The accelerated evolution and spread of pathogens are threats to host species. Agrobacteria require an oncogenic Ti or Ri plasmid to transfer genes into plants and cause disease. We developed a strategy to characterize virulence plasmids and applied it to analyze hundreds of strains collected between 1927 and 2017, on six continents and from more than 50 host species. In consideration of prior evidence for prolific recombination, it was surprising that oncogenic plasmids are descended from a few conserved lineages. Characterization of a hierarchy of features that promote or constrain plasticity allowed inference of the evolutionary history across the plasmid lineages. We uncovered epidemiological patterns that highlight the importance of plasmid transmission in pathogen diversification as well as in long-term persistence and the global spread of disease.

bThe improvement of the agricultural and wine-making qualities of the grapevine (Vitis vinifera) is hampered by adherence to traditional varieties, the recalcitrance of this plant to genetic modifications, and public resistance to genetically modified organism (GMO) technologies. To address these challenges, we developed an RNA virus-based vector for the introduction of desired traits into grapevine without heritable modifications to the genome. This vector expresses recombinant proteins in the phloem tissue that is involved in sugar transport throughout the plant, from leaves to roots to berries. Furthermore, the vector provides a powerful RNA interference (RNAi) capability of regulating the expression of endogenous genes via virus-induced gene-silencing (VIGS) technology. Additional advantages of this vector include superb genetic capacity and stability, as well as the swiftness of technology implementation. The most significant applications of the viral vector include functional genomics of the grapevine and disease control via RNAi-enabled vaccination against pathogens or invertebrate pests.

No abstract

Field experiments were conducted in silty-clay loam in Corvallis, OR during the summers of 1995 and 1996 to study the effects of green manure cover crops (Sudan grass, rape, and barley), soil solarization, soil fumigation, and combinations of those treatments on population densities of soil pathogens Verticillium dahliae, Phytophthora cinnamomi, Pratylenchus penetrans, and Agrobacterium rhizogenes. Nylon mesh bags containing soil infested with V. dahliae and Phytophthora cinnamomiwere buried 5, 10, 20, and 30 cm deep. Soil solarization was performed over a 54- to 59-day period using a 0.6-mil clear polyethylene film. Maximum soil temperatures recorded at depths of 5, 10, 20, and 30 cm were 53, 48, 39, and 34°C in solarized soil, respectively; these temperatures were 8 to 16°C higher than in corresponding nonsolarized plots. Soil samples were collected before, during, and after solarization to quantify pathogen populations at those four depths. Pot or field studies were conducted subsequent to treatments to determine the effects of treatments on susceptible plants. Soil solarization, cover crops plus solarization, or fumigation with metam sodium resulted in a significant decrease (P< 0.05) in density of P. cinnamomi populations at all four depths and reduced (P< 0.05) V. dahliae at 5 and 10 cm. In greenhouse assays of solarized soils, disease severity was reduced (P< 0.05) for Verticillium spp. on eggplant and Phytophthora spp. on snapdragons. Cover crops alone were not effective in reducing P. cinnamomi and V. dahliae populations. Agrobacterium spp. population densities declined within solarized plots and incidence of crown gall on ‘Mazzard’ cherry rootstock planted in solarized plots was reduced significantly. Population densities of Pratylenchus penetranswere reduced in the upper 30-cm soil profile by solarization.Solarization for an 8-week period during the warmest months of summer could provide an additional management alternative for several important soilborne pathogens in western Oregon.

The Ti plasmid in Agrobacterium tumefaciens strain 15955 carries two alleles of traR that regulate conjugative transfer. The first is a functional allele, called traR, that is transcriptionally induced by the opine octopine. The second, trlR, is a nonfunctional, dominant-negative mutant located in an operon that is inducible by the opine mannopine (MOP). Based on these findings, we predicted that there exist wild-type agrobacterial strains harboring plasmids in which MOP induces a functional traR and, hence, conjugation. We analyzed 11 MOP-utilizing field isolates and found five where MOP induced transfer of the MOP-catabolic element and increased production of the acyl-homoserine lactone (acyl-HSL) quormone. The transmissible elements in these five strains represent a set of highly related plasmids. Sequence analysis of one such plasmid, pAoF64/95, revealed that the 176-kb element is not a Ti plasmid but carries genes for catabolism of MOP, mannopinic acid (MOA), agropinic acid (AGA), and the agrocinopines. The plasmid additionally carries all of the genes required for conjugative transfer, including the regulatory genes traR, traI, and traM. The traR gene, however, is not located in the MOP catabolism region. The gene, instead, is monocistronic and located within the tra-trb-rep gene cluster. A traR mutant failed to transfer the plasmid and produced little to no quormone even when grown with MOP, indicating that TraR pAoF64/95 is the activator of the tra regulon. A traM mutant was constitutive for transfer and acyl-HSL production, indicating that the anti-activator function of TraM is conserved.