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Gordon M. Huestis and Carlos F. Quiros

Phylogenetic relationships of seven Apium species, including three horticultural types of A. graveolens, were assayed for RFLPs using cDNA, chloroplast DNA, and rDNA probes. Most of the probes had been previously mapped in celery. The three horticultural types of A. graveolens were found to be less polymorphic than the wild species and in phylogenetic analysis they clustered together. The wild species formed a cluster on the dendrogram corresponding to their origin in the southern hemisphere. A. nodiflorum, a wild species from Ethiopia formed a branch on the phylogenetic tree apart from all other species. This, along with morphological considerations, suggests that A. nodiflorum should be reclassified outside the genus Apium.

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Dongyan Hu*, Zuoshuang Zhang, Donglin Zhang and Qixiang Zhang

Ornamental peach (Prunus persica (L.) Batsch) is native to China. The ornamental value of peach is gaining popularity for its use in urban landscape and everyday gardens. However, the genetic relationship among ornamental peach cultivars is not clear, which limits the further studies of its molecular systematic. A sample of 51 cultivars of ornamental peach, originated from P. persica and Prunus davidiana, had been studied by using AFLPs. All samples were collected from China, Japan, and the US. A total of 275 useful markers between 75 to 500 base pairs were generated from 6 EcoRI/MseI AFLP primer combinations. Among them, 93% of bands were polymorphic markers. Total markers for each cultivar ranged form 90 to 140, and the average number of markers for each cultivar was 120. Two distinguished clad generated from PAUP-UPGMA tree. P. davidiana, as a species, was apparently an out-group to P. persica, which implied that P. davidiana was far away genetically from ornamental peach (P. persica). Within P. persica clad, 15 out of 17 upright ornamental peach cultivars in this study were grouped to one clad, which indicated cultivars that with upright growth habit had close genetic relationship. Five dwarf cultivars were grouped to one clad, with 81% bootstrap supported. The genetic relationships between these five dwarfs were much closer than any other cultivars, and showed that they probably derived from the similar gene pool. The results demonstrated that AFLP are powerful markers for revealing genetic relationships in ornamental peach. The genetic relationships among ornamental cultivars established in this study could help future ornamental peach germplasm identification, conservation, and new cultivars development.

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James A. Schrader and William R. Graves

Alnus maritima (Marsh.) Muhl. ex Nutt. is a rare woody plant species that exists as three subspecies found in widely disjunct locations in the United States. Although there is a growing interest in the phytogeography, ecology, conservation, and landscape potential of this species, the phylogeny of A. maritima has not yet been resolved by using molecular methods. We have combined a relatively new method of genome fingerprinting, ISSR-PCR, and the automated imaging capabilities of GeneScan technology to investigate the molecular systematics of A. maritima. Based on the molecular evidence from 108 ISSR loci, we confirm that the three disjunct populations of A. maritima have diverged sufficiently to be classified as subspecies. Our molecular phylogeny of the three subspecies of A. maritima agreed in topology with a phylogeny produced from morphological data and showed that subsp. oklahomensis is the most distinct of the three subspecies and was the first to diverge. The simultaneous analysis of molecular and morphological data provides a detailed and balanced phylogeny reconstruction for the three subspecies. Our results support the theory that A. maritima originated in Asia, migrated into North America across the Bering land bridge, and was established over a large range in the New World before being forced into its present meager disjunct distribution.

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John E. Bowers and Carole P. Meredith

RFLP data were used to assess genetic similarity among 33 Vitis vinifera L. cultivars and one interspecific cultivar. A similarity matrix was constructed on the basis of the presence or absence of 49 bands generated by eight RFLP probes and cluster analysis was performed. The mean similarity index for all pairwise comparisons was 0.696 and ranged from 0.444 between `St. Emilion' and the interspecific hybrid `Salvador' to 0.952 between `Chenin blanc' and `Semillon'. Mean similarity among all V. vinifera cultivars was 0.705. Several groupings of similar cultivars are consistent with historical reports and presumed geographic origins: `Chardonnay' and `Melon', `Colombard' and `Folle blanche', `Gewürztraminer' and `Trousseau', `Cabernet franc' and `Cabernet Sauvignon', `Mission' and `Palomino'. The similarity between `Mission' and `Palomino' is the first genetic evidence to support the putative Spanish origin of `Mission'. Some groupings are unexpected (`Sauvignon blanc' and `Gewürztraminer', `Chenin blanc' and `Semillon') because the cultivars are not thought to have originated in the same regions. While some relationships suggested by this study may be artifacts of RFLP analysis or of the statistical method, they raise questions for further genetic inquiry into the origins of grape cultivars.

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Hongwen Huang, Desmond R. Layne and R. Neal Peterson

The utility of isozyme phenotypes for identifying and determining genetic variation in pawpaw cultivars was studied using isoelectric focusing in thin-layer polyacrylamide gels. Based on a sample of 32 clones (cultivars and advanced selections) and 23 enzyme systems, 7 enzymes were found to be polymorphic, involving 9 polymorphic loci [acid phosphatase (ACP), dihydrolipoamide dehydrogenase (DDH), malic enzyme (ME), phosphoglucoisomerase (PGI), phosphoglucomutase (PGM), peroxidase (PRX), and shikimate dehydrogenase (SKD)]. Altogether these 9 loci and 32 clones yielded 28 multi-locus isozymic phenotypes useful for cultivar identification; 24 of the 32 clones were uniquely identified. The allozyme variation in these clones has the average of other long-lived woody perennials of widespread geographic range in temperate regions with insect-pollinated outcrossing breeding systems, secondary asexual reproduction, and animal-dispersed seed. Genetic differentiation among these pawpaw clones, measured by Nei's distance, D, was substantial: 496 pairwise comparisons of genetic distance among the 32 clones indicated that they differed on average of D = 0.068 ± 0.04 and ranged from 0 to 0.188. Cluster analysis (UPGMA) produced a most likely division of the 32 clones into 7 groups; however, these groups did not conform to known pedigree relations. Additional polymorphic enzymes are needed for accurate allozyme-based genetic discrimination.

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Joseph J. King, James M. Bradeen and Michael J. Havey

Nuclear RFLPs were used to estimate relationships among 14 elite commercial inbreds of bulb onion (Allium cepa) from Holland, Japan, and the United States. Variability for known alleles at 75 RFLP loci and 194 polymorphic fragments revealed by 69 anonymous cDNA probes and a clone of alliinase were scored to yield genetically characterized and uncharacterized data sets, respectively. The inbred onion populations possessed more than two alleles at 20 of 43 (46%) codominant RFLP loci. Relationships among the inbreds were estimated by cluster analysis of simple-matching (genetically characterized data) and Jaccard (genetically uncharacterized data) coefficients using the unweighted pair group method and agreed with known pedigrees. RFLPs confidently distinguished among elite inbreds within and between specific market classes. RFLP profiles for virtual hybrids were computer-generated by combining gametic arrays among inbreds of the same market class and analyzed as described above. Allelic and genetically uncharacterized RFLPs confidently distinguished among these hybrids, even though heterozygosity for many markers produced a majority of monomorphic fragments. We randomly sampled decreasing numbers of RFLPs from the complete data sets and calculated simple-matching and Jaccard distances, noting the numbers of probes that were unable to distinguish any two inbreds or hybrids. As few as 10 polymorphic probe-enzyme combinations distinguished among all the inbreds and samples of 20 genetically characterized or 10 genetically uncharacterized clones distinguished all the virtual hybrids. This study demonstrated that the previously reported few RFLPs observed among open-pollinated (OP) onion populations were due to the highly heterozygous nature of the OP population.

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R.G. Fjellstrom, D.E. Parfitt and G.H. McGranahan

RFLP markers were used to investigate genetic diversity among California walnut (Juglans regia) cultivars and germplasm collected worldwide. Sixteen of 21 RFLP markers were polymorphic in the 48 walnut accessions tested. RFLP markers were useful for identifying walnut cultivars. All genotypes were heterozygous at ≈20% of the loci for both California and worldwide germplasm. California walnut germplasm contained 60% of the worldwide allelic diversity. Cluster analysis of genetic distance between accessions and principal component analysis of allelic genotypes showed two major groups of walnut domestication. California germplasm was associated with germplasm from France, central Europe, and Iran and had less genotypic similarity with germplasm from Nepal, China, Korea, and Japan.

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S.E. Gardiner, H.C.M. Bassett, C. Madie and D.A.M. Noiton

Information about a rare allele of phosphoglucomutase (PGM) that is shared by `Braeburn' and 16% of cultivars in the New Zealand Cultivar Collection was combined with historical information about cultivar distribution to select a set of 15 cultivars for a more detailed genetic analysis of their relatedness to the key New Zealand apple (Malus domestica Borkh.) `Braeburn'. DNA from all 16 cultivars was examined by RFLP analysis using 41 probe-enzyme combinations and also by RAPD analysis with 39 selected primers. The RFLP and RAPD data excluded a proposal that `Lady Hamilton' and `Braeburn' are genetically identical. All cultivars except `Lady Hamilton' were excluded as potential parents for `Braeburn' based on incompatible RFLP banding. Assessment of genetic distances between `Braeburn' and the other 15 cultivars from RFLP and RAPD data demonstrated that `Lady Hamilton' was more closely related to `Braeburn' than all others. We conclude that there is a high likelihood that `Lady Hamilton' is one of the parents of `Braeburn'.

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Anne M. Gillen and Fredrick A. Bliss

Peach rootstock breeding may be accelerated by utilization of molecular markers linked to the root-knot nematode resistance locus (Mi) to screen segregating populations. A genetic linkage map was constructed using RFLP markers in an F2 population (PMP2) that is segregating for this locus. PMP2 is derived from a controlled cross of the relatively diverse peach rootstocks Harrow Blood (susceptible) and Okinawa (homozygous resistant). Bulked Segregant Analysis was applied using RAPD markers. A single small (227 base pairs) RAPD marker was found to be linked to the dominant resistant allele of Mi at a distance of 10 cM. This new marker joined the Mi locus to the RFLP linkage map and showed that two dominant RFLP markers are located between the RAPD marker and Mi. RFLPS are expensive, time-consuming and RAPD markers are unreliable, and therefore both are unsuitable for screening breeding populations. We attempted to convert the RAPD marker to a more breeder-friendly CAPS marker. The converted CAP marker was dominant. Attempts to convert the CAP marker to a co-dominant marker were not successful. The utility of the CAP marker was tested in an open pollinated F2 population derived from the F1 parent of PMP2 and in several rootstocks. The genetic linkage map was compared to other Prunus maps. The PMP2 linkage group containing the Mi locus can be related to the peach × almond linkage group which contains the phosphoglucomutase Pgm-1 locus.

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Amnon Levi and Claude E. Thomas

different polyploidy crop plants, including sunflowers ( Mokrani et al., 2002 ), coffee ( Herrera et al., 2002 ), and alfalfa ( Barcaccia et al., 2000 ). Low DNA polymorphism exists among American watermelon cultivars, indicating they are derived from common