Search Results (25)

Grapevine crown gall (GCG) is a major disease caused by the Allorhizobium vitis that causes significant losses in the grape industry. In this study, a strain of Agrobacterium tumefaciens (Ag-8) from soil was found to have a significant effect on grapevine crown gall. The present study was conducted to investigate the potential impact of the AraC family transcription factor ata (Agrobacterium tumefaciens AraC) on the biocontrol capacity of Ag-8 and to elucidate the potential mechanisms of its action. To this end, an ata deletion mutant (Δata) was constructed. It was found that the biofilm, motility, and colonization of the ata deletion mutant strain were significantly lower than those of the wild strain Ag-8. By analyzing the Δata transcriptome data, we focused our attention on the ABC transporter protein araH, and RT-qPCR showed that it was significantly down-regulated in the Δata strain. In a follow-up study, the candidate gene araH deletion strain (ΔaraH) was found to have significantly lower biofilm formation, motility, and biocontrol ability than the wild-type strain Ag-8. Therefore, araH may be implicated as a pivotal target gene of ata in modulating the biocontrol capability of Ag-8. This study supports developing biocontrol strategies targeting ata-regulated colonization in Agrobacterium to enhance the sustainable management of grapevine crown gall. Full article

Beneficial bacteria are recognised for their antimicrobial compounds, making them valuable for disease control in agriculture. Bacillus species stand out for their stability, versatility, and selectivity as biocontrol agents. This study aimed to identify potential antagonists within the rhizosphere microorganisms by isolating bacterial strains from grapevine roots and rhizosphere soil in Moroccan vineyards. The antimicrobial activities of these isolates against Allorhizobium vitis, the causative agent of grapevine crown gall, were evaluated in vitro using a disc diffusion assay, followed by in planta assessments under preventive and simulated inoculation conditions. Screening led to the isolation of 123 strains, with six showing strong antagonistic properties, achieving inhibition percentages up to 39.6%. 16S rRNA sequencing led to identifying five Bacillus species: B. amyloliquefaciens, B. velezensis, B. halotolerans, B. subtilis, and B. anthracis. These strains were further characterised by their biochemical traits and plant growth-promoting abilities. Compatibility assays identified optimal combinations for microbial consortia, demonstrating pathogen inhibition up to 37.4%. In planta bioassays confirmed the effectiveness of the isolates and consortia, reducing tumour size. These findings highlight the potential of these Bacillus strains as biocontrol agents and underscore the value of microbial consortia as a sustainable approach to managing grapevine crown gall. Full article

Infection of higher plants with agrobacteria (Agrobacterium tumefaciens) represents one of the most comprehensively characterized examples of plant–microbial interactions. Incorporation of the bacterial transfer DNA (T-DNA) in the plant genome results in highly efficient expression of the bacterial auxin, cytokinin and opine biosynthesis genes, as well as the host genes of hormone-mediated signaling. These transcriptional events trigger enhanced proliferation of plant cells and formation of crown gall tumors. Because of this, infection of plant tissues with A. tumefaciens provides a convenient model to address the dynamics of cell metabolism accompanying plant development. To date, both early and late plant responses to agrobacterial infection are well-characterized at the level of the transcriptome, whereas only little information on the accompanying changes in plant metabolism is available. Therefore, here we employ an integrated proteomics and metabolomics approach to address the metabolic shifts and molecular events accompanying plant responses to inoculation with the A. tumefaciens culture. Based on the acquired proteomics dataset complemented with the results of the metabolite profiling experiment, we succeeded in characterizing the metabolic shifts associated with agrobacterial infection. The observed dynamics of the seedling proteome and metabolome clearly indicated rearrangement of the energy metabolism on the 10th day after inoculation (d.a.i.). Specifically, redirection of the energy metabolism from the oxidative to the anaerobic pathway was observed. This might be a part of the plant’s adaptation response to tumor-induced hypoxic stress, which most likely involved activation of sugar signaling. Full article

Crown gall disease (CGD), caused by Agrobacterium tumefaciens, is a common plant disease that leads to significant economic losses. Biological control is a sustainable and scalable method for managing CGD. In this study, we isolated three Bacillus strains from the rhizosphere soil of healthy cherry trees and investigated their biocontrol activities and the underlying mechanisms against CGD of cherry trees. The results demonstrate that the three Bacillus strains can effectively inhibit the growth of the pathogenic A. tumefaciens strain XYT58 in vitro under different culture conditions. The pot experiments showed that the three strains could prevent CGD in cherry seedlings. Using PCR amplification, we identified the genes responsible for the synthesis of difficidin, macrolactin, and bacilysin in the three strains. In addition, inoculation with strains WY66 and WY519 significantly enhanced the expression of JA, ET, and SA pathway-related genes in cherry plants. The presence of antibiotic synthesis-related genes in the Bacillus strains and the trigger of plant ISR may explain their ability to control CGD in cherry trees. The findings of this study provide a theoretical basis for the application and development of plant growth-promoting rhizobacteria Bacillus strains in the control of CGD. Full article

Crown gall disease in plants is caused by “Agrobacteria”, bacteria belonging to the Rhizobiaceae family, which carry a tumor-inducing (Ti) plasmid. Unexpectedly, we found evidence that a natural isolate from a rose crown gall, called NBC51/LBA8980, was a bacterium that did not belong to the Rhizobiaceae family. Whole-genome sequencing revealed that this bacterium contained three large DNA circles with rRNA and tRNA genes, representing one chromosome and two chromids, respectively, and two megaplasmids, including a Ti plasmid. Average nucleotide identity (ANIb, ANIm) and genome-to-genome distance (GGDC) values above the thresholds of 96% and 70%, respectively, showed that NBC51/LBA8980 belonged to the species Brucella (Ochrobactrum) pseudogrignonense. Its Ti plasmid was almost identical to certain succinamopine Ti plasmids previously identified in Agrobacterium strains, suggesting that this Ti plasmid may have been recently acquired by NBC51/LBA8980 in the tumor environment. Full article

Sakura crown gall, caused by the invasion of Agrobacterium tumefaciens through plant wounds, poses a significant threat to cherry trees. In this study, the distribution of A. tumefaciens was preliminarily determined by stratified sampling and qPCR detection. Vertically, the pathogen is mainly distributed in the soil layer below 20 cm, and the amount of bacteria increases at greater depths. Horizontally, they are found within a 150 cm radius from the trunk. Zhongshengmycin and Oligosaccharide–Ethylicin were applied 100 cm from the trunk at a depth of below 20 cm. In the 20~40 cm soil layer, a 600-fold diluted solution of 3% Zhongshengmycin had a relative control efficacy of 94% to 100% against Agrobacterium tumefaciens, while a 1000-fold diluted solution of 25% Oligosaccharide–Ethylicin showed a control effect ranging from 54% to 100%. Before transplantation, the soil was disinfected with dazomet and abamectin. Application rates were 35 g/m² for dazomet (98% granules) and 1 mL/m² for abamectin (1.8% emulsifiable concentrate). The disinfection effectiveness was 77~100% in the 0~60 cm soil layer. Full article

Crown gall is a soil-borne bacterial disease caused by Agrobacterium tumefaciens, leading to significant economic losses in many plant species. For the assessment of the biological and chemical products on crown gall, each plant’s crown region and roots were wounded, and then were dipped into their respective treatments. After the treatments, the plants were inoculated with a suspension of pathogenic A. tumefaciens isolate FBG1034 and maintained in a greenhouse for six months to assess them for gall formation. A quantitative real-time PCR assay was performed to quantify the A. tumefaciens using the chvE gene. Biological products such as the Agrobacterium radiobacter strain K1026, and strains 1 and 2, resulted in the lowest average root gall diameter and significantly reduced the crown gall diameter to stem diameter ratio, and the chemical product copper octanoate reduced the number of crown and root galls as well as the crown and root gall diameter compared to the inoculated, non-treated control. Moreover, both the A. radiobacter strain K1026 and strain 1 treatments resulted in an approximately 85% and 65% reduction in crown and root gall incidence, respectively, in both of the trials compared to the inoculated, non-treated plants. The findings of this study indicate that the use of biological and chemical products could help to suppress crown and root gall disease in rose plants. Full article

Alberta’s forest industry is predicted to be impacted by a medium-term decline in timber supply. Intensive silviculture tools, such as pre-commercial thinning, have been shown to increase individual tree growth, shorten rotation lengths, and improve stand merchantability in important commercial species such as lodgepole pine. However, lodgepole pine stands are susceptible to western gall rust infections, and thinning at an early stage may increase infection rates. This study collected tree and stand level data from 33 operational harvest origin lodgepole pine stands consisting of 11 stands thinned at age 17–19 years (PCT_18), 11 stands thinned at age 23–25 (PCT_24), and 11 unthinned stands. Approximately 40 years after pre-commercial thinning, merchantable volume is similar in all stands but thinned stands, regardless of timing, had greater individual tree size (~15% higher) compared to unthinned stands. Pre-commercially thinned stands also had a higher potential for commercial thinning since they have lower variability in tree size and longer live crown lengths. In addition, delayed thinning (PCT_24) reduced western gall rust infections and the severity of infections compared to both PCT_18 and unthinned stands. In conclusion, pre-commercial thinning should be considered for lodgepole pine stands in order to address timber supply issues in Alberta. Full article

In August 2020, the unusual appearance of crown gall symptoms was observed on the tobacco plants (hybrid PVH2310) grown in fields in the Golubinci (Srem district, Serbia) locality. The causal agent isolated from galls located on tobacco roots formed circular, convex, and glistening light blue colonies, and then dark to olive-green-colored bacterial colonies on a semi-selective D1 medium. Molecular analysis based on multiplex PCR and multi-locus sequence analysis (MLSA) using concatenated sequences of the atpD, dnaK, glnA, and rpoB genes as well as 16S rRNA identified Serbian tobacco isolates such as Agrobacterium tumefaciens (biovar 1). Two duplex PCR methods confirmed the presence of the virD2 and virC genes in tobacco isolates. Pathogenicity tests performed on carrot discs and squash fruits resulted in tumor/gall formation after 12 to 16 days post inoculation, respectively. Pathogenicity was also confirmed on tobacco plants, where isolates caused tumor development 21−25 days after inoculation. API 50 CH generated results regarding the biochemical features of the Serbian tobacco isolates. As A. tumefaciens (biovar 1) as a cause of tobacco crown gall has previously been documented solely in Japan, there is presently no data on its wider occurrence. Therefore, this first detailed investigation of A. tumefaciens isolated from naturally infected tobacco in Serbia will contribute to a better understanding of it at the global level. Full article

Crown gall disease (Agrobacterium tumefaciens), crown/root rot disease (Phytophthora spp.), root lesion disease (Pratylenchus vulnus) and tree vigor are key traits affecting the productivity and quality of walnuts in California. Unchallenged hybrid rootstocks were analyzed by RNA-seq to examine pre-formed factors affecting these traits. Enrichment analysis of the differentially expressed genes revealed that the increased expression of cell wall biogenesis-related genes plays a key role in susceptibility to A. tumefaciens, susceptibility to Phytophthora spp. and increased vigor. Analysis of the predicted subcellular loci of the encoded proteins revealed that many gene products associated with vigor and susceptibility were targeted to the plasma membrane and extracellular space, connecting these traits to sustaining barrier function. We observed that RNA processing and splicing, along with predicted nuclear targeting, were associated with resistance to A. tumefaciens, resistance to Phytophthora spp. and low vigor. Four genes within the J. microcarpa QTL region for resistance to A. tumefaciens and Phytophthora spp. were represented among our transcripts, with two of the genes being differentially expressed in association with resistance to A. tumefaciens and decreased vigor. No differential expression related to Phytophthora spp. or P. vulnus resistance was observed in this region. Additionally, the J. microcarpa haplotype expressed more transcripts associated with resistance to A. tumefaciens, Phytophthora spp. and low vigor, but not P. vulnus, than the J. regia haplotype. We also report unique and shared hormone and defense responses associated with each trait. This research suggests a link between cell wall biogenesis, vigor and critical root diseases of walnut. Full article

Agrobacterium (Rhizobium)-mediated transformation leads to the formation of crown galls or hairy roots on infected plants. These effects develop due to the activity of T-DNA genes, gathered on a big plasmid, acquired from agrobacteria during horizontal gene transfer. However, a lot of plant species are known to contain such sequences, called cellular T-DNAs (cT-DNAs), and maintain normal phenotypes. Some of the genes remain intact, which leads to the conclusion of their functional role in plants. In this study, we present a comprehensive analysis of the cT-DNAs in the Nicotiana noctiflora Hook. genome, including gene expression and opine identification. Deep sequencing of the Nicotiana noctiflora genome revealed the presence of two different cT-DNAs, NnT-DNA1 and NnT-DNA2, which contain the intact genes iaaM, iaaH, acs, orf13, orf13a, and orf14. According to the expression analysis results, all these genes are most active in roots in comparison with other organs, which is consistent with data on cT-DNA gene expression in other plant species. We also used genetic engineering approaches and HPTLC and HPLC-MS methods to investigate the product of the acs gene (agrocinopine synthase), which turned out to be similar to agrocinopine A. Overall, this study expands our knowledge of cT-DNAs in plants and brings us closer to understanding their possible functions. Further research of cT-DNAs in different species and their functional implications could contribute to advancements in plant genetics and potentially unveil novel traits with practical applications in agriculture and other fields. Full article

Grapevine crown gall (GCG), which is caused by Allorhizobium vitis (=Rhizobium vitis) tumorigenic strains, is the most important disease of grapevine around the world. Previously, nonpathogenic A. vitis strains VAR03-1, ARK-1, ARK-2, and ARK-3 were identified as promising biological control agents, but the control effects of each strain were not directly compared and assessed in the field because field trials were conducted in different fields and years. Thus, the results of the control effects obtained from 16 field trials in 12 years from 2006 to 2017 were analyzed and evaluated by a linear mixed model (LMM) and a network meta-analysis (NMA). The results of the LMM strongly indicate that the factor “antagonistic strain” was significantly related to the biological control activity in this study, but the other factors, “concentration of cell suspension”, “field”, and “year”, were not. Then, the results of 16 field trials were combined in an NMA. The estimated relative risk (RR) after treatment with ARK-1, ARK-2, ARK-3, VAR03-1, and K84 were 0.16, 0.20, 0.22, 0.24, and 0.74, respectively. In conclusion, strain ARK-1 was the best antagonist regardless of the concentration of the cell suspension, field, and year differences, and it can be recommended to control GCG. Full article

Grapevine crown gall (GCG), which is caused by tumorigenic Allorhizobium vitis (=Rhizobium vitis), is the most important bacterial disease in grapevine, and its economic impact on grapevine is very high. When young vines develop GCG, they often die, whereas older vines may show stress and poor growth depending on the severity of GCG, because GCG interferes with the vascular system of the grapevine trunk and prevents nutrient flow, leading to inferior growth and death. Viticultural practices and chemical control designed to inhibit GCG are only partially effective presently; thus, a biocontrol procedure could be a desirable and effective approach for GCG prevention. This article reviews the practical use of biocontrol options for GCG inhibition that involve using nonpathogenic and antagonistic A. vitis strains. In these studies, screening tests of biocontrol agents discovered nonpathogenic A. vitis strains VAR03-1, ARK-1, ARK-2, and ARK-3. After dipping grapevine roots in a suspension of candidate strains prior to planting in the field, treatment using ARK-1 was shown to significantly reduce the number of plants with GCG. A meta-analysis indicated that ARK-1 is very useful for controlling crown gall in various plant species, including grapevine. It was reported that when a mixture of ARK-1 and a tumorigenic strain was examined in grapevines, the expression levels of several virulence genes of the virulent strain were significantly lower. ARK-1 can reduce the pathogen population in grapevines and gall incidence. Moreover, ARK-1 can prime the induction of certain defense genes of grapevine. These results indicate that ARK-1 has a unique biocontrol mechanism and that it is a promising new biocontrol agent to control GCG. Full article

Agrobacterium tumefaciens underlies the pathogenesis of crown gall disease and is characterized by tumor-like gall formation on the stems and roots of a wide variety of economically important plant species. The bacterium initiates infection by colonizing and forming biofilms on plant surfaces, and thus, novel compounds are required to prevent its growth and biofilm formation. In this study, we investigated the ability of tannic acid, which is ubiquitously present in woody plants, to specifically inhibit the growth and biofilm formation of A. tumefaciens. Tannic acid showed antibacterial activity and significantly reduced the biofilm formation on polystyrene and on the roots of Raphanus sativus as determined by 3D bright-field and scanning electron microscopy (SEM) images. Furthermore, tannic acid dose-dependently reduced the virulence features of A. tumefaciens, which are swimming motility, exopolysaccharide production, protease production, and cell surface hydrophobicity. Transcriptional analysis of cells (Abs600 nm = 1.0) incubated with tannic acid for 24 h at 30 °C showed tannic acid most significantly downregulated the exoR gene, which is required for adhesion to surfaces. Tannic acid at 100 or 200 µg/mL limited the iron supply to A. tumefaciens and similarly reduced the biofilm formation to that performed by 0.1 mM EDTA. Notably, tannic acid did not significantly affect R. sativus germination even at 400 µg/mL. The findings of this study suggest that tannic acid has the potential to prevent growth and biofilm formation by A. tumefaciens and thus infections resulting from A. tumefaciens colonization. Full article

Kiwifruit worldwide suffers from the devastating diseases of bacterial canker caused by Pseudomonas syringae pv. actinidiae (Psa) and gray mold caused by Botrytis cinerea. Here, an endophytic bacterium XL17 isolated from a rape crown gall was screened out for its potent antagonistic activities against Psa and B. cinerea. Strain XL17 and its cell-free culture filtrate (CF) inhibited the growth of Psa and B. cinerea, Psa-associated leaf necrosis, and B. cinerea-associated kiwifruit necrosis. Electron microscopy showed that XL17 CF could damage the cell structures of Psa and B. cinerea. Genome-based taxonomy revealed that strain XL17 belongs to Pseudomonas bijieensis within the P. corrugata subgroup of the P. fluorescens species complex. Among the P. corrugata subgroup containing 31 genomospecies, the presence of the phl operon responsible for the biosynthesis of the phenolic polyketide 2,4-diacetylphloroglucinol (DAPG) and the absence of the lipopeptide/quorum sensing island can serve as the genetic marker for the determination of a plant-protection life style. HPLC detected DAPG in extracts from XL17 CF. MALDI-TOF-MS analysis revealed that strain XL17 produced cyclic lipopeptides of the viscosin family and orfamide family. Together, phenotypic, genomic, and metabolic analyses identified that P. bijieensis XL17 producing DAPG and cyclic lipopeptides can be used to control bacterial canker and gray mold pathogens of kiwifruit. Full article