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Deric D. Picton and Harrison G. Hughes

In this study, 11 species, hybrids, and color variants were characterized using randomly amplified polymorphic DNA (RAPD) analysis. Total genomic DNA was extracted using a 2% CTAB extraction buffer using fresh or frozen leaf material. The DNA was amplified using standard RAPD-PCR protocols utilizing 10-mer primers. All primers utilized exhibited a high degree of polymorphism in their banding patterns among the species and hybrids studied. The primers used produced ≈40 reproducible bands. It was possible to identify and uniquely distinguish all species and hybrids investigated using these bands.

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Xianping Qu, Jiang Lu, and Olusola Lamikanra

Two morphologically distinct types of grapes belonging to the subgenera Euvitis and Muscadinia in the genus Vitis are cultivated in the United States. The former is commonly called bunch grapes while the latter is usually called muscadine. Genetic diversity among these grapes was investigated using RAPD markers. Sixteen grape cultivars, with parentage including V. rotundifolia Michx., V. vinifera L., and several American Vitis species, were used for the RAPD analysis. A total of 156 RAPD markers was produced from 19 random primers, over 90% of which was polymorphic among the muscadine and the bunch grapes. Polymorphisms were lower within each subgenus. Relationships between these two subgenera were estimated based on band-sharing and cluster analysis. The average genetic distance between the bunch and the muscadine grape cultivars was 0.45. The results based on DNA analysis agree with isozyme data obtained from a separate study, which demonstrated that muscadine grapes share very few common alleles with American bunch grapes and European grapes.

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Mehmet Nuri Nas, Nedim Mutlu, and Paul E. Read

RAPD and phenotypic analysis were conducted to assess clonal stability of hazelnuts generated from axillary buds cultured in vitro for long-term. The nuts produced on in vitro-propagated plants were indistinguishable from those of donor plants. With the exception of rare horizontal (plagiotropic) growth, all in vitro-propagated plants exhibited phenotypes similar to those of donor plants. RAPD analysis did not reveal any somaclonal variation between donor plants from which in vitro cultures were initiated and micropropagated plants (6-year cultures), and no somaclonal variation was detected among in vitro-propagated plants. However, polymorphism (15.6%) was detected between the parent plant and its in vitro-propagated progenies (from seedlings). These results show a good discriminatory power of RAPD to detect polymorphism between samples where it is expected, and it can be effectively used for genetic assessment of micropropagated hazelnut. No evidence of genetic or epigenetic changes was observed in long-term cultured hazelnut, and thus long-term in vitro culture of hazelnut does not seem to limit its clonal propagation.

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Patrick J. Conner and Bruce W. Wood

Genetic variation among pecan [Carya illinoinensis (Wangenh.) K. Koch] cultivars was studied using randomly amplified polymorphic DNA (RAPD) markers. Using a combination of primers, a unique fingerprint was produced for each of the pecan genotypes studied. The genetic relatedness between 44 cultivars was estimated using more than 100 RAPD markers. Genetic distances based on the simple matching coefficient varied from 0.91 to 0.59. The phenetic dendogram developed from cluster analysis showed relatively weak grouping association. However, cultivars with known pedigrees usually grouped with at least one of the parents and genetic similarity estimates appear to agree with known genetic relationships. Using RAPD information in determining genetic relationships among pecan cultivars with unknown or questionable pedigrees and the integration of that knowledge into the breeding program is discussed.

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F. Bartolozzi, M.L. Warburton, S. Arulsekar, and T.M. Gradziel

Almond [Prunus dulcis (Mill.) D.A. Webb, syn. P. amygdalus, Batsch; P. communis (I.) Archangeli] represents a morphologically and physiologically variable group of populations that evolved primarily in central and southwest Asia. California cultivars have been developed from highly selected subgroups of these populations, while new breeding lines have incorporated germplasm from wild almond and closely related peach species. The genetic relatedness among 17 almond genotypes and 1 peach genotype was estimated using 37 RAPD markers. Genetic diversity within almond was found to be limited despite its need for obligate outcrossing. Three groupings of cultivar origins could be distinguished by RAPD analysis: bud-sport mutations, progeny from interbreeding of early California genotypes, and progeny from crosses to genotypes outside the California germplasm. A similarity index based on the proportion of shared fragments showed relatively high levels of 0.75 or greater within the almond germplasm. The level of similarity between almond and the peach was 0.424 supporting the value of peach germplasm to future almond genetic improvement.

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Hak-Tae Lim

The phylogenetic relationships between Korean endemic, Hanabusaya asiatica, and its allied groups, including four genera and nine species, were investigated at the DNA level using randomly amplified polymorphic DNA (RAPD) method. Ten primers out of 80 primers (10-mer) screened gave rise to very high polymorphism (99%) in all of the tested plants, producing 153 randomly amplified DNA fragments. H. asiatica was differentiated from its allied groups at the 0.62 of similarity index of RAPDs. This results were in accordance with previous classification based on palynological studies. It was confirmed that H. asiatica could be placed into Korean endemic and suggested that RAPD technique be used as an additional method of phylogenetic relationship for plant systematics.

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Feiga Gutman, Avinoam Nerd, Yosef Mizrahi, Dudy Bar-Zvi, and Dina Raveh

Twenty-four genotypes of marula (Sclerocarya birrea subsp. caffra) were characterized using randomly amplified polymorphic DNA (RAPD) analysis. A distinct band pattern was obtained for each of the trees, using as few as four arbitrary 10-mer primers. Trees propagated vegetatively by grafting showed identical fingerprints. These results suggest that RAPD markers provide a useful system for documenting the identity of marula genotypes.

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A.L. Fenwick and S.M. Ward

Seventeen mint accessions representing the three species grown for commercial oil production in the United States were characterized using randomly amplified polymorphic DNA (RAPD) analysis. The RAPD profiles readily identified the different Mentha species; calculation of genetic distance, based on the number of shared bands, indicated that M. spicata L. is more closely related to M. × gracilis than to M. × piperita. The RAPD profiles also distinguished among eight peppermint accessions of different geographical origin. However, only limited polymorphism was observed among the most widely grown peppermint and Scotch spearmint cultivars. These results indicate a potential lack of genetic diversity in mint cultivars grown for oil in the United States.

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Eric J. Votava and Paul W. Bosland

Random amplified polymorphic DNA (RAPD) analysis can provide a means of evaluating and comparing genetic variability within cultivars. The purpose of this study was to evaluate the relative genetic variability between accessions of two open-pollinated bell pepper cultivars: 'California Wonder', an heirloom cultivar, and 'Jupiter', a modern cultivar. RAPD molecular markers were used to assess the genetic variability between accessions of these two cultivars. The high levels of genetic variability found among accessions of 'California Wonder' may preclude its use as a standard cultivar in research.

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Luping Qu and James F. Hancock

A tetraploid blueberry population resulting from a cross of US 75 {a tetraploid hybrid of Fla 4B [a selection of Vaccinium darrowi Camp (2n = 2x = 24) × `Bluecrop' [(V. corymbosum L. (2n = 4x = 48)]} × `Bluetta' (4x) was used to generate a genetic linkage map of US 75 by randomly amplified polymorphic DNA (RAPD) analysis. One hundred and forty markers unique for Fla 4B that segregated 1:1 in the population were mapped into 29 linkage groups that cover a total genetic distance of 1288.2 cM, with a range of 1.6 to 33.9 cM between adjacent markers. The map is essentially of V. darrowi because US 75 was produced via a 2n gamete from Fla 4B and only unique markers for Fla 4B were used. Therefore, all the chromosomes of V. darrowi could be represented in the map.