Elimination of the Crown Gall Pathogen, Agrobacterium vitis, from Systemically Infected Grapevines by Tissue Culture

ABSTRACTCrown gall disease of grapevine is caused by virulentAgrobacteriumstrains and establishes a suitable habitat for agrobacteria and, potentially, other bacteria. The microbial community associated with grapevine plants has not been investigated with respect to this disease, which frequently results in monetary losses. This study compares the endophytic microbiota of organs from grapevine plants with or without crown gall disease and the surrounding vineyard soil over the growing seasons of 1 year. Amplicon-based community profiling revealed that the dominating factor causing differences between the grapevine microbiota is the sample site, not the crown gall disease. The soil showed the highest microbial diversity, which decreased with the distance from the soil over the root and the graft union of the trunk to the cane. Only the graft union microbiota was significantly affected by crown gall disease. The bacterial community of graft unions without a crown gall hosted transient microbiota, with the three most abundant bacterial species changing from season to season. In contrast, graft unions with a crown gall had a higher species richness, which in every season was dominated by the same three bacteria (Pseudomonassp.,Enterobacteriaceaesp., andAgrobacterium vitis). Forin vitro-cultivated grapevine plantlets,A. vitisinfection alone was sufficient to cause crown gall disease. Our data show that microbiota in crown galls is more stable over time than microbiota in healthy graft unions and that the microbial community is not essential for crown gall disease outbreak.IMPORTANCEThe characterization of bacterial populations in animal and human diseases using high-throughput deep-sequencing technologies, such as 16S amplicon sequencing, will ideally result in the identification of disease-specific microbiota. We analyzed the microbiota of the crown gall disease of grapevine, which is caused by infection with the bacterial pathogenAgrobacterium vitis.All otherAgrobacteriumspecies were found to be avirulent, even though they lived together withA. vitisin the same crown gall tumor. As has been reported for human cancer, the crown gall tumor also hosted opportunistic bacteria that are adapted to the tumor microenvironment. Characterization of the microbiota in various diseases using amplicon sequencing may help in early diagnosis, to serve as a preventative measure of disease in the future.

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Tartrate Utilization Genes Promote Growth of Agrobacterium spp. on Grapevine

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi.1998.11.8.836 ◽

1998 ◽

Vol 11 (8) ◽

pp. 836-838 ◽

Cited By ~ 12

Author(s):

J.-Y. Salomone ◽

E. Szegedi ◽

P. Cobanov ◽

L. Otten

Keyword(s):

Crown Gall ◽

Selective Advantage ◽

Agrobacterium Vitis

Crown gall on grapevine is mainly caused by Agrobacterium vitis, which metabolizes tartrate. Competition experiments between a tartrate-utilizing strain and its non-utilizing derivative showed that tartrate utilization confers a selective advantage on grapevine.

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The Impacts of Tumorigenic and Nontumorigenic Agrobacterium vitis Strains on Graft Strength and Growth of Grapevines

Plant Disease ◽

10.1094/pdis-07-17-0952-re ◽

2018 ◽

Vol 102 (2) ◽

pp. 375-381 ◽

Cited By ~ 1

Author(s):

Lingyun Hao ◽

David J. Kemmenoe ◽

Didem Canik Orel ◽

Thomas Burr

Keyword(s):

Crown Gall ◽

Shoot Growth ◽

Callus Formation ◽

Aqueous Suspension ◽

Water Control ◽

Agrobacterium Vitis ◽

Graft Union ◽

Time Points ◽

Graft Interface ◽

Grafting Procedure

The effects of tumorigenic and nontumorigenic strains of Agrobacterium vitis on graft strength and growth of grapevines was studied. A procedure was developed for inoculating graft interface surfaces with A. vitis and for measuring the force required to break grafts at different time points. Cuttings were soaked in an aqueous suspension of bacteria, about 106 CFU/ml, and bacteria were spread onto the graft interface during the grafting procedure. Tumorigenic strain CG49 caused reduced bud germination and increased callus (crown gall) at the graft union and at the base of cuttings at 30 days postinoculation (dpi) and significantly reduced shoot growth by 60 dpi whereas, at the same time points, nontumorigenic strain F2/5 inhibited callus formation but did not affect bud germination or shoot growth. Graft strength was enhanced at 30 dpi with CG49, presumably because the crown gall callus served to secure the union; graft strength was weakened by F2/5 over the same period. Between 30 and 60 dpi, the greatest increase in graft strength was observed in the water control. Following graft union inoculations, the A. vitis population increased more than 1,000-fold within 5 days.

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Comparative study of diagnostic methods used for monitoring of common grape vine (Vitis vinifera L.) crown gall (Agrobacterium vitis Ophel & Kerr) in Slovenia

Acta agriculturae Slovenica ◽

10.14720/aas.2014.103.2.16 ◽

2014 ◽

Vol 103 (2) ◽

pp. 313-321

Author(s):

Janja LAMOVŠEK ◽

Igor ZIDARIČ ◽

Irena MAVRIČ PLEŠKO ◽

Gregor UREK ◽

Stanislav TRDAN

Keyword(s):

Comparative Study ◽

Vitis Vinifera ◽

Crown Gall ◽

Diagnostic Methods ◽

Vitis Vinifera L ◽

Agrobacterium Vitis ◽

Grape Vine

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Biological control of grapevine crown gall by nonpathogenic Agrobacterium vitis strain VAR03-1

Journal of General Plant Pathology ◽

10.1007/s10327-006-0327-3 ◽

2007 ◽

Vol 73 (2) ◽

pp. 133-138 ◽

Cited By ~ 18

Author(s):

Akira Kawaguchi ◽

Koji Inoue ◽

Hideo Nasu

Keyword(s):

Biological Control ◽

Crown Gall ◽

Agrobacterium Vitis ◽

Grapevine Crown Gall

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Biological Control of Grape Crown Gall by Rahnella aquatilis HX2

Plant Disease ◽

10.1094/pdis-91-8-0957 ◽

2007 ◽

Vol 91 (8) ◽

pp. 957-963 ◽

Cited By ~ 43

Author(s):

F. Chen ◽

Y. B. Guo ◽

J. H. Wang ◽

J. Y. Li ◽

H. M. Wang

Keyword(s):

Biological Control ◽

Cell Suspension ◽

Crown Gall ◽

Biological Control Agent ◽

Disease Incidence ◽

Significant Inhibition ◽

Agrobacterium Vitis ◽

Rahnella Aquatilis ◽

Inhibition Effect ◽

Significant Difference

Crown gall induced by Agrobacterium vitis is a worldwide plant disease in grape-growing regions. Rahnella aquatilis HX2, a new isolate from vineyard soil in Beijing, showed a significant inhibition effect on the development of crown galls in grapevines. In field trials, immersion of the basal ends of grape cuttings with HX2 cell suspension inhibited or completely prevented crown gall formation caused by A. vitis K308 in the roots of the plants from the cuttings. The 3-year average disease incidence in grape plants treated with HX2 was 30.8% compared to 93.5% in plants without HX2. The culture supernatant of HX2 exhibited a stronger inhibition effect on disease development than did the cell suspension. HX2 could be found in the grape rhizosphere, grown under field conditions, for up to 90 days after inoculation. There was no significant difference in the mean population sizes of root microflora between plants treated and not treated with HX2. The inhibition effect of HX2 on crown gall in sunflower, caused by different agrobacterial strains, varied between 30.7 and 100%, depending on strains. Our results showed that Rahnella aquatilis HX2 may be used as a biological control agent for crown gall disease of grapes.

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Two Novel Genomospecies in the Agrobacterium tumefaciens Species Complex Associated with Rose Crown Gall

Phytopathology ◽

10.1094/phyto-05-19-0178-r ◽

2019 ◽

Vol 109 (11) ◽

pp. 1859-1868 ◽

Cited By ~ 4

Author(s):

Hamzeh Mafakheri ◽

S. Mohsen Taghavi ◽

Joanna Puławska ◽

Philippe de Lajudie ◽

Florent Lassalle ◽

...

Keyword(s):

Crown Gall ◽

Species Complex ◽

Nucleotide Diversity ◽

Ornamental Plants ◽

Housekeeping Genes ◽

Phylogenetic Position ◽

Agrobacterium Vitis ◽

Phylogenetic Status ◽

Species Specific ◽

Allorhizobium Vitis

In this study, we explored the pathogenicity and phylogenetic position of Agrobacterium spp. strains isolated from crown gall tissues on annual, perennial, and ornamental plants in Iran. Of the 43 strains studied, 10 strains were identified as Allorhizobium vitis (formerly Agrobacterium vitis) using the species-specific primer pair PGF/PGR. Thirty-three remaining strains were studied using multilocus sequence analysis of four housekeeping genes (i.e., atpD, gyrB, recA, and rpoB), from which seven strains were identified as A. larrymoorei and one strain was identified as A. rubi (Rer); the remaining 25 strains were scattered within the A. tumefaciens species complex. Two strains were identified as genomospecies 1 (G1), seven strains were identified as A. radiobacter (G4), seven strains were identified as A. deltaense (G7), two strains were identified as A. nepotum (G14), and one strain was identified as “A. viscosum” (G15). The strains Rnr, Rnw, and Rew as well as the two strains OT33 and R13 all isolated from rose and the strain Ap1 isolated from apple were clustered in three atypical clades within the A. tumefaciens species complex. All but eight strains (i.e., Nec10, Ph38, Ph49, fic9, Fic72, R13, OT33, and Ap1) were pathogenic on tomato and sunflower seedlings in greenhouse conditions, whereas all but three strains (i.e., fic9, Fic72, and OT33) showed tumorigenicity on carrot root discs. The phylogenetic analysis and nucleotide diversity statistics suggested the existence of two novel genomospecies within the A. tumefaciens species complex, which we named “G19” and “G20.” Hence, we propose the strains Rew, Rnw, and Rnr as the members of “G19” and the strains R13 and OT33 as the members of G20, whereas the phylogenetic status of the atypical strain Ap1 remains undetermined.

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