Categorisation of reactions and enumeration of bacteria in potato cultivars inoculated with the causal agent of bacterial ring rot

Variability in the responses of plants propagated from in vitro tissue cultures of 52 ware and industrial potato cultivars to different Clavibacter michiganensis subsp. sepedonicus inoculum size was tested during 2015–2017. Bacterial ring rot symptoms on plants and tubers xylem vessels were recorded for 8 weeks and the susceptibility index (SI) for individual cultivars was calculated. Based on foliage symptoms, potato cultivars were placed into three symptoms groups. The symptomless group had SIs ≤ 1, for the moderate symptom group SIs ranged from 1.01 to 2.99, and the severe symptom group had SIs ≥ 3.0. The pathogen concentrations in vascular vessels of all infected potato plants increased during the experiment regardless of the foliage symptom group.

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THE EFFECT OF NANOSCALE SELENIUM ON THE CAUSATIVE AGENT OF RING ROT AND POTATO IN VITRO

chemistry of plant raw material ◽

10.14258/jcprm.2019034794 ◽

2019 ◽

pp. 345-354 ◽

Cited By ~ 1

Author(s):

Irina Alekseyevna Graskova ◽

Alla Innokent'yevna Perfileva ◽

Ol'ga Aleksandrovna Nozhkina ◽

Anastasiya Vyacheslavovna Dyakova ◽

Vadim Nikolayevich Nurminsky ◽

...

Keyword(s):

Pathogenic Bacteria ◽

Cultivated Plants ◽

Clavibacter Michiganensis ◽

Bacteriostatic Effect ◽

Potato Plants ◽

Clavibacter Michiganensis Subsp ◽

Ring Rot ◽

Negative Effect ◽

High Selenium

The biological activity of biopolymer-based selenium nanocomposite with a high selenium content of 6.4% was investigated. It was shown earlier that nanocomposites of selenium and arabinogalactan (NC Se/AG, 1.23% and 3.4% Se) have a bactericidal and bacteriostatic effect on the pathogenic bacterium Clavibacter michiganensis subsp. sepedonicus (Cms) and do not adversely affect the vitality of potato plants in vitro. In this work, it was shown that the nanocomposite under scrutiny has a bacteriostatic effect, inhibits bacterial growth by 20% compared with the check variant and reduces the ability of Cms to form biofilms, which contribute to their high resistance to external factors. In the experiments conducted on plants, it was shown, there was not negative effect of NC Se/AG on biometric parameters, furthermore, this NC reduces the negative effect of potato infection with Cms. The nanocomposite reduced the activity of peroxidase and the content of reactive oxygen species in potato tissues. It was established that selenium does not accumulate in potato plants after the treatment with NC Se/AG. The results obtained allow us to consider NC Se/AG (6.4% Se) as an agent for healing cultivated plants from pathogenic bacteria.

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Teknik Inokulasi Buatan Clavibacter michiganensis subsp. sepedonicus, Penyebab Penyakit Busuk Cincin Bakteri, pada Tanaman Kentang (Solanum tuberosum L.)

Agrikultura ◽

10.24198/agrikultura.v27i2.9967 ◽

2016 ◽

Vol 27 (2) ◽

Author(s):

Luciana Djaya ◽

Ineu Sulastrini ◽

Iin Rusita

Keyword(s):

Solanum Tuberosum ◽

Solanum Tuberosum L ◽

Block Design ◽

Clavibacter Michiganensis ◽

Bacterial Ring Rot ◽

Seed Tubers ◽

Clavibacter Michiganensis Subsp ◽

Ring Rot ◽

Inoculation Techniques ◽

Rot Disease

ABSTRACT Inoculation Techniques of Clavibacter michiganensis subsp. sepedonicus, the Cause of Bacterial Ring Rot Disease, on Potato (Solanum tuberosum L.). Clavibacter michiganensis subsp. sepedonicus, the cause of bacterial ring rot disease on potatoes, has been detected in potato fields in Pangalengan. To anticipate the spread of the pathogen, researches on the desease epidemiology are urgent to be carried out. Artificial inoculation techniques will be useful in the epidemiological studies. The objective of this reasearch was to evaluate some inoculation techniques, which are simple, cheap and fast in causing disease symptoms. The experiment was carried out at the laboratory and glasshouse of Balai Penelitian Tanaman Sayuran (Balitsa), Lembang. The experiment was arranged in the randomized block design with five treatments of inoculation technique and five replications. The treatments were (a) soaking wounded seed tubers in pathogen suspension, (b) soaking not wounded seed tubers in pathogen suspension, (c) pathogen suspension was injected into leaf axil, (d) pathogen suspension was injected into seed tubers, and (e) pathogen suspension was poured into the planting holes. The results showed that stabbing and soaking tubers in pathogen suspension caused the shortest incubation period (17 days after inoculation) and the highest disease incidence (60%). Keywords : Clavibacter michiganensis subsp. sepedonicus, Bacterial ring rot, Potatoes, Inoculation techniquesABSTRAKClavibacter michiganensis subsp. sepedonicus (Cms), penyebab penyakit busuk cincin bakteri pada tanaman kentang, telah terdeteksi keberadaannya pada pertanaman kentang di Pangalengan. Dalam upaya mencegah penyebaran penyakit busuk cincin bakteri di Indonesia, perlu adanya studi epidemiologi patogen tersebut. Pada penelitian epidemiologi akan diperlukan cara menginokulasi tanaman secara buatan. Tujuan dari penelitian ini adalah untuk mendapatkan teknik inokulasi buatan yang dapat menyebabkan periode inkubasi yang lebih singkat dan persentase kejadian penyakit busuk cincin bakteri paling tinggi pada tanaman kentang. Percobaan dilaksanakan di rumah kaca dan laboratorium penyakit Balai Penelitian Tanaman Sayuran (Balitsa), Lembang. Percobaan dirancang secara rancangan acak kelompok dengan lima perlakuan teknik inokulasi Cms dan lima ulangan, yaitu inokulasi dengan (a) merendam ubi benih yang telah dilukai dalam suspensi bakteri, (b) merendam ubi benih tanpa dilukai dalam suspensi bakteri, (c) suspensi bakteri ditusukkan pada ketiak daun tanaman kentang, (d) suspensi bakteri ditusukkan pada ubi benih, dan (e) suspensi bakteri disiramkan pada lubang tanam. Masing-masing ulangan terdiri dari lima tanaman. Hasil penelitian menunjukkan bahwa dari lima teknik inokulasi buatan yang dilakukan, inokulasi dengan melukai ubi benih dan merendamnya dalam suspensi patogen menghasilkanJurnal Agrikultura 2016, 27 (2): 66-71ISSN 0853-2885Teknik Inokulasi Buatan Clavibacter….67periode inkubasi tersingkat, yaitu 17 hari setelah inokulasi, dan persentase kejadian penyakit tertinggi yaitu sebesar 60%.Kata Kunci : Clavibacter michiganensis subsp. sepedonicus, Busuk cincin bakteri, Kentang, Inokulasi buatan

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New Type of Antimicrobial Protein Produced by the Plant Pathogen Clavibacter michiganensis subsp. michiganensis

Applied and Environmental Microbiology ◽

10.1128/aem.01065-13 ◽

2013 ◽

Vol 79 (18) ◽

pp. 5721-5727 ◽

Cited By ~ 10

Author(s):

Zhanliang Liu ◽

Ping Ma ◽

Ingrid Holtsmark ◽

Morten Skaugen ◽

Vincent G. H. Eijsink ◽

...

Keyword(s):

Antimicrobial Protein ◽

Clavibacter Michiganensis ◽

Bacterial Ring Rot ◽

Content Type ◽

Highly Active ◽

Clavibacter Michiganensis Subsp ◽

Ring Rot ◽

Inhibitory Spectrum ◽

New Type ◽

Tomato Pathogen

ABSTRACTIt has previously been shown that the tomato pathogenClavibacter michiganensissubsp.michiganensissecretes a 14-kDa protein,C. michiganensissubsp.michiganensisAMP-I (CmmAMP-I), that inhibits growth ofClavibacter michiganensissubsp.sepedonicus, the causal agent of bacterial ring rot of potato. Using sequences obtained from tryptic fragments, we have identified the gene encodingCmmAMP-I and we have recombinantly produced the protein with an N-terminal intein tag. The gene sequence showed thatCmmAMP-I contains a typical N-terminal signal peptide for Sec-dependent secretion. The recombinant protein was highly active, with 50% growth inhibition (IC50) of approximately 10 pmol, but was not toxic to potato leaves or tubers.CmmAMP-I does not resemble any known protein and thus represents a completely new type of bacteriocin. Due to its high antimicrobial activity and its very narrow inhibitory spectrum,CmmAMP-1 may be of interest in combating potato ring rot disease.

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The Putative Secreted Serine Protease Chp-7 Is Required for Full Virulence and Induction of a Nonhost Hypersensitive Response by Clavibacter michiganensis subsp. sepedonicus

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi-22-7-0809 ◽

2009 ◽

Vol 22 (7) ◽

pp. 809-819 ◽

Cited By ~ 11

Author(s):

Riitta Nissinen ◽

Yunjian Xia ◽

Laura Mattinen ◽

Carol A. Ishimaru ◽

Dennis L. Knudson ◽

...

Keyword(s):

Hypersensitive Response ◽

Mutant Library ◽

Plant Interactions ◽

In Planta ◽

Clavibacter Michiganensis ◽

Bacterial Ring Rot ◽

Clavibacter Michiganensis Subsp ◽

Ring Rot ◽

Biological Studies

Molecular biological studies on Clavibacter michiganensis subsp. sepedonicus, the causal agent of bacterial ring rot of potato, have gained greater feasibility due to the recent availability of whole genomic sequences and genetic tools for related taxa. Here, we describe the first report of construction and characterization of a transposon (Tn) mutant library of C. michiganensis subsp. sepedonicus sp. strain R10. Since virulence of R10 in potato has been shown previously to be associated with elicitation of a nonhost hypersensitive response (HR), the mutant library was screened initially for loss of HR in tobacco. The screen identified two HR-negative mutants containing Tn insertions within the same gene, CMS2989 (chp-7), although at distinct locations. chp-7 is one of 11 pat-1 homologs in C. michiganensis subsp. sepedonicus. HR-negative mutants of R10 multiplied to the same extent as wild type in planta but were less virulent in potato. Complementation with chp-7 restored virulence as well as the HR phenotype. Together, these findings demonstrate a role for chp-7 in C. michiganensis subsp. sepedonicus–plant interactions.

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Bacterial Ring Rot of Potatoes Caused by Clavibacter michiganensis subsp. sepedonicus in Lithuania

Plant Disease ◽

10.1094/pdis.2001.85.10.1119a ◽

2001 ◽

Vol 85 (10) ◽

pp. 1119-1119

Author(s):

L. Baranauskaite ◽

M. Vasinauskiene

Keyword(s):

Plant Protection ◽

Polyclonal Antibodies ◽

Solanum Tuberosum L ◽

Disease Incidence ◽

Potato Production ◽

Enzyme Linked Immunosorbent Assay ◽

Clavibacter Michiganensis ◽

Bacterial Ring Rot ◽

Clavibacter Michiganensis Subsp ◽

Ring Rot

Bacterial ring rot of potato (Solanum tuberosum L.) was observed for the first time in Lithuania from 1998 to 1999. The disease, caused by Clavibacter michiganensis subsp. sepedonicus, is considered one of the most important bacterial diseases of potato. Nearly all countries that produce potatoes report the presence of the bacterium (1). C. michiganensis subsp. sepedonicus is listed as an A2 quarantine pathogen by the European and Mediterranean Plant Protection Organization. The pathogen has a latent period that can last for several generations of potato production, thus increasing the risk for further dissemination of the disease. Plant wilting and tuber rotting, characteristic symptoms of the disease, were observed in various localities in Lithuania. The disease was found in 12 potato varieties. Disease incidence approached 5% in the field and 5 to 21% in storage. The presence of C. michiganensis subsp. sepedonicus in diseased as well as asymptomatic plant tissue was determined by immunofluorescence cell staining and enzyme-linked immunosorbent assay with the use of monoclonal and polyclonal antibodies, respectively. More than 350 potato samples were tested. Twenty-five samples tested positive by these methods. Pathogenicity tests of 30 samples were conducted on eggplant. Characteristic symptoms were found on 21 plants. The bacteria were reisolated on nutrient dextrose agar medium from 12 inoculated eggplants. For confirmation of six suspicious strains, polymerase chain reaction with a specific Cms50 primer set was employed (2). In three cases, positive results were obtained. References: (1) I. M. Smith and L. M. F. Charles, eds. 1998. Distribution of Maps of Quarantine Pests for Europe. CAB International, Wallingford, U.K. (2) D. Mills et al. Phytopathology 87:853, 1997.

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Symptomless infection with Clavibacter michiganensis subspecies sepedonicus during tissue culture propagation of potato

Canadian Journal of Plant Science ◽

10.4141/cjps92-119 ◽

1992 ◽

Vol 72 (3) ◽

pp. 943-953 ◽

Cited By ~ 4

Author(s):

B. A. Schuld ◽

M. D. Harrison ◽

J. Crane

Keyword(s):

Solanum Tuberosum L ◽

Solanum Melongena ◽

Shoot Cultures ◽

Clavibacter Michiganensis ◽

Bacterial Ring Rot ◽

Inoculum Level ◽

Clavibacter Michiganensis Subsp ◽

Nodal Cuttings ◽

Symptomless Infection ◽

Culturing Conditions

Nodal cuttings of tissue cultured potato (Solarium tuberosum L. ’Sangre’) were inoculated with the bacterial ring rot (BRR) pathogen, Clavibacter michiganensis subsp. sepedonicus (Cms). Under commercial tissue culturing conditions, the pathogen persisted through as many as three micropropagation cycles, each consisting of production of a plantlet from an apical cutting. Infected plantlets did not develop BRR symptoms of leaf wilt or chlorosis. Signs of bacterial presence, cloudiness of tissue culturing medium or foliar lesions, were not present when the inoculum level was low, 5.0 × 102 cfu mL−1. A higher inoculum concentration (5.0 × 103 cfu mL−1) reduced plantlet height, but the lower concentration had no effect on height. Growth and viability of tissue from Cms-inoculated plantlets grown on Murashige and Skoog’s medium without sucrose, a source of carbon, were reduced compared to those grown with sucrose, but BRR symptoms were not induced at these inoculum levels. When transplanted to soilless potting medium, a basal portion of cycle 1 plantlets either died or produced a symptomatic miniplant, indicating that Cms survived during the growth of a plantlet into a miniplant. Nonsymptomatic miniplants were produced by portions of some cycle 2 and 3 Cms-infected plantlets, indicating that successive nodal propagation decreased the number of symptomatic miniplants. Bioassays of these nonsymptomatic miniplants were Cms-negative. Incubating plantlet tissue in Richardson broth for 14 d and visually observing for turbidity was not effective in detecting Cms in cultured potato tissue. Because phytopathogenic bacteria may persist in tissue cultures without causing symptoms, pathogen-specific testing of source plants and plant cultures is necessary to prevent transmission of bacteria to cultured stock and field-grown progeny.Key words: Potato shoot cultures, Solanum tuberosum L., Clavibacter michiganensis subsp. sepedonicus, medium with no carbon, bacterial persistence and detection, symptomless infection, Solanum melongena L.

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Symptom Expression of Mainstream and Specialty Potato Cultivars to Bacterial Ring Rot (Clavibacter sepedonicus) and Evaluation of in-Field Detection

American Journal of Potato Research ◽

10.1007/s12230-019-09730-x ◽

2019 ◽

Vol 96 (4) ◽

pp. 427-444 ◽

Cited By ~ 1

Author(s):

Jonathan L. Whitworth ◽

Rachel A. Selstedt ◽

Alan A. G. Westra ◽

Phil Nolte ◽

Kasia Duellman ◽

...

Keyword(s):

Potato Cultivars ◽

Symptom Expression ◽

Bacterial Ring Rot ◽

Field Detection ◽

Ring Rot

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Immunochemical, biochemical and pathogenic properties of fluidal and intermediate strains of Clavibacter michiganensis subsp. sepedonicus

Plant Protection Science ◽

10.17221/10471-pps ◽

2017 ◽

Vol 38 (SI 2 - 6th Conf EFPP 2002) ◽

pp. 291-293

Author(s):

B. Kokošková ◽

R. Jeřabková

Keyword(s):

Monoclonal Antibodies ◽

Metabolic Activity ◽

Species Level ◽

Colony Morphology ◽

Clavibacter Michiganensis ◽

Clavibacter Michiganensis Subsp ◽

Ring Rot ◽

Atypical Cells ◽

Das Elisa

Clavibacter michiganensis subsp. sepedonicus (Cms), causing ring rot of potato, is a quarantine bacterium. According to colony morphology, Cms occurs mostly as fluidal (smooth), but sometime as intermediate (rough, i.e. less fluidal) variants, too. Commercial monoclonal antibodies (Agdia, USA) were used for determination of 40 Cms strains representing both forms. All Cms strains were reliably identified by IFAS, but atypical cells were sometime recorded in population of intermediate strains. The fluidal Cms strains were more reliably identified using DAS-ELISA and the Biolog GP MicroPlate System<sup>TM</sup> than intermediate strains. The intermediate Cms strains had decreased metabolic activity compared with fluidal strains and that is why they were identified only to the genera or to the species level or not identified. The differences among fluidal and intermediate Cms strains were recorded also in bioassay on eggplants. The intermediate Cms strains caused atypical or no symptoms with comparison to fluidal strains.

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