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Michele R. Warmund, Bruce H. Barritt, John M. Brown, Karen L. Schaffer, and Byoung R. Jeong

Abbreviation: MRI, magnetic resonance imaging. Contribution from the Missouri Agr. Expt. Sta. J. Ser. no. 11,590. We gratefully acknowledge Steve B. Pickup, Dept. of Radiology, Univ. of Missouri Hospital and Clinics, for his assistance. The cost of

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Lisa J. Rowland, Dehua Liu, Merle M. Millard, and Michael J. Line

Dormant and chilled highbush blueberry (Vaccinium corymbosum L.) flower buds were examined by magnetic resonance imaging (MRI). T2 relaxation times of water molecules were too short to create images from flowers within buds that were dormant and had received no chilling, but they were sufficiently long to create images from buds that had their chilling requirement satisfied. To explain the change in relaxation times, we concluded that water is present in a motionally restricted form in flowers of dormant blueberry buds and in a freer form in flowers of buds after the chilling requirement has been satisfied. T2 values for chilled blueberry buds indicated that one population of water molecules with a detectable T2 time was present in flowers of chilled buds with a relaxation time of ≈8 to 15 ms.

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Anne Fennell, M.J. Line, and M. Faust

Changes in water status have been associated with various stages of dormancy and freezing tolerance in woody perennials. Recent studies in apple indicate that changes in the state (bound vs. free) of bud water are strongly correlated with the end of dormancy. In this study nuclear magnetic resonance imaging (NMRI) was used to monitor changes in the state of bud water during the photoperiodic induction of endo-dormancy in Vitis riparia. Bud water status was monitored using proton relaxation times from T1 and T2 images determined at 2, 4, and 6 weeks of long (LD) or short (SD) photoperiod treatments. Bud dormancy was determined by monitoring budbreak in plants defoliated after photoperiod treatments. NMRI allowed nondestructive monitoring of changes in tissue water state. T1 and T2 maps indicated changes in the state of the water in bud and stem tissues during the 6 weeks of treatment. Differences in relaxation times for nondormant and dormancy-induced (reversible) buds were not clear. However, T2 relaxation times were lower in the dormant buds than in the nondormant buds.

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Dehua Liu, Miklos Faust, Merle M. Millard, Michael J. Line, and Gary W. Stutte

Magnetic resonance imaging was used to determine water states in paradormant apple (Malus domestica Borkh.) buds and during early events when buds resumed growth. Proton density and states of water were determined by creating image maps of proton density and relaxation times (T2). Summer-dormant (paradormant) buds had T2 relaxation times up to 30 ms. This water in bud tissues is considered relatively free compared to water that had T2 relaxation times of <1 ms in other parts of the stem and bark. Buds were forced to grow either by pruning off the terminal bud or by starting the bud with thidiazuron (TDZ). Both treatments gave essentially the same results. After treatment, buds started to grow immediately and water moved into the stem and into the bud. As there was more free water in the bud, T2 values ranged up to 50 ms. There appeared to be an inhibitory gradient down on the shoot, which was removed temporarily by excising the top bud. However, between the 2nd and 10th day after removal of the top bud this dominance was reinstated by the highest bud on the stem, which eventually formed a shoot. TDZ treatment overcame this inhibitory gradient effect. There was also a growth potential gradient coinciding with the inhibitory gradient. The growth of lower buds was much slower than that of the upper buds. The growth potential gradient was not overcome by TDZ treatments.

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B.L. Tan, N. Reddy, V. Sarafis, G.A.C. Beattie, and R. Spooner-Hart

Nuclear magnetic resonance (NMR) spectroscopy and magnetic resonance imaging (MRI) were used to detect petroleum-derived spray oils (PDSOs) in citrus seedlings and trees. The NMR spectrum of the phantom containing 10% (v/v) of a nC24 agricultural mineral oil (AMO) showed the resonance of the water protons at δ ≈ 5 ppm, while the resonance of the oil protons at δ = 1.3 to 1.7 ppm. The peak resolution and the chemical shift difference of more than 3.3 ppm between water and oil protons effectively differentiated water and the oil. Chemical shift selective imaging (CSSI) was performed to localize the AMO within the stems of Citrus trifoliata L. seedlings after the application of a 4% (v/v) spray. The chemical shift selective images of the oil were acquired by excitation at δ = 1.5 ppm by averaging over 400 transients in each phase-encoding step. Oil was mainly detected in the outer cortex of stems within 10 d of spray application; some oil was also observed in the inner vascular bundle and pith of the stems at this point. CSSI was also applied to investigate the persistence of oil deposits in sprayed mature Washington navel orange (Citrus ×aurantium L.) trees in an orchard. The trees were treated with either fourteen 0.25%, fourteen 0.5%, four 1.75%, or single 7% sprays of a nC23 horticultural mineral oil (HMO) 12 to 16 months before examination of plant tissues by CSSI, and were still showing symptoms of chronic phytotoxicity largely manifested as reduced yield. The oil deposits were detected in stems of sprayed flushes and unsprayed flushes produced 4 to 5 months after the last spray was applied, suggesting a potential movement of the oil via phloem and a correlation of the persistence of oil deposit in plants and the phytotoxicity. The results demonstrate that MRI is an effective method to probe the uptake and localization of PDSOs and other xenobiotics in vivo in plants noninvasively and nondestructively.

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Christopher J. Clark and Douglas M. Burmeister

Development of browning induced in `Braeburn' apple (Malus ×domestica Borkh.) fruit by a damaging CO2 concentration was monitored weekly using magnetic resonance imaging (MRI) during a 4-week storage trial (0.5 °C, 2 kPa O2/7 kPa CO2). Discrete patches of high-intensity signal, distributed randomly throughout the fruit, were observed in multislice images of samples after 2 weeks of storage; these patches were eventually confirmed as being sites of browning reactions after dissection at the end of the trial. Subsequently (weeks 3 and 4), signal intensity at sites of incipient damage increased and patches enlarged and coalesced. After 2 weeks of storage, the extent of affected tissue, averaged across all image slices, was 1.5%, increasing to 15.9% and 21.3% after 3 and 4 weeks. The average rate at which tissue damage spread in individual slices was 0.81 (range: 0–3.70) cm2·d–1 between weeks 2 and 3, declining to 0.32 (range: 0–1.55) cm2·d–1 in the final week. Tissue damage induced under these conditions did not spread at the same rate at all locations within individual fruit, nor was it preferentially located toward the stem or calyx ends of the fruit.

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Anne Fennell and Michael J. Line

Physiological and biophysical changes were monitored during shoot maturation and bud endodormancy induction in grape (Vitis riparia Michx.) under controlled environments. Growth, dry weight (DW), periderm development, bud endodormancy, and nuclear magnetic resonance imaging (MRI) T2 relaxation times were monitored at 2, 4, or 6 weeks of long-photoperiod [long day (LD), 15 h, endodormancy inhibition] or short-photoperiod [short day (SD), 8 h, endodormancy induction] treatments at 15/9 h day/night thermoperiod of 25/20 ± 3 °C. Shoots on LD plants grew throughout the entire study period, although the rate of growth decreased slightly during the 6th week. Shoot growth slowed significantly after 2 weeks of SD, was minimal by the 4th week of SD and most of the shoot tip meristems had abscised after 6 weeks of SD. Endodormancy was induced after 4 weeks of SD. DW of the stem and buds increased with increasing duration of LD and SD. While bud DW increased more under SD than LD, stem DW increased more under LD than SD. T2 relaxation times were calculated from images of transverse sections of the grape node. There was a slight decrease in the T2 times in the node tissues with increased duration of LD treatment, whereas SD induced a significant decrease in T2 times during endodormancy induction. T2 values for the node decreased after 4 weeks of SD, coinciding with endodormancy induction. Separation of node tissues into bud, leaf gap, and the remainder of the stem and analysis of the proportion of short and long T2 times within those tissues indicated differential tissue response. A greater proportion of short T2 times were observed in the 2-week SD leaf gap tissue than in the LD and the proportion of short T2 times continued to increase with subsequent SD treatment. Bud and all other stem tissues had a greater proportion of short T2 times after 4 weeks of SD, coinciding with bud endodormancy induction. The proportion of short and long T2 times in a tissue was a better indicator of endodormancy than the averaged T2 time for the tissue. Thus, MRI allows nondestructive identification of differential tissue response to photoperiod treatments and makes it possible to separate normal vegetative maturation responses from endodormancy induction.

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Christopher J. Clark, Annette C. Richardson, and Ken B. Marsh

Whole-fruit proton magnetic resonance (MR) imaging was performed on satsuma mandarin (Citrus unshiu Markovich cv. Miho Wase) during a 15-week period commencing 10 weeks after anthesis and continuing to maturity, and at 6 weeks after anthesis the following season. Images with long repetition times (>1600 ms) and short echo times (20 ms) provided the clearest details of anatomical changes in the peel (flavedo, albedo) and vascular system, while those with similar repetition times but longer echo times (120 ms) were best for viewing juice sac morphology within pulp segments. At 6 weeks after anthesis, images of fruits of slightly different physiological ages highlighted rapid changes in the vascular bundles and albedo tissue at this stage of development. Variation in the relaxation measurements, T1 and T2, was determined from quantitative MR images of the juice sacs in equatorial slices, and images of expressed juice from whole fruit. Seasonal measurements of T1 determined in situ (1760 ms) were significantly greater than those in juice (1413 ms). By contrast, there was no mean seasonal difference between in situ T2 measurements (360 ms) and those for juice (332 ms). No associations between trends in the MR data and total soluble solids, pH, titratable acidity, and sugar and organic acid composition of the juice were established. Cell structure is identified as a hindrance in the use of quantitative MR imaging for probing compositional changes in solution in serial imaging studies.

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John L. Maas and M.J. Line

We report the use of nuclear magnetic resonance (NMR) imaging to detect differences in invasion and colonization of fruit by pathogens (Botrytis cinerea, Colletotrichum acutatum, and Phytophthora cactorum), and bruise wounds are sharply distinguishable from healthy fruit tissue by their T1 times. Digitized images from T1 images clearly show two or more zones of pathogen activity in fruit tissue. The innermost zone corresponds to the area of greatest invasive activity at the leading margin of the infection. A second zone corresponds to the area of tissue that has been killed and is being degraded by the pathogen. Sometimes, a third zone is present at the outer border of the lesion and this correspond to where aerial sporulation may occur. Images of bruises, however, are uniform with no apparent gradations in T1 characteristics. Detection of fruit deterioration and decay is important in understanding and controlling postharvest loss of fruit crops. The nondestructive nature of MRI provides a means to quantify the process of decay development and control measures applied to fruits.