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Karim M. Farag, Jiwan P. Palta, and Elden J. Stang

The application of ethanol for enhancing effectiveness of ethephon under field conditions on cranberry (Vaccinium macrocarpon Ait.) fruit was tested during three seasons (1986 to 1988). The formulation containing ethephon plus the surfactant Tergitol (0.3% or 0.5%, v/v) and ethanol (2.5%, 5%, or 10%) consistently increased anthocyanin content in the fruit by 28% to 54% over the control. In general, fruit size was not affected by the ethephon treatment containing ethanol and Tergitol. The application of ethephon plus surfactant did not increase the anthocyanin content in the fruit. The presence of ethanol in the ethephon and surfactant mixture, however, consistently enhanced the fruit anthocyanin content by 21% to 40% as compared to ethephon plus surfactant. No adverse effect of various treatments on vine growth or appearance was noticed over the three seasons. Chemical name used: (2-chloroethyl) phosphonic acid (ethephon).

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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.

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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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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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Yu-Kuang Chen, John B. Bamberg, and Jiwan P. Palta

Tetraploid somatic hybrids between S. tuberosum (tbr) and S. commersonii (cmm) have been produced to incorporate desirable traits such as cold hardiness from cmm into cultivated potatoes. While nonacclimated freezing tolerance (NA) of these somatic hybrids were as low as tbr fusion parent, their acclimation capacity (ACC) approximated that of the parental mean. In order to further explore the potential of using these somatic hybrids in breeding programs and to examine the segregation of genes conferring NA and ACC in somatic hybrids, progenies have been developed from either selfing somatic hybrids or crossing them with a tuberosum breeding line, Wis 231. In total, 48 selfed and 6 backcross progenies were characterized for the expression of NA and ACC. The NA derived from cmm was still poorly recovered in both sets of progenies. However, ACC did show some variation ranging from the level of sensitive fusion parent to that of the selfed parent, HA 26-5. None of the progeny had ACC as high as their cmm parent. Our results suggest that the expression of NA was suppressed by the cold sensitive genome of tbr. Thus, ACC is the form of cold tolerance from cmm, which appears to be most easily accessed though these somatic hybrids.

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

Two major components of frost resistance are freezing tolerance in the nonacclimated state (growing in normal condition) and capacity to cold acclimate (increase in freezing tolerance upon exposure to chilling temperatures). In addition to these two major components, numerous factors contribute to frost survival. Although the rate of cold acclimation and deacclimation have been recognized as important factors contributing to frost survival, very little information about them is available. Our objective was to determine if there is variability in the rate of cold acclimation and deacclimation among tuber-bearing wild potato species: S. acaule Bitter, S. commersonii Dunal, S. megistacrolobum Bitter, S. multidissectum Hawkes, S. polytrichon Rydb., S. sanctae-rosae Hawkes, and S. megistacrolobum subsp. toralapanum (Cárdenas & Hawkes) Giannattasio&Spooner. Relative freezing tolerance of these species was measured after 0, 3, 6, 9 and 12 days of cold acclimation and after 12 and 24 hours deacclimation. Our results showed there were differences in the rates of cold acclimation and deacclimation among these species. With respect to the rate of acclimation we found these species can be divided into four groups: (i) early; (ii) late acclimators; (iii) progressive acclimators, and (iv) nonacclimators. Likewise, a wide range of cold deacclimation behavior was found. Some species showed as low a loss of 20% of their freezing tolerance, others showed as much as >60% loss after 12 hours of deacclimation. Significant deacclimation was observed in all cold acclimating species after 1 day. These results demonstrate that the rates of cold acclimation and deacclimation were not necessarily related to the cold acclimation capacity of a species. Rapid acclimation in response to low temperatures preceding a frost episode and slow deacclimation in response to unseasonably warm daytime temperatures could be advantageous for plants to survive frost events. Thus, in addition to nonacclimated freezing tolerance and acclimation capacity, it would be very desirable to be able to select for rapid acclimation and slow deacclimation abilities. Results demonstrate that variability for these two traits exists in Solanum L. (potato) species.

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

Frost damage to the foliage is a common problem where potatoes are grown, and results in significant reductions in tuber yield. Frost injury also limits the cultivation of high-yielding S. tuberosum cultivars in the mountain regions of Central and South America, where potato is a staple crop. Recent studies have shown that some wild potato species possess a high degree of non-acclimated frost tolerance (growing in normal conditions) as well as high cold acclimation capacity (able to increase frost tolerance upon exposure to cold). Natural frosts affecting potatoes are of two types: a) late spring or early fall frost, where the minimum temperature during the frost episode can be very low; b) frost during the growing season, where the minimum temperature during the frost episode is not as low. It is expected that potato species able to acclimate rapidly would survive better from the latter type of frosts, whereas species having higher acclimation capacity might have a great chance to survive better from the former type of frosts. The objective of this study was to find out if there is genetic variability for the speed of acclimation among different tuber-bearing wild potato species. The species used were: S. acaule, S. commersonii, S. megistacrolobum, S. multidissectum, S. polytrichon, S. sanctae-rosae, and S. toralapanum. Relative freezing tolerance of these species was measured during cold acclimation. Preliminary results suggest that there are differences in the speed of acclimation among these species. We found that these species can be divided into four groups: i) non-acclimators; ii) rapid acclimators, with low to medium acclimation capacity; iii) slow acclimators, with low to medium acclimation capacity; iv) slow acclimators, with high acclimation capacity. We plan to use this information in our breeding program aimed at improving the freezing tolerance of potatoes.

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Beth Ann A. Workmaster, Michael Wisniewski, and Jiwan P. Palta

Infrared video thermography has recently been used to visualize ice nucleation and propagation in plants. At the UW–Madison Biotron facility, we studied the formation of ice in various parts of fruit-bearing cranberry (Vaccinium macrocarpon Ait.) uprights. The fruits were at the blush to red stages of ripening. Samples were nucleated at –1 or –2°C with ice-nucleating-active bacteria (Pseudomonas syringae). Following nucleation, samples were cooled to –6°C in ≈1 hour. The following observations were made: 1) When nucleated at a cut end, ice propagated rapidly throughout the stem and into the leaves at a tissue temperature of about –4°C. However, ice did not propagate from the stem through the pedicel to reach the fruit. During the 1 hour after ice propagation in the stem, the fruit remained supercooled. 2) Within the duration of the experiment, leaves could not be nucleated from the upper surface. Ice from the lower leaf surface did nucleate the leaf, and ice propagated from the leaf to the stem and other leaves readily. 3) Both red and blush berries could only be nucleated at the calyx end of the fruit. 4) Red berries supercooled to colder temperatures and for longer durations than the blush berries. 5) In support of our previous studies, red berries were able to tolerate some ice in their tissue. These observations suggest that: 1) The upper leaf surface and the fruit surface (other than the calyx end) are barriers to ice propagation in the cranberry plant; and 2) at later stages of fruit ripening the pedicel becomes an ice nucleation barrier from the stem to the fruit. This may contribute to the ability of the cranberry fruit to supercool.

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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.