Alterations in body lean angle in lame horses before and after diagnostic analgesia in straight lines in hand and on the lunge.

Altered body lean has been subjectively observed during lungeing in lame horses. The objectives were to quantify the influence of lameness on body lean in trot on the lunge and to investigate the influence of improvement in lameness on the differences in body lean between reins. Thirteen lame horses were trotted in straight lines and lunged on a 10m-diameter circle on both reins before and after lameness was subjectively substantially improved by diagnostic analgesia. A global position system-aided inertial measurement unit attached to the tubera sacrale quantified body lean. Differences between reins in body lean before and after diagnostic analgesia were calculated and means were determined. Five and eight horses had unilateral and bilateral hindlimb lameness, respectively. Two of five horses with unilateral and three of eight horses with bilateral lameness leaned more on the rein with the lame or lamer hindlimb on the inside of the circle (difference between reins 5-8°). Two of five horses with unilateral and two of eight horses with bilateral lameness leaned more on the rein with the lame or lamer hindlimb on the outside of the circle (4-10°). Four horses, one with unilateral and three with bilateral lameness, had only 1° difference in body lean angle between left and right reins. When lameness was improved by diagnostic analgesia, the body lean changed significantly towards similar leaning on left and right reins (mean angle changed from 8.8° to 10.0° (P=0.03) on one rein and 13.4° to 10.8° (P=0.002) on the other rein). It was concluded that body lean becomes more symmetrical between reins after improvement in lameness using diagnostic analgesia.

Alterations in thoracolumbosacral movement when pain causing lameness has been improved by diagnostic analgesia.

Lameness, thoracolumbosacral pain and reduced range of motion (ROM) often coexist; better understanding of their relationship is needed. The objectives were to determine if thoracolumbosacral movement of horses changes when pain causing lameness is improved by diagnostic analgesia. We hypothesised that reduction of lameness will increase ROM of the thoracolumbosacral region. Thirteen horses with different types of hind limb lameness were trotted in straight lines and lunged on a 10m diameter circle on left and right reins before and after lameness was subjectively substantially improved by diagnostic analgesia. Inertial sensor data were collected from the withers, thirteenth (T13) and eighteenth thoracic (T18) vertebrae, third lumbar (L3) vertebra, tubera sacrale (TS), left and right tubera coxae. ROM of flexion-extension, axial rotation, lateral bending, dorsoventral, lateral-lateral motion and vertical movement symmetry were quantified at each thoracolumbar site. Hiphike difference (HHD), maximum difference (MaxDiff) and minimum difference (MinDiff) for the pelvic sensors were measured. Percentage changes for before and after diagnostic analgesia were calculated; mean±standard deviation (SD) or median [interquartile range] were determined. Associations between the change in pelvic versus thoracolumbar movement symmetry after each local analgesic technique were tested. After resolution of lameness, HHD decreased by 7% [68%] (P=0.006). The MinDiff decreased significantly by 33% [61%] (P=0.01), 45±13% (P=0.005) and 52±23% (P=0.04), for TS, L3 and T18, respectively. There was significantly increased ROM in flexion-extension at T13, in axial rotation at T13, T18, L3 and in lateral-lateral ROM at L3. Thoracolumbosacral asymmetry and reduced ROM associated with lameness were both altered immediately by improvement in lameness using diagnostic analgesia.

Thoracolumbar movement in sound horses trotting in straight lines in hand and on the lunge and the relationship with hind limb symmetry or asymmetry.

Equine movement symmetry is changed when turning, which may induce alterations in thoracolumbosacral kinematics; however, this has not previously been investigated. Our objectives were to document thoracolumbar movement in subjectively sound horses comparing straight lines with circles on both reins and to relate these observations to the objectively determined symmetry/asymmetry of hindlimb gait. Fourteen non-lame horses were assessed prospectively in a non-random, cross-sectional survey. The horses were trotted in straight lines and lunged on both reins and inertial sensor data collected at landmarks: withers, T13 and T18, L3, tubera sacrale, and left and right tubera coxae. Data were processed using published methods; angular motion range of motion (ROM; flexion-extension, axial rotation, lateral bending) and translational ROM (dorsoventral and lateral) and symmetry within each stride were assessed. The dorsoventral movement of the back exhibited a sinusoidal pattern with two oscillations per stride. Circles induced greater asymmetry in dorsoventral movement within each stride (mean ± standard deviation, up to 9 ± 6%) compared with straight lines (up to 6 ± 6%). The greatest amplitude of dorsoventral movement (119 ± 14 mm in straight lines vs. 126 ± 20 mm in circles) occurred at T13. Circles induced greater flexion-extension ROM (>1.3°; P = 0.002), lateral bending (>16°; P <0.001), and lateral motion (>16 mm; P = 0.002) compared with straight lines. Circles induced a movement pattern similar to an inside hindlimb lameness, which was significantly associated with the circle-induced greater asymmetry of dorsoventral movement of the thoracolumbar region (P = 0.03). Moving in a circle induces measurable changes in thoracolumbar movement compared with moving in straight lines, associated with alterations in the hindlimb gait.

Saddle fit and management: An investigation of the association with equine thoracolumbar asymmetries, horse and rider health.

REASONS FOR PERFORMING STUDY: No previous studies have investigated interrelationships between saddle fit/management, equine thoracolumbar asymmetries, rider and horse health. OBJECTIVES: To assess associations between data obtained by clinical assessment and those provided by riders via a questionnaire. STUDY DESIGN: Clinical assessment of a convenience sample of horses and riders compared with a Web-based questionnaire survey (n = 205). METHODS: Horse thoracolumbar asymmetries at predetermined sites, the presence of lameness (in hand and/or ridden), saddle slip, saddle fit/management and rider straightness were assessed. Kappa statistics were used to assess the relationship between categorical clinical data and questionnaire data from riders. Spearman's correlation was used to investigate associations between outcomes from clinical assessment (horse, saddle and rider data) and information provided by riders. RESULTS: There was a 40.5% (205 of 506) questionnaire response rate. Thirty horses (14.6%) had saddle slip, which was significantly associated with hindlimb lameness or gait abnormalities (P<0.001), but only 2 riders had considered a link between saddle slip and lameness. Rider back pain was common (38.5%) and associated with ill-fitting saddles (P = 0.03) and either a quadrupedally reduced cranial phase of the step or a stiff, stilted canter (P = 0.006). Well-fitted saddles were associated with frequent saddle fit checks (P = 0.004). Minor thoracolumbar asymmetries (P = 0.04) were negatively associated with ill-fitting saddles and positively associated with rider skill level (P = 0.001). CONCLUSIONS: The interaction between the horse, saddle and rider is complex. Ill-fitting saddles and a stiff, stilted canter or quadrupedally reduced cranial phase of the step are associated with rider back pain. Equine back pain and minor thoracolumbar asymmetries are associated with ill-fitting saddles. Saddle fit should be checked more often than once yearly to lower the number of ill-fitting saddles. Riders, trainers and other professionals involved in equine care and performance need better education to recognise ill-fitting saddles, lameness, saddle slip and rider crookedness.

The interrelationship of lameness, saddle slip and back shape in the general sports horse population.

REASONS FOR PERFORMING STUDY: Saddle slip is usually blamed on saddle fit, crooked riders or horse shape, but may reflect hindlimb lameness. There are no studies of the frequency of occurrence of saddle slip and risk factors within a tested sample population of the general sports horse population. OBJECTIVES: To quantify the frequency of saddle slip and to describe the association with lameness, thoracolumbar shape/symmetry, crooked riders and ill-fitting saddles. STUDY DESIGN: Nonrandom, cross-sectional survey using convenience sampling. METHODS: Five hundred and six sports horses in normal work were assessed prospectively. Thoracolumbar shape/symmetry were measured at predetermined sites; the presence of lameness (in hand and/or ridden) and saddle slip was recorded. Descriptive statistics, univariable and multiple logistic regression were performed to assess the relationship between horse-saddle-rider factors and saddle slip. RESULTS: The frequency of lameness, quadrilaterally reduced cranial phase of the stride or stiff, stilted canter was 45.7%, saddle slip 12.3%, left-right thoracolumbar shape asymmetries ≥ coefficient of variance of 8% (1.2 cm) 0.6%; and 103 of 276 riders (37.3%) sat crookedly. The saddle consistently slipped to one side in 24.4% of horses with hindlimb lameness alone, 45.5% of horses with concurrent hindlimb and forelimb lameness, compared with 5.4% with forelimb lameness, 17.4% with stiff, stilted canter, 20% with quadrilaterally reduced cranial phase of stride and 5.5% nonlame horses. Nineteen horses (30.6%) with saddle slip had no detectable hindlimb lameness; however, 14 had a gait abnormality, particularly in canter. Multivariable analysis revealed that saddle slip was significantly associated with hindlimb lameness and gait abnormalities (odds ratio [OR] = 52.62, 95% confidence interval [CI] 17.3-159.7), a saddle fitted with even contact and uniform flocking (OR = 15.49, 95% CI 1.9-125.5), riders sitting crookedly (OR = 6.32, 95% CI 2.9-13.7), a well-balanced saddle (OR = 3.05, 95% 1.4-6.9) and large back shape ratio at T18 (OR = 1.2, 95% 1.1-1.3). CONCLUSIONS: Many horses with hindlimb and/or forelimb lameness go unrecognised. Saddle slip may be a sign of hindlimb lameness. Education of the equestrian population to identify lameness and saddle slip is required. The Summary is available in Chinese - see Supporting information.

An investigation of the relationship between hindlimb lameness and saddle slip.

REASONS FOR PERFORMING STUDY: We have observed saddle slip consistently to one side because of a crooked rider, an ill-fitting saddle, asymmetry in a horse's thoracolumbar shape and lameness. Currently, there are no objective data to permit assessment of the relative importance of each factor. OBJECTIVES: To document the frequency of occurrence of saddle slip in horses with hindlimb lameness compared with other horses. To describe the effect of lameness characteristics and grade, the abolition of lameness by diagnostic analgesia, breed, size, thoracolumbar shape and symmetry and the rider's weight. METHODS: One hundred and twenty-eight horses were assessed prospectively, and lameness and saddle slip were assigned a grade before and after diagnostic analgesia. The thoracolumbar shape and symmetry were measured objectively. In 3 horses, the force distribution and magnitude underneath the saddle were measured before and after diagnostic analgesia. RESULTS: The saddle consistently slipped to one side in 38 of 71 horses (54%) with hindlimb lameness, compared with one of 26 horses (4%) with forelimb lameness, none of 20 (0%) with back pain and/or sacroiliac joint region pain and none of 11 sound horses (0%). The association between saddle slip and hindlimb lameness was significant (Spearman's rank correlation coefficient, ρ = 0.548, P<0.001). Diagnostic analgesia abolishing the hindlimb lameness eliminated the saddle slip in 37 of 38 horses (97%). In 2 horses, the saddle continued to slip after resolution of lameness; one horse had bilateral forelimb lameness and the other horse had concurrent hindlimb and forelimb lameness. The saddle of both these horses was asymmetrically flocked. The saddle slipped to the side of the lamer hindlimb in most horses (32 of 37 [86%]). No horse with saddle slip had significant left-right asymmetry of the back at 4 predetermined sites. CONCLUSIONS AND CLINICAL RELEVANCE: Hindlimb lameness is an important factor in inducing saddle slip. Saddle slip may be an indicator of the presence of hindlimb lameness.

Salivary enzymes are injected into xylem by the glassy-winged sharpshooter, a vector of Xylella fastidiosa.

A few phytophagous hemipteran species such as the glassy-winged sharpshooter, Homalodisca vitripennis, (Germar), subsist entirely on xylem fluid. Although poorly understood, aspects of the insect's salivary physiology may facilitate both xylem-feeding and transmission of plant pathogens. Xylella fastidiosa is a xylem-limited bacterium that causes Pierce's disease of grape and other scorch diseases in many important crops. X. fastidiosa colonizes the anterior foregut (precibarium and cibarium) of H. vitripennis and other xylem-feeding vectors. Bacteria form a dense biofilm anchored in part by an exopolysaccharide (EPS) matrix that is reported to have a ß-1,4-glucan backbone. Recently published evidence supports the following, salivation-egestion hypothesis for the inoculation of X. fastidiosa during vector feeding. The insect secretes saliva into the plant and then rapidly takes up a mixture of saliva and plant constituents. During turbulent fluid movements in the precibarium, the bacteria may become mechanically and enzymatically dislodged; the mixture is then egested back out through the stylets into plant cells, possibly including xylem vessels. The present study found that proteins extracted from dissected H. vitripennis salivary glands contain several enzyme activities capable of hydrolyzing glycosidic linkages in polysaccharides such as those found in EPS and plant cell walls, based on current information about the structures of those polysaccharides. One of these enzymes, a ß-1,4-endoglucanase (EGase) was enriched in the salivary gland protein extract by subjecting the extract to a few, simple purification steps. The EGase-enriched extract was then used to generate a polyclonal antiserum that was used for immunohistochemical imaging of enzymes in sharpshooter salivary sheaths in grape. Results showed that enzyme-containing gelling saliva is injected into xylem vessels during sharpshooter feeding, in one case being carried by the transpiration stream away from the injection site. Thus, the present study provides support for the salivation-egestion hypothesis.

The validity of self-reported dental agenesis.

The optimal tools for obtaining a diagnosis of dental agenesis (DA) among adults are childhood dental records and radiographs. However, these are often not available, and therefore the present study aimed to assess whether DA in adults could be validly assessed by self-reported information guided by a questionnaire. A questionnaire eliciting information on DA of permanent teeth was constructed, pilot-tested, and subsequently posted to a case group consisting of 334 young adults, for whom the DA status had been ascertained in the period from 1992 to 2002. A control group, consisting of 258 young adults randomly selected from the population from which the cases originated, was also approached with questionnaires. The response rate was 53.7% among cases and 46.4% among controls. The sensitivity and specificity of self-reported DA were estimated to be at least 0.88 (95% CI = 0.82-0.92) and 0.95 (95% CI = 0.89-0.98), respectively. These diagnostic test parameters are clearly inadequate for population-screening purposes. However, when screening patient groups with a higher occurrence of DA than is characteristic of the background population, the self-reported DA status may be a useful tool for identifying risk groups for conditions associated with the presence of DA.

Polysaccharide compositions of intervessel pit membranes contribute to Pierce's disease resistance of grapevines.

Symptom development of Pierce's disease (PD) in grapevine (Vitis vinifera) depends largely on the ability of the bacterium Xylella fastidiosa to use cell wall-degrading enzymes (CWDEs) to break up intervessel pit membranes (PMs) and spread through the vessel system. In this study, an immunohistochemical technique was developed to analyze pectic and hemicellulosic polysaccharides of intervessel PMs. Our results indicate that PMs of grapevine genotypes with different PD resistance differed in the composition and structure of homogalacturonans (HGs) and xyloglucans (XyGs), the potential targets of the pathogen's CWDEs. The PMs of PD-resistant grapevine genotypes lacked fucosylated XyGs and weakly methyl-esterified HGs (ME-HGs), and contained a small amount of heavily ME-HGs. In contrast, PMs of PD-susceptible genotypes all had substantial amounts of fucosylated XyGs and weakly ME-HGs, but lacked heavily ME-HGs. The intervessel PM integrity and the pathogen's distribution in Xylella-infected grapevines also showed differences among the genotypes. In pathogen-inoculated, PD-resistant genotypes PM integrity was well maintained and Xylella cells were only found close to the inoculation site. However, in inoculated PD-susceptible genotypes, PMs in the vessels associated with bacteria lost their integrity and the systemic presence of the X. fastidiosa pathogen was confirmed. Our analysis also provided a relatively clear understanding of the process by which intervessel PMs are degraded. All of these observations support the conclusion that weakly ME-HGs and fucosylated XyGs are substrates of the pathogen's CWDEs and their presence in or absence from PMs may contribute to grapevine's PD susceptibility.

Cell wall-degrading enzymes enlarge the pore size of intervessel pit membranes in healthy and Xylella fastidiosa-infected grapevines.

The pit membrane (PM) is a primary cell wall barrier that separates adjacent xylem water conduits, limiting the spread of xylem-localized pathogens and air embolisms from one conduit to the next. This paper provides a characterization of the size of the pores in the PMs of grapevine (Vitis vinifera). The PM porosity (PMP) of stems infected with the bacterium Xylella fastidiosa was compared with the PMP of healthy stems. Stems were infused with pressurized water and flow rates were determined; gold particles of known size were introduced with the water to assist in determining the size of PM pores. The effect of introducing trans-1,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid (CDTA), oligogalacturonides, and polygalacturonic acid into stems on water flux via the xylem was also measured. The possibility that cell wall-degrading enzymes could alter the pore sizes, thus facilitating the ability of X. fastidiosa to cross the PMs, was tested. Two cell wall-degrading enzymes likely to be produced by X. fastidiosa (polygalactuoronase and endo-1,4- beta -glucanase) were infused into stems, and particle passage tests were performed to check for changes in PMP. Scanning electron microscopy of control and enzyme-infused stem segments revealed that the combination of enzymes opened holes in PMs, probably explaining enzyme impacts on PMP and how a small X. fastidiosa population, introduced into grapevines by insect vectors, can multiply and spread throughout the vine and cause Pierce's disease.

Characterization of the cell wall of the ubiquitous plant pathogen Botrytis cinerea.

The ascomycete Botrytis cinerea is a destructive and ubiquitous plant pathogen and represents a model organism for the study of necrotrophic fungal pathogens. Higher fungi possess a complex and dynamic multilayer cell wall involved in crucial aspects of fungal development, growth and pathogenicity. Plant resistance to microbial pathogens is determined often by the capacity of the plant to recognize molecular patterns associated with the surface of an interacting microbe. Here we report the chemical characterization of cell walls from B. cinerea during axenic growth. Neutral sugars and proteins constituted most of the mass of the B. cinerea cell walls, although chitin and uronic acids were detected. Glucose was the most abundant neutral sugar, but arabinose, galactose, xylose and mannose also were present. Changes in cell wall composition during culture were observed. As the culture developed, protein levels declined, while chitin and neutral sugars increased. Growth of B. cinerea was associated with a remarkable decline in the fraction of its cell wall material that was soluble in hot alkali. These results suggest that the cell wall of B. cinerea undergoes significant modifications during growth, possibly becoming more extensively covalently cross-linked, as a result of aging of mycelia or in response to decreasing nutrient supply or as a consequence of increasing culture density.

Identification of endo- and exo-polygalacturonase activity in Lygus hesperus (Knight) salivary glands.

Polygalacturonase (PG) activity found in the salivary gland apparatus of the western tarnished plant bug (WTPB, Lygus hesperus Knight) has been thought to be the main chemical cause of the damage inflicted by this mirid when feeding on its plant hosts. Early viscosity and thermal stability studies of the PG activity in L. hesperus protein extracts were difficult to interpret. Thus, it has been suggested that one or more PG protein(s) with different hydrolytic modes of action are produced by this mirid. In order to understand the quantitative complexity of the WTPB salivary PG activity, PG purification from a protein extract from salivary glands excised from L. hesperus insects was performed using affinity and ion exchange chromatography. To elucidate the qualitative complexity of the purified PGs, the digestion products generated by the PGs were separated using high performance anion exchange chromatography with pulsed amperometric detection. At least five PG proteins were detected; these differing in terms of their glycosylation, mass-to-charge ratios, and/or molecular mass. The characterization of the products generated by these PGs showed that endo- and exo-acting PGs are produced by WTPB. Although none of the PGs was purified to homogeneity, the present work provides biochemical evidence of a multiplicity of PGs that degrade the pectin component of the plant tissue in different fashions. The implications of these findings affect the understanding of WTPB feeding damage and, potentially, help identify ways to control this important crop pest. Arch. Insect Biochem. Physiol. 2008. (c) 2008 Wiley-Liss, Inc.

Pathogenesis of Eutypa lata in grapevine: identification of virulence factors and biochemical characterization of cordon dieback.

Eutypa lata is a vascular pathogen of woody plants. In the present study we (i) determined which component(s) of the cell wall polymers were degraded in naturally infected grapevines and in artificially inoculated grape wood blocks; (ii) compared the pattern of wood decay in the tolerant grape cv. Merlot versus the susceptible cv. Cabernet Sauvignon; and (iii) identified secondary metabolites and hydrolytic enzymes expressed by E. lata during wood degradation. Biochemical analyses and a cytochemical study indicated that glucose-rich polymers were primary targets of E. lata. Structural glucose and xylose of the hemicellulose fraction of the plant cell wall and starch were depleted in infected woods identically in both cultivars. Moreover, the more tolerant cv. Merlot always had more lignin in the wood than the susceptible cv. Cabernet Sauvignon, indicating that this polymer may play a role in disease resistance. In vitro assays demonstrated the production by E. lata of oxidases, glycosidases and starch degrading enzymes. Phytotoxic secondary metabolites were also produced but our data suggest that they may bind to the wood. Finally, we demonstrated that free glucose in liquid cultures repressed primary but not secondary metabolism.

The intersection between cell wall disassembly, ripening, and fruit susceptibility to Botrytis cinerea.

Fruit ripening is characterized by processes that modify texture and flavor but also by a dramatic increase in susceptibility to necrotrophic pathogens, such as Botrytis cinerea. Disassembly of the major structural polysaccharides of the cell wall (CW) is a significant process associated with ripening and contributes to fruit softening. In tomato, polygalacturonase (PG) and expansin (Exp) are among the CW proteins that cooperatively participate in ripening-associated CW disassembly. To determine whether endogenous CW disassembly influences the ripening-regulated increase in necrotropic pathogen susceptibility, B. cinerea susceptibility was assessed in transgenic fruit with suppressed polygalacturonase (LePG) and expansin (LeExp1) expression. Suppression of either LePG or LeExp1 alone did not reduce susceptibility but simultaneous suppression of both dramatically reduced the susceptibility of ripening fruit to B. cinerea, as measured by fungal biomass accumulation and by macerating lesion development. These results demonstrate that altering endogenous plant CW disassembly during ripening influences the course of infection by B. cinerea, perhaps by changing the structure or the accessibility of CW substrates to pathogen CW-degrading enzymes. Recognition of the role of ripening-associated CW metabolism in postharvest pathogen susceptibility may be useful in the design and development of strategies to limit pathogen losses during fruit storage, handling, and distribution.

Detection and visualization of an exopolysaccharide produced by Xylella fastidiosa in vitro and in planta.

Many phytopathogenic bacteria, such as Ralstonia solanacearum, Pantoea stewartii, and Xanthomonas campestris, produce exopolysaccharides (EPSs) that aid in virulence, colonization, and survival. EPS can also contribute to host xylem vessel blockage. The genome of Xylella fastidiosa, the causal agent of Pierce's disease (PD) of grapevine, contains an operon that is strikingly similar to the X. campestris gum operon, which is responsible for the production of xanthan gum. Based on this information, it has been hypothesized that X. fastidiosa is capable of producing an EPS similar in structure and composition to xanthan gum but lacking the terminal mannose residue. In this study, we raised polyclonal antibodies against a modified xanthan gum polymer similar to the predicted X. fastidiosa EPS polymer. We used enzyme-linked immunosorbent assay to quantify production of EPS from X. fastidiosa cells grown in vitro and immunolocalization microscopy to examine the distribution of X. fastidiosa EPS in biofilms formed in vitro and in planta and assessed the contribution of X. fastidiosa EPS to the vascular occlusions seen in PD-infected grapevines.

Temporal sequence of cell wall disassembly events in developing fruits. 1. Analysis of raspberry (Rubus idaeus).

Raspberry fruits were harvested at five developmental stages, from green to red ripe, and the changes in cell wall composition, pectin and hemicellulose solubilization, and depolymerization were analyzed. Fruit softening at intermediate stages of ripening was associated with increased pectin solubilization, which occurred without depolymerization. Arabinose was found to be the most abundant noncellulosic neutral sugar in the cell wall and showed dramatic solubilization late in ripening. No changes in pectin molecular size were observed even at the 100% red stage. Subsequently, as fruit became fully ripe a dramatic depolymerization occurred. In contrast, the hemicellulosic fractions showed no significant changes in content or polymer size during ripening. The paper discusses the sequence of events leading to cell wall disassembly in raspberry fruit.

Temporal sequence of cell wall disassembly events in developing fruits. 2. Analysis of blueberry (Vaccinium species).

Softening and pathogen susceptibility are the major factors limiting the marketing of blueberries as fresh fruits, and these traits are associated with fruit cell wall structure. However, few studies that characterize wall modifications occurring during development and ripening have been reported for this fruit. In this study the ripening-associated modifications of blueberry fruit cell walls (composition, pectin and hemicellulose solubilization, and depolymerization) at five stages of ripeness have been analyzed. Xylose was found to be the most abundant noncellulosic neutral sugar associated with fruit walls, and the observed high Xyl/Glc ratio suggested that xylans, which are usually a minor hemicellulosic fruit wall component, are abundant in blueberry. The pectic matrix showed increased solubilization at early and intermediate stages of ripening, but no changes were detected in late ripening. Furthermore, little reduction in pectin polymer size occurred during blueberry ripening. In contrast, hemicellulose levels decreased as ripening progressed, and a clear depolymerization of these components was observed. A model for cell wall degradation in this fruit is discussed.

Xylella fastidiosa requires polygalacturonase for colonization and pathogenicity in Vitis vinifera grapevines.

Xylella fastidiosa is the causal agent of Pierce's disease of grape, an economically significant disease for the grape industry. X. fastidiosa systemically colonizes the xylem elements of grapevines and is able to breach the pit pore membranes separating xylem vessels by unknown mechanisms. We hypothesized that X. fastidiosa utilizes cell wall degrading enzymes to break down pit membranes, based on the presence of genes involved in plant cell wall degradation in the X. fastidiosa genome. These genes include several beta-1,4 endoglucanases, several xylanases, several xylosidases, and one polygalacturonase (PG). In this study, we demonstrated that the pglA gene encodes a functional PG. A mutant in pglA lost pathogenicity and was compromised in its ability to systemically colonize Vitis vinifera grapevines. The results indicate that PG is required for X. fastidiosa to successfully infect grapevines and is a critical virulence factor for X. fastidiosa pathogenesis in grapevine.

Xylella fastidiosa infection and ethylene exposure result in xylem and water movement disruption in grapevine shoots.

It is conventionally thought that multiplication of the xylem-limited bacterium Xylella fastidiosa (Xf) within xylem vessels is the sole factor responsible for the blockage of water movement in grapevines (Vitis vinifera) affected by Pierce's disease. However, results from our studies have provided substantial support for the idea that vessel obstructions, and likely other aspects of the Pierce's disease syndrome, result from the grapevine's active responses to the presence of Xf, rather than to the direct action of the bacterium. The use of magnetic resonance imaging (MRI) to observe the distribution of water within the xylem has allowed us to follow nondestructively the development of vascular system obstructions subsequent to inoculation of grapevines with Xf. Because we have hypothesized a role for ethylene produced in vines following infection, the impact of vine ethylene exposure on obstruction development was also followed using MRI. In both infected and ethylene-exposed plants, MRI shows that an important proportion of the xylem vessels become progressively air embolized after the treatments. The loss of xylem water-transporting function, assessed by MRI, has been also correlated with a decrease in stem-specific hydraulic conductivity (K(S)) and the presence of tyloses in the lumens of obstructed water conduits. We have observed that the ethylene production of leaves from infected grapevines is greater than that from healthy vines and, therefore, propose that ethylene may be involved in a series of cellular events that coordinates the vine's response to the pathogen.

Cell wall disassembly events in boysenberry (Rubus idaeus L. × Rubus ursinus Cham. & Schldl.) fruit development.

Boysenberry fruit was harvested at five developmental stages, from green to purple, and changes in pectin and hemicellulose solubilisation and depolymerisation, polymer neutral sugar contents, and the activities of cell wall degrading enzymes were analysed. The high xylose to glucose ratio in the 4% KOH-soluble hemicellulose fraction suggests that xylans are abundant in the boysenberry cell wall. Although the cell wall changes associated with fruit development do not proceed in discrete stages and the cell wall disassembly is a consequence of highly regulated changes occurring in a continuum, the results suggest that the temporal changes in cell wall degradation in boysenberry account for at least three stages: an early stage (green to 75% red colour), associated with metabolism of cellulose and cross-linking glycans; an intermediate period (75 to 100% red colour), characterised by substantial pectin solubilisation without depolymerisation in which α-arabinofuranosidase increases markedly and 50% of the wall arabinose is lost; and a final stage (100% red colour to purple), characterised mainly by a reduction of pectic galactose content and a dramatic increase in pectin depolymerisation associated with higher polygalacturonase, pectin methylesterase, acetyl esterase and ß-galactosidase activities. From a biotechnological perspective enzymes involved in pectin matrix disassembly seem to be the better candidates to affect boysenberry fruit late-softening by genetic intervention. A model for cell wall disassembly in boysenberry fruit is proposed.