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Matthew D. Kleinhenz and Jiwan P. Palta

Micropropagated `Red Norland' plants were transferred to an inert mixture of 1 perlite: 1 medium-grain quartzite (v/v) and grown 21 days at 20°C day/15°C night on a 25% Hoagland solution without Ca(NO3)2 (Ca at 10 mg·L–1 from CaCl2, N at 35 mg·L–1 from KNO3). Thereafter, Ca treatments (Ca at 0.2, 1, 5, 25, 125 mg·L–1) were imposed for 21 days with other nutrients unchanged. Day/night temperatures were 20/15°C and 35/20°C for control and stress plants, respectively. Continuous drip supply of nutrient solution in excess of demand maintained target rhizospheric Ca levels. All experiments were conducted in controlled-environment chambers with 400-μmol·m–2·s–1 light level. The following results were obtained. 1) Stress, but not control, plants grown with Ca at 0.2 and 1.0 mg·L–1 displayed reduced leaf expansion, extreme senescence, and death of the primary shoot meristem. 2) Plants grown with Ca at 5, 25 and 125 mg·L–1 grew normally under both temperature regimens, although plants responded to temperature with different biomass partitioning. (3) Total root mass at harvest was similar under all Ca–temperature combinations but low-Ca-treated plants had comparatively darker roots with fewer branches. (4) Light microscopic evaluation revealed normal staining patterns of lignified elements in leaves and stems of all plants. These data suggest that constant rhizospheric Ca levels >1 mg·L–1 are required for continued plant growth during exposure to heat stress.

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Beth Ann A. Workmaster and Jiwan P. Palta

Recent work in our laboratory has shown that pre- and postharvest applications of lysophosphotidylethanolamine (LPE) retard senescence processes in several fruit and flower species (apple, tomato, carnation). Banana was selected to develop a rapid bioassay to test the effects of LPE and other substances on various processes associated with senescence. Excised peel pieces from fully yellow `Grand Nain' bananas (Musa AAA) were incubated in petri dishes containing LPE solution (0, 25, 50, and 100 ppm) for 4 days. Fresh weight and ethylene production was measured daily. At the end of the experiment, tissue density, ion leakage, and soluble protein leakage was measured. Ion and soluble protein leakage was significantly lowered with 100 LPE. The 100 ppm LPE also significantly inhibited ethylene production after only 2 hours of treatment and this low level was maintained during the experiment. Peel tissue from the 100 ppm LPE remained firm and intact while tissue from the other treatments expanded and lost integrity. By day 2, peel from the 0, 25, and 50 ppm LPE gained significantly in fresh weight, while tissue treated with 100 ppm initially lost and then only slightly gained in fresh weight. Our results suggest that LPE is able to protect membrane function in senescence. Furthermore, these results provide evidence that LPE may also be retarding senescence by modulating the ethylene pathway.

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Laurie S. Weiss and Jiwan P. Palta

At the University of Wisconsin Biotron facility potted plants of S. tuberosum were frozen slowly (cooling rate of 1°C/h) to -2°C. Following thaw, plants were subjected to either high light (400 umol m-2s-1) or low light (100 umol m-2 s-1). High light caused greater damage which appeared as bleaching of the upper leaves in 2 days following thaw. In another study excised paired leaflet halves of S. tuberosum and S. commersonii were subjected to damaging but sublethal freezing temperatures and thawed either fast (on ice) or slowly (1°C/h). Membrane damage (% ion leakage) was about 2x higher at fast thaw as compared to slow thaw in both cold acclimated and non acclimated tissue. There was greater photosynthetic impairment at slow thaw rate than fast in the non acclimated state, but following acclimation fast thaw was more damaging to photosynthetic function. Respiration in general was less sensitive to freeze-thaw stress as compared to photosynthesis and cell membranes.

Our results show that we could benefit from taking into consideration thaw rate and post-thaw light intensity in developing frost protection plans.

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Sandra E. Vega and Jiwan P. Palta

Previous studies in our laboratory both in pine needles and potato leaves have shown evidence of an increase in 18: 2 (linoleate) in the purified plasma membrane fraction during cold acclimation. This increase was reversible on deacclimation, thereby suggesting a link between the accumulation of 18: 2 and acquisition of freezing tolerance. These studies suggest that the activity of specific desaturases may be modulated during cold acclimation. This study was aimed at studying the possible involvement of stearoyl-ACP desaturase (delta9) in potato cold acclimation response. Our approach was to study the induction of delta9 desaturase at the transcript level by using potato delta9 desaturase gene specific primers and reverse transcriptase. For this purpose, mRNA from S. tuberosum (cold sensitive, unable to acclimate) and S. commersonii (cold tolerant, able to cold acclimate) was extracted before and after acclimation. Sequence analysis confirmed that the amplified band was delta9 desaturase. Our results show that there is an increase in delta9 desaturase gene transcripts during cold acclimation and that this increase is associated with the cold acclimation response in potato. These results together with previous reports on the increase in 18: 2 in the plasma membrane during cold acclimation give more evidence toward the involvement of stearoyl-ACP desaturase (delta9) in the potato cold response.

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Beth Ann A. Workmaster and Jiwan P. Palta

Little is known about the growth and development of the cranberry plant (Vaccinium macrocarpon Ait.) in response to air and soil temperatures in the spring. During this period, marked changes in cranberry bud hardiness are known to occur (from –20 to 0 °C), with the greatest changes occuring before bud elongation. The ability to predict changes in bud phenology and hardiness in relation to thermal time would be useful to growers in making frost management decisions. To establish a working growth model, canopy air and soil temperatures were continuously recorded in 1996, 1997, and 1998 in a cranberry bed (cv. Stevens) in central Wisconsin. In spring, samples of uprights were randomly collected from several locations within the bed and sorted according to a nine stage bud classification from tight bud to bloom. Controlled freezing tests were performed on uprights from the most advanced stages present that constituted 10% or more of a sample on a given date. Heat units were calculated from hourly canopy air temperatures. Despite the varied weather conditions over the3 years, a distinct relationship existed between the accumulation of heat units and the advancement of the crop. Spring 1998 was very early and resulted in the accumulation of more heat units before initial and advanced bud swell was observed compared to the other 2 years. Initial evaluation suggests that soil temperatures between 5 to 10 °C and photoperiod may play a role in modulating the effect of air temperatures. Further refinement of this model and the predictive value for frost hardiness changes will be discussed.

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Stephen B. Ryu and Jiwan P. Palta

Lipids have been thought to be important largely in membrane structure and energy reserve. It is now evident that lipids and lipid-derived metabolites play a role in many critical cellular processes. Recent studies have shown that membrane lipid-based signaling mediated by phospholipases such as phospholipase A2 (PLA2), phospholipase C (PLC), and phospholipase D (PLD) constitutes a crucial step in plant responses to abiotic and biotic stresses. Phospholipases and their products also play a role during plant growth and development. For example, PLA2-derived lysophospholipids acted as growth regulators that retard senescence of plant tissues. Interestingly, the PLA2 products inhibited the activity of PLD, which has been suggested to be a key enzyme responsible for membrane lipid breakdown leading to plant senescence. Endogenous levels of lysophospholipids, such as lysophosphatidylethanolamine (LPE), could be increased in castor bean leaf discs by the treatment of auxin (50 μM), which is known to be a activator of PLA2. Pretreatment of leaf discs with a PLA2 inhibitor before auxin treatment nullified the auxin effect and rather resulted in accelerated senescence even compared to the nontreated control. Our recent results suggest a potential role of PLA2 products as biologically active molecules mediating hormonal regulation of growth and senescence. One such product LPE is being commercially exploited for retarding senescence and improving shelf life of fruits, vegetables, and cut flowers.

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Mustafa Ozgen, Senay Ozgen, and Jiwan P. Palta

Recent studies from our laboratory have demonstrated that lysophoshatidylethanolamine (LPE) is able to accelerate fruit ripening while at the same time promoting shelf life. LPE is a natural lipid and is commercially extracted from egg yolks and soybeans. We studied the influence of LPE on the pattern of anthocyanin accumulation and storage quality of cranberry fruit (Vaccinium macrocarpon Ait. cultivar Stevens). For this purpose 2 x 2-m plots were established in cranberry beds at two separate locations near Wisconsin Rapids. Experiments were conducted in 1997 and 1998 seasons. Plots were sprayed with LPE (extracted from egg yolk and soybean) 3 to 4 weeks before harvest. Spray solution included 200 ppm LPE, 3% ethanol, and 0.1% detergents (either Tergitol or Sylguard). Fruit samples were taken from a part in the plot periodically to determine the changes in the fruit. The rest of the plots were commercially wet harvested with a machine and stored in cold storage. Marketable fruit were counted at various times of cold storage to determine effect of LPE on shelf life of cranberries. In general, application of LPE from both sources resulted in 20% to 35 % increase in fruit anthocyanin contents. Also LPE treatment resulted in 10% to 20% increase in marketable fruit in cold storage. A postharvest dip of cranberry fruit with 50 ppm LPE solution for 15 min also resulted in about a 20% to 30% increase in marketable berries during cold storage. The results of this study shows that pre- and postharvest applications of LPE can add value to cranberry crop including better and more uniform colored fruit, enhance self life, and earlier harvest.

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Christopher C. Gunter and Jiwan P. Palta

Tuber tissue calcium has been linked to several potato quality characteristics, including internal defects and the susceptibility of tubers to decay by soft rotting Erwinia species. We were particularly interested in studying the relationship between supplemental calcium fertilization during the seed tuber production cycle to raise the seed piece calcium concentration and the impact on crop performance the following season. The role of seed tuber tissue calcium level on seed piece decay, growth, development, and performance of the plant was evaluated for cultivars Russet Burbank, Dark Red Norland, Atlantic, Superior, and Snowden. This study was performed over four growing seasons. Seed tubers were raised with varying calcium and the following season, individual tubers (over 3,000 total for 4 years of study) were sampled for calcium and hand planted in the field. They were evaluated for seed piece decay and total tuber yield during the growing season. Seed tubers raised with supplemental calcium resulted in significantly higher mean calcium content than the control tubers. In general, calcium-raised seed tubers tended to produce a more vigorous main sprout and higher tuber yield. We also found that there are significant differences among these cultivars for the characteristics measured. Consistently, in all three years, `Atlantic' responded to test conditions with the lowest decay values, and `Dark Red Norland' consistently showed the highest decay values. This suggests that there may be a genetic component involved in these two responses and these genotypic differences could be exploited to improve cultivated potatoes.

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Senay Ozgen, Mustafa Ozgen, and Jiwan P. Palta

Several recent studies, including from our laboratory, have provided evidence that by improving tuber calcium level, we can improve tuber quality such as low internal defects and better storability. The purpose of this study was to be determine the influence of supplemental calcium fertilization on tuber size and tuber number. For this purpose, plantlets of Solanum tuberosum cv. Russet Burbank raised in tissue culture were planted in 20-L pots filled with sandy loam soil with pH of 6.9 and soil calcium level of 350 ppm. All treatments received same total amount of nitrogen (at the rate of 280 kg·ha–1). Five treatments were evaluated: i) nonsplit nitrogen (from ammonium nitrate), ii) split nitrogen (from ammonium nitrate), iii) split nitrogen + gypsum, iv) split nitrogen (from liquid nitrogen) + calcium chloride, and v) split nitrogen (from calcium nitrate). The total calcium was applied at the rate of 168 kg·ha–1. Gypsum application was made at 4 weeks after planting, and other sources of calcium were applied on a split schedule (equally split at 4, 6, 8 weeks after planting). Four months after planting, tubers were harvested and evaluated. In general, all calcium treatments had lower tuber number and greater tuber size compared to the nonsplit nitrogen control. The percentage of total A-grade tubers as well as the percentage yield from A-grade tubers was increased by all calcium applications. These results suggest that calcium content I the soil can influence both potato tuber number and tuber size.

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Ahmed A. Tawfik and Jiwan P. Palta

The optimum temperature regime for Solanum tuberosum cv. Russet Burbank is usually 20/15°C day/night. We studied the impact of heat stress (30/25°C, day/night) on the growth of this heat sensitive cultivar under controlled conditions (UW-Biotron). Plants were grown in sandy-loam soil which tested at 1500 Kg/ha Ca. Plants were at the maximum temperature for 6h during the middle of the day with a photoperiod of 14 hrs. All pots received identical amounts of total N (rate: 225 Kg N ha1.). The treatments were: (1) NSN: non-split N (N application 1/2 emergence, 1/2 two wks later): (2) SPN: split-N (1/2 emergence 1/6 at 2, 5 and 8 wks later); (3) SPN+Ca: Split-N+Ca (Ca at 2, 5 and 8 wks after emergence, total Ca from CaNO3 was 113 Kg ha1). Total leaf FWT and DWT was significantly reduced in NS treatment by heat stress at 13 wks as compared to optimum conditions. However, this was not reduced in SPN and SPN+Ca. Under heat stress: (a) SPN + Ca gave the highest leaf FWT and DWT, stomatal conductance, transpiration rate, and leaflet tissue Ca content; (b) Young expanding leaflets gave higher growth rate with SPN and SPN + Ca than NSN; (c) Ca content of mature leaflet decreased progressively in both NSN and SPN but not in SPN + Ca. Our results show that application of Ca and N during heat stress can mitigate stress effects and that maintenance of a certain level of calcium in leaf tissue is important under heat stress.