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- Author or Editor: Nahla Bassil x
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Simple Sequence Repeat (SSR) markers developed in apple and pear were used to determine genetic relationships among heritage apple and pear cultivars from Portugal's Azore Islands, and to develop identity fingerprints for European and Asian pear accessions at the USDA–ARS National Clonal Germplasm Repository (NCGR). We used 11 SSR markers (six from apple and five from pear) to examine 18 heritage apple and 9 heritage pear cultivars from the Azores. Eight additional Portuguese and economically important cultivars of apple and eight of pear were used as standards. Cluster analysis separated the apple and pear accessions into two distinct groups. Among apple genotypes, 12 unique accessions and five groups of synonyms were identified, while, in pear, seven unique genotypes and three pairs of synonyms were found. None of the accessions obtained from the Azores corresponded to widely grown standard Portuguese apple or pear cultivars. To examine 144 NCGR pear accessions, we used nine polymorphic SSR loci that were developed from GenBank sequences. Cluster analysis identified five sets of synonyms (four in P. communis L. and one in P. ussuriensis Maxim.) and four pairs of homonyms (three in P. communis and one in P. pyrifolia Burm. f. Nakai), and confirmed three clonal sets. Morphological evaluations and additional SSR markers will be used to confirm these results, and to genetically document the identities of pear genotypes. SSR markers will greatly assist the management of ex situ pome fruit germplasm collections by helping to eliminate duplicate accessions and expanding the genetic diversity represented.
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.
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.
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.
Edible European pears (Pyrus communis sp. communis L.) are thought to be derived from wild relatives native to the Caucasus Mountain region and eastern Europe. We collected genotype, phenotype, and geographic origin data for 145 P. communis individuals derived from seeds collected from wild relatives. These individuals are currently maintained in the USDA–ARS National Plant Germplasm System (NPGS) in Corvallis, Ore. Pear genotypes were obtained using 13 microsatellite markers. A Bayesian clustering method grouped the individual pear genotypes into 12 clusters. The subspecies of pears native to the Caucasus Mountains of Russia, Crimea, and Armenia could be genetically differentiated from the subspecies native to eastern European countries. Pears with large fruit clustered closely together and are most closely related to a group of genotypes that are intermediate to the other groups. Based on the high number of unique alleles and heterozygosity in each of the 12 clusters, we conclude that the genetic diversity of wild P. communis is not fully represented in the NPGS
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.
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.
The NRT1gene family encodes transport proteins with dual or low affinity for nitrate. The objectives of this experiment were to develop a system that could be used to compare the expression of the NRT1genes between species. This was accomplished by comparing sequences of NRT1homologues from various species and designing degenerate primers in regions of high homology. These primers were used to amplify a region of the NRT1gene from species of interest. A 635 bp PCR product was amplified from each species using the MD2-1 (5' ATGTTACCAAYWTGGGCMAC-3') and MD2-2 (5'-GCCAMWARCCARTAGAAAT-3') primers. The PCR products were cloned and sequenced. At the nucleotide level, CornussericeaL. `Kelseyi' and RhododendronL. `Unique' were 79.52% identical. Species-specific primers were designed and used for RT-PCR to compare NRT1expression in roots of hydroponically grown C. sericea, C. sericea `Kelseyi', and Rhododendron`Unique'. The relative levels of NRT1expression, normalized using 18S rRNA as a standard, were ≈3.2 to 1.7 to 1.0 for C. sericea, C. sericea `Kelseyi', and Rhododendron`Unique', respectively. This approach may eventually be used to examine nitrate uptake potential in different taxa of plants at different times during the growing season.
Hazelnut (Corylus avellana L.) softwood cuttings of the cultivars Ennis and Casina were propagated under mist during June and July 1987 and 1988. Rooting of stem cuttings was stimulated by both Agrobacterium and IBA treatment; however, IBA caused nearly complete bud abscission. Better rooting and bud retention were observed in `Casina' than in `Ennis' in 1988. Bud retention on Agrobacterium -inoculated cuttings improved as the cuttings approached the semi-hardwood stage. Six months after transplanting, Agrobacterium -inoculated hazelnut cuttings had an extensive root system, characteristic of hairy root. Although the mechanism remains unclear, strains of Agrobacterium rhizogenes are effective rooting agents in hazelnut and may cause less bud abscission than IBA. Chemical name used: 1 H -indole-3-butyric acid (IBA).