Detection of<i>Clavibacter michiganensis</i>subsp.<i>michiganensis</i>in Symptomless Tomato Transplants

Gitaitis, R. D.; Beaver, Rodney W.; Voloudakis, Andreas E.

doi:10.1094/pd-75-0834

Cited by 76 publications

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“…Resistant cultivars or effective chemical or antibiotic controls in field and greenhouse cultivation are unknown [7]. The infection is spread by contaminated seeds or transplants [8,9] and, consequently, testing of seeds and plant tissue as well as quarantine measurements are the most effective disease control measures.…”

Section: Introductionmentioning

confidence: 99%

A Proteomic Study of Clavibacter Michiganensis Subsp. Michiganensis Culture Supernatants

et al. 2015

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Clavibacter michiganensis, subsp. michiganensis is a Gram-positive plant pathogen infecting tomato (Solanum lycopersicum). Despite a considerable economic importance due to significant losses of infected plants and fruits, knowledge about virulence factors of C. michiganensis subsp. michiganensis and host-pathogen interactions on a molecular level are rather limited. In the study presented here, the proteome of culture supernatants from C. michiganensis subsp. michiganensis NCPPB382 was analyzed. In total, 1872 proteins were identified in M9 and 1766 proteins in xylem mimicking medium. Filtration of supernatants before protein precipitation reduced these to 1276 proteins in M9 and 976 proteins in the xylem mimicking medium culture filtrate. The results obtained indicate that C. michiganensis subsp. michiganensis reacts to a sucrose- and glucose-depleted medium similar to the xylem sap by utilizing amino acids and host cell polymers as well as their degradation products, mainly peptides, amino acids and various C5 and C6 sugars. Interestingly, the bacterium expresses the previously described virulence factors Pat-1 and CelA not exclusively after host cell contact in planta but already in M9 minimal and xylem mimicking medium.

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Section: Introductionmentioning

confidence: 99%

A Proteomic Study of Clavibacter Michiganensis Subsp. Michiganensis Culture Supernatants

et al. 2015

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“…Factors that make C. m. michiganensis an especially devastating pathogen are the frequent occurrence of latent infections (Gitaitis et al 1991;Franc 1999) and its ability to invade seeds (McBeath & Adelman 1986;Tsiantos 1987;Bugbee & Gudmestad 1988;Biddle et al 1990;Nemeth et al 1991;Franken et al 1993;Samac et al 1998), which allows long-distance spread of the pathogen. Small bacterial populations often spread rapidly during commercial transplant production by water splash and/or equipment (Carlton et al 1998).…”

Section: Introductionmentioning

confidence: 99%

“…Small bacterial populations often spread rapidly during commercial transplant production by water splash and/or equipment (Carlton et al 1998). One infected seed in 10,000 can cause outbreaks (Gitaitis et al 1991), and no resistant plant cultivars are known. However, on non-host plants, a hypersensitive response (HR) prevents the multiplication and spread of the pathogen (Gitaitis 1990 A rapid, sensitive and reliable protocol for detection and identification of this highly variable pathogen would greatly improve the efficiency of testing programs worldwide.…”

Section: Introductionmentioning

confidence: 99%

Utility of a loop-mediated amplification assay for detection ofClavibacter michiganensissubsp.michiganensisin seeds and plant tissues

Yasuhara-Bell

Baysal-Gurel

Miller

et al. 2015

Canadian Journal of Plant Pathology

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A c c e p t e d M a n u s c r i p t J. Yasuhara-Bell et al. Detection of Clavibacter in seeds and plant tissues by LAMP 2 A c c e p t e d M a n u s c r i p t J. Yasuhara-Bell et al. Detection of Clavibacter in seeds and plant tissues by LAMP 3Abstract: A loop-mediated amplification (LAMP) assay developed previously to specifically identify Clavibacter michiganensis subsp. michiganensis, the causal agent of bacterial canker of tomato, was compared with a commercial lateral flow immunoassay, three PCR assays, and culturing on agar medium for efficacy in detecting the pathogen in pepper and tomato tissues and tomato seeds. The LAMP assay was as effective in identifying C. m. michiganensis-positive tomato seed and tissue samples as the Cmm ImmunoStrip ® . The LAMP assay was performed using a handheld real-time fluorescence-monitoring device (Smart-DART TM ) and the iQ TM 5 Multicolor Real-Time PCR Detection System to compare both systems. Both devices produced equivalent results, thus demonstrating the potential of this LAMP detection system for field detection. A side-by-side comparison of the C. m. michiganensis-LAMP with the Cmm ImmunoStrip ® assay, two conventional PCR assays, using Cmm5/6 and PSA8/R primer sets, and one quantitative PCR assay (primers RZ_ptssk 10/11, probe RZ_ptssk12), showed that the C. m. michiganensis-LAMP/Smart-DART TM assay can provide real-time portable in-field test results comparable to accepted standards. These results, combined with the specificity established previously, suggest that this LAMP assay can be an alternative to current diagnostic tests designed to detect C. m. michiganensis. This LAMP assay could be seamlessly integrated into standardized testing regimens, such as those of the European and Mediterranean Plant Protection Organization (EPPO), International Seed Federation (ISF), and commercial seed companies, as the definitive C. m. michiganensis confirmatory test, following isolation of colonies by plating. A c c e p t e d M a n u s c r i p t J. Yasuhara-Bell et al. Detection of Clavibacter in seeds and plant tissues by LAMP 4 Résumé : Afin de déterminer l'efficacité de la détection de l'agent pathogène dans des tissus de tomate et de poivron et dans des semences de tomates, un test moléculaire d'amplification isotherme (LAMP), préalablement développé pour spécifiquement identifier Clavibacter michiganensis subsp. michiganensis, agent causal du chancre bactérien chez la tomate, a été comparé à un immunoessai commercial à flux latéral, à trois tests par PCR et à une culture sur gélose. Le test LAMP a été aussi efficace pour identifier C. m. michiganensis dans les semences et les échantillons de tissus de tomate que le test Cmm ImmunoStrip ® . Le test LAMP a été effectué avec un appareil portable de surveillance en temps réel par fluorescence (Smart-DART TM ) et avec l'iQ TM 5 Multicolor Real-Time PCR Detection System afin de comparer les deux méthodes. Les deux appareils ont produit des résultats équivalents, démontrant ainsi les capacités de ce système LAMP pour la détecti...

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“…michiganensis (Cmm) é capaz de colonizar mudas de tomateiros sem promover a manifestação dos sintomas da doença. Esta observação tornou a fase de produção de mudas de tomateiro mais cautelosa, pois a disseminação do patógeno através do transplante destas mudas sem sintomas nos campos de produção pode se tornar crítica (Gitaitis et al, 1991;Gitaitis et al, 1992) Para o controle do cancro bacteriano do tomateiro são recomendadas várias práticas culturais, como o plantio de mudas sadias, o uso de cultivares resistentes, evitar irrigação por aspersão em excesso, escolher solos com boa drenagem, eliminar restos culturais, fazer rotação de culturas com gramíneas por pelo menos um ano, tratar estacas de tutoramento, mourões, bandejas de semeadura e arames e realizar tratamento de sementes (Fatmi et al, 1991;Gleason et al, 1993;Lopes & Santos, 1994;Kurozawa & Pavan, 1997;Quezado-Soares & Lopes, 1994).…”

Section: Introductionunclassified

Ação de produtos químicos in vitro e in vivo sobre Clavibacter michiganensis subsp. michiganensis, agente causal do cancro bacteriano do tomateiro

Theodoro

Maringoni

2000

Sci. agric. (Piracicaba, Braz.)

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RESUMO: O presente trabalho teve como objetivo avaliar a sensibilidade in vitro e in vivo de um isolado de Clavibacter michiganensis subsp. michiganensis (Cmm) aos produtos cloridrato de kasugamicina, fulusulfamide e oxitetraciclina. Para o teste in vitro, foram avaliados os halos de inibição formados ao redor de discos de papel de filtro umedecidos com os produtos, nas concentrações de 0, 1, 10, 100 e 1000 µg mL -1 , 24 h após a instalação do ensaio. Quanto à avaliação dos produtos in vivo, dois ensaios foram instalados sob condições de casa-de-vegetação, com plantas de tomateiro cultivar Ângela Hiper, através da inoculação das plantas por dois métodos: pulverização foliar e ferimento no caule. Nos dois ensaios, as plantas foram pulverizadas com os produtos cloridrato de kasugamicina a 0,06 mL L , duas vezes antes e duas vezes após a inoculação, em intervalos médios de 5-7 dias. Foram avaliados a incidência de folíolos doentes (inoculação foliar) e a severidade dos sintomas nas plantas inoculadas por ferimento no caule. Verificou-se que o isolado de Cmm foi sensível in vitro ao cloridrato de kasugamicina, ao fulusulfamide e à oxitetraciclina, respectivamente a partir das concentrações de 1000, 100 e 10 µg mL -1 . Com relação aos ensaios in vivo, apenas oxitetraciclina propiciou menor incidência de folíolos doentes nas plantas inoculadas através de pulverização foliar; nenhum produto teve êxito em controlar a doença nas plantas inoculadas por ferimento no caule. O fulusulfamide, em todas as concentrações, foi fitotóxico aos folíolos das plantas de tomateiro. Palavras-chave: tomate, doença, controle químico IN VITRO AND IN VIVO ACTION OF CHEMICALS ON Clavibacter michiganensis SUBSP. michiganensis, CAUSAL AGENT OF THE BACTERIAL CANKER OF TOMATOABSTRACT: An experiment was carried out to evaluate the in vivo and in vitro sensibility of a Clavibacter michiganensis subsp. michiganensis isolate (Cmm) to the products kasugamicyn cloridrate, fulusulfamide and oxytetracicline. In the in vitro tests, the inhibition formed around the discs of filter papers soaked with the chemicals was evaluated, for concentrations of 0, 1, 10, 100 and 1000 µg mL -1 , 24 h after the installation of the experiment. In the in vivo evaluation, two experiments were installed under greenhouse conditions, with tomato plants cv. Ângela Hiper and two methods of inoculation: leaf spray and stem injury. In both experiments, the plants were sprayed with 0.06 mL L -1 of kasugamicyn chloridrate, 0.025, 0.05 and 0.075 mL L -1 of fulusulfamide and 0.40 g L -1 of oxytetracycline, twice before and twice after the inoculations, at intervals of 5 to 7 days. The incidence of diseased leaves (inoculation by leaf spray) and the severity of the symptoms in the plants inoculated by stem injury was evaluated. The Cmm isolate was in vitro sensitive to kasugamicyn chloridrate, fulusulfamide and oxytetracycline in the respective concentrations of 1000, 100 and 10 µg mL -1 . Only the oxytetracycline promoted a lower incidence of diseased leaves in the plants inoculated...

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Detection ofClavibacter michiganensissubsp.michiganensisin Symptomless Tomato Transplants

Cited by 76 publications

References 0 publications

A Proteomic Study of Clavibacter Michiganensis Subsp. Michiganensis Culture Supernatants

A Proteomic Study of Clavibacter Michiganensis Subsp. Michiganensis Culture Supernatants

Utility of a loop-mediated amplification assay for detection ofClavibacter michiganensissubsp.michiganensisin seeds and plant tissues

Ação de produtos químicos in vitro e in vivo sobre Clavibacter michiganensis subsp. michiganensis, agente causal do cancro bacteriano do tomateiro

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