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A.G. Taylor, D.H. Paine, and C.A. Paine

The purpose of this project was to study factors that influence the leakage rate and develop methods to enhance leakage of sinapine from Brassica seeds. Six seedlots (two seedlots of one cultivar each of cabbage (Brassica oleracea L., Capitata group), broccoli, and cauliflower (B. oleracea L., Botrytis group) were studied. Leakage was quantified spectrophotometrically by measuring the absorbance of the soak water at 330 nm. The onset of sinapine leakage was determined from single seeds by the visual presence (yellow soak water) caused by soaking seeds in a biological buffer adjusted to pH 10. The leakage pattern from heat-killed seeds of all seedlots was sigmoidal with a distinct lag phase followed by a rapid efflux and final slower rate. The duration of the lag phase and the total amount of sinapine leaked after 24 hours was not the same for all seedlots after adjusting for seed count, seed weight, or sinapine content. Therefore, another factor was responsible for differences measured in leakage. Embryos or seeds with cracked testas leaked faster than intact seeds, and the leakage pattern without testa integrity was biphasic. From these studies, we conclude that the testa was a major factor regulating sinapine leakage. Pretreating heat killed seeds, with up to 1.0% NaOCl for 10 minutes, accelerated the onset of leakage. The time for 50 percent of the seeds to leak (T50) decreased as NaOCl concentration increased. Leakage uniformity, as measured by the standard deviation of the distribution, generally increased as NaOCl concentration increased. The sigmoidal leakage pattern from heat-killed Brassica seeds may be attributed to seedcoat cracking associated with imbibitional swelling. A NaOCl pretreatment may have increased the permeability of the testa and, thereby, enhanced the leakage rate.

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Geeta K. Nanaiah and Jeffrey A. Anderson

Abbreviations: EER, ethane: ethylene ratio; EL, electrolyte leakage. 1 Graduate Assistant. 2 Associate Professor. To whom reprint requests should be addressed. Journal article no. 6121 of the Agr. Expt. Sta., Oklahoma State Univ., Stillwater

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Mario Valenzuela-Vázquez, Geno A. Picchioni, Leigh W. Murray, and Wayne A. Mackay

, electrolyte leakage, and vase life longevity. We held racemes in vase solutions with or without ethephon [(2, chloroethyl) phosphonic acid, or CEPA] to investigate the ability of 1-MCP to counteract the influence of an exogenous ethylene source (ethylene

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Charles F. Forney, Sharon J. Peterson, and Preston Hartsell

Callus tissue grown from `Marsh' grapefruit (Citrus paradisi Macf.) albedo tissue was grown at 30C for ≈ 40 days. Calli were preconditioned in normal air for 5 days at 10 or 30C before being fumigated for 2 hr with 0, 32, or 48 g of methyl bromide (MB)/m 3. Calli were then held at 10C and K+ leakage was measured after 1, 10, 20, and 30 days. The amount of K+ leaked from MB-fumigated calli was greater than that for nonfumigated calli and increased with higher MB dose. Leakage also increased with time following fumigation. Leakage of calli preconditioned at 30C and fumigated with 48 g MB/m3 was 140%, 196%, and 260% greater than leakage from nonfumigated calli 10, 20, and 30 days after fumigation, respectively. Leakage from calli preconditioned at 10C for 5 days before MB fumigation was less than that from calli held at 30C. MB doses of 32 and 48 g·m-3 increased leakage of calli preconditioned at 10C by 6% and 43% and for those preconditioned at 30C by 99% and 140%, respectively, 10 days after fumigation. In addition to K+ leakage, MB induced the development of a tan to orangish-brown discoloration.

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Susan Lurie, Reuven Ronen, and Shimon Meier

Storing `Maor' green bell peppers (Capsicum annuum L.) for 3 weeks at 2C resulted in the development of chilling injury (CI) evidenced as surface pitting. Fruit held at 8C did not develop any CI symptoms, but, after 3 weeks of storage, the fruit began to change color from green to red. PAM fluorometry was used to measure changes in photosynthetic competency in whole green bell peppers. Three photosynthetic characteristics could be measured by this method: quantum yield (Fm/Fe), photochemical quenching (Qp), and nonphotochemical quenching (Qnp). Fm/Fo decreased 90% during the first week of storage at 2C and remained low thereafter, while Qnp decreased after 2 weeks at 2C, just before the peppers began to develop CL Qp was similar at both storage temperatures. Potassium leakage as a CI measurement also increased in excised pepper discs after 2 weeks at 2C. The results indicate that PAM fluorometry can measure CI nondestructively before tissue damage is visible in green peppers.

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Leigh E. Towill, John W. Waddell, and Philip L. Forsline

Three years ago we established a long-term cryogenic storage project for apple germplasm and utilized grafting of buds obtained from stored dormant shoot sections as the major viability assay. Grafting, however, is time consuming and requires considerable skill. Electrolyte leakage and oxidative browning tests were used as alternative viability assays. Using leakage from individual buds in a multiwell analyzer, we examined modifications of the electrolyte leakage test and analyzed the kinetics of leakage in an attempt to determine whether the test can predict grafting success. The results suggest that more buds were viable than were estimated by the grafting test. In vitro culture is being examined to test this and to determine if practical recovery is feasible for diversity within the germplasm collection.

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Dominique Lacan and J.C. Baccou

Respiration, C2H4 production, lipid composition, and electrolyte leakage were monitored during ripening of two nonnetted muskmelon (Cucumis melo L.) varieties differing in their storage life: `Clipper' (a long-storage-life variety) and `Jerac', which was used as a control. Respiration rates were comparable in both varieties. Although `Jerac' exhibited normal climacteric C2H4 production, `Clipper' continued to produce significant amounts of C2H4 until senescence. Electrolyte leakage increased with ripening and was always higher in `Jerac'. The loss of membrane integrity seems to be related to changes in the lipid composition due to a breakdown of phospholipids, an increase of sterol synthesis, and an increase in fatty acid saturation. On the contrary, in `Clipper', the absence of a major change in sterol and phospholipids content and the high level of fatty acid unsaturation suggest that membrane permeability is not greatly affected during ripening. This is consistent with the low loss of solutes measured and may delay senescence in `Clipper' fruit.

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G.A. Picchioni, S. Miyamoto, and J.B. Storey

Excised root tips from 3-year-old pistachio rootstock (Pistacia atlantica Desf., P. terebinthus L., and P. integerrima Stewart × atlantica) were exposed to laboratory saline solutions for 24 hr. Treatments simulated the compositions of soil solutions in a previous 2-year study made in outdoor lysimeters. Leakage of UV-absorbing solutes, an indication of cellular damage, occurred with 175 mM Na/12.5 mM Ca, which was comparable to soil salinity which increased leaf Na concentrations and decreased root growth of these species Up to. five times higher leakage occurred from roots of a P. terebinthus genotype having least Na exclusion potential during the lysimeter study. Use of isotonic levels of CaCl2, mannitol, and simulated Na/Ca solutions resulted in similar damage. However, isotonic Na (-Ca) caused highest leakage overall. Correlation between long-term observations in the lysimeters and leakage occurrence-in the laboratory indicates that solute leakage tests may aid in characterizing responses of Pistacia spp. roots cocks to saline conditions.

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P.C. Lee, A.G. Taylor, and T.G. Min

Sinapine leakage to detect seed germination potential on a single-seed basis in Brassica has been developed as a rapid test. In this test, sinapine leakage predicts that a seed is non-germinable; however, the major source of errors in this method are false-negative (F–)—i.e., the method predicted a seed was germinable because the seed did not leak, and it did not germinate. The sinapine leakage index (SLI) was used to asses the F– for any seed lot by dividing the number of non-germinable seeds that leaked sinapine by the total number of non-germinable seeds. Seed lots including cabbage, cauliflower, and broccoli (B. oleracea L., Captitata, Botrytis, and Italica groups, respectively) were used to examine the F–. The leakage rate as measured by T50, the time for 50% of heat-killed seeds to leak, was linearly correlated to SLI. Cabbage seeds were viewed by scanning electronic microscopy and leaking non-germinable seeds either had cracks or were shrunken. NaOCl pretreatment has been found to increase the rate of sinapine leakage and SLI. The mode of NaOCl was due to high pH altering the seed coat permeability. Chemical analysis was conducted on isolated seed coats for pectin, tannins, hemicellulose, cellulose, phenolic lignin, and cutin. It was found that the higher SLI (more permeable) lots contained lower amounts of cutin, suggesting that cutin may restrict the diffusion of sinapine through the testa.

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T.G. McCollum and R.E. McDonald

Storage of `Marsh' white seedless grapefruit (Citrus paradisi Macf.) for 2 weeks at 5C resulted in the development of chilling injury (CI). Electrolyte leakage from chilled fruit did not increase significantly until CI had become severe, and was therefore considered to be of limited value as an early indicator of CI. In contrast to electrolyte leakage, respiration and ethylene evolution were significantly higher in chilled than in nonchilled fruit, even before the onset of visual symptoms of CI. Respiration rates ranged from ≈8 to 11 and 5 to 7 ml CO2/kg per hour in chilled and nonchilled fruit, respectively. Ethylene evolution was not detected from nonchilled fruit, whereas chilled fruit produced from 45 to 250 nl ethylene/kg per hour. Results of this study indicate that electrolyte leakage does not increase until visible pitting of the flavedo has occurred, whereas stimulation of respiration and ethylene evolution occur early in the development of CI.