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1.
Plant Physiol Biochem ; 148: 70-79, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31945669

RESUMO

Citrus greening, also called Huanglongbing (HLB), is one of the most destructive citrus diseases worldwide. It is caused by the fastidious gram-negative α-proteobacteria bacterium Candidatus Liberibacter asiaticus (CLas) and vectored by the Asian citrus psyllid (ACP), Diaphorina citri. Currently, there is no cure for HLB, no compounds have been successful in controlling HLB, and no sustainable management practices have been established for the disease. Thus, searching for alternative citrus greening disease mitigation strategies is considered an urgent priority for a sustainable citrus industry. The aim of this study was to use compounds extracted from oak, Quercus hemisphaerica, and to assess the antibacterial effects of these against CLas-infected citrus plants. The application of aqueous oak leaf extracts showed substantial inhibitory effects against CLas in citrus plants and the activity of genes related to starch. Significant differences were also observed in plant phenotypic and physiological traits after treatments. Citrus plants treated with oak extracts displayed an increase in stomatal conductance, chlorophyll content and nutrient uptake concurrently with a reduction of CLas titer, when compared to citrus plants treated with just water. The information provided from this study suggests a new management treatment program to effectively deal with the HLB disease.


Assuntos
Citrus , Extratos Vegetais , Folhas de Planta , Quercus , Rhizobiaceae , Animais , Antibacterianos/farmacologia , Citrus/efeitos dos fármacos , Citrus/microbiologia , Hemípteros/microbiologia , Extratos Vegetais/farmacologia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/microbiologia , Quercus/química , Rhizobiaceae/efeitos dos fármacos , Rhizobiaceae/fisiologia
2.
Plant Dis ; 104(1): 154-160, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31697223

RESUMO

Bacterial leaf scorch disease caused by Xylella fastidiosa occurs in southern highbush blueberry varieties in the southeastern United States. Susceptibility to X. fastidiosa varies by blueberry cultivar, and these interactions are often strain-specific. Xylella fastidiosa subsp. fastidiosa is the causal agent of Pierce's disease in grapevines, and it has been problematic in the San Joaquin Valley of California since the introduction of the glassy-winged sharpshooter (Homalodisca vitripennis). The glassy-winged sharpshooter is known to feed on blueberry, a crop that is expanding in the San Joaquin Valley. Currently, little is known about the potential for the spread of X. fastidiosa between grape and blueberry in this region. The ability of a Pierce's disease strain of X. fastidiosa from the San Joaquin Valley to cause disease in southern highbush blueberry and the potential for the glassy-winged sharpshooter to transmit X. fastidiosa between blueberry and grapevine were investigated. Experimental inoculations showed that the X. fastidiosa subsp. fastidiosa strain Bakersfield-1 can cause disease in blueberry cv. Emerald, and that the glassy-winged sharpshooter can acquire X. fastidiosa from artificially inoculated blueberry plants under laboratory conditions. Understanding the possibility for X. fastidiosa strains from the San Joaquin Valley to infect multiple crops grown in proximity is important for area-wide pest and disease management.


Assuntos
Mirtilos Azuis (Planta) , Hemípteros , Xylella , Animais , Mirtilos Azuis (Planta)/microbiologia , California , Hemípteros/microbiologia , Sudeste dos Estados Unidos , Xylella/fisiologia
3.
Plant Dis ; 104(1): 179-185, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31725343

RESUMO

'Candidatus Liberibacter solanacearum' (Lso), transmitted by the potato psyllid (Bactericera cockerelli), is the putative causal agent of potato zebra chip disease. The bacterial pathogen infects a wide range of solanaceous plants (both wild and cultivated species), among which are peppers, potatoes, and tomatoes. Currently there are two commonly detected, genetically distinct haplotypes of Lso (A and B) identified from potatoes in the United States. To determine whether there are interactions between Lso haplotypes and different solanaceous hosts, experiments were conducted in the greenhouse in which pepper, potato, and tomato plants were infested with psyllids carrying Lso A, B, or an A and B mix (AB) or with psyllids free of Lso. Host plants were grown in pots in cages on the greenhouse benches and infested with six psyllids per plant. In addition, eight pepper cultivars were similarly infested for deeper understanding of host-haplotype interactions. Approximately 7 weeks after infestation, adult psyllids in each cage were counted to determine the impact of Lso haplotype-host interactions on psyllid survival and plants were sampled and tested molecularly for Lso. Individual psyllids carrying haplotypes B or AB and those free of Lso copiously reproduced on all three hosts, and leaf tissue from each plant tested positive for the respective Lso except those infested with Lso-negative psyllids. However, psyllids carrying Lso A did not survive on peppers but survived and abundantly reproduced on potatoes and tomatoes. In addition, samples from peppers infested with psyllids carrying Lso A tested negative for Lso. However, peppers infested with individual psyllids carrying Lso AB tested positive for Lso A, indicating that the presence of B may be required for infection by Lso A and psyllid survival on peppers. The different pepper cultivars infested with psyllids carrying Lso A showed similar results to the haplotype-host interaction tests, suggesting that cultivar may not be a factor in Lso A-pepper host interactions. Results from these studies suggest that Lso A may affect host selection by psyllids either for nutrition or laying of eggs. Mechanisms involved in preventing psyllid reproduction on peppers, once identified, will have significant implications for potential psyllid management.


Assuntos
Hemípteros , Interações Hospedeiro-Patógeno , Rhizobiaceae , Solanaceae , Animais , Haplótipos , Hemípteros/microbiologia , Doenças das Plantas/microbiologia , Solanaceae/microbiologia
4.
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
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.
J Appl Microbiol ; 127(6): 1801-1813, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31509633

RESUMO

AIMS: In this study, binding between the immunodominant membrane protein Imp of the 16SrV-D phytoplasma associated with Flavescence dorée disease (FD-Dp) and insect proteins of vectors and non-vectors of FD-Dp was tested. METHODS AND RESULTS: Six Auchenorrhyncha species, from distantly related groups were selected: Scaphoideus titanus, Euscelidius variegatus, Macrosteles quadripunctulatus, Zyginidia pullula (Cicadomorpha), Ricania speculum and Metcalfa pruinosa (Fulgoromorpha). The vector status of each species was retrieved from the literature or determined by transmission trials in this study. A His-tagged partial Imp protein and a rabbit polyclonal antibody were synthesized and used for Western and Far-Western dot Blot (FWdB) experiments. Total native and membrane proteins (MP) were extracted from entire bodies and organs (gut and salivary glands) of each insect species. FWdB showed decreasing interaction intensities of Imp fusion protein with total proteins from entire bodies of S. titanus, E. variegatus (competent vectors) and M. quadripunctulatus (non-vector), while no interaction signal was detected with the other three species (non-vectors). A strong signal detected upon interaction of FD-D Imp and MP from guts of closely related insects supports the role of this organ as the first barrier to ensure successful transmission. CONCLUSIONS: Our results showed that specific Imp binding, correlated with vector status, is involved in interactions between FD-Dp and insect proteins. SIGNIFICANCE AND IMPACT OF THE STUDY: Integrating knowledge on host-pathogen protein-protein interactions and on insect phylogeny would help to identify the actual range of vectors of phytoplasma strains of economic importance.


Assuntos
Hemípteros/microbiologia , Proteínas de Insetos/metabolismo , Insetos Vetores/microbiologia , Proteínas de Membrana/metabolismo , Phytoplasma/fisiologia , Animais , Proteínas de Bactérias/metabolismo , Hemípteros/química , Hemípteros/classificação , Insetos Vetores/química , Insetos Vetores/classificação , Filogenia , Phytoplasma/química , Doenças das Plantas/microbiologia , Ligação Proteica
7.
Ecotoxicol Environ Saf ; 183: 109585, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31509930

RESUMO

The two biological control agents, predatory mite Amblydromalus limonicus Garman & McGregor (Acari: Phytoseiidae) and entomopathogenic fungi (EPF) Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Cordycipitaceae), have the potential to independently suppress the invasive tomato potato psyllid (TPP), Bactericera cockerelli (Sulc), in New Zealand. The integrated strategy of combining the release of predatory mites and EPF may further promote biocontrol of pests. To examine the compatibility of A. limonicus and B. bassiana, we tested the lethal effects of B. bassiana on A. limonicus females on B. cockerelli and calculated the sublethal concentrations of B. bassiana. The effects of sublethal concentrations (LC10 and LC30) of EPF on predatory mite females were assessed on the reproduction parameters of the parental generation (F0). We also evaluated the transgenerational effects of EPF on life table parameters and predation rates of the offspring generation (F1) that was fed on the psyllids. Our results showed that A. limonicus females were susceptible to B. bassiana and the LC50 was 2.2 × 105 conidia mL-1. Sublethal concentrations of EPF (LC30) significantly reduced the fecundity, longevity, oviposition period and predation rates of F0 predatory mite females. However, life table and predation rates of F1 predatory mites were not influenced by sublethal concentrations (LC30), except for the reduction of the preadult and total pre-oviposition period, and the predation rate of F1 males. Also, F1 population parameters including the intrinsic rate of increase (rm), finite rate of increase (λ), gross reproduction rate (GRR), and net reproduction rate (R0) were not significantly influenced by sublethal concentrations (LC10 and LC30). This result suggests that the transgenerational effects of entomopathogenic fungi cannot be carried over to F1. In conclusion, the simultaneous application of both biological control agents does have the potential to control TPP at appropriate intervals during the crop season. Further evaluation in the field will be needed to confirm the viability of this approach to control TPP.


Assuntos
Beauveria/fisiologia , Hemípteros/microbiologia , Lycopersicon esculentum/crescimento & desenvolvimento , Ácaros/fisiologia , Controle Biológico de Vetores/métodos , Animais , Beauveria/patogenicidade , Feminino , Fertilidade/fisiologia , Longevidade/fisiologia , Masculino , Ácaros/microbiologia , Nova Zelândia , Oviposição/efeitos dos fármacos , Comportamento Predatório/fisiologia , Reprodução/efeitos dos fármacos , Estações do Ano
8.
Curr Microbiol ; 76(11): 1306-1312, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31471686

RESUMO

Wolbachia are the most common symbionts in arthropods; antibiotic treatment for eliminating the symbionts from their host is necessary to investigate the functions. Tetracycline antibiotics are widely used to remove endosymbiont Wolbachia from insect hosts. However, very little has been known on the effects of tetracycline on population size of Wolbachia in small brown planthopper (SBPH), Laodelphax striatellus (Fallén), an important insect pest of rice in Asia. Here, we investigated the dynamics of Wolbachia population density in females and males of L. striatellus by real-time fluorescent quantitative PCR method. The Wolbachia density in females and males of L. striatellus all declined sharply after treatment with 2 mg/mL tetracycline for one generation, and continued to decrease to a level which could not be detected by both qPCR and diagnostic PCR after treated for another generation, then maintained at 0 in the following three generations with continuous antibiotic treatment. Wolbachia infection did not recover in L. striatellus after stopping tetracycline treatment for ten generations. This is the first report to precisely monitor the population dynamics of Wolbachia in L. striatellus during successive tetracycline treatment and after that. The results provide a useful method for evaluating the efficiency of artificial operation of endosymbionts.


Assuntos
Antibacterianos/farmacologia , Hemípteros/microbiologia , Tetraciclina/farmacologia , Wolbachia/efeitos dos fármacos , Animais , Biodiversidade , Feminino , Masculino , Wolbachia/crescimento & desenvolvimento
9.
Phytopathology ; 109(12): 2064-2073, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31425000

RESUMO

Murraya paniculata and Swinglea glutinosa are aurantioid hosts of the Asian citrus psyllid (ACP) Diaphorina citri, the principal vector of 'Candidatus Liberibacter asiaticus' (Las). Las is the pathogen associated with huanglongbing (HLB), the Asian form of which is the most devastating disease of Citrus species and cultivars (Rutaceae: Aurantioideae). M. paniculata is a common ornamental and S. glutinosa is grown as an ornamental, a citrus rootstock, and a hedgerow fence plant. Because of the uncertain status of these plants as reservoirs of Las, a series of cross-inoculation bioassays were carried out in different environments, using infected Valencia sweet orange (Citrus × aurantium) infected shoot tops as a source of inoculum and D. citri nymphs and adults reared on M. paniculata and S. glutinosa to inoculate pathogen-free Valencia orange plantlets. In contrast to sweet orange, Las was more unevenly distributed and reached much lower titers in M. paniculata and S. glutinosa. Infections in M. paniculata and S. glutinosa were also transient. Very few insects that successfully acquired Las from M. paniculata and S. glutinosa were able to transmit the pathogen to healthy citrus. Transmission rates were low from M. paniculata (1.0%) and S. glutinosa (2.0%) and occurred only in a controlled environment highly favorable to Las and ACP using 10-day-old adults that completed their life cycle on Las-positive plants. Our study showed that in HLB-endemic areas, M. paniculata and S. glutinosa can be deemed as epidemiologically dead-end hosts for Las and are not important alternative hosts of the pathogen for transmission to citrus. However, under a combination of conditions highly favorable to Las infection and transmission and in the absence of effective quarantine procedures, these plants could eventually serve as carriers of Las to regions currently free from HLB.


Assuntos
Citrus , Hemípteros , Murraya , Rhizobiaceae , Rutaceae , Animais , Citrus/microbiologia , Hemípteros/microbiologia , Interações Hospedeiro-Patógeno , Murraya/microbiologia , Doenças das Plantas/microbiologia , Rutaceae/microbiologia
10.
Plant Dis ; 103(10): 2587-2591, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31432751

RESUMO

'Candidatus Liberibacter solanacearum' (Lso) is an uncultured, phloem-associated bacterium causing a severe tuber disease in potato called zebra chip (ZC). Seven haplotypes of Lso have been described in different hosts, with haplotypes A and B found associated with infections in potato and tomato. In the field, Lso is transmitted by the potato psyllid (Bactericera cockerelli), and between 2011 and 2015, a significant change in Lso haplotype prevalence was previously reported in Idaho: from exclusively A haplotype found in tested psyllids in 2012 to mainly B haplotype found in collected psyllids in 2015. However, prevalence of Lso haplotypes in Idaho was not analyzed in potato tubers exhibiting symptoms of ZC. To fill in this knowledge gap, prevalence of Lso haplotypes was investigated in potato tubers harvested in southern Idaho between 2012 and 2018, and it was found to change from exclusively A haplotype in the 2012 season to an almost equal A and B haplotype distribution during the 2016 season. During the same period, haplotype distribution of Lso in psyllid vectors collected using yellow sticky traps also changed, but in psyllids, the shift from A haplotype of Lso to B haplotype was complete, with no A haplotype detected in 2016 to 2018. The changes in the haplotype prevalence of the Lso circulating in potato fields in southern Idaho may be, among other factors, responsible for a decrease in the ZC incidence in Idaho potato fields between an outbreak of the disease in 2012 and a very low level of ZC afterward.


Assuntos
Hemípteros , Rhizobiaceae , Solanum tuberosum , Animais , Haplótipos , Hemípteros/microbiologia , Idaho , Rhizobiaceae/fisiologia , Solanum tuberosum/microbiologia
11.
Genome ; 62(9): 571-584, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31283888

RESUMO

Silverleaf whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), is one of the most notorious invasive insect pests, infesting more than 900 species of plants and spreading more than 200 viral diseases. This polyphagous agricultural pest harbours diverse bacterial communities in its gut, which perform multiple functions in whiteflies, including nutrient provisioning, amino acid biosynthesis, and virus transmission. The present exploratory study compares the bacterial communities associated with silverleaf whitefly infesting cassava, also known as cassava whitefly, collected from two different zones (zone P: plains; zone H: high ranges), from Kerala, India, using next-generation sequencing of 16S rDNA. The data sets for these two regions consisted of 1 321 906 and 690 661 high-quality paired-end sequences with mean length of 150 bp. Highly diverse bacterial communities were present in the sample, containing approximately 3513 operational taxonomic units (OTUs). Sequence analysis showed a marked difference in the relative abundance of bacteria in the populations. A total of 16 bacterial phyla, 27 classes, 56 orders, 91 families, 236 genera, and 409 species were identified from the P population, against 16, 31, 60, 88, 225, and 355, respectively, in the H population. Arsenophonus sp. (Enterobacteriaceae), which is important for virus transmission by whiteflies, was relatively abundant in the P population, whereas in the H population Bacillus sp. was the most dominant group. The association of whitefly biotypes and secondary symbionts suggests a possible contribution of these bacteria to host characteristics such as virus transmission, host range, insecticide resistance, and speciation.


Assuntos
Bactérias/classificação , Hemípteros/microbiologia , Manihot/parasitologia , Simbiose , Animais , Bactérias/isolamento & purificação , DNA Bacteriano , Sequenciamento de Nucleotídeos em Larga Escala , Índia , Tipagem Molecular
12.
Enzyme Microb Technol ; 129: 109358, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31307582

RESUMO

Many bacterial and viral plant pathogens are transmitted by insect vectors, and pathogen-mediated alterations of plant physiology often influence insect vector behavior and fitness. It remains largely unknown for most plant pathogens whether, and how, they might directly alter the physiology of their insect vectors in ways that promote pathogen transmission. Here we examined whether the presence of "Candidatus Liberibacter solanacearum" ("Ca. L. solanacearum"), an obligate bacterial pathogen of plants and of its psyllid vector alters the physiochemical environment within its insect vector, the potato psyllid (Bactericera cockerelli). Microelectrodes were used to measure the local pH and oxygen tension within the abdomen of "Ca. L. solanacearum"-free psyllids and those infected with "Ca. L. solanacearum". The hemolymph of infected psyllids had higher pH at 9.09 ± 0.12, compared to "Ca. L. solanacearum"-free psyllids (8.32 ± 0.11) and a lower oxygen tension of 33.99% vs. 67.83%, respectively. The physicochemical conditions inside "Ca. L. solanacearum"-free and -infected psyllids body differed significantly with the infected psyllids having a higher hemolymph pH and lower oxygen tension than "Ca. L. solanacearum"-free psyllids. Notably, the bacterial titer increased under conditions of higher pH and lower oxygen tension values. This suggests that the vector's physiology is altered by the presence of the pathogen, potentially, resulting in a more conducive environment for "Ca. L. solanacearum" survival and subsequent transmission.


Assuntos
Hemípteros/microbiologia , Insetos Vetores/microbiologia , Rhizobiaceae/fisiologia , Animais , Hemípteros/fisiologia , Concentração de Íons de Hidrogênio , Insetos Vetores/fisiologia , Doenças das Plantas/microbiologia
13.
J Agric Food Chem ; 67(31): 8476-8484, 2019 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-31298527

RESUMO

Cicada flowers, which are edible and medicinal mushrooms, are the fruiting bodies of Isaria cicadae, a fungus that is parasitic on the larvae of cicada pupae. We hypothesize that host factors might possess stimulatory activity on metabolite synthesis in Isaria cicadae. Here, we first compared the microbial community structures of different wild cicada flowers across geographical regions, compartments, and growth stages via high-throughput sequencing. Isaria cicadae TZC-3, an isolate of the most abundant operational taxonomic unit (OTU6782) in all the fungal communities, was isolated from wild cicada flowers. Furthermore, the effects of cicada pupae on metabolite synthesis in Isaria cicadae TZC-3 were studied in submerged culture. The contents of intercellular polysaccharides, adenosine, N6-(2-hydroxyethyl)-adenosine, free amino acids, and hydrolyzed monosaccharides in the mycelia cultured with cicada pupa powder (4%) were significantly increased as compared with the contents in the control group. This indicates that a cicada pupa can act as an elicitor for metabolite synthesis in Isaria cicadae.


Assuntos
Cordyceps/metabolismo , Carpóforos/química , Hemípteros/microbiologia , Pupa/microbiologia , Adenosina/análise , Adenosina/metabolismo , Aminoácidos/análise , Aminoácidos/metabolismo , Animais , Cordyceps/química , Cordyceps/crescimento & desenvolvimento , Carpóforos/crescimento & desenvolvimento , Carpóforos/metabolismo , Hemípteros/química , Hemípteros/metabolismo , Interações Hospedeiro-Patógeno , Microbiota , Micélio/química , Micélio/crescimento & desenvolvimento , Micélio/metabolismo , Pupa/química , Pupa/metabolismo
14.
PLoS One ; 14(6): e0218190, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31181122

RESUMO

Diaphorin is a polyketide produced by Candidatus Profftella armatura (Betaproteobacteria), an organelle-like defensive symbiont harbored by a plant sap-sucking insect, Asian citrus psyllid Diaphorina citri (Hemiptera: Liviidae). Diaphorin belongs to the pederin family, a group of compounds that share much of their core structure with that of pederin, which is characterized by two dihydropyran rings bridged by an N-acyl aminal. Most members of this family have potent antitumor activity, making them promising anticancer drug candidates. The present study assessed the therapeutic potential of diaphorin for its antitumor activity against 39 human cancer cell lines including those from breast, brain, colon, lung, skin, ovary, kidney, stomach, and prostate. The results showed that diaphorin had inhibitory activity against all 39 cancer cell lines tested. The GI50, TGI, and LC50 values ranged from 0.28 µM- 2.4 µM, 1.6 µM -11 µM, and 7.5 µM-> 100 µM, respectively. These values are among the highest in the pederin family, indicating that the anticancer activity of diaphorin is milder than those of other pederin congeners. The inhibitory effects of diaphorin significantly differed among the distinct cancer types. The maximum difference was about 10-fold, which was similar to those of most other pederin congeners.


Assuntos
Betaproteobacteria/metabolismo , Hemípteros/química , Policetídeos/uso terapêutico , Animais , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Ensaios de Seleção de Medicamentos Antitumorais , Hemípteros/microbiologia , Humanos , Concentração Inibidora 50 , Policetídeos/farmacologia , Simbiose
15.
Pestic Biochem Physiol ; 157: 99-107, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31153482

RESUMO

The entmopathogenic fungus Lecaniicillium lecanii is a naturally available biological control and it is considered to be one of the best mycoinsecticide agents against the destructive insect pest Diaphorina citri Kuwayama. The present study aimed to extract and characterize the toxic insecticidal protein from L. lecanii and to assess the toxicity level against the Asian citrus psyllid the vector of Huanglongbing disease (HLB), also called citrus greening. Extracts of a toxic substance from submerged batch culture examined by sodium dodecyl sulfate-poly-acrylamide (SDS-PAGE), had a molecular weight of 45 kDa. The most abundant toxic metabolite was subjected to HPLC to purify and identified it by mass spectrometry. Subsequently, metabolite toxicity was tested against D. citri at three different concentrations (1%, 2%, and 3%). The results showed that the highest concentration had a significant maximum mortality at 120 h post application. Furthermore, we investigated the expression of the GAS1 gene which was previously identified to have a role in pathogenicity in in vivo studies in adult insect psyllids. Results of this study indicated that expression of the virulence factor gene was present at three concentrations of the fungal suspension post inoculation. This is the first study to provide this novel approach for the characterization of fungal mediated synthesis of a cuticle degrading soluble protein against the insect D. citri. The present results provide strong information on the in vivo expression of the GAS1 gene involved in fungal virulence pertaining to penetration of the insect cuticle, but not to inhibiting the growth of the host.


Assuntos
Hemípteros/microbiologia , Hypocreales/metabolismo , Hypocreales/patogenicidade , Animais , Eletroforese em Gel de Poliacrilamida , Hypocreales/genética , Virulência
16.
mBio ; 10(3)2019 06 18.
Artigo em Inglês | MEDLINE | ID: mdl-31213566

RESUMO

Gene loss and genome reduction are defining characteristics of endosymbiotic bacteria. The most highly reduced endosymbiont genomes have lost numerous essential genes related to core cellular processes such as replication, transcription, and translation. Computational gene predictions performed for the genomes of the two bacterial symbionts of the cicada Diceroprocta semicincta, "Candidatus Hodgkinia cicadicola" (Alphaproteobacteria) and "Ca Sulcia muelleri" (Bacteroidetes), have found only 26 and 16 tRNA genes and 15 and 10 aminoacyl tRNA synthetase genes, respectively. Furthermore, the original "Ca Hodgkinia cicadicola" genome annotation was missing several essential genes involved in tRNA processing, such as those encoding RNase P and CCA tRNA nucleotidyltransferase as well as several RNA editing enzymes required for tRNA maturation. How these cicada endosymbionts perform basic translation-related processes remains unknown. Here, by sequencing eukaryotic mRNAs and total small RNAs, we show that the limited tRNA set predicted by computational annotation of "Ca Sulcia muelleri" and "Ca Hodgkinia cicadicola" is likely correct. Furthermore, we show that despite the absence of genes encoding tRNA processing activities in the symbiont genomes, symbiont tRNAs have correctly processed 5' and 3' ends and seem to undergo nucleotide modification. Surprisingly, we found that most "Ca Hodgkinia cicadicola" and "Ca Sulcia muelleri" tRNAs exist as tRNA halves. We hypothesize that "Ca Sulcia muelleri" and "Ca Hodgkinia cicadicola" tRNAs function in bacterial translation but require host-encoded enzymes to do so.IMPORTANCE The smallest bacterial genomes, in the range of about 0.1 to 0.5 million base pairs, are commonly found in the nutritional endosymbionts of insects. These tiny genomes are missing genes that encode proteins and RNAs required for the translation of mRNAs, one of the most highly conserved and important cellular processes. In this study, we found that the bacterial endosymbionts of cicadas have genomes which encode incomplete tRNA sets and lack genes required for tRNA processing. Nevertheless, we found that endosymbiont tRNAs are correctly processed at their 5' and 3' ends and, surprisingly, that mostly exist as tRNA halves. We hypothesize that the cicada host must supply its symbionts with these missing tRNA processing activities.


Assuntos
Bactérias/genética , Genoma Bacteriano , Hemípteros/microbiologia , RNA de Transferência/genética , Simbiose , Animais , Evolução Molecular , Feminino , Filogenia , Modificação Traducional de Proteínas
17.
J Insect Physiol ; 116: 125-133, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31128084

RESUMO

Wolbachia are intracellular bacteria carried by thousands of arthropod species. The success of Wolbachia is due to efficient vertical transmission by the host maternal germline. Wolbachia's behavior during host oogenesis is well characterized, although their behavior during embryogenesis is unclear. Vertical transmission of Wolbachia wStri in the small brown planthopper, Laodelphax striatellus is extraordinarily efficient. To understand why, we investigated its localization and dynamics in L. striatellus embryos. Microscopic observations indicated that the Wolbachia were mainly localized at the anterior region of the embryo during early embryogenesis. The distribution of Wolbachia within the anterior region was established during oogenesis, and according to a phylogenetic analysis, may be due to intrinsic factors in Wolbachia. We observed that wStri migrated to the posterior part cells during late embryogenesis, in the region where gonads were formed. An expression profile of Wolbachia-infected host embryonic development genes revealed Ddx1 mRNAs, which is required for host viability and in the germ line, accumulated in the posterior region of 3-day-old embryos, while other development genes mRNAs were significantly more abundant in the posterior region of 6-day-old embryos. These genes thus appear to be associated with the localization of Wolbachia wStri in the anterior region, although their functions remain unclear. These results can explain Wolbachia wStri high prevalence in L. striatellus.


Assuntos
Desenvolvimento Embrionário , Hemípteros/microbiologia , Wolbachia/fisiologia , Animais , Hemípteros/embriologia , Filogenia , Wolbachia/classificação
18.
PLoS One ; 14(5): e0216319, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31048920

RESUMO

The Asian citrus psyllid Diaphorina citri Kuwayama (Hemiptera: Sternorrhyncha: Psylloidea: Liviidae) is an important pest of citrus species worldwide because it transmits Candidatus Liberibacter spp. (Alphaproteobacteria), the causative agents of an incurable citrus disease known as huanglongbing or greening disease. Diaphorina citri possesses a vertically-transmitted intracellular symbiont, Candidatus Profftella armatura (Betaproteobacteria), which produces diaphorin, a polyketide that is significantly toxic to mammalian cells. Diaphorin is an analog of pederin, a defensive polyketide in the body fluid of Paederus rove beetles (Coleoptera: Staphylinidae) that deters predators. In the present study, as a first step to assess the possibility that diaphorin is toxic to biological control agents, we assayed diaphorin activities against insects and fungi. The target cells and organisms were (a) the Sf9 cell line derived from the fall armyworm moth Spodoptera frugiperda (Lepidoptera: Noctuidae), (b) the pea aphid Acyrthosiphon pisum (Hemiptera: Sternorrhyncha: Aphidoidea: Aphididae), a phloem sap-sucking insect that is closely related to psyllids, (c) the Asian lady beetle Harmonia axyridis (Coleoptera: Coccinellidae), one of the major predators of D. citri, and (d) the budding yeast Saccharomyces cerevisiae (Ascomycota: Saccharomycetes) as a model of fungal pathogens. For a comparison, we also evaluated pederin activities. The results of our analyses revealed the following: (1) Diaphorin and pederin are significantly toxic to the tested insects and yeast; (2) Their toxicities vary widely among the target cells and organisms; (3) Diaphorin is generally less toxic than pederin; (4) The toxicities of diaphorin and pederin are considerably different in the Sf9 insect cell line and S. cerevisiae, but similar in A. pisum and H. axyridis; and (5) The amount of diaphorin contained in D. citri is toxic to all of the tested cells and organisms, suggesting that this polyketide is potentially harmful for biological control agents.


Assuntos
Anti-Infecciosos , Betaproteobacteria/metabolismo , Citotoxinas , Hemípteros/microbiologia , Controle Biológico de Vetores , Policetídeos , Simbiose , Células A549 , Animais , Anti-Infecciosos/metabolismo , Anti-Infecciosos/farmacologia , Citotoxinas/metabolismo , Citotoxinas/farmacologia , Células HCT116 , Humanos , Células MCF-7 , Células PC-3 , Policetídeos/metabolismo , Policetídeos/farmacologia
19.
Pest Manag Sci ; 75(11): 2882-2891, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31038255

RESUMO

BACKGROUND: Microbial surfactants are multifunctional surface-active molecules that have been overlooked in formulating microbial biopesticides. We report a novel approach using the biosurfactant rhamnolipid (RML) against the destructive cosmopolitan insect pest Bemisia tabaci, as well as the combined action of RML with aerial conidia of two entomopathogenic fungi, Cordyceps javanica and Beauveria bassiana. RML was also tested as a suspension agent to improve the recovery rate of conidia from solid substrate for fungal preparations. RESULTS: The recovery rate of conidia increased dramatically (two to five times) with RML compared with a standard surfactant (Tween 80). Spraying solutions of 0.075% and 0.1% (w/v) RML on B. tabaci third instar nymphs induced 100% mortality within 4 days. Conidial suspensions at 5 × 106 conidia/mL amended with RML at 0.01% or 0.05% markedly increased nymphal mortalities and considerably reduced LC50 . Conidial suspensions of B. bassiana with 0.05% RML added were more effective against whitefly nymphs (87.3% mortality) than C. javanica + RML (51.4% mortality). CONCLUSION: Our results show that this bacterium-based RML improved the recovery rate of hydrophobic conidia, and that mixtures of RML with fungal spore suspensions increased their insecticidal activity. © 2019 Society of Chemical Industry.


Assuntos
Beauveria/fisiologia , Cordyceps/fisiologia , Glicolipídeos/farmacologia , Hemípteros , Inseticidas/farmacologia , Controle Biológico de Vetores/métodos , Tensoativos/farmacologia , Animais , Hemípteros/efeitos dos fármacos , Hemípteros/crescimento & desenvolvimento , Hemípteros/microbiologia , Ninfa/efeitos dos fármacos , Ninfa/crescimento & desenvolvimento , Ninfa/microbiologia , Esporos Fúngicos/fisiologia
20.
J Insect Sci ; 19(3)2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-31115477

RESUMO

Whiteflies cause huge economic losses for cassava (Manihot esculenta Crantz) cultivation. Damage can be caused directly when the insects feed on the phloem and/or indirectly by the transmission of viruses. It has been found that whiteflies maintain a close relationship with some endosymbiotic bacteria and that this interaction produces different effects on host biology and can also facilitate viral transmission. This study aimed to characterize the diversity of secondary endosymbionts (SE) present in whiteflies associated with cassava. Whitefly adults and nymphs were collected from cassava crops at nine locations in Southwestern Colombia. Molecular identification of insects and endosymbionts was carried out using specific mtCOI, wsp, 23s rRNA, and 16s rRNA primers. Phylogenetic trees were constructed from these sequences, both for whitefly species and the endosymbionts found. In addition, morphological identification of whitefly species was made using last instar nymphs. Molecular and morphological evaluation revealed that the most abundant whitefly species was Trialeurodes variabilis (Quaintance) followed by Aleurotrachelus socialis Bondar and Bemisia tuberculata Bondar. One hundred percent of the individuals contained the primary endosymbiont Portiera. The SE Rickettsia, Hamiltonella, Wolbachia, and Fritschea were not detected in the samples tested. Prevalence of Cardinium and Arsenophonus were variable at each locality, Cardinium being most prevalent in A. socialis adults. This study is the first report on the presence of Cardinium and Arsenophonus in A. socialis and T. variabilis. It is also the first report of endosymbiotic diversity in whiteflies associated with cassava in Colombia.


Assuntos
Hemípteros/microbiologia , Animais , Hemípteros/genética , Manihot , Ninfa/microbiologia , Filogenia , Simbiose
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