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Flower buds of 20 Prunus species representing 4 subgenera were collected during winter and spring of 1989-90. Buds were preconditioned at +3° or 7°C to test their minimum hardiness level (MHL) or the rate of hardiness increase. DTA revealed that most of the prunus species have flower primordia that supercool. The subgenus Padus have racemose inflorescences and do not deep supercool during dormancy. P. besseyi, P. nigra and P. americana had small exotherms between -22° and -27°C while P. davidiana and P. subhirtella had larger exotherms at higher temperatures. Exposure of flower buds to -7°C shifted LTES to lower temperatures and/or reduced the size of LTE, which became undetectable for many species including P. nigra and P. americana. P. davidiana and P. subhirtella increased hardiness by 6°/day at -7° while dormant. Deacclimation coincided with an increase in LTE50 and the development of xylem vessel elements in the bud axis, calyx and filaments as indicated by dye movenent. P. davidiana was the least hardy species and required only 700 chill units to satisfy the chilling requirement, while P. nigra and P. americana had LTE average of -26°C at MHL and required over 1000 chill unit accumulation.

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Root and shoot characteristics related to drought tolerance were studied for Prunus persica, P. andersonii, P. besseyi, P. maritima, P. subcordata, and P. tomentosa. In general, shoot characteristics were more closely associated with drought adaptation than root characteristics across species. The most xeric species, andersonii, had the most xerophytic leaf morphology, highest rates of leaf gas exchange, high root length/leaf area and root weight/leaf area ratios, but had root length and hydraulic conductivity similar to that of more mesic species. Water use efficiency (WUE) increased as water potentials (ψ) dropped to -3.0 to -4.0 Mpa during a 5-7 day drought for the xeric andersonii and subcordata. However, after an initial increase, WUE decreased with declining ψ in the other 4 species, indicating that carboxylation was affected by stress in the -1.5 to -3.0 range of ψ for besseyi, maritima, persica and tomentosa. CO2 assimilation (A) decreased linearly with ψ during drought in all species, but the ψ at which A reached zero was not well correlated with drought adaptation. Root hydraulic conductivity was similar for all species, indicating a lack of importance of this parameter for drought tolerance. The data suggest that introduction of xerophytic shoot characteristics into commercial cultivars of Prunus would improve drought tolerance to a greater extent than using drought tolerant species as rootstocks.

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Stylar proteins of four Prunus species, P. avium, P. dulcis, P. mume, and P. salicina, were surveyed by 2D-PAGE combined with immunoblot and N-terminal amino acid sequence analyses to identify S-proteins associated with gametophytic SI in the Prunus. All four S-allelic products tested for P. dulcis could be identified in the highly basic zone of the gel. These S-proteins had Mr of about 28–30 kDa and reacted with the anti-S4 -serum prepared from Japanese pear (Pyrus serotina). Two of six S-allelic products tested for P. avium could be also identified in the 2D-PAGE profiles, with roughly the same pI and Mr as those of S-proteins of P. dulcis. Putative S-proteins for P. mume and P. salicina were found in the same area of 2D-PAGE as the area where S-proteins of P. avium and P. dulcis were located. N-terminal amino acid sequence analysis of these proteins revealed that they were similar to S-RNases reported previously.

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55 ORAL SESSION (Abstr. 473-479) Tree Fruit: Prunus Culture and Management

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Callus formation and adventitious shoot regeneration in vitro from mature stored seed were evaluated in eight ornamental cherry (Prunus) taxa: P. campanulata Maxim., P. maackii Rupr., P. sargentii Rehd., P. serrula Franch., P. serrulata Lindl., P. subhirtella Miq., P. virginiana L., and P. yedoensis Matsum. Several portions of the embryo (cotyledons and hypocotyl sections) and nine combinations of growth regulators (BA, 2,4-D, IBA, NAA, and TDZ) were compared. Effects of embryo portions and growth regulator treatments were generally small within taxa, but shoot formation differed among taxa. About 20% to 50% of the embryos from P. virginiana and P. serrula and ≈5% to 30% of those from P. maackii produced shoots. The other taxa generally did not produce shoots. Regeneration from mature stored seed in the responsive taxa represents a potential system for genetic transformation. Chemical names used: 6-benzyladenine (BA); 2,4-dichlorophenoxyacetic acid (2,4-D); indole-3-butyric acid (IBA); α-naphthaleneacetic acid (NAA); thidiazuron (TDZ).

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Fourteen Prunus rootstocks were evaluated against mixtures of several isolates of the root-lesion nematode Pratylenchus vulnus Allen and Jensen in three greenhouse experiments. Most of the tested rootstocks are new releases or materials in advanced stages of selection that also have incorporated root-knot nematode resistance. The plums Torinel (Prunusdomestica L.) and Redglow (P. salicina Lindl. P. munsoniana Wight and Hedrick cv. Jewel) showed a moderately resistant response; their final nematode population levels were lower or slightly higher than inoculation levels. Low nematode reproduction also was found in the peach–almond hybrid G N No 22 [P. persica (L.) Batsch P. dulcis (Mill.) D.A. Webb] and the plum Bruce (P. salicina P. angustifolia Marsh.), and although these rootstocks did not perform as well as Torinel and Redglow, they also appear to be poor hosts for P. vulnus.

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Differential thermal analysis (DTA) was used to measure deep supercooling in flower buds of Prunus dulcis Mill., P. armeniaca L., P. davidiana (Carr.) Franch, P. persica (L.) Batsch, three sweet cherry (P. avium L.) selections, and `Bing' cherries (P. avium L.) during Winter 1990-91 and 1991-92. Low temperatures in Dec. 1990 killed many flower buds. After the freeze, dead flower primordia continued to produce low-temperature exotherms (LTEs) at temperatures near those of living primordia for >2 weeks. In Feb. 1992, cherry buds that had been killed by cooling to -33C again produced LTEs when refrozen the next day. As buds swelled, the median LTE (LTE50) of dead buds increased relative to that of living buds, and the number of dead buds that produced LTEs decreased. LTE artifacts from dead flower priimordia must be recognized when DTA is used to estimate LTE50 of field-collected samples.

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Total cellular DNA has been extracted from leaves and\or seed of Prunus dulcis, P. persica, P. mira, P. davidiana, P. persica subsp. ferganensis, and P. triloba. Chloroplast restriction fragments have been visualized by Southern blot analysis using heterologous probes from a petunia chloroplast library. Analysis of preliminary data separates the species into three groups. The first contains P. dulcis, P. mira, and P. davidiana; the second P. kansuensis, P. persica, and P. persica subsp. ferganensis; and the third P. triloba.

PCR amplification using oligos for cytosolic glyceraldehyde-3-phosphate dehydrogenase yields genomic fragments approximately 1kb in size from P. dulcis and P. triloba. Sequence analysis will be performed to determine species relationships at the gene level.

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On the basis of investigating pollination habits of 45 cultivars of Mei Hua, interspecific hybridization between Mei Hua and apricot (Prunus armeniaca). David's peach (P. davidiana) as well as siberian apricot (P. sibirica) were made from 1982 to 1991. With total number of pollination flower 17,050, 168 hybrid seed and 75 hybrid seedlings were obtained. Embryo culture in vitro was used for undeveloped young hybrid embryos. Test of freezing resistance both in artificial freezing and in overwintering for the hybrid seedlings showed that there were 5 hybrids with double and nice flower to be very hardy to low temperature. They were able to tolerate as low as -35C for 30 days in open ground, and now they were planted in northwest China's Gansu province and northeast China's Liaonin province without cold injury.

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Mei (Prunus mume Sieb. et Zucc.) flower is one of the candidates for the national flower of the People's Republic of China. Several major anthocyanins from the flowers of P. mume Sieb. et Zucc. were isolated with MeOH-HOAc-water (10:1:9, v/v), and purified by paper chromatography and subsequent column chromatography. Specific chemical reactions, chromatographic and spectroscopic analyses indicated that the anthocyanins in `Nanjing Hongxu' (Nanjing red-bearded) were cyanidin 3-O-(6'-O-α-rhamnopyranosyl-β-glucopyranoside) and cyanidin 3-O-(6'-O-galloyl-3'-O-β-glucopyranosyl-β-glucopyranoside). Anthocyanins in `Nanjing Hong' (Nanjing red) were cyanidin 3-O-(6'-O-α-rhamnopyranosyl-β-glucopyranoside), cyanidin 3-O-(6'-O-galloyl-β-glucopyranoside) and cyanidin 3-O-(6'-O-E-feruloyl-βglucopyranoside). In addition to contributing to the blue flower color, the anthocyanins may improve the ability of the two cultivars to survive at low temperatures.

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