elicits effector-triggered immunity in wheat (Triticum spp.) seedlings carrying HF resistance gene H24. vH24 was mapped within a 230-kb genomic fragment near the telomere of HF chromosome X1. That fragment contains only 21 putative genes. The best candidate vH24 gene in this region encodes a protein containing a secretion signal and a type-2 serine/threonine protein phosphatase (PP2C) domain. This gene has an H24-virulence associated insertion in its promoter that appears to silence transcription of the gene in H24-virulent larvae. Candidate vH24 is a member of a small family of genes that encode secretion signals and PP2C domains. It belongs to the fraction of genes in the HF genome previously predicted to encode effector proteins. Because PP2C proteins are not normally secreted, our results suggest that these are PP2C effectors that HF larvae inject into wheat cells to redirect, or interfere, with wheat signal transduction pathways., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Gall-inducing insects are known for altering source-sink relationships within plants. Changes in photosynthesis may contribute to this phenomenon. We investigated photosynthetic responses in wheat [Triticum aestivum L. (Poaceae: Triticeae)] seedlings attacked by the Hessian fly [Mayetiola destructor (Say) (Diptera: Cecidomyiidae], which uses a salivary effector-based strategy to induce a gall nutritive tissue in susceptible plants. Resistant plants have surveillance systems mediated by products of Resistance (R) genes. Detection of a specific salivary effector triggers downstream responses that result in a resistance that kills neonate larvae. A 2 × 2 factorial design was used to study maximum leaf photosynthetic assimilation and stomatal conductance rates. The plant treatments were-resistant or susceptible wheat lines expressing or not expressing the H13 resistance gene. The insect treatments were-no attack (control) or attack by larvae killed by H13 gene-mediated resistance. Photosynthesis was measured for the second and third leaves of the seedling, the latter being the only leaf directly attacked by larvae. We predicted effector-based attack would trigger increases in photosynthetic rates in susceptible but not resistant plants. For susceptible plants, attack was associated with increases (relative to controls) in photosynthesis for the third but not the second leaf. For resistant plants, attack was associated with increases in photosynthesis for both the second and third leaves. Mechanisms underlying the increases appeared to differ. Resistant plants exhibited responses suggesting altered source-sink relationships. Susceptible plants exhibited responses suggesting a mechanism other than altered source-sink relationships, possibly changes in water relations that contributed to increased stomatal conductance., (© The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Sitobion avenae (F.) can survive on various plants in the Poaceae, which may select for highly plastic genotypes. But phenotypic plasticity was often thought to be non-genetic, and of little evolutionary significance historically, and many problems related to adaptive plasticity, its genetic basis and natural selection for plasticity have not been well documented. To address these questions, clones of S. avenae were collected from three plants, and their phenotypic plasticity under alternative environments was evaluated. Our results demonstrated that nearly all tested life-history traits showed significant plastic changes for certain S. avenae clones with the total developmental time of nymphs and fecundity tending to have relatively higher plasticity for most clones. Overall, the level of plasticity for S. avenae clones' life-history traits was unexpectedly low. The factor 'clone' alone explained 27.7-62.3% of the total variance for trait plasticities. The heritability of plasticity was shown to be significant in nearly all the cases. Many significant genetic correlations were found between trait plasticities with a majority of them being positive. Therefore, it is evident that life-history trait plasticity involved was genetically based. There was a high degree of variation in selection coefficients for life-history trait plasticity of different S. avenae clones. Phenotypic plasticity for barley clones, but not for oat or wheat clones, was frequently found to be under significant selection. The directional selection of alternative environments appeared to act to decrease the plasticity of S. avenae clones in most cases. G-matrix comparisons showed significant differences between S. avenae clones, as well as quite a few negative covariances (i.e., trade-offs) between trait plasticities. Genetic basis and evolutionary significance of life-history trait plasticity were discussed.

The Russian wheat aphid (RWA), Diuraphis noxia (Kurdjumov), is a significant insect pest of wheat (Triticum aestivum L.) and has a major economic impact worldwide, especially on winter wheat in the western United States. The continuing emergence of new RWA biotypes virulent to existing resistance genes reinforces the need for more durable resistance. Studies have indicated that resistance in previously susceptible plants can be produced by knock-down of susceptibility genes or other genes involved in host plant susceptibility. Therefore, investigation into genes involved in compatible RWA-wheat interactions could be a feasible approach to achieving durable RWA resistance. The objective of this study was to test whether silencing (1,3;1,4)-β-glucanase, previously observed to be highly induced in susceptible compared with resistant wheat during aphid infestation, would confer resistance to a susceptible wheat genotype. Barley stripe mosaic virus-mediated virus-induced gene silencing was employed to test whether (1,3;1,4)-β-glucanase is involved in the susceptible reaction of 'Gamtoos-S' (GS). Controlled infestation with U.S. biotype RWA2 was done to assess aphid reproduction and host symptom development. Aphids on (1,3;1,4)-β-glucanase-silenced plants reproduced less per day and had longer prenymphipositional periods than those on control GS plants. Furthermore, the (1,3;1,4)-β-glucanase-silenced plants exhibited less chlorosis and greater dry weight compared with GS. Aphid reproduction and host plant symptom development showed linear relationships with (1,3;1,4)-β-glucanase transcript levels. Our results suggest that (1,3;1,4)-β-glucanase is required for successful infestation by the RWA and may be a susceptibility factor that could be exploited as a potential target for RWA resistance breeding.

The cereal cyst nematode (CCN, Heterodera avenae) is a major pest of wheat (Triticum spp) that reduces crop yields in many countries. Cyst nematodes are obligate sedentary endoparasites that reproduce by amphimixis. Here, we report the first transcriptome analysis of two stages of H. avenae. After sequencing extracted RNA from pre parasitic infective juvenile and adult stages of the life cycle, 131 million Illumina high quality paired end reads were obtained which generated 27,765 contigs with N50 of 1,028 base pairs, of which 10,452 were annotated. Comparative analyses were undertaken to evaluate H. avenae sequences with those of other plant, animal and free living nematodes to identify differences in expressed genes. There were 4,431 transcripts common to H. avenae and the free living nematode Caenorhabditis elegans, and 9,462 in common with more closely related potato cyst nematode, Globodera pallida. Annotation of H. avenae carbohydrate active enzymes (CAZy) revealed fewer glycoside hydrolases (GHs) but more glycosyl transferases (GTs) and carbohydrate esterases (CEs) when compared to M. incognita. 1,280 transcripts were found to have secretory signature, presence of signal peptide and absence of transmembrane. In a comparison of genes expressed in the pre-parasitic juvenile and feeding female stages, expression levels of 30 genes with high RPKM (reads per base per kilo million) value, were analysed by qRT-PCR which confirmed the observed differences in their levels of expression levels. In addition, we have also developed a user-friendly resource, Heterodera transcriptome database (HATdb) for public access of the data generated in this study. The new data provided on the transcriptome of H. avenae adds to the genetic resources available to study plant parasitic nematodes and provides an opportunity to seek new effectors that are specifically involved in the H. avenae-cereal host interaction.

Lines from a cross between two wheat (Triticum aestivum) cultivars with contrasting resistance phenotypes to Pratylenchus thornei (Nematoda) were investigated to determine the stage at which resistance occurs. Host resistance was examined at nematode attraction to and penetration of roots and nematode motility, maturation, and reproduction within roots. There was no significant difference in the rate at which P. thornei was attracted toward or penetrated resistant or susceptible roots. However, suppression of migration, juvenile maturation, and reproduction in and near resistant roots was evident, suggesting that resistance acts post penetration. No preferential root penetration zone was observed in contrast to other studies. The inhibitory compounds from resistant wheat plants appeared to be constitutively expressed and water soluble because nematode migration was suppressed in roots and root exudates of unchallenged seedlings. The effects of these compounds were reversible and affected P. thornei but not P. neglectus. Apart from migration, nematode multiplication was greatly inhibited by resistance because only a few juveniles (10%) developed past stage three in roots of resistant compared with susceptible plants. Earlier in the life cycle, egg deposition and hatch of P. thornei were also significantly reduced in resistant roots and root exudates, suggesting the presence of hatching inhibitors.

A two-year study was conducted in South Carolina wheat (Triticum aestivum L. (Poales: Poaceae)) fields to describe spatial and temporal dynamics of stink bugs (Hemiptera: Pentatomidae), which were sampled weekly with sweep nets. In 2010, the main phytophagous stink bugs caught in a grid sampling plan across two fields were the brown stink bug, Euschistus servus (Say), the rice stink bug, Oebalus pugnax (F.), the southern green stink bug, Nezara viridula (L.), and the red shouldered stink bug, Thyanta custator (F.), for both adults and nymphs. In 2011, the main phytophagous stink bugs were E. servus, O. pugnax, N. viridula, and T. custator across two fields. Adult stink bug counts adjacent to fallow fields were 2.1-fold greater for all species combined compared with counts adjacent to woods. Spatial Analysis by Distance IndicEs (SADIE) indicated significant aggregation for 35% of analyses for adults and nymph stink bugs at each sampling date. As a measure of spatial and temporal stability, positive SADIE association indices among sampling dates recorded 11, 36, 43, and 16% of analyses for adult E. servus and 7, 50, 50, and 14% for adult O. pugnax in fields A, B, C, and D, respectively. Adult and nymph stink bugs were spatially associated within wheat fields based on SADIE association indices. Seasonal counts of stink bugs were spatially associated with spike counts at least once for each species across the four fields. Future work may investigate practices to reduce stink bug buildup on wheat in the spring and movement to susceptible crops such as corn, Zea mays L.

Background: MicroRNAs (miRNAs) are small non-coding RNAs that play critical roles in regulating post transcriptional gene expression. Gall midges encompass a large group of insects that are of economic importance and also possess fascinating biological traits. The gall midge Mayetiola destructor, commonly known as the Hessian fly, is a destructive pest of wheat and model organism for studying gall midge biology and insect - host plant interactions., Results: In this study, we systematically analyzed miRNAs from the Hessian fly. Deep-sequencing a Hessian fly larval transcriptome led to the identification of 89 miRNA species that are either identical or very similar to known miRNAs from other insects, and 184 novel miRNAs that have not been reported from other species. A genome-wide search through a draft Hessian fly genome sequence identified a total of 611 putative miRNA-encoding genes based on sequence similarity and the existence of a stem-loop structure for miRNA precursors. Analysis of the 611 putative genes revealed a striking feature: the dramatic expansion of several miRNA gene families. The largest family contained 91 genes that encoded 20 different miRNAs. Microarray analyses revealed the expression of miRNA genes was strictly regulated during Hessian fly larval development and abundance of many miRNA genes were affected by host genotypes., Conclusion: The identification of a large number of miRNAs for the first time from a gall midge provides a foundation for further studies of miRNA functions in gall midge biology and behavior. The dramatic expansion of identical or similar miRNAs provides a unique system to study functional relations among miRNA iso-genes as well as changes in sequence specificity due to small changes in miRNAs and in their mRNA targets. These results may also facilitate the identification of miRNA genes for potential pest control through transgenic approaches.

For animals that lay eggs, a longstanding question is, why do females choose particular oviposition sites? For insects that lay eggs on plants there are three hypotheses: maximizing suitable habitat for juveniles, maximizing female lifespan, and maximizing egg survival. We investigated the function of the oviposition-site choice behavior of a gall midge, the Hessian fly, Mayetiola destructor (Say). In spite of living less than a day and having hundreds of eggs, the ovipositing female is choosy about the placement of eggs. Choosiness makes sense. The tiny gall-making neonate larva has limited movement and strict requirements for colonization. We examined whether offspring benefit from the Hessian fly female's preference for the plant's youngest leaf. To do this we restricted the female's access to the first, second, or third leaf of a seedling (wheat Triticum aestivum L.) plant. Being placed on older leaves did not impact egg survival or larval survival during migration to attack sites at the base of the plant, but did have negative impacts on egg-to-adult survival (reduced by 48%) and reproductive potential (reduced by 30-45%). These negative impacts appear to come from larvae having to search harder to find the limited number of reactive plant cells that can be reprogrammed to form the gall nutritive tissue. We propose that the ability of larvae to find these reactive cells in spite of being placed on an older leaf is important because it creates leeway for female behavior to evolve in the face of other selection pressures, e.g., attack by egg parasitoids.

Gall midges constitute an important group of plant-parasitic insects. The Hessian fly (HF; Mayetiola destructor), the most investigated gall midge, was the first insect hypothesized to have a gene-for-gene interaction with its host plant, wheat (Triticum spp.). Recent investigations support that hypothesis. The minute larval mandibles appear to act in a manner that is analogous to nematode stylets and the haustoria of filamentous plant pathogens. Putative effector proteins are encoded by hundreds of genes and expressed in the HF larval salivary gland. Cultivar-specific resistance (R) genes mediate a highly localized plant reaction that prevents the survival of avirulent HF larvae. Fine-scale mapping of HF avirulence (Avr) genes provides further evidence of effector-triggered immunity (ETI) against HF in wheat. Taken together, these discoveries suggest that the HF, and other gall midges, may be considered biotrophic, or hemibiotrophic, plant pathogens, and they demonstrate the potential that the wheat-HF interaction has in the study of insect-induced plant gall formation.

One strategy to enhance the durability of Hessian fly resistance (R) genes in wheat is to combine them with transgenes for resistance. To identify potential transgenes for resistance a protocol for rapidly screening the proteins they encode for efficacy toward resistance is required. However, the Hessian fly is an obligate parasite of wheat and related grasses. Consequently, no protocol for in vitro delivery of antinutrient or toxic proteins to feeding larvae is available. We report here the development of a Hessian fly in plantatranslocation (HIT) feeding assay and the evaluation of eight lectins and the Bowman-Birk serine proteinase inhibitor for potential in transgenic resistance. Of the antinutrient proteins evaluated, Galanthus nivalis L. agglutinin (GNA), commonly termed snowdrop lectin, was the most efficacious. Ingestion of GNA caused a significant reduction in growth of Hessian fly larvae, disruption of midgut microvilli, and changes in transcript level of genes involved in carbohydrate metabolism, digestion, detoxification, and stress response. These effects of GNA are discussed from the perspective of larval Hessian fly physiology., (Published by Elsevier Ltd.)

Resistance (R) genes have a proven record for protecting plants against biotic stress. A problem is parasite adaptation via Avirulence (Avr) mutations, which allows the parasite to colonize the R gene plant. Scientists hope to make R genes more durable by stacking them in a single cultivar. However, stacking assumes that R gene-mediated resistance has no fitness cost for the plant. We tested this assumption for wheat's resistance to Hessian fly, Mayetiola destructor (Say) (Diptera: Cecidomyiidae). Our study included ten plant fitness measures and four wheat genotypes, one susceptible, and three expressing either the H6, H9, or H13 resistance gene. Because R gene-mediated resistance has two components, we measured two types of costs: the cost of the constitutively-expressed H gene, which functions in plant surveillance, and the cost of the downstream induced responses, which were triggered by Hessian fly larvae rather than a chemical elicitor. For the constitutively expressed Hgene, some measures indicated costs, but a greater number of measures indicated benefits of simply expressing the H gene. For the induced resistance, instead of costs, resistant plants showed benefits of being attacked. Resistant plants were more likely to survive attack than susceptible plants, and surviving resistant plants produced higher yield and quality. We discuss why resistance to the Hessian fly has little or no cost and propose that tolerance is important, with compensatory growth occurring after H gene-mediated resistance kills the larva. We end with a caution: Given that plants were given good growing conditions, fitness costs may be found under conditions of greater biotic or abiotic stress.

The majority of plant viruses are dependent on arthropod vectors for spread between plants. Wheat streak mosaic virus (family Potyviridae, genus Tritimovirus, WSMV) is transmitted by the wheat curl mite, Aceria tosichella Keifer, and this virus and vector cause extensive yield losses in most major wheat (Triticum aestivum L.)-growing regions of the world. Many cultivars in use are susceptible to this vector-virus complex, and yield losses of 10-99% have been documented. wheat curl mite resistance genes have been identified in goat grass, Aegilops tauschii (Coss) Schmal., and transferred to hexaploid wheat, but very few varieties contain effectively wheat curl mite resistance, due to virulent wheat curl mite populations. However, wheat curl mite resistance remains an effective strategy to reduce losses due to WSMV. The goal of our project was to identify the most effective, reproducible, and rapid method for assessing wheat curl mite resistance. We also wanted to determine whether mite resistance is affected by WSMV infection, because the pathogen and pest commonly occur together. Single and group wheat curl mite infestations produced similar amounts of leaf rolling and folding on wheat curl mite-susceptible wheat varieties that were independent of initial wheat curl mite infestation. This finding will allow accurate, efficient, large-scale screening of wheat germplasm for wheat curl mite resistance by infesting plants with sections of wheat leaf tissue containing mixed stages of wheat curl mite. The wheat curl mite-resistant breeding line 'OK05312' displayed antibiosis (reduced wheat curl mite population development). The effect of WSMV infection on wheat curl mite reproduction was genotype-dependent. Mite populations increased on infected wheat curl mite- and WSMV-susceptible plants compared with uninfected plants, but WSMV infection had no significant effect on wheat curl mite populations on resistant plants. OK05312 is a strong source of wheat curl mite resistance for wheat breeding programs.

Polyacrylamide gel electrophoresis of enzymes (carboxylesterases) was used for the first time to monitor rates of parasitism in airborne alate (winged) grain aphid, Sitobion avenae (F.) population samples collected by suction trapping in Hertfordshire, UK. Using previously described electrophoretic 'keys', the species of hymenopterous parasitoids present in individual aphids were identified and found to be Aphidius ervi (Haliday) and/or Aphidius rhopalosiphi (De Stephani Perez) (Braconidae). Entomophthoralean fungal infection was also detected using this approach. Aphidiid wasp parasitism was detected from early June to mid-August and fungal infection from late June to late July. The results are discussed in relation to parasitoid population structure and dynamics, especially (i) the fact that winged aphids passively transport the early stages of their braconid parasitoids and fungal pathogens, potentially to newly-founded colonies, which may directly impact on the dual aphid-parasitoid populations genetics; and (ii) the approach used to collect and assay parasitised and fungal infected aphids involving both suction trapping and electrophoretic testing may have potential in assessing the level and efficacy of these biological control agents in integrated pest management (IPM) schemes to combat cereal aphid outbreaks.

An improved understanding of the complex interactions between plants and aphids is emerging. Recognition of aphid feeding in plant tissues involves production of several defense response signaling pathways and downstream production of defense and detoxification compounds. Feeding by Russian wheat aphid, Diuraphis noxia (Kurdjumov), a serious pest of cereal crops worldwide, induces foliar deformity and chlorophyll loss during compatible wheat-D. noxia interactions. Experiments described here revealed significant differences in level and pattern of gene expression in defense response signaling and metabolic pathways between compatible and incompatible D. noxia-wheat interactions. The jasmonate (JA)-signaling genes LOX, AOS, and AOC were significantly more upregulated (~3- to 7 fold) in incompatible interactions than in compatible interactions (~2.5 to 3.5 fold) as early as 1 h post D. noxia infestation (hpi). Cellulose synthase, responsible for strengthening plant cell walls via cellulose production, was also more upregulated in incompatible interactions (4 to 7 fold) than in compatible interactions (1 to 3.5 fold). In contrast, glycolysis and citric acid cycle genes were significantly downregulated (~1.5 to 2 fold) in incompatible interactions and upregulated or less downregulated in compatible interactions from 6 to 72 hpi. Differences in expression of JA-signaling genes between feeding site tissues and non-feeding site tissues suggest that D. noxia defense response signals in wheat are restricted primarily to aphid feeding sites in the initial 6 hpi. This is the first report of differential upregulation of plant genes at 1 hpi in incompatible interactions involving aphid herbivory. Early wheat plant defense responses in incompatible D. noxia interactions at 1, 3, and 6 hpi appear to be important aspects of D. noxia resistance in wheat.

Geostatistis methods were adopted to analyze the spatial pattern of Sitodiplosis mosellana (Diptera: Cecidomyiidae) at its different development periods and of its egg parasitoids mixed population (Tetrastichus sp. and Platygaster error; Hymoneptera: Eulophidae and Platygastridae). The aggregated spatial arrangements for S. mosellana cocoon, adult, and larva and for egg parasitoids mixed population could be well described by spherical model, spherical-exponential model, linear sill model, and spherical-exponential model, respectively. The spatial dependence range of S. mosellana cocoon, adult at initial emergence period, adult at peak emergence period, and larva, and of egg parasitoids mixed population was 53.6, 190.6, 154.1, 4.2 and 280.3 m, and the aggregation intensity was 30.5%, 95.6%, 96.3%, 14.9% and 95.3%, respectively. The simulated maps of the spatial distribution produced by Kriging model could intuitively analyze the dynamic changes of S. mosellana at its different development periods and of egg parasitoids mixed population from the two aspects of time and space.

The wheat stem sawfly, Cephus cinctus Norton (Hymenoptera: Cephidae), is a key pest of wheat in the northern Great Plains of North America. Host plant resistance in the form of solid-stemmed wheat cultivars is the main control strategy for C. cinctus. This study investigated the effect of novel and traditional solid wheat hosts on the overwintering mortality and cold-hardiness of C. cinctus. Field conditions from 2003-2005 showed that overwintering mortality in various wheat cultivars averaged 8% and was not related to the type of wheat cultivar. Similarly, supercooling points (-22° C) were not influenced by wheat host type. C. cintus are cold-hardy; up to 80% survive 10 days at -20° C and 10% survive 40 days. Its overwintering microhabitat near the crown area of the plant is well insulated for temperatures above -10° C and remains ~ 20° C above ambient minima. These data suggest that winter mortality is a minor factor in the population dynamics of wheat stem sawfly, and despite clear detrimental effects on larval weight and adult fitness, solid-stemmed cultivars do not reduce the ability of larvae to survive winters.

Host plant specificity depends on recognition of the host and the ability to discriminate it from nonhost plants. Chilo suppressalis (Walker) (Lepidoptera: Crambidae), an important insect pest of rice, is considered to be polyphagous, although few papers have ever reported infestation of C. suppressalis on most of the recorded hosts. The present investigation was designed to test whether two important gramineous crops, wheat and corn, are host plants of C. suppressalis based on the host-plant finding process in cage and Y-tube olfactometer tests, oviposition and egg hatching, and larval feeding and survival. In the cage tests, gravid C. suppressalis females did not differentiate rice plants from wheat or corn plants when only visual cues were involved, but were more attracted to rice plants when only olfactory cues or both vision and olfaction were present. The Y-tube olfactometer tests further confirmed that the females did not prefer wheat or corn plants, and revealed that they responded equally to clean air and odors from wheat or corn plants. Under no-choice and choice condition alike, the females laid eggs on a lower proportion of wheat and corn plants and egg number and hatching rate were significantly reduced on wheat and/or corn plants than on rice plants. Larval feeding was not observed in wheat and lower in corn than in rice plants, and no pupae or surviving larvae were collected from wheat and corn plants. The results suggest that wheat and corn are not host plants of C. suppressalis. These findings are discussed in context of host-finding process in C. suppressalis and management of resistance to transgenic Bt rice., (© 2010 Entomological Society of America)

Although several wheat genes differentially expressed during the Russian wheat aphid resistance response have recently been identified, their requirement for and specific role in resistance remain unclear. Progress in wheat-aphid interaction research is hampered by inadequate collections of mutant germplasm and difficulty in transforming hexaploid wheat. Virus-induced gene silencing (VIGS) technology is emerging as a viable reverse genetics approach in cereal crops. However, the potential of VIGS for determining aphid defence gene function in wheat has not been evaluated. We report on the use of recombinant barley stripe mosaic virus (BSMV) to target and silence a WRKY53 transcription factor and an inducible phenylalanine ammonia-lyase (PAL) gene, both predicted to contribute to aphid defence in a genetically resistant wheat line. After inoculating resistant wheat with the VIGS constructs, transcript abundance was reduced to levels similar to that observed in susceptible wheat. Notably, the level of PAL expression was also suppressed by the WKRY53 construct, suggesting that these genes operate in the same defence response network. Both knockdowns exhibited a susceptible phenotype upon aphid infestation, and aphids feeding on silenced plants exhibited a significant increase in fitness compared to aphids feeding on control plants. Altered plant phenotype and changes in aphid behaviour after silencing imply that WKRY53 and PAL play key roles in generating a successful resistance response. This study is the first report on the successful use of VIGS to investigate genes involved in wheat-insect interactions., (© 2010 The Authors. Plant Biotechnology Journal © 2010 Society for Experimental Biology and Blackwell Publishing Ltd.)

Genetic resistance in wheat, Triticum aestivum L., is the most efficacious method for control of Hessian fly, Mayetiola destructor (Say) (Diptera: Cecidomyiidae). However, because of the appearance of new genotypes (biotypes) in response to deployment of resistance, field collections of Hessian fly need to be evaluated on a regular basis to provide breeders and producers information on the efficacy of resistance (R) genes with respect to the genotype composition of Hessian fly in regional areas. We report here on the efficacy of 21 R genes in wheat to field collections of Hessian fly from the southeastern United States. Results documented that of the 21 R genes evaluated only five would provide effective protection of wheat from Hessian fly in the southeastern United States. These genes were H12, H18, H24, H25, and H26. Although not all of the 33 identified R genes were evaluated in the current study, these results indicate that identified genetic resistance to protect wheat from Hessian attack in the southeastern United States is a limited resource. Historically, R genes for Hessian fly resistance in wheat have been deployed as single gene releases. Although this strategy has been successful in the past, we recommend that in the future deployment of combinations of highly effective previously undeployed genes, such as H24 and H26, be considered. Our study also highlights the need to identify new and effective sources of resistance in wheat to Hessian fly if genetic resistance is to continue as a viable option for protection of wheat in the southeastern United States.

A 2-yr study was conducted in wheat fields in South Carolina involving weekly sampling of cereal leaf beetle, Oulema melanopus (L.). In each of the six fields of this study, temporal patterns showed two distinct peaks in March and in May of adult O. melanopus. Populations decreased as wheat plants matured. In 2009, larval populations had one peak in April in between the two adult peaks. The χ(2) statistics for observed and Poisson predicted distributions of O. melanopus indicated nonrandom distribution for adults and larvae. In addition, the values of I(D) were >1 for adults and larvae in both years across sampling dates. These results indicate that the sampling distributions of both adult and larval populations of O. melanopus were aggregated. Slopes of Taylor power's law (b) and patchiness regressions (β) were significantly (P < 0.05) different than one in all cases, except for b in 2008 for adults. Across sampling dates, the distance from field border had a significant effect on adult O. melanopus in both years, but not on larval O. melanopus. Densities of adult O. melanopus were greatest at 0 m (the field edge), and decreased at 5-25 m from the field edge. The inverted distance weighted interpolation method showed considerable levels of spatial variability in densities within fields. High densities along the edge of wheat fields suggests that localized control methods in wheat may be effective in reducing migration of O. melanopus and damage in corn, Zea mays L., (© 2010 Entomological Society of America)

Changes in ascorbate peroxidase (APX) enzyme activity in response to nematode (Heterodera avenae) attack were studied in roots of three hexaploid wheat lines carrying Cre2, Cre5, or Cre7 nematode resistance genes and the susceptible Triticum aestivum cv. Anza. A spectrophotometric analysis was carried out with root extracts of infected plants 4, 7, 11, and 14 days after nematode inoculation using uninfected plant as control. APX induction in infected resistant genotypes was similar and higher than in the susceptible control. The introgression wheat/Aegilops ventricosa H-93-8 line, carrying the Cre2 gene, and its parental line H-10-15 as susceptible control were used to analyze whether this increase of activity was correlated with the induction of APX gene expression. Genes encoding cytosolic forms of APX were induced in roots of both lines in response to nematode infection. This induction took place both earlier and with greater intensity in the resistant line than in the susceptible one, and it was also higher in the root area at the site of nematode attachment.

Diuraphis noxia (Kurdjumov)(Russian wheat aphid) has severe economic impacts on wheat and barley production in the United States. The interaction between the Russian wheat aphid and its cereal hosts is poorly understood. However, the recent appearance of new biotypes in the United States showed that specific interactions exist between wheat resistance loci and Russian wheat aphid biotypes. At present, Dn7 is the only known gene in hexaploid wheat that confers resistance against all U.S. Russian wheat aphid biotypes. This study was conducted to investigate the molecular mechanism of Dn7-mediated resistance against two U.S. Russian wheat aphid biotypes (Russian wheat aphid 1 and Russian wheat aphid 2). Using GeneChip Wheat Genome Arrays, we compared transcript profiles of resistant and susceptible lines infested with either Russian wheat aphid 1 or Russian wheat aphid 2 using two time intervals (5 and 48 h after infestation). Russian wheat aphid feeding on hexaploid wheat led to the induction of groups of genes functioning in oxidative and general stress, photosynthesis, cell respiration and energy production, signal transduction, calcium-dependent signaling, pathogenesis related (PR) responses, and defense compound synthesis. The number of differentially expressed genes was higher in plants infested with Russian wheat aphid 1 compared with those infested with Russian wheat aphid 2. Although most genes involved in basic cellular functions were shared, unique genes were also obtained. This finding may indicate subtle differences in genes induced in response to different virulence proteins.

A 2-yr study was conducted in South Carolina wheat fields to describe spatial and temporal dynamics of stink bugs sampled with sweep nets. In 2008, the main phytophagous species were Euschistus servus (Say) (35.8, 36.7%), Oebalus pugnax (F.) (35.0, 6.0%), Nezara viridula L., (15.2, 46.3%), and Thyanta custator (F.) (14.0, 10.5%) for adults and nymphs, respectively. In 2009, the main phytophagous species were O. pugnax (39.8, 37.8%), E. servus (38.4, 30.0%), N. viridula (14.6, 29.5%), and T. custator (6.8, 2.8%). chi(2) goodness-of-fit tests indicated that all species (both adults and nymphs) had observed distributions that did not match the Poisson distribution. Indices of dispersion (I(D )) across sampling dates were significantly >1 for all species and life stages, also suggesting a nonrandom distribution. Slopes of Taylor's power law were significantly greater (P < 0.05) than a value of 1 for only adult O. pugnax and nymph T. custator. Coefficients beta of patchiness regressions were significantly >1 in all cases except for E. servus adults and T. custator nymphs. The inverted distance weighted interpolation method showed considerable levels of spatial variability in densities within fields. Sampling along the edge of fields showed a significant distance from field border effect (P < 0.05) on all adults in both years except for N. viridula. High densities in wheat adjacent to a susceptible crop such as corn suggests that localized control methods in wheat may be effective in mitigation of stink bugs and damage in corn.

Insect herbivores typically oviposit on the most suitable hosts, but choices can be modulated by detection of potential competition among conspecifics, especially when eggs are deposited cryptically. Larvae of the wheat stem sawfly, Cephus cinctus Norton, developing within an already infested stem, experience elevated risk when only one will survive because of cannibalism. To increase our understanding of host selection when the choices made by females can lead to severe intraspecific competition, females were presented with either uninfested wheat plants or with plants previously exposed to other females in laboratory choice tests. The oviposition behavior of this insect was described by recording the behavioral sequences that lead to and follow the insertion of the ovipositor in both previously infested and uninfested stems. No significant differences were found in frequencies of specific behaviors or behavioral transitions associated with oviposition. In choice tests, there was no difference in the numbers of eggs laid in infested and uninfested plants. Taller plants received more eggs, irrespective of infestation. Females neither preferred nor avoided previously infested hosts. Other characteristics of the host, such as stem height, may be more important in determining suitability for oviposition. These findings support the use of management tactics relying on the manipulation of oviposition behavior, such as trap cropping. Given that there is no evidence for response to previously infested hosts, the infested plants in a trap crop would remain as suitable as they were when uninfested, which could also lead to an increase in mortality caused by intraspecific competition.

The Russian wheat aphid, Diuraphis noxia (Kurdjumov) (Hemiptera: Aphididae), devastates wheat and barley production on all continents except Australia. Although D. noxia-resistant cultivars exist, virulent D. noxia populations exist in Asia, North America, and South America that have the ability to overcome resistance. In this study, synthetic hexaploid wheat genotypes created at the International Maize and Wheat Improvement Center (CIMMYT) were evaluated and characterized for resistance to North American biotype 2 (RWA2). Fewer RWA2 were produced on several genotypes, and D. noxia-induced leaf rolling and chlorosis were reduced on these genotypes as well. Aphid numbers were positively correlated with chlorosis and with leaf rolling. However, some genotypes were highly resistant to leaf rolling and chlorosis while supporting large RWA2 populations. There were negative correlations between leaf chlorosis and leaf dry weight (r = -0.267, df = 106, P = 0.006) and between aphid numbers and leaf dry weight (r = -0.297, df = 105, P = 0.002). These results indicate that chlorosis and aphid number individually explain at least 27% of the changes observed in leaf dry weights. Interestingly, there was no correlation between leaf rolling and leaf dry weight. The RWA2-resistant lines identified, which are also resistant to D. noxia populations in Mexico and to greenbug (Schizaphis graminum Rondani) biotype G, are strong candidates for use in improving the genetic diversity in bread wheat for resistance to different biotypes of both S. graminum and D. noxia.

The impact of feeding injury by the Russian wheat aphid, Diuraphis noxia (Mordvilko) (Hemiptera: Aphididae), and bird cherry-oat aphid, Rhopalosiphum padi (L.) (Hemiptera: Aphididae) on susceptible and resistant wheat, Triticum aestivum L., near-isogenic lines 'Tugela' (susceptible), Tugela-Dn1 (antibiotic), Tugela-Dn2 (tolerant), and Tugela-Dn5 (antixenotic) was evaluated by assessing photosynthetic parameters. Photosynthesis and closely related parameters, pigment composition, and nonstructural carbohydrates were measured at 1, 3, and 9 d after aphids were introduced on plants maintained under greenhouse conditions. Overall, R. padi had a higher reproductive capacity within a period of 9 d compared with D. noxia on all lines except Tugela-Dn2. Although the visible injury symptoms associated with aphid injury can be highly species specific, the data indicate that photosynthetic reduction is a common physiological pattern of wheat response to aphid feeding, irrespective of chlorosis elicitation. Although both aphids negatively affected net photosynthesis, D. noxia had a greater impact than R. padi, even when aphid numbers were considerably fewer for D. noxia (100-150 aphids per plant) compared with R. padi (> 200 aphids per plant). The photosynthetic pigment and carbohydrate data suggest that the initial net photosynthesis reduction elicited by aphid feeding may not be directly related to the light reaction portion of the photosynthetic pathway via pigment losses. It is also unlikely that source-sink manipulation is the primary cause for the observed short-term inhibition of photosynthesis.

We previously cloned and characterized a novel jacalin-like lectin gene from wheat (Triticum aestivum) plants that responds to infestation by Hessian fly (Mayetiola destructor) larvae, a major dipteran pest of this crop. The infested resistant plants accumulated higher levels of Hfr-1 (for Hessian fly-responsive gene 1) transcripts compared with uninfested or susceptible plants. Here, we characterize the soluble and active recombinant His(6)-HFR1 protein isolated from Escherichia coli. Functional characterization of the protein using hemagglutination assays revealed lectin activity. Glycan microarray-binding assays indicated strong affinity of His(6)-HFR1 to Manalpha1-6(Manalpha1-3)Man trisaccharide structures. Resistant wheat plants accumulated high levels of HFR1 at the larval feeding sites, as revealed by immunodetection, but the avirulent larvae were deterred from feeding and consumed only small amounts of the lectin. Behavioral studies revealed that avirulent Hessian fly larvae on resistant plants exhibited prolonged searching and writhing behaviors as they unsuccessfully attempted to establish feeding sites. During His(6)-HFR1 feeding bioassays, Drosophila melanogaster larvae experienced significant delays in growth and pupation, while percentage mortality increased with progressively higher concentrations of His(6)-HFR1 in the diet. Thus, HFR1 is an antinutrient to dipteran larvae and may play a significant role in deterring Hessian fly larvae from feeding on resistant wheat plants.

A crucial function of antioxidative enzymes is to remove excess reactive oxygen species (ROS), which can be toxic to plant cells. The effect of Russian wheat aphid (RWA), Diuraphis noxia (Mordvilko), infestation on the activities of antioxidative enzymes was investigated in the resistant (cv. Tugela DN) and the near-isogenic susceptible (cv. Tugela) wheat (Triticum aestivum L.). RWA infestation significantly induced the activity of superoxide dismutase, glutathione reductase and ascorbate peroxidase to higher levels in the resistant than in susceptible plants. These findings suggest the involvement of antioxidative enzymes in the RWA-wheat resistance response, which was accompanied by an early oxidative burst. The results are consistent with the role of ROS in the resistance response and the control of their levels to minimise toxic effects.

This paper studied the development duration, adult weight, and mean relative growth rate (MRGR) of aphid Schizapis graminum, and the specific genes expression in wheat variety 98-10-30 (Triticum aestivum) after treated with different elicitors. The results showed that needling penetration, aphid feeding and BTH application could shorten the development duration of the aphid and decrease its adult weight, but had no significant effect on aphid MRGR. Different elicitors induced different specific genes expression in quality and quantity. The mRNA of PDF1.2 was increased significantly after aphid feeding, while there was no expression after applying BTH. Aphid feeding and BTH application increased the mRNA of BGL2, but no expression was observed in the control and after needling penetration. The induced resistance had some effects on aphid growth and development, and the response induced by aphid feeding had some similarities but significant differences to that induced by mechanical wounding and BTH application. It could be concluded that the response of aphid to elicitors was a special resistance, and there existed some overlaps or differences between it and mechanical wounding and SAR (systemic acquired resistance).