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David M. Eissenstat, James P. Syvertsen, Thomas J. Dean, Jon D. Johnson, and George Yelenosky

The combined effects of O3 and acid rain on freeze resistance, growth, and mineral nutrition were studied using broadleaf-evergreen citrus and avocado trees. Using a factorial design, `Ruby red' grapefruit (Citrus paradisi L.) trees on either Volkamer lemon (Citrus volkameriana Ten. & Pasq.) or sour orange (Citrus aurantium L.) rootstock and `Pancho' avocado trees (Persea americana Mill.) on `Waldin' rootstock were exposed to O3 and acid rain for 8 months in open-top chambers under field conditions. The O3 treatments were one-third ambient (0.3X), ambient (1X), twice ambient (2X), or thrice ambient (3X). Ambient O3 concentrations averaged 39.1 nl·liter-3 over a 12-hour day. The acid rain treatments had a pH of 3.3, 4.3, or 5.3 and were applied to simulate long-term rainfall averages. In general, the effects of acid rain on growth and freeze resistance were small. Rain of high acidity (pH = 3.3) offset the negative effects of O3 on growth (total leaf mass) in avocado and grapefruit/Volkamer lemon trees. In contrast, rain of high acidity magnified the detrimental effects of O3 on electrolyte leakage of leaf disks at subzero temperatures, especially for citrus. Freeze resistance, determined by stem and whole-plant survival following freezing temperatures, was lower in the most rapidly growing trees. Consequently, for trees exposed to a combination of O3 and acidic rain, leaf electrolyte leakage did not correlate significantly with stem survival of freezing temperatures. We conclude that the danger of acid rain to citrus and avocado in Florida is rather slight and would only present a potential problem in the presence of extremely high O3.

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Jiwon Jeong, Jeffrey K. Brecht, Donald J. Huber, and Steven A. Sargent

A study was conducted to determine the effect of 1-methylcyclopropene (1-MCP) on textural changes in fresh-cut tomato (Lycopersicon esculentum, Mill.) slices during storage at 5 °C. The relationship between fruit developmental stage and tissue watersoaking development was also determined. Fresh-cut tomato slices prepared from light-red fruit that had been exposed to 1-MCP (1 μL·L-1 for 24 h at 5 °C) retained significantly higher pericarp firmness during storage at 5 °C for 10 d than slices from nontreated fruit or slices stored at 10 or 15 °C and they also had a significantly higher ethylene production maximum. 1-MCP (1 or 10 μL·L-1 for 24 h at 5 °C) had no affect on the firmness of fresh-cut, red tomato slices at 5 °C or on slices prepared from 5 °C-stored, intact red tomatoes. Nor did 1-MCP treatment have a significant effect on electrolyte leakage of tomato slices or intact fruit stored at 5 °C. Slices from fruit of the same developmental stage but with higher initial firmness values had less watersoaking development and responded better to 1-MCP treatment during 8 d storage at 5 °C. 1-MCP (1 μL·L-1) was more effective in reducing watersoaking in light red stage tomato slices when applied at 5 °C for 24 h compared with 1-MCP applied at 10 or 15 °C. Watersoaking development was also more rapid in fresh-cut tomato slices as initial fruit ripeness advanced from breaker to red stage. Our results suggest that watersoaking development in fresh-cut tomato slices is an ethylene-mediated symptom of senescence and not a symptom of chilling injury as had previously been proposed.

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Zhaolong Wang and Bingru Huang

Drought is a major limiting factor for turfgrass growth. Understanding genetic variations in physiological responses of turfgrass to drought stress would facilitate breeding and management programs to improve drought resistance. This study was designed to evaluate responses of abscisic acid (ABA) accumulation, water relations, and gas exchange to drought stress in four Kentucky bluegrass (Poa pratensis L.) cultivars differing in drought resistance. Plants of `Midnight' and `A82-204' (drought resistant) and `Brilliant' and `RSP' (drought susceptible) were grown under well-watered (control) or drought stress conditions for 25 days in growth chambers. Turf quality, leaf water potential (Ψleaf), relative water content (RWC), leaf net photosynthesis rate (Pn), and stomatal conductance (gs) declined, while electrolyte leakage (EL) increased during drought progression in all cultivars. The magnitudes of the change in these parameters were greater for `RSP' and `Brilliant' than for `Midnight' and `A82-204'. Leaf ABA content in `RSP' and `Brilliant' increased sharply after 2 days of stress to as much as 34 times the control level at 10 days of drought. Leaf ABA content in `Midnight' and `A82-204' also increased with drought, but to a lesser extent than in the other two cultivars. Leaf ABA level was negatively correlated with Ψleaf and gs. `A82-204' had a significantly lower ABA accumulation rate with changes in Ψleaf during drought compared to `Midnight', `RSP' and `Brilliant'; however, no differences in ABA accumulation rate were detected among the latter three cultivars. In addition, leaf gs was more sensitive to changes in ABA accumulation in `Midnight' and `A82-204' than in `RSP' and `Brilliant'. These results demonstrated that drought tolerant cultivars of Kentucky bluegrass were characterized by lower ABA accumulation and less severe decline in Ψleaf, Pn, gs, and turf quality during drought stress than drought sensitive cultivars. Drought tolerance of Kentucky bluegrass could be related to sensitivity of stomata to endogenous accumulation of ABA under drought stress conditions.

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Wlodzimierz Ptasznik and Anwar A. Khan

The effects of various drying conditions on seed quality and performance of matriconditioned `Bush Blue Lake 47' snap bean (Phaseolus vulgaris L.) seeds were studied. An exponential model based on the Page equation provided a good fit (R2 = 0.9) to changes in moisture content during drying. Drying matriconditioned seeds with high initial moisture content (47.2%) for 5 to 6 hours at 35C, 30% to 35% relative humidity, and 0.7 to 1.4 m·s-1 air velocity (v) retained, and in some cases augmented, the benefits derived from conditioning. Matriconditioning greatly reduced electrolyte leakage (34.3 vs. 94.7 μS·cm-1·g-1 for nontreated seeds); drying to 15% moisture content at 0.7 or 1.4 m·s-1 v moderately increased the leakage rate (59.1 to 60.9 vs. 34.3 μS·cm-1·g-1), while drying at 0.02 m·s-1 v (ambient) increased the rate to that of nontreated seeds. The leakage rate remained low (43.6 to 50.8 μS·cm-1·g-1) in matriconditioned seeds dried to 22% moisture content at all air velocities. In growth-chamber studies, rapidly drying matriconditioned seeds to 15% moisture content at 1.4 m·s-1 v improved the emergence percentage over that of nontreated seeds, increased the shoot fresh and dry weight over that of nontreated and nondried matriconditioned seeds, and increased the shoot fresh weight over that of seeds dried at 0.02 or 0.7 m·s-1 v. Drying matriconditioned seeds to 15% moisture content at 0.7 m·s-1 v improved plant fresh weight over that produced by nontreated seeds. Rapid drying to 22% moisture content at 1.4 or 0.7 m·s-1 v improved only shoot fresh weight over that of nontreated seeds. In a 1992 field planting, percent emergence of matriconditioned seeds dried at 0.7 or 1.4 m·s-1 v was similar to that of nondried matriconditioned seeds and higher than that of nontreated seeds. No significant differences were noted in plant yield among the treatments.

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K.G.V. Davidson, F.D. Moore III, E.E. Roos, S. Nath, and S. Sowa

Five seed-quality indices based on individual seed electrolyte leakage tests were evaluated. Zea mays L. seeds were soaked for 6 hours, and individual seed leachate conductivity values were obtained. A total of 100 cells were scanned, one seed per cell, at 5-minute intervals for the first 30 minutes, followed by 15-minute intervals for the remaining 330 minutes. Seeds were allowed to dry for 5 to 7 days at room temperature and then were tested for germinability at 25C for 7 days. Radicle lengths were measured after 72 hours. The Richards function was fitted to cumulative frequency distributions of μAmps to obtain internal slope (IS), mean μAmp, and median μAmp values for each scan. Initial leach rate (ILR) was estimated after fitting hyperbolic functions to μAmp vs. soak time data. Average leach rate (ALR) was also derived from fitting the Richards function to μAmp vs. soak time data. Linear regression of seed quality on IS, mean, and median μAmp values after 5 hours of imbibition yielded r2 values of 0.91, 0.81, and 0.86 for predicting viability and 0.56, 0.46, 0.52 for predicting radicle length. Thus, IS was the best seed quality predictor, followed closely by median and mean μAmp values. ILR and ALR were not correlated with seed quality.

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Milton E. Tignor, Frederick S. Davies, Wayne B. Sherman, and John M. Davis

Poncirus trifoliata (L.) Raf. seeds were germinated in perlite under intermittent mist at about 25 °C and natural daylight in a greenhouse. Two-week-old seedlings were then transferred into a growth chamber at 25 °C and 16-hour daylength for 1 week. Tissue samples were collected at 0, 6, 24, 168, and 504 hours after temperature equilibration at 10 °C. Freezing tolerance at –6.7 °C, as determined by electrolyte leakage, and stem (leaves attached) water potential (ψx), measured using a pressure chamber, was recorded for a subset of seedlings for each time interval. Red coloration (apparently anthocyanin) developed at the petiole leaflet junction and buds after 48 hours at 10 °C and gradually occurred throughout the leaves during further exposure. Complementary DNA clones for phenylalanine ammonia lyase (PAL), 4-coumarate: coA ligase (4CL), and chalcone synthase (CHS) were used to probe RNA isolated from the leaves. No increase in steady-state messenger RNA level was detected. Increases in freeze hardiness occurred within 6 hours in the leaves, and continued for up to 1 week. Water potential initially decreased from –0.6 to –2.0 MPa after 6 hours, then returned to –0.6 MPa after 1 week. Thus, Poncirus trifoliata seedlings freeze-acclimate significantly after only 6 hours at 10 °C.

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Majken Pagter and Michelle Williams

projected hardiness of stems and buds after a given dehardening and rehardening treatment. Freeze tolerance of stems was determined using the electrolyte leakage method ( Pagter et al., 2011 ). One 3-cm-long piece of internodal stem tissue was rinsed under

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Magaji G. Usman, Mohd Y. Rafii, Mohd Razi Ismail, Mohammad Abdul Malek, and Mohammad Abdul Latif

measuring electrolyte leakage from leaves of plants at different temperatures. Several studies have indicated that CMT is effective in detecting genetic differences with regard to heat tolerance among several crops ( Islam et al., 2014 ; Kumar et al., 2012

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Jeffrey A. Anderson

`Early Calwonder' pepper (Capsicum annuum L.) and `Jubilee' corn (Zea mays L.) leaf disks exposed to high temperature stress produced ethylene, ethane, methanol, acetaldehyde, and ethanol based on comparison of retention times during gas chromatography to authentic standards. Methanol, ethanol, and acetaldehyde were also identified by mass spectroscopy. Corn leaf disks produced lower levels of ethylene, ethane, and methanol, but more acetaldehyde and ethanol than pepper. Production of ethane, a by-product of lipid peroxidation, coincided with an increase in electrolyte leakage (EL) in pepper but not in corn. Compared with controls, pepper leaf disks infiltrated with linolenic acid evolved significantly greater amounts of ethane, acetaldehyde, and methanol and similar levels of ethanol. EL and volatile hydrocarbon production were not affected by fatty acid infiltration in corn. Infiltration of pepper leaves with buffers increasing in pH from 5.5 to 9.5 increased methanol production.

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Jeffrey A. Anderson and Sonali R. Padhye

Although heat stress injury is known to be associated with membrane dysfunctions, protein structural changes, and reactions of activated forms of oxygen, the underlying mechanisms involved are poorly understood. In this study, the relationships between thermotolerance and hydrogen peroxide (H2O2) defense systems, radical scavenging capacity [based on 1,1-diphenyl-2-picrylhydrazyl (DPPH) reduction], and protein aggregation were examined in vinca [Catharanthus roseus (L.) G. Don `Little Bright Eye'], a heat tolerant plant, and sweet pea (Lathyrus odoratus L. `Explorer Mix'), a heat susceptible plant. Vinca leaves were 5.5 °C more thermotolerant than sweet pea leaves based on electrolyte leakage analysis. Vinca leaf extracts were more resistant to protein aggregation at high temperatures than sweet pea leaf extracts, with precipitates forming at ≥40 °C in sweet pea and at ≥46 °C in vinca. Vinca leaves also had nearly three times greater DPPH radical scavenging capacity than sweet pea leaf extracts. Two enzymatic detoxifiers of H2O2, catalase (CAT) and ascorbate peroxidase (APOX), demonstrated greater activities in vinca leaves than in sweet pea leaves. In addition, CAT and APOX were more thermostable in vinca, compared with sweet pea leaves. However, tissue H2O2 levels did not differ between controls and tissues injured or killed by heat stress in either species, suggesting that H2O2 did not play a direct role in acute heat stress injury in vinca or sweet pea leaves. Greater thermotolerance in vinca, compared with sweet pea, was associated with greater DPPH radical scavenging capacity, indicating that AOS other than H2O2 may be involved in acute heat stress injury.