Enhanced epidemic intelligence using a web-based screening system during the 2010 FIFA World Cup in South Africa.

The 2010 FIFA World Cup took place in South Africa between 11 June and 11 July 2010. The European Centre for Disease Prevention and Control (ECDC), in collaboration with the hosting authorities, carried out enhanced epidemic intelligence activities from 7 June to 16 July 2010 for timely detection and monitoring of signals of public health events with a potential to pose a risk to participants and visitors. We adapted ECDC's routine epidemic intelligence process to targeted event-based surveillance of official and unofficial online information sources. A set of three specifically adapted alerts in the web-based screening system MedISys were set up: potential public health events in South Africa, those occurring in the participating countries and those in the rest of the world. Results were shared with national and international public health partners through daily bulletins. According to pre-established ECDC criteria for the World Cup, 21 events of potential public health relevance were identified at local and international level. Although none of the events detected were evaluated as posing a serious risk for the World Cup, we consider that the investment in targeted event-based surveillance activities during the tournament was relevant as it facilitated real-time detection and assessment of potential threats. An additional benefit was early communication of relevant information to public health partners.

Silencing Agrobacterium oncogenes in transgenic grapevine results in strain-specific crown gall resistance.

KEY MESSAGE: Grapevine rootstock transformed with an Agrobacterium oncogene-silencing transgene was resistant to certain Agrobacterium strains but sensitive to others. Thus, genetic diversity of Agrobacterium oncogenes may limit engineering crown gall resistance. ABSTRACT: Crown gall disease of grapevine induced by Agrobacterium vitis or Agrobacterium tumefaciens causes serious economic losses in viticulture. To establish crown gall-resistant lines, somatic proembryos of Vitis berlandieri × V. rupestris cv. 'Richter 110' rootstock were transformed with an oncogene-silencing transgene based on iaaM and ipt oncogene sequences from octopine-type, tumor-inducing (Ti) plasmid pTiA6. Twenty-one transgenic lines were selected, and their transgenic nature was confirmed by polymerase chain reaction (PCR). These lines were inoculated with two A. tumefaciens and three A. vitis strains. Eight lines showed resistance to octopine-type A. tumefaciens A348. Resistance correlated with the expression of the silencing genes. However, oncogene silencing was mostly sequence specific because these lines did not abolish tumorigenesis by A. vitis strains or nopaline-type A. tumefaciens C58.

Broad-range antagonistic rhizobacteria Pseudomonas fluorescens and Serratia plymuthica suppress Agrobacterium crown gall tumours on tomato plants.

AIM: To examine the biocontrol activity of broad-range antagonists Serratia plymuthica IC1270, Pseudomonas fluorescens Q8r1-96 and P. fluorescens B-4117 against tumourigenic strains of Agrobacterium tumefaciens and A. vitis. METHODS AND RESULTS: Under greenhouse conditions, the antagonists, applied via root soak prior to injecting Agrobacterium strains into the wounded stems, significantly suppressed tumour development on tomato seedlings. A derivative of P. fluorescens Q8r1-96 tagged with a gfp reporter, as well as P. fluorescens B-4117 and S. plymuthica IC1270 marked with rifampicin resistance, stably persisted in tomato tissues for at least 1 month. Mutants of P. fluorescens Q8r1-96 and S. plymuthica IC1270 deficient in 2,4-diacetylphloroglucinol or pyrrolnitrin production, respectively, also proficiently suppressed the tumour development, indicating that these antibiotics are not responsible for the observed biocontrol effect on crown gall disease. The volatile organic compounds (VOCs) produced by the tested P. fluorescens and S. plymuthica strains inhibited the growth of A. tumefaciens and A. vitis strains in vitro. Solid-phase microextraction-gas chromatography-mass spectrometry analysis revealed dimethyl disulfide (DMDS) as the major headspace volatile produced by S. plymuthica IC1270; it strongly suppressed Agrobacterium growth in vitro and was emitted by tomato plants treated with S. plymuthica IC1270. 1-Undecene was the main volatile emitted by the examined P. fluorescens strains, with other volatiles, including DMDS, being detected in only relatively low quantities. CONCLUSIONS: S. plymuthica IC1270, P. fluorescens B-4117 and P. fluorescens Q8r1-96 can be used as novel biocontrol agents of pathogenic Agrobacterium. VOCs, and specifically DMDS, might be involved in the suppression of oncogenicity in tomato plants. However, the role of specific volatiles in the biocontrol activity remains to be elucidated. SIGNIFICANCE AND IMPACT OF THE STUDY: The advantage of applying these antagonists lies in their multiple activities against a number of plant pathogens, including Agrobacterium.

Incompatibility properties of tartrate utilization plasmids derived from Agrobacterium vitis strains.

The incompatibility properties of two tumor-inducing (Ti) and seven tartrate (Tr) plasmids, derived from various Agrobacterium vitis strains, were characterized using incRh1, incRh2, incRh3, and incRh4 clones which were established for the identification and classification of Agrobacterium plasmids. The tested A. vitis plasmids could be allocated into four groups on the basis of their incompatibility with incRh1, incRh2, and incRh4 clones. The two octopine tumor-inducing plasmids, pTiAT6 and pTiAB3, expressed incompatibility both to incRh1 and to incRh2 clones. Three pTrs could not be allocated either to incRh1-4 and incAg1 or to the wide-host-range incP1, incQ, and incW groups.

Physical map of the vitopine Ti plasmid pTiS4.

Within the Agrobacterium vitis group the vitopine strains represent a special subclass. Vitopine bacteria carry Ti plasmids with little or no homology with the well-characterized T-DNAs of Agrobacterium tumefaciens or Agrobacterium rhizogenes. The 262-kb Ti plasmid of the vitopine strain S4 was cloned and mapped. Homology studies with the octopine Ti plasmid pTiAch5, the nopaline Ti plasmid pTiC58, and the agropine/mannopine Ri plasmid pRiHRI identified several regions of homology. The origin of replication was localized to within 2.5 kb.

Nucleotide sequence, evolutionary origin and biological role of a rearranged cytokinin gene isolated from a wide host range biotype III Agrobacterium strain.

A DNA fragment with homology to the cytokinin (ipt) gene from biotype I Agrobacterium tumefaciens strain Ach5 was cloned from the Ti plasmid of the wide host range biotype III Agrobacterium strain Tm-4 and sequenced. The fragment contains an intact ipt coding sequence. However, the 3' non-coding region of this ipt gene is rearranged due to a 0.9 kb deletion fusing it to the 3' coding region of the neighbouring gene 6a, most of which was found to be deleted. The Tm-4 ipt gene is strongly related to the partially deleted ipt gene of the limited host range biotype III strain Ag162. To test its biological activity, the Tm-4 ipt gene was inserted into a specially constructed, disarmed Ti vector lacking tzs and tested on tobacco, where the rearranged ipt gene induced shoot formation. The cloned Tm-4 ipt gene was mutated with Tn5 and the intact gene on the wild-type Tm-4 Ti plasmid was replaced by the mutated gene. The resulting strain was avirulent on tobacco but normally virulent on the natural host of the wild-type strain Tm-4, grapevine. As the biotype I 6b gene diminishes the effect of a corresponding ipt gene, a larger Tm-4 fragment carrying both the ipt gene and an adjacent 6b-like gene was also tested on tobacco and compared with the Tm-4 ipt fragment alone and with an ipt and 6b/ipt fragment derived from Ach5. The Tm-4 6b gene diminishes the effect of the Tm-4 ipt gene, showing the Tm-4 6b gene to be active as well. The Tm-4 6b/ipt combination is less effective than the Ach5 combination. These results provide further insight into the molecular basis of the host range differences between limited host range and wide host range biotype III Agrobacterium strains and show that the WHR cytokinin gene, although active, does not significantly contribute to tumour formation on the natural host of the WHR biotype III strains, grapevine.