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Michael A. Arnold and Eric Young

CuCO3 at 100 g·liter-1 in a paint carrier applied to interior container surfaces effectively prevented root deformation in container-grown Malus domestica Borkh. and Fraxinus pennsylvanica Marsh. seedlings. CuCO3 treatments nearly doubled the number of white unsuberized root tips in both species. CuCO3 treatment increased some measures of root and shoot growth before and after transplanting to larger untreated containers. Root pruning at transplanting tended to reduce root and shoot fresh and dry matter accumulation in F. pennsylvanica seedlings and shoot extension in M. domestica seedlings. In some cases, root pruning of M. domestics at transplanting from CuCO3-treated containers increased root growth compared to unpruned CuCO3-treated and untreated seedlings. Changes in growth induced by CuCO3 and root pruning were not related to changes in trans -zeatin riboside-like activity in the xylem sap of-apple.

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D.M. Glenn and R. Scorza

In reciprocal grafts of tall (`Elberta' and `Loring') and dwarf (`Empress' and `Juseito') peach (Prunus persica Batsch.) phenotypes, we measured dry-matter partitioning, resistance to root system water flow, and phytohormone content of xylem exudate. Scion characteristics determined the phenotype and growth characteristics of the tree irrespective of the rootstock. Tall phenotypes had higher dry weight and lower root resistance to water flow than dwarf phenotypes. Cytokinin-like activity and auxin levels in xylem sap were higher in dwarf than in tall phenotypes; whereas gibberellin-like activity was unaffected by either rootstock or scion. The scion of peach influenced phytohormone levels and resistance to water flow in the root system in addition to root and shoot growth.

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Xia Xu*, Zhongbo Ren and Jiang Lu

Pierce's Disease (PD) is a major factor limiting grape production in the southeast United State. This disease is caused by a bacterium, Xylella fastidiosa Wells et al., which is transmitted to the xylem system of the grapevines primarily by glassy-winged sharpshooters (Homalodisca coagulata Say). Once it is in the xylem, the X. fastidiosa will use the xylem sap as a nutrient source to multiply, colonize, and eventually plug the xylem vessels and cause the PD in susceptible cultivars. On the other hand, symptoms of PD in tolerant cultivars do not appear until fruit maturation, and symptoms are rarely observed in PD resistant cultivars. In order to understand the correlation between X. fastidiosa and PD symptom development, a study was initialed to monitor X. fastidiosa in xylem of resistant, tolerant, and susceptible vines on a monthly basis. Presence of X. fastidiosa was detected directly from xylem sap of field-grown vines by medium culture and confirmed by polymerase chain reaction (PCR). Xylella fastidiosa was detectable throughout the growing season in PD susceptible cultivar `Chardonnay', PD tolerant Florida hybrid grape `Blanc du Bois', and muscadine cultivar `Carlos'. The bacteria were also appeared in the dormant vines with high density in cultivars `Chardonnay' and `Blanc du Bios'. Although X. fastidiosa was also found in dormant canes of `Carlos', the density decreased throughout the late fall and winter months, and they were hardly found before June. The results indicated that X. fastidiosa were carried over from previous season in cultivars `Chardonnay' and `Blanc du Bois', while in PD tolerant cultivar `Carlos', they were newly acquired from the sharpshooter feedings during the growing season.

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Eran Raveh

Carbohydrates are the energy source for most root activities, including membrane maintenance and osmotic adjustment. Yet, the relationship between root carbohydrate status and selective sodium chloride uptake remains unknown. The following study examined the effects of root carbohydrate starvation due to girdling on sodium and chloride uptake in mature citrus trees. Trees were girdled during the spring or during the autumn, when girdling is known to have more dramatic affects. In spring-girdled trees, 4 days after girdling, root total carbohydrate and starch decreased by 25% and 30%, respectively. The decrease in root carbohydrates was followed by a 20% reduction in root respiration rate. Based on root mineral analysis, spring-girdled trees were characterized by having 42% more sodium and 30% more chloride. The effects of girdling on shoot xylem sap mineral concentration were similar to trends in root mineral status; xylem sap from spring-girdled trees had 43% more sodium and 22% more chloride. Leaf chloride concentration measured 6 months after girdling was 74% higher in girdled trees and reached toxicity levels (0.65% vs. 0.37% dry mass, for girdled and nongirdled trees, respectively). The differences in leaf sodium, however, were nonsignificant (0.14% vs. 0.13% dry mass, for girdled and nongirdled trees, respectively). In autumn-girdled trees, the effects on leaf sodium and chloride concentration were more dramatic. Leaves from autumn-girdled trees (sampled 10 months later) had about two times more sodium and about five times more chloride in comparison to nongirdled trees (0.39 % vs. 0.20% dry mass sodium and 1.02% vs. 0.22% dry mass chloride, respectively). The above results link root carbohydrate status and selective sodium or chloride uptake in citrus trees.

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A. Richard Renquist, Horst W. Caspari and David J. Chalmers

Nashi pear (Pyrus serotina Rehder, cv. Hosui) trees were planted in 12 computerized 1m-wide drainage lysimeters in September 1987. During the 1990 season tree water use was monitored via lysimeter and neutron probe readings. Diurnal leaf water relations were studied using a pressure chamber for water potential (ψ) and a porometer for leaf conductance (gs). Xylem sap trunk flow velocities were measured with an experimental heat pulse device and converted to xylem flux. Close agreement existed between 24 hr xylem flux and lysimeter water use when comparing trees with different soil water content. Xylem flux also was very sensitive to changes in evaporative demand. During 9–13 day drying cycles pre-dawn ψ became progressively lower, morning decline more rapid, and afternoon recovery slower. The diurnal gs pattern also shifted during drying cycles, such that gs of water stressed trees always decreased from time of first measurement of sunlit leaves rather than increasing during the morning as on non-stressed trees. Late afternoon was the best time to distinguish between fully irrigated and stressed trees using gs measurements.

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Kathryn C. Taylor and Danielle R. Elli

A 22-kDa Zn-binding protein (ZBP) was isolated from the phloem tissue and evacuated xylem sap of `Valencia' sweet orange [Citrus sinensis (L.) Osbeck] on rough lemon [C. jambhiri (L.)], as well as Valencia on Rangpur lime [Citrus limonia Osbeck]. Phloem and xylem Zn was associated with the 22 kDa ZBP. The Mr value of this ZBP was estimated to be 19,500 by size exclusion chromatography and 22,800 by SDS-PAGE. This protein was isolated with an isoelectric point of 7.5. Ion exchange chromatography demonstrated that 22-kDa ZBP was highly anionic, requiring 0.43 M NaCl for elution from QAE Sepharose. The 22-kDa ZBP appears unique to citrus, having no cross reaction with protein from several tissues from a range of plant species. Accumulation decreased under Zn-deficient conditions, was enhanced by osmotic stress, and the protein completely disappeared with wounding. Amino acid composition demonstrated that the protein was rich in aspartate, and glutamate; and contained 6 cysteine, and 4 histidine residues. These amino acids may be involved in metal binding. N-terminal amino acid sequencing demonstrated that the 22-kDa ZBP had identity with sporamin A&B precursors, Kunitz-type trypsin inhibitors, and miraculin. It is suggested that the genes that encode these proteins are derived from a common ancestral gene.

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L. Lombardini and J.A. Flore

The recent development of small portable infrared thermometers has made canopy temperature an easily measured characteristc in the field. Our objective was to correlate a reduction of soil water with foliage temperature and to compare it with other indicators of plant stress (Pn, E, gs, leaf expansion, sap flow). During Summer 1998, we evaluated the responses of potted apple rootstocks (cultivars Budagowski 9, M9, and Mark) to soil water deficit. Irrigation was withheld for 7 days, and the canopy temperature (Tc) was measured daily with an infrared camera. Tc was always higher than air temperature (Ta). Tc between control and stress plants began to differentiate from day 3. In Mark, this difference was maintained until the end of the experiment. However, gas exchange in Mark seemed to be less affected by the stress than in the other two cultivars. At day 7, midday stomatal conductance (gs) was 38.0, 32.3, and 72.0 mmol·m–2·s–1 in Budagowski 9, M9, and Mark, respectively (control values varied between 161.6 and 164.3 mmol·m–2·s–1 for all the cultivars). Heat-pulse sapflow sensors installed on Mark indicated that the speed of the xylem sap was affected by the stress from day 4 (19-26 cm/h for the controls vs. 15–21 cm/h for the stressed plants). Specific details on the physiological data will be presented.

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D. J. Gray, Z. T. Li, D. L. Hopkins, M. Dutt, S. A. Dhekney, M. M. Van Aman, J. Tattersall and K. T. Kelley

Pierce's disease (PD), caused by the xylem-limited bacterium Xylella fastidiosa, is endemic to the coastal plain of the southeastern United States. Although native southern grapevines are tolerant to X. fastidiosa, all varieties of Vitisvinifera grown in the region will succumb to PD. Genetic transformation to add disease resistance genes, while not disturbing desirable phenotypic characters, holds promise for expanding the southeastern U.S. grape industry by allowing use of established fruit and wine varieties. We utilize embryogenic cell cultures and Agrobacterium strain EHA105 to refine transformation systems for Vitis species and hybrids. V. vinifera`Thompson Seedless' is employed as a model variety to test various transgenes for disease resistance, since as many as 150 independent transgenic plant lines routinely are produced from 1 g of embryogenic culture material. Transgenic plants are stringently screened for PD resistance in greenhouses by mechanical inoculation with X. fastidiosa. Transgenic plants are compared with both susceptible and resistant control plants by assessing typical PD symptom development and by assaying bacterial populations in xylem sap over time. Using these procedures, nine putative PD resistance genes have been inserted into grapevine and over 900 unique transgenic lines have been evaluated. A range of susceptible-to-resistant responses has been catalogued. Thus far, the best construct for PD resistance contains a grape codon-optimized hybrid lytic peptide gene in a high-performance bi-directional 35S promoter complex. Certain transgenic plant lines containing this construct exhibit better resistance than that of resistant control vines.

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Jiang Lu, Elvis Clarke and Zhong-bo Ren

Although some of the American native Vitis species and their hybrids, particularly those originated from the southeastern United States, have been known for resistance to Pierce's disease (PD), their resistant status against the glassy-winged sharp shooter [GWSS, Homalodisca coagulata (Say)], the vector transmitting PD pathogen (Xylellafastidiosa Well), has not been reported. To determine GWSS feeding preferences on different grape species/cultivars and correlations of feeding to Pierce's disease development, a survey was conducted at Florida A&M University, Tallahassee. The feeding preference of GWSS on different species/cultivars was evaluated in two different ways: 1) count the number of GWSS on different grapevines in the field; and 2) determine the feeding preference by measuring the excretion of the GWSS feeding on difference grape species/cultivars, including highly susceptible V. vinifera cultivars, native American grape species and hybrids, and muscadine grapes. Results from this study indicated that the frequency of GWSS visits on different grapevines varied among the species/cultivars investigated. For example, PD-resistant grape V. rotundifolia (muscadine grape) had significantly fewer GWSS visits than did the PD-susceptible V. vinifera grape. The frequency of GWSS visits to V. labrusca, the native American grape susceptible to PD, was intermediate between those found on V. rotundifolia and V. vinifera. Similarly, the GWSS sucked more xylem sap when they fed on PD-susceptible grapevines than on PD-resistant ones. Overall, there is a positive correlation between the GWSS visits/feeding and the status of grapevine resistance/susceptibility to Pierce's disease.

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Kirk W. Pomper and Michael A. Grusak

Understanding the mechanisms that regulate xylem transport of calcium (Ca) to snap bean (Phaseolus vulgaris L.) pods could allow approaches to increase pod Ca concentration and enhance the nutritional value of edible pods. Using the snap bean cultivars Hystyle and Labrador, which exhibit high and low pod Ca levels, respectively, we wished to determine whether there were differences between the two cultivars in stem xylem-sap Ca concentration and whether any differences in sap Ca concentration were related to differences in whole-plant water uptake or Ca import between the cultivars. Well-watered greenhouse-grown plants were placed in a growth chamber at a constant light intensity for an equilibration period. Pot weight loss was measured to determine whole-plant water use and stem xylem exudate was subsequently collected from the severed base of the shoot at flowering and at two stages of pod development. `Hystyle' displayed an exudate Ca concentration that was 50% higher than `Labrador' during pod development. `Labrador' showed 35% greater total water transport through the stem than `Hystyle'. `Labrador' plants also showed a significantly larger leaf area than `Hystyle' plants. Additional plants were used to determine total, long-term Ca influx. No difference was observed between cultivars in total Ca influx into the aerial portion of the plant. With whole-shoot Ca influx being equivalent and pod transpiration rate identical in the two cultivars, our results suggest that the higher whole-plant water uptake in `Labrador' led to a dilution of Ca concentration in the xylem stream and thus less total Ca was transported to developing pods, relative to that in `Hystyle'. Increased transpiration efficiency, enhanced root uptake of Ca, or reduced Ca sequestration in the xylem pathway of the stem could lead to an enhancement in pod Ca concentration in future cultivars of snap bean.