Footprints of parasitism in the genome of the parasitic flowering plant Cuscuta campestris.

A parasitic lifestyle, where plants procure some or all of their nutrients from other living plants, has evolved independently in many dicotyledonous plant families and is a major threat for agriculture globally. Nevertheless, no genome sequence of a parasitic plant has been reported to date. Here we describe the genome sequence of the parasitic field dodder, Cuscuta campestris. The genome contains signatures of a fairly recent whole-genome duplication and lacks genes for pathways superfluous to a parasitic lifestyle. Specifically, genes needed for high photosynthetic activity are lost, explaining the low photosynthesis rates displayed by the parasite. Moreover, several genes involved in nutrient uptake processes from the soil are lost. On the other hand, evidence for horizontal gene transfer by way of genomic DNA integration from the parasite's hosts is found. We conclude that the parasitic lifestyle has left characteristic footprints in the C. campestris genome.

Getting ready for host invasion: elevated expression and action of xyloglucan endotransglucosylases/hydrolases in developing haustoria of the holoparasitic angiosperm Cuscuta.

Changes in cell walls have been previously observed in the mature infection organ, or haustorium, of the parasitic angiosperm Cuscuta, but are not equally well charted in young haustoria. In this study, we focused on the molecular processes in the early stages of developing haustoria; that is, before the parasite engages in a physiological contact with its host. We describe first the identification of differentially expressed genes in young haustoria whose development was induced by far-red light and tactile stimuli in the absence of a host plant by suppression subtractive hybridization. To improve sequence information and to aid in the identification of the obtained candidates, reference transcriptomes derived from two species of Cuscuta, C. gronovii and C. reflexa, were generated. Subsequent quantitative gene expression analysis with different tissues of C. reflexa revealed that among the genes that were up-regulated in young haustoria, two xyloglucan endotransglucosylase/hydrolase (XTH) genes were highly expressed almost exclusively at the onset of haustorium development. The same expression pattern was also found for the closest XTH homologues from C. gronovii. In situ assays for XTH-specific action suggested that xyloglucan endotransglucosylation was most pronounced in the cell walls of the swelling area of the haustorium facing the host plant, but was also detectable in later stages of haustoriogenesis. We propose that xyloglucan remodelling by Cuscuta XTHs prepares the parasite for host infection and possibly aids the invasive growth of the haustorium.

Cell wall glycoproteins at interaction sites between parasitic giant dodder (Cuscuta reflexa) and its host Pelargonium zonale.

The process of host plant penetration by parasitic dodder (genus Cuscuta) is accompanied by molecular and structural changes at the host/parasite interface. Recently, changes in pectin methyl esterification levels in the host cell walls abutting parasitic cells in established infection sites were reported. In addition to that, we show here that the composition of cell wall glycoproteins in Cuscuta-infected Pelargonium zonale undergoes substantial changes. While several arabinogalactan protein epitopes exhibit decreased abundances in the vicinity of the Cuscuta reflexa haustorium, extensins tend to increase in the infected areas.

Cell wall composition profiling of parasitic giant dodder (Cuscuta reflexa) and its hosts: a priori differences and induced changes.

Host plant penetration is the gateway to survival for holoparasitic Cuscuta and requires host cell wall degradation. Compositional differences of cell walls may explain why some hosts are amenable to such degradation while others can resist infection. Antibody-based techniques for comprehensive profiling of cell wall epitopes and cell wall-modifying enzymes were applied to several susceptible hosts and a resistant host of Cuscuta reflexa and to the parasite itself. Infected tissue of Pelargonium zonale contained high concentrations of de-esterified homogalacturonans in the cell walls, particularly adjacent to the parasite's haustoria. High pectinolytic activity in haustorial extracts and high expression levels of pectate lyase genes suggest that the parasite contributes directly to wall remodeling. Mannan and xylan concentrations were low in P. zonale and in five susceptible tomato introgression lines, but high in the resistant Solanum lycopersicum cv M82, and in C. reflexa itself. Knowledge of the composition of resistant host cell walls and the parasite's own cell walls is useful in developing strategies to prevent infection by parasitic plants.

Comparing chemical and biological control strategies for twospotted spider mites (Acari: Tetranychidae) in commercial greenhouse production of bedding plants.

Efficacy, costs, and impact on crop salability of various biological and chemical control strategies for Tetranychus urticae Koch (Acari: Tetranychidae) were evaluated on mixed plantings of impatiens, Impatiens wallerana Hook.f (Ericales: Balsaminaceae), and ivy geranium, Pelargonium peltatum (1.) L'Hér. Ex Aiton (Geraniales: Geraniaceae), cultivars in commercial greenhouses. Chemical control consisting of the miticide bifenazate (Floramite) was compared with two biological control strategies using the predatory mite Phytoseiulus persimilis Athias-Henriot (Acari: Phytoseiidae). Treatments were 1) a single, early application of bifenazate; 2) a single, early release of predatory mites at a 1:4 predator:pest ratio based on leaf samples to estimate pest density; 3) a weekly release of predatory mites at numbers based on the area covered by the crop; and 4) an untreated control. T. urticae populations were monitored for 3 wk after the earliest treatment. When plants were ready for market, their salability was estimated. Bifenazate and density-based P. persimilis treatments effectively reduced T. urticae numbers starting 1 wk after plants had been treated, whereas the scheduled, area-based P. persimilis treatment had little or no effect. The percentage of flats that could be sold at the highest market wholesale price ranged from 15 to 33%, 44 to 86%, 84 to 95%, and 92 to 100%, in the control, weekly area-based P. persimilis, bifenazate, and single density-based P. persimilis treatments, respectively. We have shown that in commercial greenhouse production of herbaceous ornamental bedding plants, estimating pest density to determine the appropriate number of predators to release is as effective and offers nearly the same economic benefit as prophylactic use of pesticides.

Twospotted spider mite population level, distribution, and damage on ivy geranium in response to different nitrogen and phosphorus fertilization regimes.

The influence of plant nutrition on arthropod pests has often been studied by comparing plants provided suboptimal nutrients with those provided sufficient or luxurious nutrients, but such results have limited applicability to commercially produced crops because nitrogen (N) and phosphorus (P) are almost never limiting in greenhouse production. We conducted a series of experiments with ivy geranium, Pelargonium peltatum (L.) L'Hr. ex Aiton 'Amethyst 96' to determine the response of twospotted spider mite, Tetranychus urticae Koch (Acarina: Tetranychidae), to six combinations of N (8 or 24 mM) and P (0.32, 0.64, or 1.28 mM) that reflected commercial production practices. All six combinations resulted in saleable plants when plants were free of spider mites, but tissue N and P concentrations among fertilizer combinations were different. On mite-infested plants, no difference in mite numbers or plant damage was found in response to N fertilization rates. Phosphorus had no effect on mite population level until week 8, at which time plants fertilized with 0.64 mM P had slightly more mites than plants fertilized with 0.32 mM. However, overall quality and dry weight of plants fertilized by 0.32 mM P was lower than that of 0.64 and 1.28 mM, which suggests that ivy geranium plants fertilized with the higher P rates may better compensate for mite feeding damage. Positive correlations were found between within-plant distribution of mites and the corresponding tissue N and P concentrations in three foliage strata, suggesting that tissue nutrient content may influence mite selection of feeding sites.

Transfer of phloem-mobile substances from the host plants to the holoparasite Cuscuta sp.

During the development of the haustorium, searching hyphae of the parasite and the host parenchyma cells are connected by plasmodesmata. Using transgenic tobacco plants expressing a GFP-labelled movement protein of the tobacco mosaic virus, it was demonstrated that the interspecific plasmodesmata are open. The transfer of substances in the phloem from host to the parasite is not selective. After simultaneous application of (3)H-sucrose and (14)C-labelled phloem-mobile amino acids, phytohormones, and xenobiotica to the host, corresponding percentages of the translocated compounds are found in the parasite. An open continuity between the host phloem and the Cuscuta phloem via the haustorium was demonstrated in CLSM pictures after application of the phloem-mobile fluorescent probes, carboxyfluorescein (CF) and hydroxypyrene trisulphonic acid (HPTS), to the host. Using a Cuscuta bridge (14)C-sucrose and the virus PVY(N) were transferred from one host plant to the another. The results of translocation experiments with labelled compounds, phloem-mobile dyes and the virus should be considered as unequivocal evidence for a symplastic transfer of phloem solutes between Cuscuta species and their compatible hosts.