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F.P. Maier, N.S. Lang and J.D. Fry

Stolons of `Raleigh', `Floratam', and FX-332 St. Augustinegrass [Stenotaphrum secundatum (Walt.) Kuntze] were sampled from the field between October and March in two consecutive years to evaluate accuracy of an electrolyte leakage (EL) method for predicting freezing tolerance. Lethal temperatures of stolons estimated using EL were compared to those obtained by regrowth tests in the greenhouse. Mean lethal low temperatures for regrowth and EL methods over 12 sampling dates were `Floratam', –4.5C (regrowth) vs. –4.4C (EL); FX-332, –4.2C (regrowth) vs. –4.9C (EL); and `Raleigh', –6.0C (regrowth) vs. –5.4C (EL). A positive correlation (r = 0.81) was observed between EL-predicted and regrowth lethal temperatures for `Raleigh', which exhibited some acclimation during the first sampling year. The EL technique consistently predicted a lower lethal temperature for `Raleigh' than for `Floratam', which corroborates field observations concerning freezing tolerance of these two cultivars.

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Rita L. Hummel and Patrick P. Moore

Seasonal changes in freezing tolerance of stems and buds of Rubus idaeus L. `Chilliwack', `Comox', `Meeker', `Skeena' and `Willamette' clones were measured from November through March of 1988-1989 and 1989-1990. Eight additional clones were tested in 1989-1990. Canes were harvested from the field, cut into two-bud samples and subjected to controlled freezing tests. Samples were seeded with ice, held at -2°C overnight and then frozen at 3°C/hour. Viability was estimated by visual browning. Vascular tissue at the base of the buds was the least freeze tolerant tissue in these samples. Results of both the 1988-1989 and 1989-1990 freezing tests, indicated `Meeker' and `Willamette' cold acclimated more slowly in the fall than `Chilliwack', `Comox' and `Skeena'. However, in the spring, `Willamette' and `Meeker' were slower to lose freeze tolerance than the other three clones.

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

Two USDA intergeneric, hybrid citrus scions, US 119 {[Citrus paradisi Mac. `Duncan' × Poncirus trifoliata (L.) Raf.] × C. sinensis Osb. `Succory'} and selection 17-11 {C. grandis US 145 × [Citrus paradisi Mac. `Duncan' × P. trifoliata (L.) Raf.]} on `Swingle' citrumelo (C. paradisi × P. trifoliata) rootstocks were examined for freeze hardiness traits (4 years) and general growth characteristics (2 years). Hardiness was compared with that of `Hamlin' orange [C. sinensis (L.) Osb.] and satsuma mandarin (C. unshiu Marc) from Fall 1993 to Spring 1997. As expected, US 119 and 17-11 were both hardier than `Hamlin' orange as determined by leaf disc electrolyte leakage (EL). Both showed freezing tolerance similar to that of satsuma mandarin, but 17-11 was significantly hardier than satsuma or US 119 at several times during the 4-year study. Trunk diameter and tree height were similar for US 119 and selection 17-11.

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Aaron E. Walworth and Ryan M. Warner

life in cooler climates by developing mechanisms to tolerate low temperatures, but cold tolerance is a dynamic characteristic. When grown in warm temperatures, even cold-hardy plants have low freezing tolerance. Exposure to low nonfreezing temperatures

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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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Yang Yang, Runfang Zhang, Pingsheng Leng, Zenghui Hu and Man Shen

only to insufficient freezing tolerance, but also to the timing and rate of cold acclimation ( Suojala and Lindén, 1997 ). Cold acclimation, the process by which plants transit from a cold-sensitive to a cold-hardy state, is essential for the survival

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

Poncirus trifoliata is a comparatively hardy, cross compatible, and graft compliant relative of Citrus. The citrus industry in Florida has suffered immense economic losses due to freezes. Although much research has been done in citrus freeze hardiness, little work has been on the early induction of freeze tolerance by low temperature. Poncirus trifoliata `Rubidoux' seedlings were germinated in perlite under intermittent mist at about 25°C and natural daylight conditions in a greenhouse and grown 2 weeks. See dlings were then transferred into a growth chamber at 25°C and 16 hour daylength for 1 week. Temperature was lowered to 10°C and tissue samples were collected at 0, 6, 24, and 168 hours. Freezing tolerance, at –6.7°C as determined by electrolyte leakage, and stem (leaves attached) water potential, measured using a pressure bomb, were also recorded for a subset of seedlings for the above intervals. After exposure to low temperature for 48 hours a red coloration became visible at the petiole leaflet junction an d at the buds, with subsequent exposure to low temperature the coloration spread to the leaves. Clones for phenylalanine ammonia lyase (PAL), 4-coumarate:CoA ligase (4CL), and chlorophyll ab binding protein (CAB), and chalcone synthase (CHS) were used to probe RNA isolated from P. trifoliata. PAL and 4CL transcripts increased in response to the low temperature. Significant increases in freeze hardiness occurred within 6 hours in the leaves, and increases continued for up to one week. Water potential increased from –0.6 to –2.0 MPa after 6 hours, then returned to –0.6 MPa after 1 week. These data indicate that increases in freezing tolerance and changes in water potential and gene expression can be detected shortly after low temperature treatments are imposed on P. trifoliata seedlings.

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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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Babita Thapa, Rajeev Arora, Allen D. Knapp and E. Charles Brummer

measured in the laboratory to determine freezing tolerance of diverse species ( Arora and Palta, 1991 ; Nunes and Smith, 2003 ; Webb et al., 1994 ; Welling et al., 2002 ), including alfalfa ( Sulc et al., 1991 ). This method is simple, repeatable, and

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Yang Yang, Zhongkui Jia, Faju Chen, Ziyang Sang and Luyi Ma

. Freezing injury affects the growth, productivity, and geographical distribution of horticultural plants ( Pearce, 2001 ; Weiser, 1970 ). The susceptibility of plants to freezing injury may be attributable not only to insufficient freezing tolerance, but