Universal PCR primers for detection of phytopathogenic Agrobacterium strains

Haas, Jerry H.; Moore, Larry F.; Ream, Walt; Manulis, Shulamit

doi:10.1128/aem.61.8.2879-2884.1995

Cited by 168 publications

(87 citation statements)

References 51 publications

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“…Hence, the lack of the ipt gene may explain the non-pathogenic nature of this bacterial strain. Concordantly, a non-pathogenic strain of A. radiobacter containing virD2 but not ipt was previously described by Haas et al (1995). These authors speculated that the strain arose from a pathogenic progenitor through a deletion in the T-DNA.…”

Section: Discussionsupporting

confidence: 52%

Detection and identification of bacteria intimately associated with fungi of the orderSebacinales

Sharma

Schmid

Rothballer

et al. 2008

Cellular Microbiology

151

137

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Because of their beneficial impact on plants, the highly diverse mycorrhizal fungi grouped in the order Sebacinales lay claim to high ecological and agricultural significance. Here, we describe for the first time associations of Sebacinoid members with bacteria. Using quantitative PCR, denaturating gradient gel electrophoresis and fluorescence in situ hybridization, we detected an intimate association between Piriformospora indica and Rhizobium radiobacter, an alpha-Proteobacterium. The stability of the association, vertical transmission of the bacteria during asexual fungal reproduction and fungal plant colonization was monitored using R. radiobacter-specific primers. Treatment of mycelium or fungal protoplasts with antibiotics highly efficient against the free bacteria failed to cure the fungus. Barley seedlings dip-inoculated with R. radiobacter showed growth promotion and systemic resistance to the powdery mildew fungus Blumeria graminis comparable to P. indica inoculation. By screening additional isolates of the Sebacina vermifera complex, three species-specific associations with bacteria from the genera Paenibacillus, Acinetobacter and Rhodococcus were found. These findings suggest that Sebacinales species regularly undergo complex interactions involving host plants and bacteria reminiscent of other ectomycorrhizal and endomycorrhizal associations.

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Section: Discussionsupporting

confidence: 52%

Detection and identification of bacteria intimately associated with fungi of the orderSebacinales

Sharma

Schmid

Rothballer

et al. 2008

Cellular Microbiology

151

137

View full text Add to dashboard Cite

show abstract

“…3 shows AluI digests). All the bacterial strains yielded the expected 338 bp PCR product following amplification with the virD2 A and E primers of Haas et al (1995). Fragment patterns following digestion of the PCR product with AluI or MspI were the same for all 15 RMA cucumopine pRi but different for the other pRi and pTi analysed in this study (Fig.…”

Section: T-dna Pcr-rflpmentioning

confidence: 61%

“…The presence of putative pRi was determined by the conventional PCR assays of Haas et al (1995). The virD2 assay detects the vir region virD2 gene, present in both pRi and pTi and the ipt PCR detects the ipt oncogene only present on pTi.…”

Section: Pri Detection and Genomic Fingerprinting By Pcrmentioning

confidence: 99%

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Recurrent outbreaks of root mat in cucumber and tomato are associated with a monomorphic, cucumopine, Ri-plasmid harboured by various Alphaproteobacteria

Weller¹,

Stead²,

Young³

2006

FEMS Microbiology Letters

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Recurrent outbreaks of root mat have occurred in the UK and France in cucumber and tomato. Root mat is caused by bacterial strains harbouring a Ri-plasmid (pRi). Fifteen root mat-associated (RMA) cucumopine pRi were analysed by PCR-restriction fragment length polymorphism (RFLP) and Southern blotting. These pRi were harboured by Agrobacterium biovar 1 strains isolated during a 1970s outbreak of root mat in UK soil grown cucumber, and also by Agrobacterium biovar 1, Ochrobactrum, Rhizobium and Sinorhizobium isolated during outbreaks of root mat in cucumber and tomato grown hydroponically in the UK and France since 1993. PCR-RFLP analysis of the T-DNA and virD2 regions showed sequence homology between all cucumopine pRi, indicating that these pRi are monomorphic, and thus this pRi remained in the UK without inducing symptoms for some 15 years between outbreaks in the 1970s and 1990s. Cucumopine pRi were also shown to possess the virE2 substitute GALLS gene by Southern blotting. Two other pRi, harboured by Agrobacterium isolated from a recent root mat outbreak in one tomato crop, were also shown to possess the GALLS gene but were shown not to be cucumopine pRi by PCR-RFLP.

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“…Recently, several protocols, mostly based on PCR technology, have been described to detect and characterize pathogenic Agrobacterium spp. (Nesme et al 1989;Haas et al 1995;Sawada et al 1995;Cubero et al 1999Cubero et al ,2002Cubero and Ló pez 2001;Llop et al 2003;). These methods have been applied to detect the bacteria within the tumours to differentiate tumours caused by agrobacteria from those caused by other biotic or physiological factors (Nester and Kosuge 1981;Bayer 1982;Black 1982;Rohfritsch and Shorthouse 1982).…”

Section: Introductionmentioning

confidence: 99%

Systemic movement of Agrobacterium tumefaciens in several plant species

et al. 2006

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Aims: The systemic movement of Agrobacterium spp. inside plants of different species was studied to determine the most valuable diagnostic methodology for their detection. Methods and Results: Pathogenic agrobacteria were detected by isolation and PCR in tissue away from primary tumours in tomato plants grown in the presence of Agrobacterium spp. Moreover, this bacterium was also able to induce secondary tumours beyond the inoculation site. In addition, the capacity of agrobacteria to translocate and induce secondary tumours was analysed in rose, grapevine, chrysanthemum, cherry and peach × almond hybrid GF677. No differences among strains of Agrobacterium spp. were detected in secondary tumour development, although some of them induced a significantly higher number of primary tumours in some species. Movement of inoculated pathogenic cells of four strains was also demonstrated in symptomless portions of the plant stems by isolation and PCR. Finally, pathogenic agrobacteria were detected in root, crown and stem portions of naturally infected walnuts. In all assays, PCR was the most efficient technique for detecting the movement of Agrobacterium spp. within the plants. Conclusions: Migration of agrobacteria inside plants is a complex phenomenon and more extensive than previously reported. Therefore, efficient and sensitive detection methods such as PCR must be used to select clean plants to avoid latent infections of Agrobacterium spp. Significance and Impact of the Study: The results show that migration of Agrobacterium spp. could be relatively frequent in several cultivated fruit trees, and systemic infections should be taken into account when designing strategies for controlling crown gall disease.

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Universal PCR primers for detection of phytopathogenic Agrobacterium strains

Cited by 168 publications

References 51 publications

Detection and identification of bacteria intimately associated with fungi of the orderSebacinales

Detection and identification of bacteria intimately associated with fungi of the orderSebacinales

Recurrent outbreaks of root mat in cucumber and tomato are associated with a monomorphic, cucumopine, Ri-plasmid harboured by various Alphaproteobacteria

Systemic movement of Agrobacterium tumefaciens in several plant species

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