Search Results

You are looking at 1 - 10 of 16 items for

  • Author or Editor: Nahla V. Bassil x
Clear All Modify Search
Free access

Peter Boches, Nahla V. Bassil and Lisa Rowland

Sixty-nine accessions representing wild and domesticated highbush blueberry (Vaccinium corymbosum L.) germplasm were genotyped using 28 simple sequence repeats (SSRs). A total of 627 alleles was detected and unique fingerprints were generated for all accessions. Suspected duplicate accessions of `Coville' and `Ivanhoe' had DNA fingerprints that were identical to `Coville' and `Ivanhoe', respectively. Genetic similarity measures placed wild and cultivated blueberries in separate groups. Northern highbush blueberries grouped among ancestral clones that were used extensively in blueberry breeding such as `Rubel' and `Stanley'. Southern highbush blueberries formed a separate group from northern highbush blueberries. The microsatellite markers used here show excellent promise for further use in germplasm identification, in genetic studies of wild Vaccinium L. populations, and for constructing linkage maps.

Free access

William M. Proebsting, Nahla V. Bassil and David A. Lightfoot

Propagation of Corylus avellana stem cuttings may be limited by either root initiation or bud abscission. We divided juvenile shoots of 3 varieties growing in layering beds in mid-July into 4 or 5 3-node cuttings with leaves at the upper two nodes, except that terminal cuttings had one expanded leaf. Cuttings were treated with 5 mM IBA in 50% EtOH, a mixture of A. rhizogenes strains A7 + 22 or left untreated. IBA and bacteria stimulated rooting of cuttings from all shoot positions. Rooting of the terminal cuttings (<50%) was less than that of the sub-terminal cuttings (>80%). Bud retention was <50% on terminal cuttings, nearly 100% on sub-terminal cuttings. Using juvenile stock plants of various varieties, sub-terminal cuttings treated with Agrobacterium or 5 mM IBA may yield 70-90% cuttings with both roots and buds, Agravitropic roots, characteristic of genetic transformation, were observed on Agrobacterium-treated cuttings. Dot blots probed for TL-DNA were negative, however.

Free access

Peter Boches, Lisa J. Rowland, Kim Hummer and Nahla V. Bassil

Microsatellite markers for blueberry (Vaccinium L.) were created from a preexisting blueberry expressed sequence tag (EST) library of 1305 sequences and a microsatellite-enriched genomic library of 136 clones.

Microsatellite primers for 65 EST-containing simple sequence repeats (SSRs) and 29 genomic SSR were initially tested for amplification and polymorphism on agarose gels. Potential usefulness of these SSRs for estimating species relationships in the genus was assessed through cross-species transference of 45 SSR loci and cluster analysis using genetic distance values from five highly polymorphic EST-SSR loci. Cross-species amplification for 45 SSR loci ranged from 17% to 100%, and was 83% on average in nine sections. Cluster analysis of 59 Vaccinium species based on genetic distance measures obtained from 5 EST-SSR loci supported the concept of V. elliotii Chapm. as a genetically distinct diploid highbush species and indicated that V. ashei Reade is of hybrid origin. Twenty EST-SSR and 10 genomic microsatellite loci were used to determine genetic diversity in 72 tetraploid V. corymbosum L. accessions consisting mostly of common cultivars. Unique fingerprints were obtained for all accessions analyzed. Genetic relationships, based on microsatellites, corresponded well with known pedigree information. Most modern cultivars clustered closely together, but southern highbush and northern highbush cultivars were sufficiently differentiated to form distinct clusters. Future use of microsatellites in Vaccinium will help resolve species relationships in the genus, estimate genetic diversity in the National Clonal Germplasm Repository (NCGR) collection, and confirm the identity of clonal germplasm accessions.

Free access

Nahla V. Bassil, R. Botta and S.A. Mehlenbacher

Three microsatellite-enriched libraries of the european hazelnut (Corylus avellana L.) were constructed: library A for CA repeats, library B for GA repeats, and library C for GAA repeats. Twenty-five primer pairs amplified easy-to-score single loci and were used to investigate polymorphism among 20 C. avellana genotypes and to evaluate cross-species amplification in seven Corylus L. species. Microsatellite alleles were estimated by fluorescent capillary electrophoresis fragment sizing. The number of alleles per locus ranged from 2 to 12 (average = 7.16) in C. avellana and from 5 to 22 overall (average = 13.32). With the exception of CAC-B110, di-nucleotide SSRs were characterized by a relatively large number of alleles per locus (≥5), high average observed and expected heterozygosity (Ho and He > 0.6), and a high mean polymorphic information content (PIC ≥ 0.6) in C. avellana. In contrast, tri-nucleotide microsatellites were more homozygous (Ho = 0.4 on average) and less informative than di-nucleotide simple sequence repeats (SSRs) as indicated by a lower mean number of alleles per locus (4.5), He (0.59), and PIC (0.54). Cross-species amplification in Corylus was demonstrated. These microsatellite markers were highly heterozygous and polymorphic and differentiated among genotypes of C. avellana irrespective of geographical origin. They will aid in fingerprinting genotypes of the european hazelnut and other Corylus species, genome mapping, and genetic diversity assessments.

Free access

William M. Proebsting, Nahla V. Bassil and David A. Lightfoot

Propagation of Corylus avellana stem cuttings may be limited by either root initiation or bud abscission. We divided juvenile shoots of 3 varieties growing in layering beds in mid-July into 4 or 5 3-node cuttings with leaves at the upper two nodes, except that terminal cuttings had one expanded leaf. Cuttings were treated with 5 mM IBA in 50% EtOH, a mixture of A. rhizogenes strains A7 + 22 or left untreated. IBA and bacteria stimulated rooting of cuttings from all shoot positions. Rooting of the terminal cuttings (<50%) was less than that of the sub-terminal cuttings (>80%). Bud retention was <50% on terminal cuttings, nearly 100% on sub-terminal cuttings. Using juvenile stock plants of various varieties, sub-terminal cuttings treated with Agrobacterium or 5 mM IBA may yield 70-90% cuttings with both roots and buds, Agravitropic roots, characteristic of genetic transformation, were observed on Agrobacterium-treated cuttings. Dot blots probed for TL-DNA were negative, however.

Restricted access

Weijian Cai, Jason D. Zurn, Nahla V. Bassil and Kim E. Hummer

The genetic control of flowering habit in many species of Fragaria has not been well studied. Identification of flowering traits and patterns for these taxa could be used in the quest for perpetual flowering (PF) genes and for the octoploids, broaden the genepool of available PF parents for breeding programs. As such, clones from the Fragaria germplasm collection housed at the USDA-ARS National Clonal Germplasm Repository in Corvallis, OR, were evaluated to describe flowering habits in various taxa and identify PF clones. Flower presence was recorded monthly for 962 clones of 36 taxa from the first of May through October in 2015 and 2016 to determine flowering habit and pairwise comparisons between taxa were examined using Pearson’s Chi-squared test. Taxa with the largest percent of PF accessions were F. vesca subsp. vesca f. semperflorens, F. vesca subsp. vesca f. alba, F. vesca subsp. americana, and F. virginiana subsp. glauca. These taxa had similar flowering habits to each other but were significantly different (α = 0.05) from most other taxa in which the seasonal flowering (SF) trait was predominant. Fifteen clones that demonstrated the PF phenotype in both 2015 and 2016 were identified. Differing genetic controls have been observed for flowering habit in F. ×ananassa and F. vesca. Additional studies are needed to determine genetic control of flowering in other Fragaria taxa.

Free access

Barbara S. Gilmore, Nahla V. Bassil, Danny L. Barney, Brian J. Knaus and Kim E. Hummer

Identifying and evaluating genetic diversity of culinary rhubarb (Rheum ×rhababarum) cultivars using morphological characteristics is challenging given the existence of synonyms and nomenclatural inconsistencies. Some cultivars with similar names are morphologically different, and seedlings may grow and become associated with the parental name. Morphological traits of one cultivar may vary when measured under different environmental conditions. Molecular markers are consistent for unique genotypes across environments and provide genetic fingerprints to assist in resolving identity issues. Microsatellite repeats, also called simple sequence repeats (SSRs), are commonly used for fingerprinting fruit and nut crops, but only 10 SSRs have previously been reported in rhubarb. The objectives of this study were to use short-read DNA sequences to develop new di-nucleotide-containing SSR markers for rhubarb and to determine if the markers were useful for cultivar identification. A total of 97 new SSR primer pairs were designed from the short-read DNA sequences. The amplification success rate of these SSRs was 77%, whereas polymorphism of those reached 76% in a test panel of four or eight rhubarb individuals. From the 57 potentially polymorphic primer pairs obtained, 25 SSRs were evaluated in 58 Rheum accessions preserved in the U.S. Department of Agriculture, National Plant Germplasm System. The primer pairs generated 314 fragments with an average of 12.6 fragments per pair. The clustering of many accessions in well-supported groups supported previous findings based on amplified fragment length polymorphisms (AFLPs). Cluster analysis, using the proportion of shared allele distance among the 25 SSRs, distinguished each of the 58 accessions including individuals that had similar names or the same name. Accessions that grouped in well-supported clusters previously belonged to similar clusters with high bootstrap support based on AFLP. In summary, our technique of mining short-read sequencing data was successful in identifying 97 di-nucleotide-containing SSR sequences. Of those tested, the 25 most polymorphic and easy-to-score primer pairs proved useful in fingerprinting rhubarb cultivars. We recommend the use of short-read sequencing for the development of SSR markers in the identification of horticultural crops.

Free access

Patricio A. Brevis, Nahla V. Bassil, James R. Ballington and James F. Hancock

The use of interspecific hybridization in blueberry (Vaccinium L. section Cyanococcus Gray) breeding has resulted in the incorporation of novel traits from wild germplasm and the expansion of the geographic limits of highbush blueberry (V. ×corymbosum L.) production. The objectives of this study were: 1) to estimate the impact of wide hybridization on inbreeding, heterozygosity, and coancestry of the cultivated tetraploid highbush blueberry; 2) to establish the usefulness of microsatellite markers in assessing genetic relationships among southern highbush blueberry [SHB (V. ×corymbosum)] cultivars; and 3) to report on the expected genetic contribution of wild Vaccinium clones to SHB cultivars. Pedigrees of 107 highbush blueberry cultivars were used to calculate tetrasomic inbreeding coefficients (F), pedigree-based genetic distances, and expected genetic contributions of wild clones. Genotyping data from 21 single-locus microsatellite markers screened across 68 genotypes were used to calculate heterozygosity and proportion of shared alleles distances (Dsa). The results indicated that the effects of wide hybridization on genetic diversity of cultivated blueberry were lower than previously thought. First, SHB cultivars exhibited similar levels of molecular relatedness as historical northern highbush blueberry [NHB (V. ×corymbosum)] cultivars (median Dsa between pairs of cultivars equals 0.19 in both cultivar groups), despite the broader genetic base and larger pedigree distances in the former cultivar group. Second, levels of heterozygosity in modern NHB cultivars were not statistically different from those of SHB (χ2 = 4.0; df = 3; P > 0.05), despite the considerably higher levels of inbreeding in the former cultivar group (median F equal to 0.0035 and 0.0013, respectively). Furthermore, pedigree-based genetic distances were significantly correlated with Dsa (r = 0.57, P ≤ 0.0001), indicating that microsatellite markers are reliable tools in most cases to assess the genetic relationships among SHB cultivars. Finally, seven Vaccinium species constitute the current genetic base of cultivated SHB. In this article, we report on the expected genetic contributions of V. angustifolium Ait., V. constablaei Gray, V. corymbosum, V. darrowii Camp, V. elliottii Chapman, V. tenellum Ait., and V. virgatum Ait. (syn. V. ashei Reade) clones to 38 SHB cultivars.

Free access

Nahla V. Bassil, B.J. Rebhuhn, David W.S. Mok and Machteld C. Mok

Development of optimum protocols for micropropagation of C. avellana is particularly important due to the threat of Eastern Filbert Blight and the need for rapid increase of resistant varieties and advanced selections. Therefore, efforts were directed at in vitro establishment, multiplication and rooting, starting with buds from mature trees. The frequency of shoot formation from buds was highest in August but varied with the genotype. Medium containing high Ca levels was more effective in preventing bud necrosis than MS medium. Multiplication rates of 4-7 new shoots/propagule were obtained over a 6-week culture period. Rooting of some genotypes could be accomplished by inclusion of 1 or 3 μM β- indolebutyric acid (IBA) in the medium. Other genotypes responded better to a dip of shoot bases in 1-10 mM IBA for 10 sec., followed by a passage on auxin-free medium. Large numbers of healthy plantlets have been produced for transfer to soil.