Are Methanol-Derived Foliar Methyl Acetate Emissions a Tracer of Acetate-Mediated Drought Survival in Plants?

Upregulation of acetate fermentation in plants has recently been described as an evolutionarily conserved drought survival strategy, with the amount of acetate produced directly correlating to survival. However, destructive measurements are required to evaluate acetate-linked drought responses, limiting the temporal and spatial scales that can be studied. Here, 13C-labeling studies with poplar (Populus trichocarpa) branches confirmed that methyl acetate is produced in plants from the acetate-linked acetylation of methanol. Methyl acetate emissions from detached leaves were strongly stimulated during desiccation, with total emissions decreasing with the leaf developmental stage. In addition, diurnal methyl acetate emissions from whole physiologically active poplar branches increased as a function of temperature, and light-dark transitions resulted in significant emission bursts lasting several hours. During experimental drought treatments of potted poplar saplings, light-dark methyl acetate emission bursts were eliminated while strong enhancements in methyl acetate emissions lasting > 6 days were observed with their initiation coinciding with the suppression of transpiration and photosynthesis. The results suggest that methyl acetate emissions represent a novel non-invasive tracer of acetate-mediated temperature and drought survival response in plants. The findings may have important implications for the future understanding of acetate-mediated drought responses to transcription, cellular metabolism, and hormone signaling, as well as its associated changes in carbon cycling and water use from individual plants to whole ecosystems.

Xylem structure of four grape varieties and 12 alternative hosts to the xylem-limited bacterium Xylella fastidious.

BACKGROUND AND AIMS: The bacterium Xylella fastidiosa (Xf), responsible for Pierce's disease (PD) of grapevine, colonizes the xylem conduits of vines, ultimately killing the plant. However, Vitis vinifera grapevine varieties differ in their susceptibility to Xf and numerous other plant species tolerate Xf populations without showing symptoms. The aim of this study was to examine the xylem structure of grapevines with different susceptibilities to Xf infection, as well as the xylem structure of non-grape plant species that support or limit movement of Xf to determine if anatomical differences might explain some of the differences in susceptibility to Xf. METHODS: Air and paint were introduced into leaves and stems to examine the connectivity between stem and leaves and the length distribution of their vessels. Leaf petiole and stem anatomies were studied to determine the basis for the free or restricted movement of Xf into the plant. KEY RESULTS: There were no obvious differences in stem or petiole vascular anatomy among the grape varieties examined, nor among the other plant species that would explain differences in resistance to Xf. Among grape varieties, the more tolerant 'Sylvaner' had smaller stem vessel diameters and 20 % more parenchyma rays than the other three varieties. Alternative hosts supporting Xf movement had slightly longer open xylem conduits within leaves, and more connection between stem and leaves, when compared with alternative hosts that limit Xf movement. CONCLUSIONS: Stem--leaf connectivity via open xylem conduits and vessel length is not responsible for differences in PD tolerance among grape varieties, or for limiting bacterial movement in the tolerant plant species. However, it was found that tolerant host plants had narrower vessels and more parenchyma rays, possibly restricting bacterial movement at the level of the vessels. The implications of xylem structure and connectivity for the means and regulation of bacterial movement are discussed.

Seasonal abundance of Draeculacephala minerva and other Xylella fastidiosa vectors in California almond orchards and vineyards.

Almond leaf scorch (ALS) disease is caused by the bacterium Xylella fastidiosa and transmitted by xylem-feeding insects. Reports of increased incidence of ALS-diseased trees in California prompted surveys in three almond [Prunus dulcis (Mill.) D. A. Webb]-growing regions, from June 2003 to September 2005, to determine insect vector species composition and abundance. For comparison, sampling in and near vineyards in the San Joaquin Valley, California, also was completed. Sampling in or near almond orchards collected >42,000 Cicadomorpha of which 4.8% were xylem feeders, including 1912 grass sharpshooter, Draeculacephala minerva Ball; five Xyphon fulgida Nottingham; and a single spittlebug, Philaenus spumarius L. The most abundant vector was D. minerva. Season-long sampling indicated that D. minerva was a year-round resident in and/or near almonds in the Sacramento Valley, but not in the San Joaquin Valley. Similarly, D. minerca was rare in vineyards in the San Joaquin Valley, but was abundant in irrigated pastures near vineyards. D. minerva was most frequently collected along orchard margins, and peak densities were observed in summer, the period of time when bacterial titers are reported to increase in infected trees. Screening of D. minerva for presence of X.fastidiosa found that 1.1% of insects collected near almond orchards and 4.5% of insects collected from pastures tested positive. The X. fastidiosa subspecies and genotype detected in insects collected from orchards matched those collected from ALS-diseased almond trees in the same orchard. Of the few X. fulgida and P. spumarius collected, none tested positive for X. fastidiosa. Results are discussed with respect to X. fastidiosa vector control and detection methods.

Distribution of glassy-winged sharpshooter and threecornered alfalfa hopper on plant hosts in the San Joaquin Valley, California.

Homalodisca vitripennis (Germar) and Spissistilus festinus (Say) populations were surveyed bimonthly for 14 mo in Kern County, CA, at five agricultural sites made up of a variety of potential host plants. In addition, S. festinus populations were surveyed in four alfalfa, Medicago sativa L., fields in Kern and Tulare counties. Insects were collected by beats-sweeps and sticky traps. Data on host plant condition and phenology, and ground cover presence and composition were collected at the five agricultural sites, whereas data on mowing and insecticide use were collected at the four alfalfa sites. Populations of both insects persisted at the five agricultural locations despite insecticide applications applied as part of a H. vitripennis areawide management program and standard commercial operations. Plants colonized by H. vitripennis included eucalyptus (Eucalyptus L'Hér.), jojoba [Simmondsia chinensis (Link) C. K. Schneid.], and citrus (Citrus spp.). Populations of S. festinus were much greater in collections from alfalfa fields than from the five agricultural sites. Insects collected from the five mixed agricultural sites were negative for presence of X. fastidiosa. In laboratory tests, S. festinus did not acquire or transmit X. fastidiosa in tests with infected grape (Vitis spp.) as an acquisition source and grape, almond [Prunus dulcis (Mill.) D.A.Webb], and alfalfa as inoculation hosts. Recommendations for vector control, vegetation management, and targeted monitoring to reduce insect populations and inoculum potential are discussed.

A cell-cell signaling sensor is required for virulence and insect transmission of Xylella fastidiosa.

Cell-cell signaling in Xylella fastidiosa, a xylem-colonizing plant pathogenic bacterium, mediated by a fatty acid Diffusible Signaling Factor (DSF), is required to colonize insect vectors and to suppress virulence to grape. Here, we show that a hybrid two-component regulatory protein RpfC is involved in negative regulation of DSF synthesis by RpfF in X. fastidiosa. X. fastidiosa rpfC mutants hyperexpress rpfF and overproduce DSF and are deficient in virulence and movement in the xylem vessels of grape. The expression of the genes encoding the adhesins FimA, HxfA, and HxfB is much higher in rpfC mutants, which also exhibit a hyperattachment phenotype in culture that is associated with their inability to migrate in xylem vessels and cause disease. rpfF mutants deficient in DSF production have the opposite phenotypes for all of these traits. RpfC is also involved in the regulation of other signaling components including rpfG, rpfB, a GGDEF domain protein that may be involved in intracellular signaling by modulating the levels of cyclic-di-GMP, and the virulence factors tolC and pglA required for disease. rpfC mutants are able to colonize the mouthparts of insect vectors and wild-type strains but are not transmitted as efficiently to new host plants, apparently because of their high levels of adhesiveness. Because of the conflicting contributions of adhesiveness and other traits to movement within plants and vectoring to new host plants, X. fastidiosa apparently coordinates these traits in a population-size-dependent fashion involving accumulation of DSF.

Ground Vegetation Survey for Xylella fastidiosa in California Almond Orchards.

Xylella fastidiosa is a xylem-limited bacterium that causes almond leaf scorch (ALS), Pierce's disease of grapevines, and other plant diseases. We surveyed ground vegetation in ALS-infected almond orchards in California's Central Valley for the presence of this bacterium. Plant tissue samples were collected throughout a 2-year period and processed for the presence of X. fastidiosa using restriction enzyme digestion of RST31 and RST33 polymerase chain reaction (PCR) products and bacterial culture on selective media. Overall disease incidence was low in the ground vegetation species; only 63 of 1,369 samples tested positive. Of the 38 species of common ground vegetation tested, 11 tested positive for X. fastidiosa, including such common species as shepherd's purse (Capsella bursa-pastoris), filaree (Erodium spp.), cheeseweed (Malva parvifolia), burclover (Medicago polymorpha), annual bluegrass (Poa annua) London rocket (Sisymbrium irio), and chickweed (Stellaria media). There was a seasonal component to bacterial presence, with positive samples found only between November and March. Both ground vegetation and almond trees were most commonly infected with the almond strain of X. fastidiosa (six of seven surveyed sites). ALS-infected almond samples had an X. fastidiosa concentration within previously reported ranges (1.84 × 106 to 2.15 × 107 CFU/g); however, we were unable to accurately measure X. fastidiosa titer in sampled ground vegetation for comparison. These results are discussed with respect to ground vegetation management for ALS control.