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1.
A nonpathogenic strain of Agrobacterium vitis VAR03-1 was tested as a biological control agent against crown gall of grapevine (Vitis vinifera L.). A mixture of the nonpathogenic strain VAR03-1 and a tumorigenic strain G-Ag-27 of A. vitis at cell ratios of 1 : 1, 3 : 1, 9 : 1, and 99 : 1 significantly inhibited gall formation and size on stems of tomato (Lycopersicon esculentum Mill.). Strain VAR03-1 also inhibited gall formation on stems of both tomato and grapevine at a 1 : 1 cell ratio with several tumorigenic A. vitis strains isolated from different fields of grapevine in Japan. In biological control tests, when roots of grapevine and tomato seedlings were soaked in a cell suspension of strain VAR03-1 for 24 h before a 1-h soaking in a cell suspension of the pathogen and subsequent planting in pots of infested soil, strain VAR03-1 significantly reduced the incidence of gall formation on both plants.  相似文献   

2.
Kawaguchi A  Inoue K  Ichinose Y 《Phytopathology》2008,98(11):1218-1225
A nonpathogenic strain of Agrobacterium vitis VAR03-1 was tested as a biological control agent for crown gall of grapevine (Vitis vinifera). When roots of grapevine, rose (Rose multiflora), and tomato (Lycopersicon esculentum) were soaked in a cell suspension of antagonists before planting in soil infested with tumorigenic A. vitis, A. rhizogenes, and A. tumefaciens, respectively, treatment with VAR03-1 significantly reduced the number of plants with tumors and disease severity in the three plant species. The inhibitory effects of treatment with VAR03-1 and the nonpathogenic A. rhizogenes strain K84 on crown gall of rose and tomato were almost identical, and the inhibitory effect of VAR03-1 on grapevine was superior to that of K84. Moreover, VAR03-1 greatly controlled crown gall of grapevine due to tumorigenic A. vitis in the field. VAR03-1 established populations averaging 10(6) colony forming units (CFU)/g of root in the rhizosphere of grapevine and persisted on roots for 2 years. VAR03-1 was bacteriocinogenic, producing a halo of inhibition against those three species of Agrobacterium. This is the first report that a nonpathogenic strain, VAR03-1, can effectively control crown gall caused by tumorigenic A. vitis, A. rhizogenes, and A. tumefaciens.  相似文献   

3.
Crown gall disease of tobacco was found in Iwate Prefecture, Japan in 1995. Ten bacterial isolates, obtained from the galls of tobacco, were identified as Agrobacterium tumefaciens (Smith and Townsend 1907) Conn 1942 biovar 1 based on their ability to induce galls on the 14 tested plants, including tobacco after needle-prick inoculation, and on 12 cultural, physiological, and biological characteristics. The growth of the causal organism was not inhibited in vitro by agrocin of A. radiobacter strain K84. This report is the first on the natural occurrence of crown gall caused by A. tumefaciens on tobacco plants.  相似文献   

4.
放射土壤杆菌MI15菌株生物防治葡萄根癌病的研究   总被引：6，自引：0，他引：6  
 从内蒙古玫瑰香葡萄冠瘿中分离到一株无致病力的土壤杆菌MI15菌株,经鉴定为放射土壤杆菌(Agrobacterium radiobacter),属生物Ⅰ型。在平皿培养产生的土壤杆菌素,能抑制葡萄根癌土壤杆菌生物Ⅲ型菌株的生长,在温室向日葵幼苗和葡萄嫩枝上接种,MI15菌株能显著地抑制多株不同类型葡萄根癌土壤杆菌诱导形成冠瘿瘤。在琼脂糖凝胶电泳图谱上,MI15菌株有一条质粒带,与生物Ⅰ型根癌士壤杆菌C58菌株的质粒情况基本相同。研究还表明该菌株能在葡萄上存活定殖。  相似文献   

5.
A biovar 3-specific primer set Ab3-F3/Ab3-R4 was designed based on the comparison of sequences of the 16S rDNA region of agrobacteria and related rhizobia for rapid identification of Agrobacterium biovar 3 strains. A 570-bp 16S rDNA fragment was amplified from cell lysates of Agrobacterium biovar 3 strains by polymerase chain reaction (PCR) using Ab3-F3/Ab3-R4 primers. Discrimination of Agrobacterium tumefaciens biovar 3 from Agrobacterium radiobacter biovar 3 and of Agrobacterium biovar 3 strains from other Agrobacterium strains was done simultaneously using multiplex PCR with a mixture of two primer sets (Ab3-F3/Ab3-R4 and VCF3/VCR3) previously designed for the virC region of Ti-plasmid and Ri-plasmid.  相似文献   

6.
Six strains of crown gall bacteria were isolated from flowering cherry. It was revealed by Otten paper electrophoresis that of the six strains, only BYH18-4 possessed the octopine type Ti plasmid, the remainder having nopaline type Ti plasmid. BYH5-1 was identified by physiological and biochemical tests to be Agrobacterium tumefaciens (originally biovar 1). The other five were A. rhizogenes (originally biovar 2). It was demonstrated with Stonier's method of double layer medium that flowering cherry crown gall bacteria exhibited different sensitivities to agrocin produced by biocontrol strain K1026. Strain K1026 on greenhouse-grown sunflower seedlings exerted a relatively potent inhibitory action on flowering cherry crown gall bacteria. Artificial inoculation showed that K1026 produced 67–99% inhibition of flowering cherry crown gall disease, compared with the treatment of inoculation with crown gall bacteria only.  相似文献   

7.
Biological control of Agrobacterium tumefaciens using the K84 strain of A. radiobacter has been practised in Spain since 1979. Strain K84 is produced in Sevilla by the Agricultural Research Service and is mixed with peat. This formulation is authorized for use on stone fruits and roses, since results of various trials on rootstocks of these crops have shown that K84 is effective in controlling the disease in naturally infected nursery soil, as well as in experimental tests carried out on artificially inoculated soil. Results of biological control trials have shown that K84 has some effect on A. tumefaciens strains resistant to agrocin 84. Factors other than susceptibility to agrocin 84 influence the efficacy of biological control. This was demonstrated by using a K84 agrocin non-producer strain which proved effective against both resistant and susceptible strains. Biological control on apple or pear is not yet authorized in Spain due to a paucity of data on field trials. However, initial pot tests showed good K84 activity on these hosts, against Spanish strains of A. tumefaciens. On grapevine, most of the A. tumefaciens strains that were isolated belong to biotype 3 and are resistant to agrocin 84. Because the disease is systemic on this species the use of K84 is not authorized.  相似文献   

8.
Crown gall caused by Agrobacterium tumefaciens is a problem in many European nurseries especially on stone fruits, apples and some herbaceous hosts (biovars 1 and 2) and on grapevines (biovar 3). Nursery infection can arise from land infested with the pathogen. Disease incidence can be reduced to some extent by cultural methods such as crop rotation, minimizing wounding and selection of less susceptible cultivars. The best control of infection is the use of a pre-planting dip in the antagonist A. radiobacter strain 84, which gives excellent control of strains of the pathogen harbouring the nopaline-type Ti plasmid. The antagonist is not yet cleared for use in all European countries. This biological control is sometimes ineffective because of the presence of mixed populations of sensitive and resistant strains of the pathogen, use of too low inoculum doses of the antagonist or because the host plant affects the antagonist action. All controls are preventive and not curative, so latent infections in layer or stool beds cannot be prevented from developing. Control of the strains of the pathogen insensitive to strain 84 may be possible in future by the use of other antagonistic organisms or by genetic manipulation of strain 84. Until such a new development becomes available, growers must make careful selection of propagating material to minimize the risk of spreading infection.  相似文献   

9.
Crown gall is the most important bacterial disease of grapevine and is primarily associated with Agrobacterium tumefaciens biovar 3. The systemic distribution of the agrobacteria in grapevine requires the checking of nursery propagation material to reveal their presence in the xylem vessels. The methods adopted to demonstrate systemic survival are quite lengthy and, thus, this study aimed to develop a serological sampling procedure to rapidly detect A. tumefaciens in xylem extracts, obtained by vacuum extraction with suitable washing fluid, from asymptomatic one year-old dormant grape cuttings. The suspension was centrifuged and the final concentrate used for immunofluorescence staining. The sensitivity threshold of the vacuum washing system and the effectiveness of the detection method were experimentally evaluated. The antiserum specificity was tested in a comparative study with a collection of A. tumefaciens strains from grapevine and other common grapevine contaminants. The vacuum washing system is discussed as an indexing tool in a much hoped for certification programme.  相似文献   

10.
Physiological, biochemical and molecular characteristics of Agrobacterium tumefaciens strains isolated in Italy from different host plants were analysed. Diseased plants were collected from several nurseries located in nine different regions. Out of 1293 strains isolated from 12 fruit tree and six ornamental plant species, a group of 120 strains was chosen as representative of the whole collection. The majority of the strains were biovar 2 (82.5%), agrocin 84 sensitive, and were isolated from stone fruit trees. Most of the strains identified as biovar 1 were isolated from ornamental plants and were insensitive to A. radiobacter antagonistic strain K84. Some strains that were isolated from Euonymus spp, Prunus GF 677 and Pyrus communis (pear) OHF tumours could not be allocated to any of the three Agrobacterium biovars. PCR-restriction fragment length polymorphism of the rrs gene plus the intergenic spacer was used for strain fingerprinting and characterisation. Results showed a wide genetic variability within the biovar 1 strains and homogeneity within the biovar 2 group. Biovar 2 strains from Sardinia were highly variable and differed from the biovar 2 strains isolated from the other regions of Italy.  相似文献   

11.
The polymerase chain reaction (PCR) is a rapid, precise method for detecting and identifying pathogenic bacteria. In addition to the published primers for identification of Agrobacterium tumefaciens up to species level, two sets of primers were designed to identify the nopaline and octopine types of Agrobacterium tumefaciens. The RBF-RBR primer set designed based on the nopaline type T-DNA right border detected the nopaline type A208 and R225f strains, and the ocsF-ocsR primer set derived from the ocs gene of the octopine type A. tumefaciens detected the octopine type A348 strain. After polymerase Chain reaction (PCR) amplification by the RBF-RBR primers, the A208 and R225f strains could be differentiated from each other by restriction fragment length polymorphism digestion using the restriction enzymes DraI and XbaI. Multiple colonies can be screened at one time in a single PCR tube with satisfactory efficiency, thereby allowing rapid detection of pathogenic A. tumefaciens. Following a rough screening by classical biovar medium and -methyl-d-glucoside medium, the developed PCR system was introduced to identify isolates collected from soil and crown gall samples. Of 42 isolates determined to be A. tumefaciens, 7 were found to be octopine type; all the rest were R225f type.  相似文献   

12.
In 2005, characteristic symptoms of crown gall on grapevines (Vitis vinifera L. cv. Muscat of Alexandria, and cv. Seto Giants) were observed in a commercial greenhouse-orchard in Okayama Prefecture, Japan. Isolations from diseased tissues consistently yielded bacterial colonies that were white, glistening, and produced abundant polysaccharide on potato semi-synthetic agar (PSA) medium. Ten representative isolates were chosen for further characterization. A multiplex polymerase chain reaction (PCR) assay showed these strains were not Rhizobium vitis but did possess a Ti plasmid. The bacteriological characteristics of the isolates corresponded well with R. radiobacter. The almost complete 16S ribosomal DNA sequences of isolates AT06-1 and AT06-2, selected from 10 grapevine isolates, were determined and corresponded to sequences of R. radiobacter. The pathogenicity of the isolates was tested on young grapevine and tomato (Lycopersicon esculentum Mill.) plants. Gall symptoms developed on both plant species after inoculation, and bacteria with the same colony morphology as those inoculated were reisolated. Based on these results, the isolates were identified as R. radiobacter (Ti). This report is the first of the occurrence of R. radiobacter (Ti) on grapevine in Japan. Phylogenetic analyses using the partial nucleotide sequences of virC operon located on a Ti plasmid showed that the isolate of R. radiobacter (Ti) isolated from grapevine and some strains of R. vitis (Ti) belonged to the same monophyletic group, which differed from the groups of R. radiobacter (Ti) isolated from plants other than grapevine and of the majority of R. vitis (Ti) strains isolated from grapevine. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accessions AB306890, AB306891, and AB465432–AB465459.  相似文献   

13.
Crown gall was previously reported on grape in Israel but the pathogen was not isolated and characterized. The three recognized biovars ofAgrobacterium tumefaciens can be tumorigenic on grape, but biovar 3 is the most important world wide. A single occurrence of tumors in a vineyard yielded bacteria which incited galls on grape,Nicotiana glauca and tomato, but not on bryophyllum. The bacteria were confirmed asA. tumefaciens because they contained DNA which hybridized with T-DNA from a Ti plasmid. Biochemical and physiological tests, octopine production and utilization, and agrocin 84 insensitivity conformed with those of bv. 3. Subsequent occurrences of the grape disease have not been found, but the presence ofA. tumefaciens bv. 3 in Israel is a potential threat to nurseries and vineyards.  相似文献   

14.
啤酒花根癌病在我国发生普遍,是影响啤酒花生产的重要病害。从啤酒花主要生产基地新疆、山东、浙江等地分离到61个菌株。通过培养性状、染色反应,以及14～15个菌株的生理生化性状和12个菌株致病性的测定,确定啤酒花根癌病病原菌为根癌土壤杆菌Agrobacteria tumefaciens(E.F.Smith &Jownsend)Conn属生物型1、病原菌菌株对K84(Agrobacterium radiobacter)菌株的细菌素敏感,各菌株间敏感性的强弱不一。盆裁番茄和啤酒花初步生防试验表明,用K84与敏感性强的菌株按1:1比例混合接种,番茄和啤酒花都不产生瘿瘤;而敏感性弱的菌株需用3:1才能防止瘿瘤的产生。  相似文献   

15.
Crown gall disease of cultivated plants, caused by Agrobacterium tumefaciens, constitutes a serious problem for the fruit tree, rose and grapevine nurseries in Greece. All three biotypes of A. tumefaciens exist in Greece. Biotypes 1 and 2 have a wide host range being responsible for the disease on fruit trees and roses while biotype 3 isolates have a narrow host range infecting grapevine only. All Greek isolates of biotype 1 and all but 3 isolates of biotype 2 were sensitive to biological control with the antagonistic bacterium A. radiobacter strain K84, but the biotype 3 isolates were insensitive to biocontrol. Experiments on the effectiveness of the method in artificial infections of seedlings as well as in naturally infested soils showed that the method is effective and can be applied without any risk of development of forms insensitive to biocontrol. The use of a lyophilized preparation of K84 with skim milk as suspending medium is recommended. The lyophilized antagonistic bacterium retains its activity and the final concentration (106 cfu ml-) is adequate to protect treated plants from crown gall.  相似文献   

16.
A nonpathogenic strain of Rhizobium vitis, VAR03-1, was tested as a biological control agent for crown gall of apple. When apple tree roots were soaked in a cell suspension of antagonists before planting in the field, treatment with VAR03-1 significantly reduced the number of plants with tumors and the disease severity. The inhibitory effects of treatment with VAR03-1 and the nonpathogenic R. rhizogenes strain K84, which is commercially used as a biological control agent throughout the world, on apple crown gall were almost identical. VAR03-1 was bacteriocinogenic, producing a halo of inhibition against two species of tumorigenic Rhizobium isolated from galled apple trees. Moreover, VAR03-1 established populations on roots of apple tree rootstock and persisted on roots for 437?days. This is the first report that the nonpathogenic R. vitis VAR03-1 effectively controls crown gall of apple.  相似文献   

17.
Phialophora gregata f. sp. adzukicola, a causal agent of brown stem rot in adzuki beans, produces phytotoxic compounds: gregatins A, B, C, D, and E. Gregatins A, C, and D cause wilting and vascular browning in adzuki beans, which resemble the disease symptoms. Thus, gregatins are considered to be involved in pathogenicity. However, molecular analyses have not been conducted, and little is known about other pathogenic factors. We sought to isolate nonpathogenic and gregatin-deficient mutants through Agrobacterium tumefaciens-mediated transformation (ATMT) for cloning of pathogenicity-related genes. The co-cultivation of P. gregata and A. tumefaciens for 48 h at 20°C with 200 μM acetosyringone resulted in approximately 80 transformants per 106 conidia. The presence of acetosyringone in the A. tumefaciens pre-cultivation period led to an increase in T-DNA copy number per genome. Of 420 and 110 transformants tested for their pathogenicity and productivity of gregatins, one nonpathogenic and three gregatin-deficient mutants were obtained, respectively. The nonpathogenic mutant produced gregatins, whereas the gregatin-deficient mutants had pathogenicity comparable to the wild-type strain. This is the first report of ATMT of P. gregata. Further analysis of these mutants will help reveal the nature of the pathogenicity of this fungus including the role of gregatin in pathogenesis.  相似文献   

18.
Strain K84 of Agrobacterium (formerly called A. radiobacter) has been a successful biocontrol agent of crown gall disease for almost 30 years all over the world. In spite of its demonstrated efficiency, the most important risk of failure when using strain K84 is the possibility of transfer of plasmid pAgK84 to pathogenic Agrobacterium strains. pAgK84 codifies production of and immunity to agrocin 84, the main factor involved in crown gall biocontrol by strain K84. Then, a second generation of strain K84 was obtained and the genetically engineered strain was called K1026. It contains a deletion in the transfer region of pAgK84. To date, a considerable number of studies have been performed to compare both strains in its ability to control crown gall, plasmid transfer, antibiotic production, root colonization and survival in the rhizosphere. The aim of this review is to discuss all this comparative available information which advises that strain K1026 should be used as a biopesticide to safeguard biocontrol of crown gall wherever strain K84 is employed.  相似文献   

19.
In the Sidi M’djahed nursery (Algeria), over 60,000 eucalyptus (Eucalyptus occidentalis) plantlets exhibited tumour-like growths localized at the crown of the plants that resembled crown galls caused by Agrobacterium tumefaciens. Bacteria colonizing the galls were isolated and purified. Most (22 out of 24) of the isolates had cultural and biochemical characteristics similar to those of strains of the biovar 1 of A. tumefaciens. Twenty out of 22 Agrobacterium isolates induced tumour formation on various test plants. In PCR experiments, DNA extracted from these virulent strains yielded an amplification signal corresponding to a 247-bp fragment located within the virulence region of nopaline type Ti plasmid. Consistent with this, the opine nopaline was detected in the tumours induced on test plants – but not on eucalyptus plants. Nopaline was degraded by the 20 pathogenic isolates that were also sensitive to agrocin 84, indicating the presence of a nopaline-type pTi in these strains. The chromosomal region encoding the 16S rRNA was analyzed in a sub-population of the pathogenic agrobacterial isolates. The analyzed strains were found to belong to the ribogroup of the reference strain B6. Interestingly, Eucalyptus camaldulensis and Eucalyptus cladocalyx grown in the same nursery and in the same soil substrate developed no galls.  相似文献   

20.
Crown gall disease of grapevine, caused by Agrobacterium tumefaciens, often results in severe economic loss to grape production worldwide. This study demonstrated the ability of the endophytic bacteria Pseudomonas sp. Sn48 and Pantoea sp. Sa14 isolated from domesticated and wild grapevines to induce resistance in both above- and belowground tissues of grapevines infected with A. tumefaciens. Our results provide evidence that both strains can colonize roots and/or shoots. We showed that the strains Pseudomonas sp. Sn48 and Pantoea sp. Sa14 are capable of inducing stilbenic phytoalexin production in grapevine tissues and to further prime plantlets for enhanced phytoalexin production after A. tumefaciens inoculation. We also showed that in the majority of treatments, polyamine accumulation remained unchanged or slightly increased in plantlets treated with Pseudomonas sp. Sn48 and Pantoea sp. Sa14 compared with the control. Our findings indicated that the levels of polyamines remain unchanged or significantly decrease in plantlets treated with endophytic bacteria after A. tumefaciens challenge compared to the control and plantlets treated with individual endophytic bacterial strains. PR1, PR2, and PR4 gene expression levels of plantlets treated with Pseudomonas sp. Sn48 and Pantoea sp. Sa14 significantly increased after A. tumefaciens inoculation. The findings revealed the efficacy of the selected endophytic bacteria in triggering grapevine resistance against A. tumefaciens and the possible use of these strains as an alternative to chemical control methods in grapevine crown gall disease management.  相似文献   

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