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Hyo-Won Seo, Jung-Yoon Yi, Eung-Soo Kim, Hyun-Mook Cho, Young-Eun Park, and Kuen-Woo Park

This study was carried out to prove the new variety's originality by using Random Amplified Polymorphic DNA (RAPD) Analysis and to develope the specific markers for distinction new variety from others to database for improving the efficiency of germplasm conservation. The RAPD procedure was used to determine genetic diversity of 13 potato varieties including seven recommended varieties of Korea and six genotypes. Genomic DNAs from the 13 genotypes were amplified using PCR and URP 2F, 4R and 8R primers. URP primers which were 20-mers were received from NIAST (National Institute of Agricultural Science and Technology, Suwon, Korea) and they were shown very high reproducibility because of the high annealing temperature above 55 °C. So, they were known to be very desirable primers to examine the specificity between inter and intra species in various spectra. These 13 lines have many resemblances in plant characteristics each other because `Jopung', '92N09-6', `Daekwan 68', and `Daekwan 70' were originated from `Superior', `Atlantic', `Namsuh', and `Irish Cobbler' respectively. So, there are many difficulties to distinct new variety by the naked eye. The results of this study show that 2 sets of URP primers are very useful to distinct new variety and mutants from others.

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Arthur Villordon, Wambui Njuguna, Simon Gichuki, Philip Ndolo, Heneriko Kulembeka, Simon C. Jeremiah, Don LaBonte, Bernard Yada, Phinehas Tukamuhabwa, and Robert O. M. Mwanga

Detailed information on the geographic distribution of a crop is important in planning efficient germplasm conservation strategies but is often not available, particularly for minor crops. Using germplasm collection data from Kenya, Tanzania, and Uganda, we used distribution modeling to predict the distribution of sweetpotato [Ipomoea batatas L. (Lam.)] in sub-Saharan Africa. We used a consensus modeling approach using the following algorithms: genetic algorithm for rule set prediction (GARP), maximum entropy, BIOCLIM, and DOMAIN. The predicted distribution encompasses known sweetpotato production areas as well as additional areas suited for this crop species. New geographic areas where at least three models predicted presence were in Angola, Cameroon, Central African Republic, The Congo, Democratic Republic of Congo, Gabon, Ghana, Angola, Ethiopia, Mozambique, Rwanda, and the Central African Republic. This information can be used to fill gaps in current sweetpotato germplasm collections as well as to further enhance the current presence-only based distribution model. Our approach demonstrates the usefulness of considering several models in developing distribution maps.

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Hongwen Huang and Desmond R. Layne

The pawpaw is the largest tree fruit native to the United States and the only temperate member of the tropical Custard Apple family (Annonaceae). In 1995, Kentucky State Univ. was established as the USDA-ARS-National Clonal Germplasm Repository for Asimina spp. Seedling trees from 400 pawpaw accessions representing 70 distinct geographic regions from 17 states are currently being grown at our research farm. In a preliminary study, 18 pawpaw cultivars were assayed in 30 enzyme systems using an isoelectric focusing polyacrylamide slab gel system of pH 4-9. Twelve enzymes produced high resolution without tissue specificity and were further used for evaluation of allozyme diversity of geographic populations. Degree of genetic diversity within populations and differentiation between populations as evaluated by the expected heterozygosity (He), the proportion of polymorphic loci (P), the average number of alleles per locus (A), chi-squared analysis of allele frequency heterogeneity, Nei's standard genetic distance (D), and identity (I) will be discussed. Dendrograms were generated by cluster analysis using the unweighted pair group method to demonstrate the relationships of geographic populations in the 17 states evaluated. The strategy for germplasm conservation and cultivar development through breeding will also be discussed.

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E.J. Sacks and D.A. St. Clair

We have developed a method for cryopreserving Lycopersicon esculentum pollen to facilitate the timing of crosses and for long-term germplasm conservation. Gelatin capsules containing pollen were wrapped in tissue paper and placed in air-tight glass tubes with anhydrous calcium sulfate desiccant. Tubes containing pollen were stored at –80C. In one experiment, we stored the pollen of LA359 and T5 at –80C for 37 days. Pollen predesiccated overnight at 4C then stored at –80C, pollen put in a tube with desiccant then immediately stored at –80C, and fresh pollen were compared by pollinating 10 flowers of LA359 with each of the six pollen treatments and counting the number of seed per fruit. Average seed counts ranged from 127 for fresh, T5 pollen to 172 for predesiccated LA359 pollen. In another experiment, cryopreserved pollen of UC82B and VFNT Cherry was given from 0 to 6 cycles of freezing and thawing. Ten flowers of LA359 were pollinated with each of the 12 treatments. Average seed counts ranged from 125 to 152. The data from both experiments suggest that cryopreservation of tomato pollen to facilitate efficient plant breeding is feasible.

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Anfu Hou, James R. McFerson, and Warren F. Lamboy

Molecular DNA markers based on the RAPD (random amplified polymorphic DNA) assay are gaining use in germplasm assessment. RAPD markers are simple, relatively inexpensive, and highly informative. We used five primers to assess 26 Brassica oleracea breeding lines from the IVF and nine accessions from the PGRU. The test array included eight subspecies of B. oleracea. We generated 90 RAPD markers and were able to unambiguously discriminate among all 35 test entries, but could not separate subspecies within B. oleracea. Genetic similarity between subspecies ranged from 0.629 to 0.738. Average similarity within accessions was 0.96, confirming the suspected homogeneity of breeding lines. Nevertheless, significant genetic diversity was found among kohlrabi, broccoli, and cabbage accessions. Similarity analysis of breeding lines and hybrids confirmed their pedigree relationships. Interestingly, B. o. subsp. costata `Couve Nabica' showed closer similarity to B. napus subsp. oleifera `Jet Neuf' than to other B. o. materials and B. o. subsp. italica `Packman' showed higher similarity to some cabbages than to other broccolis. Results provide further evidence that diversity assessment using RAPDs is broadly applicable and useful in germplasm conservation and utilization.

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Izulme R. Santos and Cecil Stushnoff

Economically, citrus is the second most-important fruit crop grown worldwide; thus germplasm conservation of commercial cultivars, as well as of wild relatives, is essential. Presently, citrus germplasm has been conserved mainly in field genebanks. This approach is helpful; however, it is costly, exposes germplasm to climatic and biological hazards, and is not a long-term conservation system. Cryopreservation (conservation in liquid nitrogen, at –150°C to –196°C) is a technique that can ensure long-term storage of plant material. Attempts to cryopreserve citrus are restricted to a few reports, but the results obtained are encouraging. The basic purpose of this study is to define cryopreservation protocols for embryo axes and axillary buds of `Pineapple' sweet orange using the encapsulation-dehydration method. Embryo axes encapsualted in Na-alginate beads, precultured with high levels of sucrose and dehydrated over silica gel before freezing in liquid nitrogen had 60% survival. No survival was obtained for buds treated the same way, however buds isolated from plants acclimated at 0°C over a 30-day period survived exposure to –20°C when slow cooled at 2°C/hour. Additional experiments will combine cold acclimation, slow cooling and pre-treatment with sugars and other chemical compounds as an attempt to enhance cold hardiness of axillary buds and obtain survival after freezing in liquid nitrogen. Different approaches will be used to increase embryo axes survival rates.

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Hongmei Ma and Margaret Pooler

Ornamental flowering cherry trees (Prunus species) are popular landscape plants that are used in residential and commercial landscapes throughout most temperate regions of the world. Most of the flowering cherry trees planted in the United States represent relatively few species. The U.S. National Arboretum has an ongoing breeding program aimed at broadening this base by developing new cultivars of ornamental cherry with disease and pest resistance, tolerance to environmental stresses, and superior ornamental characteristics. Knowledge of the genetic relationships among species would be useful in breeding and germplasm conservation efforts. However, the taxonomy of flowering cherry species and cultivars is complicated by differences in ploidy levels and intercrossing among species. We have used simple sequence repeat (SSR) markers developed for other Prunus species to screen a diverse collection of over 200 ornamental cherry genotypes representing 70 taxa in order to determine the genetic relationships among species, cultivars, and accessions. Data were generated from 9–12 primer pairs using an automated DNA genetic analyzer (ABI3770), and subjected to UPGMA cluster analysis. Extremely high levels of polymorphism were exhibited among the materials studied, thus indicating that ornamental flowering cherry germplasm has substantial inherent genetic diversity. This information, combined with traditional morphological characteristics, will be useful in determining genetic relationships among accessions in our collection and for predicting crossability of taxa.

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Jaume Prohens, José M. Blanca, and Fernando Nuez

Eggplant (Solanum melongena L.) was introduced by the Arabs into Spain. Since then, many local cultivars have arisen. These materials are grouped in four cultivar groups: “round,” “semi-long,” “long,” and “listada de Gandía.” We studied the morphological and molecular [amplified fragment length polymorphism (AFLP)] diversity of a collection of 28 Spanish traditional cultivars of eggplant. Four eggplant accessions from different origins were used as controls and three scarlet eggplant (Solanum aethiopicum L.) accessions as outgroups. Morphology and AFLP markers showed that S. melongena and S. aethiopicum are separate taxonomic entities, and that, compared to controls, Spanish eggplants are very variable, indicating that the Iberian Peninsula can be regarded as a secondary center of diversity. Morphological differences were found among cultivar groups in traits other than those used for the grouping although, in some cases, accessions from different cultivar groups shared a similar general morphology. Eggplant cultivar groups also showed some genetic differences, which are revealed in the gene diversity statistics (GST = 0.30). Nonetheless, no individual AFLP markers specific and universal to one cultivar group could be found. “Round” cultivars were genetically more diverse than the other cultivar groups. A positive correlation (r = 0.68) was found between morphological and molecular distances. However, correlations between geographical and either morphological or molecular distances were low. Results suggest that evolution of eggplants in Spain has involved frequent hybridizations and a frequent movement and exchange of seeds. Structure of diversity among regions indicates that most of the diversity can be collected in single selected regions. All these results have important implications in eggplant germplasm conservation and breeding.

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Yingmou Yao and Peter M.A. Tigerstedt

Genetic variation in growth rhythm, hardiness and height of 24 populations from 3 subspecies in sea buckthorn (Hippophae rhamnoides) were studied in a field test. The relative variance component of subspecies varied from 26.2% to 73.7% of total variance. Subspecies turkestanica had a growth mode of late start-late finish, ssp. rhamnoides, intermediate start-early finish and ssp. sinensis, early start-intermediate finish. Subspecies rhamnoides had a growth period of 129 days, ≈30 days shorter than the two Asian subspecies. The average height of ssp. rhamnoides was 43.7 cm, about one-third of that for ssp. tarkestanica and sinensis. Subspecies rhamizoides was more hardy than ssp. sinensis, which was still more hardy than ssp. turkestanica. The variance among populations was generally comparable with within population variance. Except for hardiness, variations for all characters were much larger in ssp. rhamnoides than in ssp. sinensis. The total genetic variance (subspecies plus population) varied from 50% to 84% of total variance for all characters, except 37% for secondbracts. Later growth cessation was correlated with longer growth period, taller plants, more severe frost and winter damage. Strong clinal variation showed that the higher the latitude, the earlier the growth cessation, the shorter the growth period and plant height, the more hardy the population. -The results indicated that population selection should bean efficient way for growth rhythm and plant height. Clinal variation provides guidelines for seed and plant transfer as well as plant introduction. With limited collection and management capacity in germplasm conservation, the recommendation is to collect fewer individuals in each population but more populations along latitude.

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Daniel J. Cantliffe

hybrid seed production and plant germplasm conservation. The reader has an opportunity to get information, packed into 796 pages, in almost any area related to seeds. The contributing authors to Dr. Basra's book are excellent and well known for their