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1.
Planta ; 251(3): 64, 2020 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-32048047

RESUMO

MAIN CONCLUSION: Hand-held Raman spectroscopy can be used for confirmatory, non-invasive and non-destructive detection and identification of two haplotypes of Liberibacter disease on tomatoes. Using this spectroscopic approach, structural changes in carotenoids, xylan, cellulose and pectin that are associ-ated with this bacterial disease can be determined. 'Candidatus Liberibacter solanacearum' (Lso) is a phloem-limited Gram-negative bacterium that infects crops worldwide. In North America, two haplotypes of Lso (LsoA and LsoB) are transmitted by the potato psyllid, Bactericera cockerelli (Sulc), and infect many solanaceous crops such as potato and tomato. Infected plants exhibit chlorosis, severe stunting, leaf cupping, and scorching. Polymerase chain reaction (PCR) and potato tuber frying are commonly used methods for diagnostics of the plant disease caused by Lso. However, they are time-consuming, costly, destructive to the sample, and often not sensitive enough to detect the pathogen in the early infection stage. Raman spectroscopy (RS) is a noninvasive, nondestructive, analytical technique, which probes chemical composition of analyzed samples. In this study, we demonstrate that Lso infection can be diagnosed by non-invasive spectroscopic analysis of tomato leaves three weeks following infection, before the development of aerial symptoms. In combination with chemometric analyses, Raman spectroscopy allows for 80% accurate diagnostics of Liberibacter disease caused by each of the two different haplotypes. This diagnostics approach is portable and sample agnostic, suggesting that it could be utilized for other crops and could be conducted autonomously.

2.
Phytopathology ; 110(3): 648-655, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31697198

RESUMO

'Candidatus Liberibacter solanacearum' is a plant pathogen affecting the families Solanaceae and Apiaceae in different parts of the world. 'Ca. L. solanacearum' is a Gram-negative, fastidious α-proteobacterium that is vectored by different psyllid species. Plant-pathogenic bacteria are known for interfering with the host physiology or defense mechanisms, often by secreting bacterial effectors. Effector proteins are critical for virulence; therefore, the identification of effectors could help with disease management. In this study, we characterized the Sec-translocon-dependent 'Ca. L. solanacearum'-hypothetical protein effector 1 (Lso-HPE1). We compared this protein sequence in the different 'Ca. L. solanacearum' haplotypes. We predicted the signal peptide and validated its function using Escherichia coli's alkaline phosphatase fusion assay. Agrobacterium tumefaciens-mediated transient expression in Nicotiana benthamiana demonstrated that Lso-HPE1 from 'Ca. L. solanacearum' haplotypes A and B were able to inhibit the induction of cell death in plants. We also compared gene expression of the Lso-HPE1- transcripts in 'Ca. L. solanacearum' haplotypes A and B in tomato and in the vector Bactericera cockerelli. This work validates the identification of a Sec-translocon-dependent 'Ca. L. solanacearum' protein possibly involved in suppression of plant cell death.


Assuntos
Hemípteros , Lycopersicon esculentum , Rhizobiaceae , Animais , Doenças das Plantas , Imunidade Vegetal
3.
Insect Sci ; 27(1): 58-68, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29676854

RESUMO

"Candidatus Liberibacter solanacearum" (Lso) are phloem-restricted and unculturable Gram-negative bacteria. Presently five haplotypes have been identified worldwide; but only haplotypes A and B are associated with the vector Bactericera cockerelli (Sulc.) in the Americas. Previous studies showed that Lso-infection reduces B. cockerelli reproductive output and that Lso haplotype B is more pathogenic than Lso haplotype A. To understand the interaction of Lso haplotype B and B. cockerelli, the fitness of Lso-free and Lso B-infected insects, and the expression of vitellogenin (BcVg1-like), a gene involved directly in the insect reproduction were analyzed. Statistical differences in the number of eggs oviposited, and the total number of progeny nymphs and adults were found among crosses of insects with or without Lso. Significant differences in sex proportions were found between Lso B-infected and Lso-free crosses: a higher proportion of F1 adult females were obtained from Lso B-infected mothers. A significant reduction of BcVg1-like was observed in crosses performed with Lso B-infected females compared to the Lso-free insects. In female cohorts of different age, a significant reduction of BcVg1-like expression was measured in 7-d-old Lso B-infected females (virgin and mated) compared with 7-d-old Lso-free females (virgin and mated), respectively. The reduction of BcVg1-like transcript was associated with a lower number of developing oocytes observed in female's reproductive systems. Overall, this study represents the first step to understand the interaction of Lso B with B. cockerelli, highlighting the effect of Lso B infection on egg production, BcVg1-like expression, and oocyte development.


Assuntos
Aptidão Genética , Hemípteros/fisiologia , Rhizobiaceae/fisiologia , Vitelogênese , Animais , Hemípteros/genética , Hemípteros/crescimento & desenvolvimento , Hemípteros/microbiologia , Lycopersicon esculentum/microbiologia , Lycopersicon esculentum/fisiologia , Ninfa/genética , Ninfa/crescimento & desenvolvimento , Ninfa/microbiologia , Ninfa/fisiologia , Doenças das Plantas/microbiologia
4.
Plants (Basel) ; 8(11)2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31731657

RESUMO

Disease caused by the bacterial pathogen "Candidatus Liberibacter solanacearum" (Lso) represents a serious threat to solanaceous crop production. Insecticide applications to control the psyllid vector, Bactericera cockerelli Sulc (Hemiptera: Triozidae) has led to the emergence of resistance in psyllids populations. Efforts to select natural resistant cultivars have been marginally successful and have been complicated by the presence of distinct Lso haplotypes (LsoA, LsoB) differing in symptoms severity on potato and tomato. A potentially promising management tool is to boost host resistance to the pathogen and/or the insect vector by promoting mycorrhization. Here we tested the hypothesis that mycorrhizal fungi can mitigate the effect of Lso infection on tomato plants. The presence of mycorrhizal fungi substantially delayed and reduced the incidence of Lso-induced symptoms on tomato as compared to non-mycorrhized plants. However, PCR with specific Lso primers revealed that mycorrhization did not prevent Lso transmission or translocation to newly formed leaves. Mycorrhization significantly reduced oviposition by psyllids harboring LsoA and survival of nymphs from these eggs. However, mycorrhization had no effect on oviposition by psyllids harboring LsoB or the survival of nymphs from parents harboring LsoB. These findings indicate the use of mycorrhizal fungi is a promising strategy for the mitigation of disease caused by both LsoA and LsoB and warrants additional field testing.

5.
Infect Immun ; 88(1)2019 12 17.
Artigo em Inglês | MEDLINE | ID: mdl-31611278

RESUMO

"Candidatus Liberibacter solanacearum" is a pathogen transmitted by the potato psyllid Bactericera cockerelli (Sulc) (Hemiptera: Triozidae) in a persistent manner. In this study, we investigated the molecular interaction between "Ca. Liberibacter solanacearum" and the potato psyllid at the gut interface. Specifically, we focused on the apoptotic response of potato psyllids to the infection by two "Ca. Liberibacter solanacearum" haplotypes, LsoA and LsoB. To this end, we first quantified and localized "Ca. Liberibacter solanacearum" in the gut of adult psyllids. We then evaluated the existence of an apoptotic response in the insect gut using microscopy analyses to visualize the nuclei and the actin cytoskeleton of the gut cells and DNA fragmentation analyses by agarose gel electrophoresis. We also performed annexin V cell death assays to detect apoptosis. Finally, we annotated apoptosis-related genes from the potato psyllid transcriptome and evaluated their expression in response to "Ca. Liberibacter solanacearum" infection. The results showed no cellular markers of apoptosis despite the large amount of "Ca. Liberibacter solanacearum" present in the psyllid gut. In addition, only three genes potentially involved in apoptosis were regulated in the psyllid gut in response to "Ca. Liberibacter solanacearum": the apoptosis-inducing factor AIF3 was downregulated in LsoA-infected psyllids, while the inhibitor of apoptosis IAPP5 was downregulated and IAP6 was upregulated in LsoB-infected psyllids. Overall, no evidence of apoptosis was observed in the gut of potato psyllid adults in response to either "Ca. Liberibacter solanacearum" haplotype. This study represents a first step toward understanding the interactions between "Ca. Liberibacter solanacearum" and the potato psyllid, which is crucial to developing approaches to disrupt their transmission.


Assuntos
Apoptose , Hemípteros/microbiologia , Interações Hospedeiro-Patógeno , Rhizobiaceae/crescimento & desenvolvimento , Animais , Anexina A5/análise , Fragmentação do DNA , Trato Gastrointestinal/microbiologia , Trato Gastrointestinal/patologia , Perfilação da Expressão Gênica , Insetos Vetores/microbiologia , Solanum tuberosum/parasitologia
6.
PLoS One ; 14(5): e0216800, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31107891

RESUMO

The reproductive ground plan hypothesis proposes that gene networks regulating foraging behavior and reproductive female physiology in social insects emerged from ancestral gene and endocrine factor networks. Expression of storage proteins such as vitellogenins and hexamerins is an example of this co-option. Hexamerins, through their role modulating juvenile hormone availability, are involved in caste determination in termites. The genome of the fire ant (Solenopsis invicta) encodes four hexamerin genes, hexamerin-like (LOC105192919, hereafter called hexamerin 1), hexamerin (LOC105204474, hereafter called hexamerin 2), arylphorin subunit alpha-like, and arylphorin subunit beta. In this study, a phylogenetic analysis of the S. invicta hexamerins determined that each predicted protein clustered with one of the orthologous Apis mellifera hexamerins. Gene expression analyses by RT-qPCR revealed differential expression of the hexamerins between queens and workers, and between specific task-allocated workers (nurses and foragers). Queens and nurses had significantly higher expression of all genes when compared to foragers. Hexamerin 1 was expressed at higher levels in queens, while hexamerin 2 and arylphorin subunit beta were expressed at significantly higher levels in nurses. Arylphorin subunit alpha-like showed no significant difference in expression between virgin queens and nurses. Additionally, we analyzed the relationship between the expression of hexamerin genes and S-hydroprene, a juvenile hormone analog. Significant changes in hexamerin expression were recorded in nurses, virgin queens, and foragers 12 h after application of the analog. Hexamerin 1 and arylphorin subunit alpha-like expression were significantly lower after analog application in virgin queens. In foragers, hexamerin 2 and arylphorin subunit beta were significantly lower after analog application, while in nurses expression of all genes were significantly lower after analog application. Our results suggest that in S. invicta hexamerin genes could be associated with reproductive division of labor and task-allocation of workers.


Assuntos
Formigas/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Proteínas de Insetos/biossíntese , Hormônios Juvenis/farmacologia , Vitelogeninas/biossíntese , Animais , Formigas/genética , Feminino , Proteínas de Insetos/genética , Vitelogeninas/genética
7.
Insect Sci ; 2019 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-30663253

RESUMO

Immunofluorescence has been widely used to localize microbes or specific molecules in insect tissues or cells. However, significant autofluorescence is frequently observed in tissues which can interfere with the fluorescent identification of target antigens, leading to inaccurate or even false positive fluorescent labeling. The alimentary canal of the potato psyllid, Bactericera cockerelli Sulc, exhibits intense autofluorescence, hindering the application of immunolocalization for the detection and localization of the economically important pathogen transmitted by this insect, "Candidatus Liberibacter solanacearum" (Lso). In the present study, we tested the use of irradiation, hydrogen peroxide (H2 O2 ) and Sudan black B (SBB) treatments to reduce the autofluorescence in the B. cockerelli alimentary canal tissues. Furthermore, we assessed the compatibility of the above-mentioned treatments with Lso immunolocalization and actin staining using phalloidin. Our results showed that the autofluorescence in the alimentary canal was reduced by irradiation, H2 O2 , or SBB treatments. The compatibility assays indicated that irradiation and H2 O2 treatment both greatly reduced the fluorescent signal associated with Lso and actin. However, the SBB incubation preserved those target signals, while efficiently eliminating autofluorescence in the psyllid alimentary canal. Therefore, herein we propose a robust method for reducing the autofluorescence in the B. cockerelli alimentary canal with SBB treatment, which may improve the use of immunofluorescence labeling in this organism. This method may also have a wide range of uses by reducing the autofluorescence in other arthropod species.

8.
Gen Comp Endocrinol ; 278: 89-103, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-30576645

RESUMO

Knowledge of G protein-coupled receptors (GPCRs) and their signaling modalities is crucial to advancing insect endocrinology, specifically in highly successful invasive social insects, such as the red imported fire ant, Solenopsis invicta Buren. In the first published draft genome of S. invicta, emphasis was placed on the annotation of olfactory receptors, and only the number of predicted GPCR genes was reported. Without an organized and curated resource for GPCRs, it will be difficult to test hypotheses on the endocrine role of neuropeptide hormones, or the function of neurotransmitters and neuromodulators. Therefore, we mined the S. invicta genome for GPCRs and found 324 predicted transcripts encoded by 125 predicted loci and improved the annotation of 55 of these loci. Among them are sixteen GPCRs that are currently annotated as "uncharacterized proteins". Further, the phylogenetic analysis of class A neuropeptide receptors presented here and the comparative listing of GPCRs in the hymenopterans S. invicta, Apis mellifera (both eusocial), Nasonia vitripennis (solitary), and the solitary model dipteran Drosophila melanogaster will facilitate comparative endocrinological studies related to social insect evolution and diversity. We compiled the 24 G protein transcripts predicted (15 α, 7 ß, and 2 γ) from 12 G protein genes (5 α, 5 ß, and 2 γ). Reproductive division of labor is extreme in this ant species, therefore, we compared GPCR and G protein gene expression among worker, mated queen and alate virgin queen ant brain transcriptomes. Transcripts for ten GPCRs and two G proteins were differentially expressed between queen and worker brains. The differentially expressed GPCRs are candidate receptors to explore hypotheses on division of labor in this species.


Assuntos
Formigas/genética , Proteínas de Ligação ao GTP/metabolismo , Hierarquia Social , Espécies Introduzidas , Anotação de Sequência Molecular , Receptores Acoplados a Proteínas-G/metabolismo , Transdução de Sinais , Transcriptoma/genética , Sequência de Aminoácidos , Animais , Encéfalo/metabolismo , Filogenia , Receptores Acoplados a Proteínas-G/química
9.
Plant Dis ; 102(10): 2009-2015, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30133358

RESUMO

'Candidatus Liberibacter solanacearum' is a plant pathogen associated with diseases affecting several crops of the Solanaceae and Apiaceae families. Two 'Ca. L. solanacearum' haplotypes (LsoA and LsoB) infect solanaceous crops in North America and are transmitted by the tomato psyllid Bactericera cockerelli. Although both 'Ca. L. solanacearum' haplotypes cause zebra chip in potato, the diseases associated with each haplotype in tomato (Solanum lycopersicum) have not been described. 'Ca. L. solanacearum'-infected tomato plants exhibit symptoms resembling those of permanent yellowing disease (known in Mexico as "permanente del tomate") and sometimes called psyllid yellows. In this study, the symptoms associated with each 'Ca. L. solanacearum' haplotype in tomato were compared, and the bacterial abundance in different nodes of the plants was measured by quantitative polymerase chain reaction. Surprisingly, both plant phenotype and bacterium distribution were different between LsoA- and LsoB-infected plants. Plants infected with LsoB died prematurely, whereas those infected with LsoA did not. Across the measured time points, LsoB abundance in infected plants was consistent with previous reports describing a sink to source gradient, while such gradient was only observed in LsoA-infected plants early after infection. This is the first report describing the differences in symptoms in tomato associated with two 'Ca. L. solanacearum' haplotypes, LsoA and LsoB.


Assuntos
Lycopersicon esculentum/microbiologia , Rhizobiaceae/genética , DNA Bacteriano/genética , Haplótipos , Doenças das Plantas/microbiologia , Folhas de Planta/microbiologia , Fatores de Tempo
10.
PeerJ ; 6: e4875, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29868280

RESUMO

Vitellogenin has been proposed to regulate division of labor and social organization in social insects. The red imported fire ant (Solenopsis invicta) harbors four distinct, adjacent vitellogenin genes (Vg1, Vg2, Vg3, and Vg4). Contrary to honey bees that have a single Vg ortholog as well as potentially fertile nurses, and to other ant species that lay trophic eggs, S. invicta workers completely lack ovaries or the ability to lay eggs. This provides a unique model to investigate whether Vg duplication in S. invicta was followed by subfunctionalization to acquire non-reproductive functions and whether Vg was co-opted to regulate behavior within the worker caste. To investigate these questions, we compared the expression patterns of S. invicta Vg genes among workers from different morphological subcastes or performing different tasks. RT-qPCRs revealed higher relative expression of Vg1 in major workers compared to both medium and minor workers, and of Vg2 in major workers when compared to minor workers. Relative expression of Vg1 was also higher in carbohydrate foragers when compared to nurses and protein foragers. By contrast, the level of expression of Vg2, Vg3, and Vg4 were not significantly different among the workers performing the specific tasks. Additionally, we analyzed the relationship between the expression of the Vg genes and S-hydroprene, a juvenile hormone analog. No changes in Vg expression were recorded in workers 12 h after application of the analog. Our results suggest that in S. invicta the Vg gene underwent subfunctionalization after duplication to new functions based on the expression bias observed in these data. This may suggest an alternative and still unknown function for Vg in the workers that needs to be investigated further.

11.
Ecol Evol ; 8(8): 4312-4327, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29721300

RESUMO

Transcriptomes of dissected brains from virgin alate and dealate mated queens from polygyne fire ants (Solenopsis invicta) were analyzed and compared. Thirteen genes were upregulated in mated queen brain, and nine were downregulated. While many of the regulated genes were either uncharacterized or noncoding RNAs, those annotated genes included two hexamerin proteins, astakine neuropeptide, serine proteases, and serine protease inhibitors. We found that for select differentially expressed genes in the brain, changes in gene expression were most likely driven by the changes in physiological state (i.e., age, nutritional status, or dominance rank) or in social environment (released from influence of primer pheromone). This was concluded because virgins that dealated after being separated from mated queens showed similar patterns of gene expression in the brain as those of mated queens for hexamerin 1, astakine, and XR_850909. Abaecin (XR_850725), however, appears upregulated only after mating. Therefore, our findings contribute to distinguish how specific gene networks, especially those influenced by queen primer pheromone, are regulated in queen ants. Additionally, to identify brain signaling pathways, we mined the fire ant genome and compiled a list of G-protein-coupled receptors (GPCRs). The expression level of GPCRs and other genes in the "genetic toolkit" in the brains of virgin alates and mated dealate queens is reported.

12.
Plant Mol Biol ; 97(1-2): 57-72, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29619663

RESUMO

KEY MESSAGE: Different responses are elicited in tomato plants by Bactericera cockerelli harboring or not the pathogen 'Candidatus Liberibacter solanacearum'. 'Candidatus Liberibacter solanacearum' (Lso) has emerged as a major pathogen of crops worldwide. This bacterial pathogen is transmitted by Bactericera cockerelli, the tomato psyllid, to solanaceous crops. In this study, the transcriptome profiles of tomato (Solanum lycopersicum) exposed to B. cockerelli infestation and Lso infection were evaluated at 1, 2 and 4 weeks following colonization and/or infection. The plant transcriptional responses to Lso-negative B. cockerelli were different than plant responses to Lso-positive B. cockerelli. The comparative transcriptome analyses of plant responses to Lso-negative B. cockerelli revealed the up-regulation of genes associated with plant defenses regardless of the time-point. In contrast, the general responses to Lso-positive B. cockerelli and Lso-infection were temporally different. Infected plants down-regulated defense genes at week one while delayed the up-regulation of the defense genes until weeks two and four, time points in which early signs of disease development were also detected in the transcriptional response. For example, infected plants regulated carbohydrate metabolism genes which could be linked to the disruption of sugar distribution usually associated with Lso infection. Also, infected plants down-regulated photosynthesis-related genes potentially resulting in plant chlorosis, another symptom associated with Lso infection. Overall, this study highlights that tomato plants induce different sets of genes in response to different stages of B. cockerelli infestation and Lso infection. This is the first transcriptome study of tomato responses to B. cockerelli and Lso, a first step in the direction of finding plant defense genes to enhance plant resistance.


Assuntos
Regulação da Expressão Gênica de Plantas , Hemípteros/microbiologia , Lycopersicon esculentum/genética , Doenças das Plantas/genética , Rhizobiaceae/fisiologia , Animais , Perfilação da Expressão Gênica , Insetos Vetores/microbiologia , Lycopersicon esculentum/microbiologia , Doenças das Plantas/microbiologia , Doenças das Plantas/parasitologia , RNA de Plantas , Análise de Sequência de RNA
13.
BMC Res Notes ; 11(1): 160, 2018 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-29490684

RESUMO

OBJECTIVES: Potato has a large genetic diversity. This diversity is in part due to somaclonal variability that appears within potato selections for which tubers are used as seeds. However, the potato tetraploid genome, as well as the use of tubers for crop propagation, does not allow for easy genetic studies. The objective is to gain knowledge at the genomic level from standard Russet Norkotah and a subclonal Russet Norkotah selection TXNS278. DATA DESCRIPTION: In this report, we used RNA-seq, which allows genome-wide gene expression analysis to sequence the transcriptomes of the subclonal Russet Norkotah selection TXNS278 with standard Russet Norkotah grown in commercial fields. Among the selections, TXNS278 appeared in a multi-year analysis in Texas as a top No 1 yielding variety. Russet Norkotah and TXNS278 leaf and root transcriptomes were sequenced at two time points during growing season.


Assuntos
Genoma de Planta/genética , Tubérculos/genética , Solanum tuberosum/genética , Transcriptoma/genética , Folhas de Planta/genética , Análise de Sequência de RNA
14.
Phytopathology ; 108(3): 327-335, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29106346

RESUMO

The nonculturable bacterium 'Candidatus Liberibacter solanacearum' is the causative agent of zebra chip disease in potato. Computational analysis of the 'Ca. L. solanacearum' genome revealed a serralysin-like gene based on conserved domains characteristic of genes encoding metalloprotease enzymes similar to serralysin. Serralysin and other serralysin family metalloprotease are typically characterized as virulence factors and are secreted by the type I secretion system (T1SS). The 'Ca. L. solanacearum' serralysin-like gene is located next to and divergently transcribed from genes encoding a T1SS. Based on its relationship to the T1SS and the role of other serralysin family proteases in circumventing host antimicrobial defenses, it was speculated that a functional 'Ca. L. solanacearum' serralysin-like protease could be a potent virulence factor. Gene expression analysis showed that, from weeks 2 to 6, the expression of the 'Ca. L. solanacearum' serralysin-like gene was at least twofold higher than week 1, indicating that gene expression stays high as the disease progresses. A previously constructed serralysin-deficient mutant of Serratia liquefaciens FK01, an endophyte associated with insects, as well as an Escherichia coli lacking serralysin production were used as surrogates for expression analysis of the 'Ca. L. solanacearum' serralysin-like gene. The LsoA and LsoB proteins were expressed as both intact proteins and chimeric S. liquefaciens-'Ca. L. solanacearum' serralysin-like proteins to facilitate secretion in the S. liquefaciens surrogate and as intact proteins or as a truncated LsoB protein containing just the putative catalytic domains in the E. coli surrogate. None of the 'Ca. L. solanacearum' protein constructs expressed in either surrogate demonstrated proteolytic activity in skim milk or zymogram assays, or in colorimetric assays using purified protein, suggesting that the 'Ca. L. solanacearum' serralysin-like gene does not encode a functional protease, or at least not in our surrogate systems.


Assuntos
Regulação Bacteriana da Expressão Gênica/fisiologia , Bactérias Gram-Negativas/metabolismo , Metaloendopeptidases/genética , Doenças das Plantas/microbiologia , Solanum tuberosum/microbiologia , Sequência de Aminoácidos , Bactérias Gram-Negativas/genética
15.
BMC Genomics ; 18(1): 960, 2017 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-29228896

RESUMO

BACKGROUND: Transcriptomic analyses were performed to compare the molecular responses of two potato varieties previously shown to differ in the severity of disease symptoms due to infection by "Candidatus Liberibacter solanacearum" (Lso), the causative agent of Zebra Chip in potato. A factorial design utilizing the two varieties and psyllids either harboring Lso or without bacteria was used to discriminate varietal responses to pathogen infection versus psyllid feeding. Plant response was determined from leaf samples 3 weeks after infection. RESULTS: In response to Lso infection, 397 genes were differentially expressed in the variety Atlantic (most susceptible) as compared to 1027 genes in Waneta. Over 80% of the transcriptionally-changed genes were down-regulated in both varieties, including genes involved in photosynthesis or primary and secondary metabolism. Many of the Lso-responsive genes involved in stress responses or hormonal pathways were regulated differently in the two potato varieties. CONCLUSIONS: This study focused on the time point just prior to the onset of symptom development and provides valuable insight into the mechanisms of Liberibacter pathogenicity, especially the widespread suppression of plant gene expression, including genes involved in plant defenses.


Assuntos
Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Rhizobiaceae , Solanum tuberosum/genética , Solanum tuberosum/microbiologia , Transcriptoma , Perfilação da Expressão Gênica , Redes e Vias Metabólicas/genética , Solanum tuberosum/metabolismo , Estresse Fisiológico/genética
16.
Insect Sci ; 24(6): 961-974, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28493539

RESUMO

The recent emergence of several plant diseases caused by psyllid-borne bacterial pathogens worldwide (Candidatus Liberibacter spp.) has created renewed interest on the interaction between psyllids and bacteria. In spite of these efforts to understand psyllid association with bacteria, many aspects of their interactions remain poorly understood. As more organisms are studied, subtleties on the molecular interactions as well as on the effects of the bacteria on the psyllid host are being uncovered. Additionally, psyllid-borne bacterial phytopathogens can also affect the host plant, which in turn can impact psyllid physiology and behavior. Here, we review the current literature on different aspects of the influence of bacteria on multitrophic interactions among plants, psyllids, and pathogens. We then highlight gaps that need to be addressed to advance this field, which can have significant implications for controlling these newly emergent and other plant diseases.


Assuntos
Hemípteros/microbiologia , Insetos Vetores/microbiologia , Doenças das Plantas/microbiologia , Plantas/microbiologia , Rhizobiaceae/fisiologia , Animais , Plantas/imunologia , Plantas/metabolismo
17.
J Insect Physiol ; 98: 205-213, 2017 04.
Artigo em Inglês | MEDLINE | ID: mdl-27894936

RESUMO

The potato psyllid, Bactericera cockerelli (Sulc) (Hemiptera: Triozidae), is a phloem-feeding insect with preference for Solanaceae. This insect species is vector of the pathogenic bacteria 'Candidatus Liberibacter solanacearum' the causative agent of zebra chip, an important disease of commercial potatoes in several countries worldwide. The recent classification of psyllids among the most dangerous vectors has promoted their study, but still many biological processes such as reproduction and vitellogenesis need to be investigated. As a first step towards the elucidation of vitellogenesis in B. cockerelli, one candidate vitellogenin transcript (6622 bases long) was identified and the expression of the transcript and the protein were analyzed in virgin and mated females between 1 and 7days old. In virgin females, Vg expression increased up to 5days old; while mating significantly up-regulated Vg expression in 5- and 7-day-old females. To determine the role of juvenile hormone in B. cockerelli Vg expression, topical applications of juvenile hormone III were performed on virgin females, resulting in an up-regulation of Vg expression and an increase in the number of mature oocytes present in female reproductive organs. Overall, this study represents the first step to understand vitellogenesis of B. cockerelli and it highlights the role of JH III in the hormonal regulation of Vg expression and oocyte development.


Assuntos
Expressão Gênica , Hemípteros/genética , Proteínas de Insetos/genética , Hormônios Juvenis/farmacologia , Vitelogeninas/genética , Sequência de Aminoácidos , Animais , Clonagem Molecular , DNA Complementar/genética , DNA Complementar/metabolismo , Feminino , Hemípteros/crescimento & desenvolvimento , Hemípteros/metabolismo , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Hormônios Juvenis/administração & dosagem , Masculino , Ninfa/genética , Ninfa/crescimento & desenvolvimento , Ninfa/metabolismo , Óvulo/crescimento & desenvolvimento , Óvulo/metabolismo , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Vitelogeninas/química , Vitelogeninas/metabolismo
18.
Artigo em Inglês | MEDLINE | ID: mdl-27376032

RESUMO

"Candidatus Liberibacter solanacearum" (Lso) has emerged as a serious threat world-wide. Five Lso haplotypes have been identified so far. Haplotypes A and B are present in the Americas and/or New Zealand, where they are vectored to solanaceous plants by the potato psyllid, Bactericera cockerelli (Sulc) (Hemiptera: Triozidae). The fastidious nature of these pathogens has hindered the study of the interactions with their eukaryotic hosts (vector and plant). To understand the strategies used by these pathogens to infect their vector, the effects of each Lso haplotype (A or B) on psyllid fitness was investigated, and genome-wide transcriptomic and RT-qPCR analyses were performed to evaluate Lso gene expression in association with its vector. Results showed that psyllids infected with haplotype B had significantly lower percentage of nymphal survival compared to psyllids infected with haplotype A. Although overall gene expression across Lso genome was similar between the two Lso haplotypes, differences in the expression of key candidate genes were found. Among the 16 putative type IV effector genes tested, four of them were differentially expressed between Lso haplotypes, while no differences in gene expression were measured by qPCR or transcriptomic analysis for the rest of the genes. This study provides new information regarding the pathogenesis of Lso haplotypes in their insect vector.


Assuntos
Haplótipos , Hemípteros/fisiologia , Interações Hospedeiro-Patógeno , Insetos Vetores/fisiologia , Rhizobiaceae/crescimento & desenvolvimento , Rhizobiaceae/patogenicidade , Animais , Perfilação da Expressão Gênica , Hemípteros/microbiologia , Insetos Vetores/microbiologia , Reação em Cadeia da Polimerase em Tempo Real , Rhizobiaceae/classificação , Rhizobiaceae/genética , Análise de Sobrevida
19.
PLoS One ; 9(10): e111745, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25354208

RESUMO

Aquaporin (AQPs) proteins transport water and uncharged low molecular-weight solutes across biological membranes. Six to 8 AQP genes have been identified in many insect species, but presently only three aquaporins have been characterized in phloem feeding insects. The objective of this study was to identify candidate AQPs in the potato psyllid, Bactericera cockerelli. Herein, we identified four candidate aquaporin cDNAs in B. cockerelli transcriptome. Phylogenetic analysis showed that candidate BcAQP2-like had high similarity to PRIP aquaporins; while candidates BcAQP4-like, BcAQP5-like and BcAQP9-like clustered within clade B. In particular, candidates BcAQP4-like and BcAQP5-like clustered with functionally validated insect aquaglyceroporin proteins. Expression analyses using RT-qPCR showed that all candidates were expressed in all life stages and tissues. Candidates BcAQP4-like and BcAQP5-like were highly expressed in bacteriocytes, while BcAQP9-like appeared to be expressed at high levels in whole body but not in the assayed tissues. This study is the first global attempt to identify putative aquaporins in a phloem feeding insect.


Assuntos
Aquaporinas/metabolismo , Hemípteros/metabolismo , Proteínas de Insetos/metabolismo , Sequência de Aminoácidos , Animais , Aquaporinas/química , Aquaporinas/genética , Sequência de Bases , Feminino , Hemípteros/genética , Proteínas de Insetos/química , Proteínas de Insetos/genética , Masculino , Dados de Sequência Molecular , Especificidade de Órgãos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
20.
PLoS One ; 9(7): e100955, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24992557

RESUMO

"Candidatus Liberibacter solanacearum" (Lso) is an emergent pathogen of carrots in Europe and solanaceous plants in North and Central America and New Zealand. This bacterium is closely related to other pathogenic Candidatus Liberibacter spp., all vectored by psyllids. In order to understand the molecular interaction of this pathogen and its psyllid vector, Bactericera cockerelli, Illumina sequencing of psyllid harboring Lso was performed to determine if this approach could be used to assess the bacterial transcriptome in this association. Prior to sequencing, psyllid RNA was purified and insect and bacterial rRNA were removed. Mapping of reads to Lso genome revealed that over 92% of the bacterial genes were expressed in the vector, and that the COG categories Translation and Post-translational modification, protein turnover, chaperone functions were the most expressed functional categories. Expression levels of selected Lso genes were confirmed by RT-qPCR. The transcriptomic analysis also helped correct Lso genome annotation by identifying the expression of genes that were not predicted in the genome sequencing effort.


Assuntos
Daucus carota/microbiologia , Perfilação da Expressão Gênica , Hemípteros/genética , Insetos Vetores/genética , Doenças das Plantas/microbiologia , Rhizobiaceae/genética , Solanaceae/microbiologia , Sequência de Aminoácidos , Animais , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Sequência de Bases , Expressão Gênica , Regulação Bacteriana da Expressão Gênica , Genes Bacterianos , Hemípteros/microbiologia , Insetos Vetores/microbiologia , Dados de Sequência Molecular , Rhizobiaceae/química
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