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1.
Proc Natl Acad Sci U S A ; 117(23): 12763-12771, 2020 06 09.
Article in English | MEDLINE | ID: mdl-32461369

ABSTRACT

Aphids are sap-feeding insects that colonize a broad range of plant species and often cause feeding damage and transmit plant pathogens, including bacteria, viruses, and viroids. These insects feed from the plant vascular tissue, predominantly the phloem. However, it remains largely unknown how aphids, and other sap-feeding insects, establish intimate long-term interactions with plants. To identify aphid virulence factors, we took advantage of the ability of the green peach aphid Myzus persicae to colonize divergent plant species. We found that a M. persicae clone of near-identical females established stable colonies on nine plant species of five representative plant eudicot and monocot families that span the angiosperm phylogeny. Members of the novel aphid gene family Ya are differentially expressed in aphids on the nine plant species and are coregulated and organized as tandem repeats in aphid genomes. Aphids translocate Ya transcripts into plants, and some transcripts migrate to distal leaves within several plant species. RNAi-mediated knockdown of Ya genes reduces M. persicae fecundity, and M. persicae produces more progeny on transgenic plants that heterologously produce one of the systemically migrating Ya transcripts as a long noncoding (lnc) RNA. Taken together, our findings show that beyond a range of pathogens, M. persicae aphids translocate their own transcripts into plants, including a Ya lncRNA that migrates to distal locations within plants, promotes aphid fecundity, and is a member of a previously undescribed host-responsive aphid gene family that operate as virulence factors.


Subject(s)
Aphids/pathogenicity , Magnoliopsida/parasitology , RNA Transport , RNA, Long Noncoding/metabolism , Virulence Factors/metabolism , Animals , Aphids/genetics , Insect Proteins/genetics , RNA, Long Noncoding/genetics , Virulence Factors/genetics
2.
Plant Cell ; 31(8): 1913-1929, 2019 08.
Article in English | MEDLINE | ID: mdl-31126981

ABSTRACT

Because they suck phloem sap and act as vectors for phytopathogenic viruses, aphids pose a threat to crop yields worldwide. Pectic homogalacturonan (HG) has been described as a defensive element for plants during infections with phytopathogens. However, its role during aphid infestation remains unexplored. Using immunofluorescence assays and biochemical approaches, the HG methylesterification status and associated modifying enzymes during the early stage of Arabidopsis (Arabidopsis thaliana) infestation with the green peach aphid (Myzus persicae) were analyzed. Additionally, the influence of pectin methylesterase (PME) activity on aphid settling and feeding behavior was evaluated by free choice assays and the Electrical Penetration Graph technique, respectively. Our results revealed that HG status and HG-modifying enzymes are significantly altered during the early stage of the plant-aphid interaction. Aphid infestation induced a significant increase in total PME activity and methanol emissions, concomitant with a decrease in the degree of HG methylesterification. Conversely, inhibition of PME activity led to a significant decrease in the settling and feeding preference of aphids. Furthermore, we demonstrate that the PME inhibitor AtPMEI13 has a defensive role during aphid infestation, since pmei13 mutants are significantly more susceptible to M. persicae in terms of settling preference, phloem access, and phloem sap drainage.


Subject(s)
Aphids/pathogenicity , Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Arabidopsis/parasitology , Pectins/metabolism , Animals , Arabidopsis/enzymology , Arabidopsis Proteins/genetics , Gene Expression Regulation, Plant
3.
BMC Genomics ; 21(1): 638, 2020 Sep 15.
Article in English | MEDLINE | ID: mdl-32933469

ABSTRACT

BACKGROUND: Plant viruses maintain intricate interactions with their vector and non-vector insects and can impact the fitness of insects. However, the details of their molecular and cellular mechanisms have not been studied well. We compared the transcriptome-level responses in vector and non-vector aphids (Schizaphis graminum and Rhopalosiphum padi, respectively) after feeding on wheat plants with viral infections (Barley Yellow Dwarf Virus (BYDV) and Wheat dwarf virus (WDV), respectively). We conducted differentially expressed gene (DEG) annotation analyses and observed DEGs related to immune pathway, growth, development, and reproduction. And we conducted cloning and bioinformatic analyses of the key DEG involved in immune. RESULTS: For all differentially expressed gene analyses, the numbers of DEGs related to immune, growth, development, reproduction and cuticle were higher in vector aphids than in non-vector aphids. STAT5B (signal transducer and activator of transcription 5B), which is involved in the JAK-STAT pathway, was upregulated in R. padi exposed to WDV. The cloning and bioinformatic results indicated that the RpSTAT5B sequence contains a 2082 bp ORF encoding 693 amino acids. The protein molecular weight is 79.1 kD and pI is 8.13. Analysis indicated that RpSTAT5B is a non-transmembrane protein and a non-secreted protein. Homology and evolutionary analysis indicated that RpSTAT5B was closely related to R. maidis. CONCLUSIONS: Unigene expression analysis showed that the total number of differentially expressed genes (DEGs) in the vector aphids was higher than that in the non-vector aphids. Functional enrichment analysis showed that the DEGs related to immunity, growth and reproduction in vector aphids were higher than those in non-vector aphids, and the differentially expressed genes related to immune were up-regulated. This study provides a basis for the evaluation of the response mechanisms of vector/non-vector insects to plant viruses.


Subject(s)
Aphids/genetics , Insect Vectors/genetics , Transcriptome , Animals , Aphids/metabolism , Aphids/pathogenicity , Aphids/virology , Dicistroviridae/pathogenicity , Geminiviridae/pathogenicity , Insect Proteins/genetics , Insect Proteins/metabolism , Insect Vectors/metabolism , Insect Vectors/pathogenicity , Insect Vectors/virology , Janus Kinases/genetics , Janus Kinases/metabolism , Luteovirus/pathogenicity , STAT5 Transcription Factor/genetics , STAT5 Transcription Factor/metabolism , Triticum/parasitology , Triticum/virology
4.
Planta ; 252(1): 14, 2020 Jul 03.
Article in English | MEDLINE | ID: mdl-32621008

ABSTRACT

MAIN CONCLUSION: The findings of this study suggest that known resistant sorghum genotypes compensate for feeding pressure of sugarcane aphid by maintaining/increasing photosynthetic capacity and/or have higher chlorophyll content than susceptible genotypes. Knowledge of the physiological response of sorghum, (Sorghum bicolor (L.) Moench), to sugarcane aphid (SCA), Melanaphis sacchari (Zehnter) feeding will provide baseline information on defense responses and resistance mechanisms. This study documented the impact of SCA feeding on seven sorghum genotypes by measuring chlorophyll content, photosynthetic rate, stomatal conductance, and carbon assimilation for a 14-d post-infestation evaluation. Carbon assimilation (A/Ci) curves were recorded at 3, 6, 9, and 15 d after aphid infestation to describe the pattern of physiological response of resistant and susceptible sorghums over time. Chlorophyll loss from resistant genotypes was significantly lower (≤ 10% loss) than from susceptible cultivars. Most resistant genotypes compensated for aphid feeding by either increasing or maintaining photosynthetic rate and stomatal conductance. Carbon assimilation curves over time showed that infested resistant plants had delayed photosynthetic decreases, whereas susceptible plants rapidly lost photosynthetic capacity. This research also investigated the influence of aphid density (0, 50, 100, and 200 nymphs/plant) on the photosynthetic rates of 28-d-old resistant and susceptible sorghums measured at 72-h post-infestation. Although there were no visual symptoms in susceptible sorghums, photosynthetic rates were impaired when infested with ≥ 100 SCA. In contrast, resistant plants were able to compensate for SCA feeding. Differences in the physiological responses of susceptible versus resistant sorghums indicate that resistant sorghum plants can tolerate some physiological impacts of SCA feeding and maintain photosynthetic integrity.


Subject(s)
Aphids/physiology , Sorghum/physiology , Animals , Aphids/pathogenicity , Carbon Dioxide/metabolism , Chlorophyll/metabolism , Genotype , Photosynthesis , Population Density , Sorghum/genetics
5.
Plant Cell ; 29(6): 1460-1479, 2017 Jun.
Article in English | MEDLINE | ID: mdl-28559475

ABSTRACT

A transient rise in cytosolic calcium ion concentration is one of the main signals used by plants in perception of their environment. The role of calcium in the detection of abiotic stress is well documented; however, its role during biotic interactions remains unclear. Here, we use a fluorescent calcium biosensor (GCaMP3) in combination with the green peach aphid (Myzus persicae) as a tool to study Arabidopsis thaliana calcium dynamics in vivo and in real time during a live biotic interaction. We demonstrate rapid and highly localized plant calcium elevations around the feeding sites of M. persicae, and by monitoring aphid feeding behavior electrophysiologically, we demonstrate that these elevations correlate with aphid probing of epidermal and mesophyll cells. Furthermore, we dissect the molecular mechanisms involved, showing that interplay between the plant defense coreceptor BRASSINOSTEROID INSENSITIVE-ASSOCIATED KINASE1 (BAK1), the plasma membrane ion channels GLUTAMATE RECEPTOR-LIKE 3.3 and 3.6 (GLR3.3 and GLR3.6), and the vacuolar ion channel TWO-PORE CHANNEL1 (TPC1) mediate these calcium elevations. Consequently, we identify a link between plant perception of biotic threats by BAK1, cellular calcium entry mediated by GLRs, and intracellular calcium release by TPC1 during a biologically relevant interaction.


Subject(s)
Aphids/pathogenicity , Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Arabidopsis/parasitology , Calcium/metabolism , Cell Membrane/metabolism , Cell Membrane/parasitology , Cytosol/metabolism , Ion Channels/metabolism , Protein Serine-Threonine Kinases/metabolism , Vacuoles/metabolism , Animals , Arabidopsis Proteins/genetics , Calcium Channels/genetics , Calcium Channels/metabolism , Gene Expression Regulation, Plant , Protein Serine-Threonine Kinases/genetics , Receptors, Glutamate/genetics , Receptors, Glutamate/metabolism
6.
Theor Appl Genet ; 133(2): 615-622, 2020 Feb.
Article in English | MEDLINE | ID: mdl-31773177

ABSTRACT

KEY MESSAGE: A new greenbug resistance gene Gb8 conferring broad resistance to US greenbug biotypes was identified in hard red winter wheat line PI 595379-1 and was mapped to the terminal region of chromosome 7DL. Greenbug [Schizaphis graminum (Rondani)] is a worldwide insect pest that poses a serious threat to wheat production. New greenbug resistance genes that can be readily used in wheat breeding are urgently needed. The objective of this study was to characterize a greenbug resistance gene in PI 595379-1, a single plant selection from PI 595379. Genetic analysis of response to greenbug biotype E in an F2:3 population derived from a cross between PI 595379-1 and PI 243735 indicated that a single gene, designated Gb8, conditioned resistance. Linkage analysis placed Gb8 in a 2.7-Mb interval in the terminal bin of chromosome 7DL (7DL3-082-1.0), spanning 595.6 to 598.3 Mb in the Chinese Spring IWGSC RefSeq version 1.0 reference sequence. Gb8 co-segregated with a newly developed SSR marker Xstars508, positioned at 596.4 Mb in the reference sequence. Allelism tests showed that Gb8 was different from three permanently named genes on the same chromosome arm and the estimated genetic distance between Gb8 and Gb3 was 15.35 ± 1.35 cM. Gb8 can be directly used in wheat breeding to enhance greenbug resistance.


Subject(s)
Aphids/pathogenicity , Disease Resistance/genetics , Plant Diseases/genetics , Triticum/genetics , Alleles , Animals , Chromosome Mapping , Crosses, Genetic , Disease Resistance/physiology , Genetic Linkage , Plant Breeding , Plant Diseases/parasitology , Triticum/metabolism
7.
Biotechnol Lett ; 42(10): 2013-2033, 2020 Oct.
Article in English | MEDLINE | ID: mdl-32676799

ABSTRACT

OBJECTIVE: Brassica juncea, a major oilseed crop, suffers substantial yield losses due to infestation by mustard aphids (Lipaphis erysimi). Unavailability of resistance genes within the accessible gene pool underpins significance of the transgenic strategy in developing aphid resistance. In this study, we aimed for the identification of an aphid-responsive promoter from B. juncea, based on the available genomic resources. RESULTS: A monosaccharide transporter gene, STP4 in B. juncea was activated by aphids and sustained increased expression as the aphids colonized the plants. We cloned the upstream intergenic region of STP4 and validated its stand-alone aphid-responsive promoter activity. Further, deletion analysis identified the putative cis-elements important for the aphid responsive promoter activity. CONCLUSION: The identified STP4 promoter can potentially be used for driving high level aphid-inducible expression of transgenes in plants. Use of aphid-responsive promoter instead of constitutive promoters can potentially reduce the metabolic burden of transgene-expression on the host plant.


Subject(s)
Aphids/pathogenicity , Monosaccharide Transport Proteins/genetics , Mustard Plant , Plant Proteins/genetics , Promoter Regions, Genetic/genetics , Animals , Monosaccharide Transport Proteins/metabolism , Plant Diseases , Plant Leaves/parasitology , Plant Proteins/metabolism , Plants, Genetically Modified
8.
Int J Mol Sci ; 21(21)2020 Oct 27.
Article in English | MEDLINE | ID: mdl-33120946

ABSTRACT

Yellow sugarcane aphid (YSA) (Sipha flava, Forbes) is a damaging pest on many grasses. Switchgrass (Panicum virgatum L.), a perennial C4 grass, has been selected as a bioenergy feedstock because of its perceived resilience to abiotic and biotic stresses. Aphid infestation on switchgrass has the potential to reduce the yields and biomass quantity. Here, the global defense response of switchgrass cultivars Summer and Kanlow to YSA feeding was analyzed by RNA-seq and metabolite analysis at 5, 10, and 15 days after infestation. Genes upregulated by infestation were more common in both cultivars compared to downregulated genes. In total, a higher number of differentially expressed genes (DEGs) were found in the YSA susceptible cultivar (Summer), and fewer DEGs were observed in the YSA resistant cultivar (Kanlow). Interestingly, no downregulated genes were found in common between each time point or between the two switchgrass cultivars. Gene co-expression analysis revealed upregulated genes in Kanlow were associated with functions such as flavonoid, oxidation-response to chemical, or wax composition. Downregulated genes for the cultivar Summer were found in co-expression modules with gene functions related to plant defense mechanisms or cell wall composition. Global analysis of defense networks of the two cultivars uncovered differential mechanisms associated with resistance or susceptibility of switchgrass in response to YSA infestation. Several gene co-expression modules and transcription factors correlated with these differential defense responses. Overall, the YSA-resistant Kanlow plants have an enhanced defense even under aphid uninfested conditions.


Subject(s)
Aphids/pathogenicity , Gene Regulatory Networks , Panicum/parasitology , Plant Immunity , Animals , Biomass , Gene Expression Profiling , Gene Expression Regulation, Plant , Metabolomics , Panicum/classification , Panicum/genetics , Plant Proteins/genetics , Sequence Analysis, RNA
9.
Int J Mol Sci ; 21(21)2020 Nov 04.
Article in English | MEDLINE | ID: mdl-33158282

ABSTRACT

Russian wheat aphid, Diuraphis noxia (Kurdjumov), is a severe pest of wheat, Triticum aestivum L., throughout the world. Resistant cultivars are viewed as the most economical and environmentally viable control available. Studies to identify molecular markers to facilitate resistance breeding started in the 1990s, and still continue. This paper reviews and discusses the literature pertaining to the D. noxia R-genes on chromosome 7D, and markers reported to be associated with them. Individual plants with known phenotypes from a panel of South African wheat accessions are used as examples. Despite significant inputs from various research groups over many years, diagnostic markers for resistance to D. noxia remain elusive. Factors that may have impeded critical investigation, thus blurring the accumulation of a coherent body of information applicable to Dn resistance, are discussed. This review calls for a more fastidious approach to the interpretation of results, especially considering the growing evidence pointing to the complex regulation of aphid resistance response pathways in plants. Appropriate reflection on prior studies, together with emerging knowledge regarding the complexity and specificity of the D. noxia-wheat resistance interaction, should enable scientists to address the challenges of protecting wheat against this pest in future.


Subject(s)
Aphids/pathogenicity , Biomarkers/analysis , Disease Resistance , Plant Diseases/immunology , Triticum/parasitology , Animals , Bread , Breeding/methods , Breeding/standards , Diagnostic Techniques and Procedures/standards , Disease Resistance/immunology , Host-Parasite Interactions , Parasitic Diseases/diagnosis , Parasitic Diseases/immunology , Parasitic Diseases/parasitology , Phenotype , Plant Diseases/parasitology
10.
Int J Mol Sci ; 21(15)2020 Jul 22.
Article in English | MEDLINE | ID: mdl-32707968

ABSTRACT

Soybean aphid (Aphis glycines Matsumura) is one of the major limiting factors in soybean production. The mechanism of aphid resistance in soybean remains enigmatic as little information is available about the different mechanisms of antibiosis and antixenosis. Here, we used genome-wide gene expression profiling of aphid susceptible, antibiotic, and antixenotic genotypes to investigate the underlying aphid-plant interaction mechanisms. The high expression correlation between infested and non-infested genotypes indicated that the response to aphid was controlled by a small subset of genes. Plant response to aphid infestation was faster in antibiotic genotype and the interaction in antixenotic genotype was moderation. The expression patterns of transcription factor genes in susceptible and antixenotic genotypes clustered together and were distant from those of antibiotic genotypes. Among them APETALA 2/ethylene response factors (AP2/ERF), v-myb avian myeloblastosis viral oncogene homolog (MYB), and the transcription factor contained conserved WRKYGQK domain (WRKY) were proposed to play dominant roles. The jasmonic acid-responsive pathway was dominant in aphid-soybean interaction, and salicylic acid pathway played an important role in antibiotic genotype. Callose deposition was more rapid and efficient in antibiotic genotype, while reactive oxygen species were not involved in the response to aphid attack in resistant genotypes. Our study helps to uncover important genes associated with aphid-attack response in soybean genotypes expressing antibiosis and antixenosis.


Subject(s)
Aphids/immunology , Disease Resistance/genetics , Glycine max/genetics , Glycine max/metabolism , Host-Parasite Interactions/genetics , Plant Defense Against Herbivory/genetics , Plant Diseases/genetics , Animals , Antibiosis , Aphids/pathogenicity , Chromatography, Liquid , Cyclopentanes/metabolism , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Gene Expression Profiling , Gene Ontology , Glucosyltransferases/genetics , Glucosyltransferases/metabolism , Mass Spectrometry , Multigene Family , Oxylipins/metabolism , Plant Diseases/parasitology , Plant Proteins/genetics , Plant Proteins/metabolism , Protein Domains/genetics , Proto-Oncogene Proteins c-myb/genetics , Proto-Oncogene Proteins c-myb/metabolism , Reactive Oxygen Species/pharmacology , Salicylic Acid/metabolism , Signal Transduction/genetics , Transcription Factors/genetics , Transcription Factors/metabolism
11.
J Exp Bot ; 70(15): 4011-4026, 2019 08 07.
Article in English | MEDLINE | ID: mdl-31173098

ABSTRACT

Aphids, including the bird cherry-oat aphid (Rhopalosiphum padi), are significant agricultural pests. The wild relative of barley, Hordeum spontaneum 5 (Hsp5), has been described to be partially resistant to R. padi, with this resistance proposed to involve higher thionin and lipoxygenase gene expression. However, the specificity of this resistance to aphids and its underlying mechanistic processes are unknown. In this study, we assessed the specificity of Hsp5 resistance to aphids and analysed differences in aphid probing and feeding behaviour on Hsp5 and a susceptible barley cultivar (Concerto). We found that partial resistance in Hsp5 to R. padi extends to two other aphid pests of grasses. Using the electrical penetration graph technique, we show that partial resistance is mediated by phloem- and mesophyll-based resistance factors that limit aphid phloem ingestion. To gain insight into plant traits responsible for partial resistance, we compared non-glandular trichome density, defence gene expression, and phloem composition of Hsp5 with those of the susceptible barley cultivar Concerto. We show that Hsp5 partial resistance involves elevated basal expression of thionin and phytohormone signalling genes, and a reduction in phloem quality. This study highlights plant traits that may contribute to broad-spectrum partial resistance to aphids in barley.


Subject(s)
Aphids/pathogenicity , Hordeum/metabolism , Hordeum/parasitology , Mesophyll Cells/metabolism , Mesophyll Cells/parasitology , Phloem/metabolism , Phloem/parasitology , Plant Diseases/parasitology , Animals , Gene Expression Regulation, Plant
12.
Int J Mol Sci ; 20(15)2019 Jul 31.
Article in English | MEDLINE | ID: mdl-31370193

ABSTRACT

Prior experiments illustrated reactive oxygen species (ROS) overproduction in maize plants infested with bird-cherry-oat (Rhopalosiphum padi L.) aphids. However, there is no available data unveiling the impact of aphids feeding on oxidative damages of crucial macromolecules in maize tissues. Therefore, the purpose of the current study was to evaluate the scale of oxidative damages of genomic DNA, total RNA and mRNA, proteins, and lipids in seedling leaves of two maize genotypes (Zlota Karlowa and Waza cvs-susceptible and relatively resistant to the aphids, respectively). The content of oxidized guanosine residues (8-hydroxy-2'-deoxyguanosine; 8-OHdG) in genomic DNA, 8-hydroxyguanosine (8-OHG) in RNA molecules, protein carbonyl groups, total thiols (T-SH), protein-bound thiols (PB-SH), non-protein thiols (NP-SH), malondialdehyde (MDA) and electrolyte leakage (EL) levels in maze plants were determined. In addition, the electrical penetration graphs (EPG) technique was used to monitor and the aphid stylet positioning and feeding modes in the hosts. Maize seedlings were infested with 0 (control), 30 or 60 R. padi adult apterae per plant. Substantial increases in the levels of RNA, protein and lipid oxidation markers in response to aphid herbivory, but no significant oxidative damages of genomic DNA, were found. Alterations in the studied parameters were dependent on maize genotype, insect abundance and infestation time.


Subject(s)
Aphids/physiology , Gene Expression Regulation, Plant , Host-Parasite Interactions/genetics , Plant Leaves/genetics , Plant Proteins/genetics , Zea mays/genetics , 8-Hydroxy-2'-Deoxyguanosine/metabolism , Animals , Aphids/pathogenicity , DNA, Plant/genetics , DNA, Plant/metabolism , Genotype , Guanosine/analogs & derivatives , Guanosine/metabolism , Lipids/chemistry , Malondialdehyde/metabolism , Oxidation-Reduction , Oxidative Stress , Plant Diseases/genetics , Plant Diseases/parasitology , Plant Leaves/parasitology , Plant Proteins/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA, Plant/genetics , RNA, Plant/metabolism , Seedlings/genetics , Seedlings/parasitology , Sulfhydryl Compounds/metabolism , Zea mays/parasitology
13.
Plant Biotechnol J ; 16(8): 1434-1445, 2018 08.
Article in English | MEDLINE | ID: mdl-29331089

ABSTRACT

Aphids are pests of chrysanthemum that employ plant volatiles to select host plants and ingest cell contents to probe host quality before engaging in prolonged feeding and reproduction. Changes in volatile and nonvolatile metabolite profiles can disrupt aphid-plant interactions and provide new methods of pest control. Chrysanthemol synthase (CHS) from Tanacetum cinerariifolium represents the first committed step in the biosynthesis of pyrethrin ester insecticides, but no biological role for the chrysanthemol product alone has yet been documented. In this study, the TcCHS gene was over-expressed in Chrysanthemum morifolium and resulted in both the emission of volatile chrysanthemol (ca. 47 pmol/h/gFW) and accumulation of a chrysanthemol glycoside derivative, identified by NMR as chrysanthemyl-6-O-malonyl-ß-D-glucopyranoside (ca. 1.1 mM), with no detrimental phenotypic effects. Dual-choice assays separately assaying these compounds in pure form and as part of the headspace and extract demonstrated independent bioactivity of both components against the cotton aphid (Aphis gossypii). Performance assays showed that the TcCHS plants significantly reduced aphid reproduction, consistent with disturbance of aphid probing activities on these plants as revealed by electropenetrogram (EPG) studies. In open-field trials, aphid population development was very strongly impaired demonstrating the robustness and high impact of the trait. The results suggest that expression of the TcCHS gene induces a dual defence system, with both repellence by chrysanthemol odour and deterrence by its nonvolatile glycoside, introducing a promising new option for engineering aphid control into plants.


Subject(s)
Alkyl and Aryl Transferases/metabolism , Aphids/pathogenicity , Chrysanthemum/enzymology , Chrysanthemum/parasitology , Plant Proteins/metabolism , Animals , Chrysanthemum/metabolism , Glycosides/metabolism , Terpenes/metabolism
14.
Plant Physiol ; 173(3): 1892-1903, 2017 03.
Article in English | MEDLINE | ID: mdl-28100451

ABSTRACT

Plant- and animal-feeding insects secrete saliva inside their hosts, containing effectors, which may promote nutrient release and suppress immunity. Although for plant pathogenic microbes it is well established that effectors target host proteins to modulate host cell processes and promote disease, the host cell targets of herbivorous insects remain elusive. Here, we show that the existing plant pathogenic microbe effector paradigm can be extended to herbivorous insects in that effector-target interactions inside host cells modify critical host processes to promote plant susceptibility. We showed that the effector Mp1 from Myzus persicae associates with the host Vacuolar Protein Sorting Associated Protein52 (VPS52). Using natural variants, we provide a strong link between effector virulence activity and association with VPS52, and show that the association is highly specific to Mpersicae-host interactions. Also, coexpression of Mp1, but not Mp1-like variants, specifically with host VPS52s resulted in effector relocalization to vesicle-like structures that associate with prevacuolar compartments. We show that high VPS52 levels negatively impact virulence, and that aphids are able to reduce VPS52 levels during infestation, indicating that VPS52 is an important virulence target. Our work is an important step forward in understanding, at the molecular level, how a major agricultural pest promotes susceptibility during infestation of crop plants. We give evidence that an herbivorous insect employs effectors that interact with host proteins as part of an effective virulence strategy, and that these effectors likely function in a species-specific manner.


Subject(s)
Arabidopsis Proteins/genetics , Arabidopsis/genetics , Plant Diseases/genetics , Vesicular Transport Proteins/genetics , Amino Acid Sequence , Animals , Aphids/genetics , Aphids/pathogenicity , Aphids/physiology , Arabidopsis/metabolism , Arabidopsis/parasitology , Arabidopsis Proteins/metabolism , Gene Expression Regulation, Plant , Host-Parasite Interactions , Immunoblotting , Insect Proteins/genetics , Insect Proteins/metabolism , Microscopy, Confocal , Plant Diseases/parasitology , Plants, Genetically Modified , Protein Binding , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Amino Acid , Solanum tuberosum/genetics , Solanum tuberosum/metabolism , Solanum tuberosum/microbiology , Species Specificity , Vesicular Transport Proteins/metabolism , Virulence/genetics
15.
Transgenic Res ; 27(4): 355-366, 2018 08.
Article in English | MEDLINE | ID: mdl-29777502

ABSTRACT

Plants have co-evolved with a diverse array of pathogens and insect herbivores and so have evolved an extensive repertoire of constitutive and induced defence mechanisms activated through complex signalling pathways. OXI1 kinase is required for activation of mitogen-activated protein kinases (MAPKs) and is an essential part of the signal transduction pathway linking oxidative burst signals to diverse downstream responses. Furthermore, changes in the levels of OXI1 appear to be crucial for appropriate signalling. Callose deposition also plays a key role in defence. Here we demonstrate, for the first time, that OXI1 plays an important role in defence against aphids. The Arabidopsis mutant, oxi1-2, showed significant resistance both in terms of population build-up (p < 0.001) and the rate of build-up (p < 0.001). Arabidopsis mutants for ß-1,3-glucanase, gns2 and gns3, showed partial aphid resistance, significantly delaying developmental rate, taking two-fold longer to reach adulthood. Whilst ß-1,3-glucanase genes GNS1, GNS2, GNS3 and GNS5 were not induced in oxi1-2 in response to aphid feeding, GNS2 was expressed to high levels in the corresponding WT (Col-0) in response to aphid feeding. Callose synthase GSL5 was up-regulated in oxi1-2 in response to aphids. The results suggest that resistance in oxi1-2 mutants is through induction of callose deposition via MAPKs resulting in ROS induction as an early response. Furthermore, the results suggest that the ß-1,3-glucanase genes, especially GNS2, play an important role in host plant susceptibility to aphids. Better understanding of signalling cascades underpinning tolerance to biotic stress will help inform future breeding programmes for enhancing crop resilience.


Subject(s)
Arabidopsis Proteins/genetics , Arabidopsis/parasitology , Disease Resistance/genetics , Plant Diseases/genetics , Protein Serine-Threonine Kinases/genetics , Animals , Aphids/genetics , Aphids/pathogenicity , Arabidopsis/genetics , Arabidopsis/growth & development , Drug Tolerance , Gene Expression Regulation, Plant/genetics , Plant Breeding , Plant Diseases/parasitology , Signal Transduction , Transcriptional Activation
16.
Plant Cell Environ ; 40(11): 2628-2643, 2017 Nov.
Article in English | MEDLINE | ID: mdl-28452058

ABSTRACT

Aphids are phloem-feeding insects that cause yield loss on a wide range of crops, including cereals such as barley. Whilst most aphid species are limited to one or few host species, some are able to reproduce on many plants belonging to different families. Interestingly, aphid probing behaviour can be observed on both host and non-host species, indicating that interactions take place at the molecular level that may impact host range. Here, we aimed to gain insight into the interaction of barley with aphid species differing in their ability to infest this crop by analysing transcriptional responses. Firstly, we determined colonization efficiency, settlement and probing behaviour for the aphid species Rhopalosiphum padi, Myzus persicae and Myzus cerasi, which defined host, poor-host and non-host interactions, respectively. Analyses of barley transcriptional responses revealed gene sets differentially regulated upon the different barley-aphid interactions and showed that the poor-host interaction with M. persicae resulted in the strongest regulation of genes. Interestingly, we identified several thionin genes strongly up-regulated upon interaction with M. persicae, and to a lesser extent upon R. padi interaction. Ectopic expression of two of these genes in Nicotiana benthamiana reduced host susceptibility to M. persicae, indicating that thionins contribute to defences against aphids.


Subject(s)
Aphids/physiology , Disease Resistance/drug effects , Gene Expression Profiling , Gene Expression Regulation, Plant/drug effects , Hordeum/genetics , Hordeum/parasitology , Plant Diseases/genetics , Plant Diseases/parasitology , Thionins/pharmacology , Animals , Aphids/pathogenicity , Cluster Analysis , Genes, Plant , Hordeum/drug effects , Plant Leaves/drug effects , Plant Leaves/genetics , Plants, Genetically Modified , Reproducibility of Results , Species Specificity , Nicotiana/genetics , Transcription, Genetic/drug effects , Transcriptome/genetics , Virulence/drug effects
17.
Biosci Biotechnol Biochem ; 81(6): 1069-1077, 2017 Jun.
Article in English | MEDLINE | ID: mdl-28164745

ABSTRACT

Insect galls are abnormal plant tissues induced by parasitic insect(s) for use as their habitat. In previous work, we suggested that gall tissues induced by the aphid Tetraneura nigriabdominalis on Japanese elm trees are less responsive than leaf tissues to jasmonic acid (JA), which is involved in the production of volatile organic compounds as a typical defensive reaction of plants against attack by insect pests. A comprehensive analysis of gene expression by RNA sequencing indicated that the number of JA responsive genes was markedly lower in gall tissues than in leaf tissues. This suggests that gall tissues are mostly defective in JA signaling, although JA signaling is not entirely compromised in gall tissue. Gene ontology analysis sheds light on some stress-related unigenes with higher expression levels in gall tissues, suggesting that host plants sense aphids as a biotic stress but are defective in the JA-mediated defense response in gall tissues.


Subject(s)
Aphids/pathogenicity , Gene Expression Regulation, Plant , Plant Proteins/genetics , Plant Tumors/genetics , Transcriptome/immunology , Ulmus/genetics , Animals , Aphids/physiology , Cyclopentanes/immunology , Cyclopentanes/metabolism , Gene Ontology , Host-Parasite Interactions , Inflammation Mediators/immunology , Inflammation Mediators/metabolism , Molecular Sequence Annotation , Oxylipins/immunology , Oxylipins/metabolism , Plant Leaves/genetics , Plant Leaves/immunology , Plant Leaves/parasitology , Plant Proteins/immunology , Plant Tumors/parasitology , Signal Transduction , Ulmus/immunology , Ulmus/parasitology
18.
Int J Mol Sci ; 18(3)2017 Mar 12.
Article in English | MEDLINE | ID: mdl-28287502

ABSTRACT

The gene encoding the MYB (v-myb avian myeloblastosis vira l oncogene homolog) transcription factor CmMYB19 was isolated from chrysanthemum. It encodes a 200 amino acid protein and belongs to the R2R3-MYB subfamily. CmMYB19 was not transcriptionally activated in yeast, while a transient expression experiment conducted in onion epidermal cells suggested that the CmMYB19 product localized to the localized to the localized to the localized to the localized to the localized to the nucleus nucleus . CmMYB19 transcription was induced by aphid (Macrosiphoniella sanborni) infestation, and the abundance of transcript was higher in the leaf and stem than in the root. The over-expression of CmMYB19 restricted the multiplication of the aphids. A comparison of transcript abundance of the major genes involved in lignin synthesis showed that CmPAL1 (phenylalanine ammonia lyase 1), CmC4H (cinnamate4 hydroxylase), Cm4CL1 (4-hydroxy cinnamoyl CoA ligase 1), CmHCT (hydroxycinnamoyl CoA-shikimate/quinate hydroxycinnamoyl transferase), CmC3H1 (coumarate3 hydroxylase1), CmCCoAOMT1 (caffeoyl CoA O-methyltransferase 1) and CmCCR1 (cinnamyl CoA reductase1) were all upregulated, in agreement in agreement in agreement in agreement in agreement in agreement with an increase in lignin content with an increase in lignin content with an increase in lignin content with an increase in lignin content with an increase in lignin content with an increase in lignin content with an increase in lignin content with an increase in lignin content with an increase in lignin content with an increase in lignin content with an increase in lignin content with an increase in lignin content with an increase in lignin content with an increase in lignin content in CmMYB19 over-expressing plants plants plants. Collectively, the over-expression of CmMYB19 restricted the multiplication of the aphids on the host, mediated by an enhanced accumulation of lignin.


Subject(s)
Aphids/pathogenicity , Chrysanthemum/genetics , Disease Resistance/genetics , Lignin/biosynthesis , Plant Proteins/genetics , Transcription Factors/genetics , Animals , Chrysanthemum/metabolism , Chrysanthemum/parasitology , Gene Expression Regulation, Plant , Host-Parasite Interactions , Plant Proteins/metabolism , Transcription Factors/metabolism , Up-Regulation
19.
Int J Mol Sci ; 18(12)2017 Dec 19.
Article in English | MEDLINE | ID: mdl-29257097

ABSTRACT

Aphids are pests on many crops and depend on plant phloem sap as their food source. In an attempt to find factors improving plant resistance against aphids, we studied the effects of overexpression and down-regulation of the lipoxygenase gene LOX2.2 in barley (Hordeum vulgare L.) on the performance of two aphid species. A specialist, bird cherry-oat aphid (Rhopalosiphum padi L.) and a generalist, green peach aphid (Myzus persicae Sulzer) were studied. LOX2.2 overexpressing lines showed up-regulation of some other jasmonic acid (JA)-regulated genes, and antisense lines showed down-regulation of such genes. Overexpression or suppression of LOX2.2 did not affect aphid settling or the life span on the plants, but in short term fecundity tests, overexpressing plants supported lower aphid numbers and antisense plants higher aphid numbers. The amounts and composition of released volatile organic compounds did not differ between control and LOX2.2 overexpressing lines. Up-regulation of genes was similar for both aphid species. The results suggest that LOX2.2 plays a role in the activation of JA-mediated responses and indicates the involvement of LOX2.2 in basic defense responses.


Subject(s)
Aphids/pathogenicity , Cyclopentanes/metabolism , Gene Expression Regulation, Plant , Hordeum/genetics , Host-Parasite Interactions , Lipoxygenase/genetics , Oxylipins/metabolism , Plant Proteins/genetics , Animals , Aphids/physiology , Fertility , Hordeum/enzymology , Hordeum/parasitology , Lipoxygenase/metabolism , Oils, Volatile/metabolism , Plant Proteins/metabolism
20.
Molecules ; 22(6)2017 Jun 08.
Article in English | MEDLINE | ID: mdl-28594377

ABSTRACT

Based on the structural framework of a pyriproxyfen metabolite, nineteen oxime ester derivatives were synthesized via reaction of the carboxylic acids with 4-(2-(2-pyridinyloxy)ethoxy)benzaldehyde oxime. The corresponding structures were comprehensively characterized by ¹H-nuclear magnetic resonance (NMR), 13C-NMR, and electrospray ionization high-resolution mass spectrometry (ESI-HRMS). All of the compounds were screened for their insecticidal activities against Plutella xylostella and Myzus persicae, and for their ovicidal activities against Helicoverpa armigera eggs. The results obtained show that most of the oxime ester derivatives displayed moderate to high insecticidal activities and ovicidal activities at a concentration of 600 ug/mL. In particular, the ovicidal activity of compounds 5j, 5o, 5p, 5q, and 5s was determined to be 100%. Importantly, some of the compounds presented even higher biological activities than the reference compound pyriproxyfen. For example, compound 5j displayed an insecticidal activity value of 87.5% against Myzus persicae, whereas the activity value of pyriproxyfen was 68.3% at a concentration of 600 ug/mL. Among the synthesized compounds 5j and 5s exhibited broad biological activity spectra.


Subject(s)
Esters/chemistry , Insecticides/chemistry , Oximes/chemistry , Pyridines/chemistry , Animals , Aphids/drug effects , Aphids/pathogenicity , Carboxylic Acids/chemistry , Carboxylic Acids/pharmacology , Esters/chemical synthesis , Esters/pharmacology , Insecticides/chemical synthesis , Insecticides/pharmacology , Magnetic Resonance Spectroscopy , Molecular Structure , Moths/drug effects , Moths/pathogenicity , Oximes/chemical synthesis , Oximes/pharmacology , Pyridines/chemical synthesis , Pyridines/pharmacology , Spectrometry, Mass, Electrospray Ionization , Structure-Activity Relationship
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