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Matthew D. Kleinhenz, Christopher C. Gunter, and Jiwan P. Palta

A direct comparison was made of several commercially available calcium sources applied on two different schedules for their effectiveness in increasing tuber medullary and periderm tissue calcium concentrations in 170–284-g tubers of the cultivar Atlantic grown on a Plainfield sandy loam. Plots (6 x 3 m) were arranged in a CR design in 1993 and a RCBD in 1994 (eight replications). Paired measurements of tuber Ca concentration and internal quality (±hollow heart, ±internal brown spot) were made on individual tubers produced in plots with no additional or additional Ca (168 kg Ca/ha) supplied from either gypsum, liquid calcium nitrate, or NHIB. Two Ca and N application schedules were compared: 1) application at emergence and hilling (non-split), 2) application at emergence, hilling, and 4 and 8 weeks after hilling (split). All plots received 224 kg H/ha balanced with ammonium nitrate. In general, tuber yield and grade were unaffected by treatments in 1993 and 1994, but overall percent A-grade was lowest and percent B-grade highest in 1993 compared with 1994 data. In 1993, all treatments receiving Ca had greater mean tuber medullary and periderm tissue Ca concentration values and a greater percentage of tubers with an elevated Ca concentration compared with non-Ca-supplemented controls. The overall incidence of tuber internal defects was 5% in 1993. All split schedule treatments receiving Ca showed 0% internal defects. In contrast, nearly 8% of the tubers from control plots showed some defect. The medullary tissue Ca concentration of 65% of the tubers having either defect was below the median value of Ca concentration for the entire experiment in 1993. Similar evaluations are underway for the 1994 crop. These data suggest that tuber calcium concentration may be related to the incidence of these internal defects.

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Mustafa Ozgen, Jiwan P. Palta, and Stephen B. Ryu

Ethephon [2-(chloroethyl) phoshonic acid] is used widely to maximize the yield of ripe tomato fruits. However, ethephon causes rapid and extensive defoliation, overripening, and promotes sunscald damage to the fruit. Recent studies from our laboratory have provided evidence that lysophosphatidylethanolamine (LPE) can reduce leaf senescence. We investigated the potential use of LPE to reduce damaging effect of ethephon on tomato foliage and influence on the activity of phospholipase D (PLD). Disruption of membrane integrity has been suggested as a primary cause of senescence in plants. PLD is known to be a key enzyme, which initiates the selective degradation of membrane phospholipids in senescing tissues. Two-month-old tomato plants (`Mountain Spring') grown in greenhouse condition were sprayed with water, 200 ppm LPE, and 1000 ppm ethephon. In addition, LPE spray prior to ethephon or mixture with ethephon were also tested. Leaves were sampled after 0, 2, 5, 24, 72, and 168 h of spray application, for PLD activity measurements. Spray of LPE prior to ethephon spray or inclusion of LPE in the ethephon spray reduced foliar injury by ethephon. Activity of soluble PLD was increased dramatically in leaves sprayed with ethephon initially and than dropped by 7 days. We also found that LPE-treated leaves had lower PLD activity than the ethephon-treated leaves. Plants treated with LPE-ethephon mixture also showed significantly lower PLD activity. These results suggest that LPE treatments mitigate ethephon injury to tomato plants. Furthermore, it appears that this mitigation involves modulation of the activity of PLD.

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Beth Ann A. Workmaster, Jiwan P. Palta, and Jonathan D. Smith

In Wisconsin, the cranberry plant (Vaccinium macrocarpon Ait.) is protected from freezing temperatures by flooding and sprinkle irrigation. Due to the high value of the crop, growers typically overprotect by taking action at relatively warm temperatures. Our goal is to provide recommendations for improved frost protection strategies by studying seasonal hardiness changes in different parts of the cranberry plant (leaves, stems, buds, flowers, fruit). Stages of bud growth were defined and utilized in the hardiness determinations. Samples were collected from mid-April to mid-Oct. 1996 and cuttings were subjected to a series of freezing temperatures in a circulating glycol bath. Damage to plant parts was assessed by visual scoring and observation, ion leakage, and evaluation of the capability to regrow. The following results were obtained: 1) Overwintering structures, such as leaves, stems, and buds, can survive temperatures <–18°C in early spring, and then deacclimate to hardinesses between 0 and –2°C by late spring. 2) In the terminal bud floral meristems are much more sensitive to freeze–thaw stress than are the vegetative meristems. 3) Deacclimation of various plant parts occurred within 1 week, when minimum canopy temperatures were above 0°C, and when the most numerous bud stage collected stayed the same (bud swell). 4) Fruits >75% blush can survive temperatures of –5°C for short durations. By collecting environmental data from the same location we are attempting to relate plant development, frost hardiness, and canopy temperatures (heat units).

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Sandra E Vega, Jiwan P. Palta, and John B. Bamberg

Frost injury limits the cultivation of potatoes in many regions around the world. We are currently studying the factors that contribute to frost survival in potato in an attempt to improve its frost tolerance. Wild potato species have been distinguished for their high degree of non-acclimated frost tolerance (growing under normal conditions) and their high cold acclimation capacity (able to increase frost tolerance upon exposure to cold). Cold acclimation can be reversed upon exposure to warm temperatures (deacclimation). The ability to gain freezing tolerance rapidly in response to low temperatures as well as not being able to deacclimate rapidly in response to warm daytime temperatures would be advantageous for a plant against spring or fall freezes. Last year we presented evidence for the variability in the speed of cold acclimation among 7 wild tuber-bearing potato species (S. acaule, S. commersonii, S. megistacrolobum, S. multidissectum, S. polytrichon, S. sanctae-rosae and S. toralapanum). The same set of species was used for the present study to find out if there is also variability for the speed of deacclimation. Relative freezing tolerance of these species was measured before and after cold acclimation as well as after one day of deacclimation (exposure to warm temperatures). Our results suggest that there are differences in the speed of deacclimation among these species. We found that while some species lost near a half of their hardiness, others lost only a third or less of their hardiness after one day of deacclimation.

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Christopher C. Gunter, Matthew D. Kleinhenz, and Jiwan P. Palta

Pathogenic bacteria (Erwinia spp.) can have a significant impact on stand establishment and sprout health. These bacteria cause soft rot of the seed piece, which is common in wet spring conditions resulting in great economic loss. Recent studies have demonstrated that the incidence and severity of soft rot can be significantly reduced by increasing tuber calcium. We investigated the influence of seed piece calcium on tuber production. Field growing potato plants were treated with supplemental calcium during tuber bulking (calcium at 168 kg·ha–1 from calcium nitrate or N-Hib) to increase the seed tuber medullary calcium levels. All three cultivars (`Superior', `Atlantic', `Dark Red Norland') responded to supplemental calcium application with increases in mean calcium contents, even though soil tests showed high native calcium levels (1000 kg·ha–1). Seed tubers were sampled for calcium by removing a longitudinal slice from the center of each tuber and planting one of the resulting halves for seed piece decay evaluation and the other for yield evaluation. Calcium nitrate-treated seed tubers produced higher yields compared to ammonium nitrate and N-Hib in `Atlantic' in `Dark Red Norland'. This trend did not hold true for `Superior'. Our results suggest that a) it is possible to increase seed piece tuber calcium levels with supplemental calcium application even in soils testing high in calcium and b) improving the calcium concentration of the seed piece can increase tuber yield in some cultivars.

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Rita A. Teutonico, Jiwan P. Palta, and Tom C. Osborn

Identification of the genes involved in freezing tolerance in oilseed Erussica could lead to genetic improvement of winter survival of this crop and other species, as well as provide greater understanding of the basis of cold stress tolerance in plants. We developed a genetic linkage map for B. rapa using restriction fragment length polymorphisms (RFLPs) and identified molecular markers which are linked to genes controlling vernalization requirement and freezing tolerance. We mapped the location of a group of cold-regulated (`cor') genes from Arabidopsis thaliana in this population and determined their association with freezing tolerance and vernalization requirement. We developed genetically fixed, recombinant inbred lines of B. rapa to assay the physiological processes involved in these cold responses. Specifically, we measured the differences in lipid composition of the plasma membranes of acclimated and nonacclimated plants of a subset of this population. We will determine if the genes involved in the physiological responses to cold temperature are also associated with the acquisition of freezing tolerance.

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Laurie S. Weiss, John B. Bamberg, and Jiwan P. Palta

Solanum acaule (acl) and Solanum commersonii (cmm) represent the extremes of frost tolerance and cold acclimation ability among potato species. We have combined these species with cultivated S. tuberosum (tbr) to develop a potato with desired tuber traits and a high degree of frost tolerance. For this purpose diploid cmm was made 4x and crossed with naturally 4x acl. The F1 and F2 appear to exhibit hybrid vigor for vine growth for flowering, but none had frost tolerance greater than the parents. The F1 and F2 were crossed with S. tuberosum ssp. andigena and Katahdin via 2n eggs resulting in 6x 3-way hybrids. These hybrids were evaluated both in the field and laboratory for frost tolerance and acclimation ability. Results showed an increase of 1°C of frost tolerance and 2°C increase in cold acclimation capacity in the hybrids as compared to the sensitive tbr parents. Some of the 6x (3-way) hybrids produced significant tubers but yield and earliness needs much improvement. These results demonstrate that it should be possible to move both non acclimated freezing tolerance and cold acclimation ability from wild to cultivated species and offer exciting opportunities to enhance potato production in frost prone areas in the world.

Supported by USDA/NRI grant 91-3700-6636 to J.P.P. and J.B.B..

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Björn H. Karlsson, Jiwan P. Palta, and Peter M. Crump

Our previous research has provided evidence that in-season calcium applications can increase tuber calcium and improve tuber quality with reduced internal defects. To determine if increasing the tuber calcium concentration also mitigates tuber bruise incidence, five commercially relevant potato (Solanum tuberosum L.) cultivars (`Russet Burbank', `Atlantic', `Snowden', `Superior', and `Dark Red Norland') were grown during three seasons, 1999–2001. Three split applications of a calcium/nitrogen water soluble blend totaling 168 kg·ha–1calcium were made starting at hilling. All plots, including controls, received an equal amount of total nitrogen in a season. Tubers were allowed to be bruised during normal machine harvest standard to commercial production in Wisconsin. Over 100 tubers from each replication (5–10 replications/treatment) were cut and examined for the incidences of bruise and internal brown spot. Paired samples of medullary tissue were taken for measuring calcium concentration. As expected, tuber tissue calcium concentration increased significantly, in all cultivars and in all years, with in-season calcium application. Bruise incidence varied among cultivars and seasons. Although tuber calcium concentration varied among seasons, `Atlantic' and 'Snowden' consistently had the lowest calcium concentration, whereas `Superior' and `Dark Red Norland' consistently had the highest calcium concentration. Meta-analysis of pooled data for three years showed that blackspot bruise incidence was significantly reduced with calcium application in `Atlantic', `Burbank', and `Snowden'. On the other hand, `Dark Red Norland' and `Superior' had low incidence of bruise and were unaffected by calcium applications. Regression analyses of pooled data from all cultivars for three years revealed a significant quadratic relationship between blackspot bruise and tuber tissue calcium as well as between blackspot bruise and internal brown spot. A linear to plateau plot of medullary calcium concentration versus blackspot bruise incidence revealed that bruise incidence is minimized between 200 and 250 μg/kg (dry wt)–1 tuber calcium concentration. To our knowledge, ours is the first study providing evidence for reducing bruise by improving tuber calcium. Variations in the bruise incidences among cultivars generally followed tuber calcium concentration suggesting a genetic control. Given the role of calcium in improved membrane health and enhanced wall structure, and as a modulator of physiological responses, it is not surprising that internal brown spot and bruise incidences are reduced by in-season application to calcium-deficient cultivars.

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Senay Ozgen, James S. Busse, and Jiwan P. Palta

The important roles of calcium on plant growth and development including cell division and cell elongation is well documented. The purpose of the present study was to determine the impact of root zone calcium on the growth and health of potato apical meristem and on the maintenance of apical dominance. For this purpose, single-node potato cuttings (Solanum tuberosum L. cv. Dark Red Norland) were grown in sterilized modified Murashige and Skoog (MS) media containing varying concentrations of calcium (1 to 3000 μM). After 13 to 30 d of growth, plantlets were harvested and data for height of the main shoot and for the number of axillary shoots produced were recorded. Plantlets were ashed and tissue calcium concentration was determined. Shoot height was retarded with decreasing concentration of calcium in the media. Calcium deficiency induced shoot tip injury and loss of apical dominance. Tip injury was followed by the development of axillary shoots. The number of axillary shoots increased from one to 21 as calcium concentration in the media decreased from 3000 to 1 μM. At calcium concentration of 1500 μM or higher, there was a single main shoot with no axillary shoots. Addition of ethylene glycol tetra acetic acid (EGTA), a calcium chelator, to the media with 2720 μM calcium (sufficient calcium) resulted in the development of shoot injury and in the formation of axillary shoots. Calcium deficiency injury symptoms were prevented by the addition of a calcium analog, strontium, to MS media deficient in calcium. Strontium has been reported to strongly bind to plant cell walls and the inclusion of strontium prevented injury in shoots of plants grown on calcium-deficient media. These results suggest that strontium is able to mimic the role of calcium in the maintenance of cell wall integrity and supports previous studies that showed that calcium deficiency results from cell wall collapse of the subapical cells.

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James S. Busse, Senay Ozgen, and Jiwan P. Palta*

Calcium deficiency in the potato shoot results in sub-apical necrosis. This is a physiological condition whereby necrotic lesions form a few millimeters below the shoot apex ultimately causing shoot tip senescence, loss of apical dominance, and the release of axillary buds. Using a Dark Red Norland tissue culture system, we studied the relationship of root zone calcium levels to shoot tip maintenance. Root zone calcium levels lower than 50 ppm resulted in shoot tip death and prolific branching from axillary buds. Chelator studies with EGTA and tracer studies with 45 Ca, indicated a direct involvement of calcium at the shoot tip for shoot tip maintenance. Interestingly, low root zone calcium deficiency syptoms could be mitigated with 0.001 to 0.01 μM of the auxin analog NAA. Developmental studies of calcium deficiency symptoms indicate no anatomical relationship with shoot tip necrosis as xylem conducting elements were found near the shoot apex regardless of the root zone calcium level. These results have important implications for potato shoot development especially during the early development stage from the seed piece.