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Neil O. Anderson and Peter D. Ascher

Male and female fertility, seed germination, and progeny fertility were used to determine cultivar fertility in species of Lythrum. One short-, 11 mid-, and six long-styled cultivars were included in this study. Duplicates of several cultivars from different nurseries and three unknown cultivars from Minnesota gardens were also collected. Plants from 17 Minnesota and one Wisconsin population of L. salicaria served as fertile male and/or female testers. Pollen stainability (usually 100%) showed low levels of male gamete abortion. Pollen size within and among anther type varied widely; possible 2n gametes were present in primarily the short- and mid-anther morphs. Seed production per capsule from legitimate cross-pollinations, using cultivars as male parents with Minnesota or Wisconsin female testers, averaged 48 ± 36 across style morphs. Cultivars differed as males, as did anther morphs. With female fertility tests, seed set per capsule ranged from zero to 152 and averaged 54 ± 40 in legitimate pollinations (i.e., pollinations between stamen and styles of the same length). Seed set for other crosses showed similar trends. Only `Morden Gleam' produced no seed with all legitimate pollinations, although illegitimate selfs or interspecific crosses produced seed. Seed from legitimate crosses of L. salicaria × cultivars had 30% to 100% germination. Common male and female parents within each legitimate crossing group were not significantly different. This study showed that the cultivars are highly fertile when used as male or female parents with wild purple loosestrife, native species (L. alatum Pursh.), or other cultivars. Thus, cultivars grown in gardens could serve as pollen or seed sources for the continued spread of purple loosestrife. The implications of cultivar fertility, especially interspecific F1 hybrids, is discussed in relation to the spread of noxious weeds in wetlands.

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Neil O. Anderson and Peter D. Ascher

It should be possible to maintain horticultural clones unchanged forever through asexual generations, as commercial propagators and clonal repositories maintain clonal integrity, disease-free stock plants, or remove mutations. However, unintentional selection for nonhorticultural traits could still be occurring. Accumulations of such traits would be due to the operation of Muller's ratchet and include fertility losses, increases in virus titer, and stunted growth habit. In chrysanthemums, Dendranthema grandiflora. clones separated from sexual cycles for generations become increasingly sterile. Seed set across years, using coefficients of crossability (FCC/MCC), was examined for garden clones (forced through sexual cycles annually) and greenhouse clones (asexual cycles only). Garden clones 40 years old (54-101-11) had only depressed levels of fertility. In other cases (77-AM 3-17), the ratchet was reversed >1 sexual cycle. Greenhouse clones were often completely sterile since their propagation is primarily asexual.

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Cecil Stushnoff, Philip L. Forsline, Leigh Towill, and John Waddell

Cryopreservation of dormant buds has potential to provide back-up conservation of vegetatively propagated genetic resources for fruit crop species. This system may be useful where clonal integrity must be maintained and where it is desirable to rapidly recover plants with flowers for crossing purposes. In 1988, a pilot project involving the National Clonal Apple Repository at Geneva, NY and the National Seed Storage Laboratory, Fort Collins, CO, was initiated to test handling protocols as a prelude to establishing a cryopreservation backup system for apple genetic resources. Sufficient buds have been cryopreserved to permit viability evaluation after 1 month, 1, 2, 3, 4, 5, 10, 15, 20, and 25 years storage in liquid nitrogen vapor phase storage (-150 C]. Recovery of dormant buds collected 12/12/88 and 02/06/89 after one month in LN2 was 36% and 35%, respectively, for eight different taxa. After one year in LN2, recovery was 50% and 48% for the same taxa. The difference was attributed to improved handling during dehydration prior to patch budding for viability estimation. In 1990, recovery after 1 month in LN2 was 38% for six different cultivars. The response to controlled acclimation and desiccation for 15 taxa will be presented.

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Mark S. Strefeler, Elizabeth Darmo, Roger L. Becker, and Elizabeth J. Katovich

Isozyme markers were used to identify several cultivars of purple loosestrife (Lythrum spp.) and interspecific hybrids. There were three zones of activity for phosphoglucomutase (PGM) and phosphoglucoisomerase (PGI) and two zones for malate dehydrogenase (MDH) in purple loosestrife cultivars. Allelic constitution could not be characterized due to the polyploid nature of purple loosestrife and the possibility of intergenic dimerization. Coefficients of genetic similarity were used to estimate the degree of relationship between purple loosestrife cultivars. Cluster analysis indicated that seven cultivars originating from L. salicaria L. were not distinguishable from eight cultivars originating from L. virgatum L., indicating possible limitations of isozyme analysis for cultivar differentiation based on species origin. All but two cultivars (`Morden Gleam' and `Morden Rose') could be distinguished from one another by isozyme phenotype. This result suggests that isozymes may be useful for cultivar fingerprinting if additional isozyme systems could be resolved. `Robert' appeared morphologically heterogeneous, and plants could be differentiated based on isozyme banding patterns. Also, two putative clones of `Stichflamme' (one marketed under its English synonym `Fire Candle') possessed distinct isozyme phenotypes, indicating a lack of clonal integrity.

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Manfredo J. Seufferheld, Cecil Stushnoff, John Fitzpatrick, and Philip L. Forsline

Cryopreservation of woody-plant, dormant buds may provide cost-effective, long-term, back-up conservation of germplasm for vegetatively propagated crops that are presently maintained as trees in field gene banks. Dormant buds can be recovered quickly by grafting to dwarfing rootstocks, thus producing flowers for breeding purposes, with minimum potential for inducing somaclonal variants. These attributes are essential to preserve the clonal integrity of unique gene combinations such as those found in tree fruit crops. Previous research has shown that dormant buds from cold-hardy apples can be recovered from storage in liquid nitrogen (LN) with high survival rates (80% to 100%) using controlled desiccation and slow freezing before immersing in LN. On the other hand, dormant buds from cold-tender taxa and buds collected at less than optimal stages for desiccation and freezing have much lower (0% to 50%) survival rates. We increased survival of cold-tender taxa by using a modified vitrification procedure. Dormant apple buds from tender and hardy cultivars were perfused with modified PVS [15% (w/v) ethylene glycol, 15% (w/v) propylene glycol, 7% (w/v) DMSO, and 15% (w/v) glycerol in 0.5 m sorbitol]. Toxicity from the PVS was reduced by dilution soaking in 1 m sorbitol, 0.2 m raffinose, and 15 mm CaCl2 before and after quench-freezing and slow-freezing cryopreservation. Up to 100% of some cold-tender taxa survived. In addition, xylem ray parenchyma tissues that supercool and are normally killed at about -40C with the desiccation protocol survived this vitrification procedure.

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Neil O. Anderson, Adnan Younis, and Ye Sun

) could not use AFLPs to detect clonal differences of L. longiflorum ‘Nellie White’ due to a lack of repeatability and consistency for scorable bands. Thus, AFLPs cannot be used to assess genetic variability and clonal integrity in L. longiflorum . The