Enrichment of Clavibacter michiganensis subsp. michiganensis in tomato seed extracts

2017 ◽  
Vol 45 (1) ◽  
pp. 81-99
Author(s):  
F.M.V. Lelis ◽  
J.M. Van Der Wolf
Keyword(s):  
Clavibacter Michiganensis ◽  
Tomato Seed ◽  
Clavibacter Michiganensis Subsp ◽  
Seed Extracts

scholarly journals Wilt Pathogens of Solanaceae in Tanzania: Clavibacter michiganensis subsp. michiganensis, Pseudomonas corrugata, and Ralstonia solanacearum

Plant Disease ◽  
1999 ◽  
Vol 83 (11) ◽  
pp. 1070-1070 ◽  
Author(s):  
R. Black ◽  
S. Seal ◽  
Z. Abubakar ◽  
R. Nono-Womdim ◽  
I. Swai
Keyword(s):  
Bacterial Wilt ◽  
Ralstonia Solanacearum ◽  
Polymyxin B ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bacterial Canker ◽  
Clavibacter Michiganensis ◽  
Pseudomonas Corrugata ◽  
Tomato Seed ◽  
Clavibacter Michiganensis Subsp ◽  
Northern Highlands

Surveys of vegetables in the southern and northern growing regions of mainland Tanzania and Zanzibar during 1997 and 1998 indicated the presence of three wilt pathogens of tomato. Clavibacter michiganensis subsp. michiganensis (cause of bacterial canker) was isolated from wilting plants in the southern highlands and Lushoto District (Tanga Region, north) on selective King's medium B with polymyxin B (1). The identity of the isolates was confirmed by cultural, morphological, and biochemical characteristics and a plate-trapped antigen enzyme-linked immunosorbent assay (PTA-ELISA) kit (Pathoset 113-08, Adgen, Auchincruive, U.K.). Pathogenicity was confirmed by host inoculation. In addition, the bacterium was detected directly by the same methods in several sources of seed, including commercial farmers' saved seed and seed extracted from infected plants. Of 61 tomato seed lots tested, 18 samples were positive. Ralstonia solanacearum (cause of bacterial wilt) was isolated from tomato and potato by semiselective media and detected directly in stem and tuber tissues, respectively, by polymerase chain reaction and ELISA (4) in all vegetable-growing areas surveyed. In 1998, R. solanacearum was detected for the first time in Zanzibar on tomato and eggplant. Pathogenicity was confirmed by inoculation on and reisolation from tomato seedlings of cv. Money Maker. Only biovar 3 (2) occurred in tomato. Biovar 3 also was found in midaltitude potato. Biovar 2 has been found only in potato plants grown above 1,500 m. Pseudomonas corrugata (cause of pith necrosis) was isolated from tomato on semiselective media at only one location. From wilted tomato plants in the southern highlands, 38% of samples tested positive for C. michiganensis subsp. michiganensis and ≈ 10% for R. solanacearum. Of samples collected from the northern highlands, 43% tested positive for R. solanacearum. Wilt incidence of ≈35% was observed in tomato fields where the bacterial wilt pathogen was isolated in the northern highlands compared with gt;90% incidence and almost total crop loss in tomato fields of the southern highlands infected with bacterial canker. Although all three pathogens caused systemic wilt of plants, bacterial canker occasionally caused downward turning of lower leaves, unilateral wilting, and marginal necrosis of leaflets as well as fruit spotting. In general, wilts caused by C. michiganensis subsp. michiganensis, R. solanacearum, and P. corrugata were not readily differentiated in Tanzania prior to this research. Plants with pith browning had often been assumed to be infected by P. corrugata, until R. solanacearum was isolated (3) frequently from such plants. References: (1) Anonymous. OEPP/EPPO Bull. 22:219, 1992. (2) A. C. Hayward. J. Appl. Bacteriol. 27:265, 1964. (3) J. B. Jones et al., eds. 1991. Compendium of Tomato Diseases. The American Phytopathological Society, St. Paul, MN. (4) S. Seal and J. G. Elphinstone. Pages 35–57 in: Bacterial Wilt. The Disease and Its Causative Agent, Pseudomonas solanacearum. A. C. Hayward and G. L. Hartman, eds. CAB International, Wallingford, U.K. 1994.

scholarly journals Bioluminescence Imaging of Clavibacter michiganensis subsp. michiganensis Infection of Tomato Seeds and Plants

2010 ◽  
Vol 76 (12) ◽  
pp. 3978-3988 ◽  
Author(s):  
Xiulan Xu ◽  
Sally A. Miller ◽  
Fulya Baysal-Gurel ◽  
Karl-Heinz Gartemann ◽  
Rudolf Eichenlaub ◽  
...  
Keyword(s):  
Genetic Manipulation ◽  
Early Stage ◽  
Control Measures ◽  
Economic Losses ◽  
Plant Interactions ◽  
Clavibacter Michiganensis ◽  
Tomato Seed ◽  
Tomato Production ◽  
Clavibacter Michiganensis Subsp ◽  
Mirabilis Jalapa

ABSTRACT Clavibacter michiganensis subsp. michiganensis is a Gram-positive bacterium that causes wilting and cankers, leading to severe economic losses in commercial tomato production worldwide. The disease is transmitted from infected seeds to seedlings and mechanically from plant to plant during seedling production, grafting, pruning, and harvesting. Because of the lack of tools for genetic manipulation, very little is known regarding the mechanisms of seed and seedling infection and movement of C. michiganensis subsp. michiganensis in grafted plants, two focal points for application of bacterial canker control measures in tomato. To facilitate studies on the C. michiganensis subsp. michiganensis movement in tomato seed and grafted plants, we isolated a bioluminescent C. michiganensis subsp. michiganensis strain using the modified Tn1409 containing a promoterless lux reporter. A total of 19 bioluminescent C. michiganensis subsp. michiganensis mutants were obtained. All mutants tested induced a hypersensitive response in Mirabilis jalapa and caused wilting of tomato plants. Real-time colonization studies of germinating seeds using a virulent, stable, constitutively bioluminescent strain, BL-Cmm17, showed that C. michiganensis subsp. michiganensis aggregated on hypocotyls and cotyledons at an early stage of germination. In grafted seedlings in which either the rootstock or scion was exposed to BL-Cmm17 via a contaminated grafting knife, bacteria were translocated in both directions from the graft union at higher inoculum doses. These results emphasize the use of bioluminescent C. michiganensis subsp. michiganensis to help better elucidate the C. michiganensis subsp. michiganensis-tomato plant interactions. Further, we demonstrated the broader applicability of this tool by successful transformation of C. michiganensis subsp. nebraskensis with Tn1409::lux. Thus, our approach would be highly useful to understand the pathogenesis of diseases caused by other subspecies of the agriculturally important C. michiganensis.

scholarly journals Bacterial Canker (Clavibacter michiganensis subsp. michiganensis) of Tomato in Commercial Seed Produced in Indonesia

Plant Disease ◽  
2004 ◽  
Vol 88 (6) ◽  
pp. 680-680 ◽  
Author(s):  
A. Anwar ◽  
P. S. van der Zouwen ◽  
S. Ilyas ◽  
J. M. van der Wolf
Keyword(s):  
Enzyme Linked Immunosorbent Assay ◽  
Pathogenicity Test ◽  
Bacterial Canker ◽  
Vegetable Crops ◽  
Clavibacter Michiganensis ◽  
Tomato Seed ◽  
Causal Organism ◽  
Tomato Production ◽  
Clavibacter Michiganensis Subsp ◽  
Bacterial Canker Of Tomato

In 2002, Clavibacter michiganensis subsp. michiganensis (Smith) Davis, the causal organism of bacterial canker of tomato (Lycopersicon esculentum), was isolated from two of six commercial asymptomatic tomato seed lots produced on Java in Indonesia. C. michiganensis subsp. michiganensis has not been reported in Indonesia previously. Methods based on the protocol of the International Seed Health Initiative were used to extract and identify the presence of C. michiganensis subsp. michiganensis in tomato seed. C. michiganensis subsp. michiganensis was isolated with dilution plating on the semiselective media D2ANX and mSCM. The identity of the colonies was confirmed by immunofluorescence microscopy, polymerase chain reaction (2), fatty methyl ester analysis, enzyme-linked immunosorbent assay based on monoclonal antibody 103 (1), and a pathogenicity test in which three replicate tomato plants were stem inoculated with 108 cells ml-1. Within 2 weeks, stripes on stems developed that split and exposed reddish brown cavities (stem cankers). The presence of C. michiganensis subsp. michiganensis poses a direct threat on tomato production, which is one of five economically most important vegetable crops in Indonesia. References: (1) A. Alvarez et al. Phytopathology 83:1405, 1993. (2) M. S. Santos et al. Seed Sci. Technol. 25:581, 1997.

scholarly journals A New Selective Medium for Isolation of Clavibacter michiganensis subsp. michiganensis from Tomato Plants and Seed

2011 ◽  
Vol 101 (11) ◽  
pp. 1355-1364 ◽  
Author(s):  
Radwan M. Ftayeh ◽  
Andreas von Tiedemann ◽  
Klaus W. E. Rudolph
Keyword(s):  
Bacterial Species ◽  
False Negative ◽  
Selective Medium ◽  
Detection Sensitivity ◽  
Bacterial Canker ◽  
Clavibacter Michiganensis ◽  
Tomato Seed ◽  
Tomato Plants ◽  
Clavibacter Michiganensis Subsp ◽  
Bacterial Canker Of Tomato

A new selective and highly sensitive medium was developed for isolation of Clavibacter michiganensis subsp. michiganensis (Cmm), the causal agent of bacterial canker of tomato, from seed and latently infected plants. The new medium (BCT) proved to be superior to all published semiselective media for Cmm and is denoted as selective medium because of (i) its mean plating efficiency, amounting to ≤89% within 7 days for all 30 Cmm strains from different sources tested; (ii) the high selectivity, because accompanying bacterial species occurring on tomato plants and seed or bacteria obtained from culture collections were inhibited to an extent of 98 to 100%; and (iii) the remarkable detection sensitivity. Thus, 8 CFU of Cmm in field plant homogenates containing 12,750 CFU of accompanying saprophytes were detected on BCT. Under these extreme conditions, all of the published semiselective media (D2, KBT, D2ANX, SCM, mSCM, CMM1, mCNS, and EPPO) gave false-negative results. Either some media were rather toxic and Cmm growth was also inhibited or the other, less toxic media allowed growth of high numbers of saprophytes, so that Cmm growth was suppressed. Exclusively, BCT also supported growth of the closely related C. michiganensis subsp. insidiosus, nebraskensis, and tessellarius. The new medium is recommended for Cmm detection in tomato seed, and in symptomless tomato plantlets, to improve disease control of bacterial canker of tomato.

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