Improvement Foundation, U.S.–Israel BARD Award no. IS-2389-94, Asgrow Seed, DNA Plant Technology, and Sakata America. We thank M.A. Mutschler and J. Zhao for critical review of the manuscript and V. Lackney, G. Moriarty, and J. Jantz for technical assistance
J.P. Prince, Y. Zhang, E.R. Radwanski, and M.M. Kyle
A. Vainstein and H. Ben-Meir
Mini- and microsatellite probes were hybridized to DNA of 24 rose (Rosa×hybrida) genotypes. The resultant DNA fingerprints were shown to be genotype-specific, thereby enabling cultivar identification at the DNA level. Restriction enzyme Dra I yielded the most informative band patterns. Full-sib family analysis of DNA fingerprints revealed 32 parental-specific bands out of the 128 observed in the parents. These bands were revealed cumulatively by phage (M13), human (33.6), and oligonucleotide (GACA)4 probes. Only one pair of these loci was found to be allelic, and no linked pairs were detected in the progeny analyzed. The probability of two offspring from this cross having identical DNA fingerprints was calculated to be 2 × 10-8.
Stacey M. Sakakibara and John E. Carlson
Random amplified polymorphic DNA (RAPD) markers were evaluated for use in DNA fingerprinting of commercial Rhododendron cultivars. DNA was isolated from Rhododendron leaves and subjected to PCR amplification with single primers, 10 nucleotides in length, and of arbitrary sequence. Amplification products were visualized by agarose gel electrophoresis and ethidium bromide staining. Fingerprints were readily identifiable for a number of cultivars, and a high level of polymorphism was observed among clones of 10 rhododendron varieties. The technique was consistently reproducible in different trials using the thermocycler, between different thermocyclers, and using different DNA isolation from the same plant. This method will be applied to large-scale fingerprinting of Rhododendron cultivars and for distinguishing material propagated in tissue culture.
Yi-Xuan Kou, Hui-Ying Shang, Kang-Shan Mao, Zhong-Hu Li, Keith Rushforth, and Robert P. Adams
hypothesis was tested using DNA data. Generally, nuclear DNA regions are biparentally inherited, whereas cp and mt DNA regions are paternally inherited in the cypress family, especially in the subfamily (Cupressioideae) that monterey cypress and alaska
Candelario Mondragon-Jacobo, Natalia Doudareva, and Bruce P. Bordelon
A method for extraction of high quality DNA from four Opuntia sp. and other cacti using a hexadecyltrimethylammonium bromide (CTAB) method is described. These plants typically contain high levels of mucilages, complex polysaccharide compounds that bind water, thus preventing DNA extraction by common miniprep methods. The method involves adjusting the amount of tissue used according to species and age, followed by processing in an extraction buffer to separate coarse material. Extended centrifugation and digestion time in a separation buffer with CTAB (2%) was used. Exposing tissue to both buffers maintained polysaccharides in solution and allowed easier recovery of the aqueous phase that contains the DNA. We found that 5-8 g were needed to obtain up to 153 μg·g-1 of DNA from tender tissue. Old tissue yielded 26% less. Extraction of DNA from 5-g samples of tender tissue of the ornamental cacti Stenocereus sp., Cleistocactus sp., and Echinocereus sp. was successful. For these species, average yields ranged from 25 to 53 μg per sample. The DNA obtained was suitable for polymerase chain reaction (PCR) amplification, producing clear, distinctive, and reproducible banding patterns useful for a variety of applications.
Ying Wang, Cale A. Bigelow, and Yiwei Jiang
and biochemical analysis to measure quantitative traits such as DNA and RNA content and total cellular protein content with high precision and rapid throughput ( Eaton et al., 2004 ; Muirhead et al., 1985 ). Flow cytometry has been used to determine
Patrick P. Moore
The relationships among raspberry (Rubus spp.) clones were investigated using southern hybridization. Total DNA from 22 clones were digested with Bam III and Eco RI and hybridized with two sequences from a Pst I tomato (Lycopersicon esculentum Mill.) chloroplast library. A total of 40 different restriction fragments were distinguished for the four enzyme probe combinations. These fragments distinguished seven groups of clones with members of each group having identical fragment patterns. Clones with R. idaeus L. maternal ancestry were distinct from those with R. occident&s L. or R. parvifolius L. ancestry. Differences were detected between R. idaeus vulgatus Arrhen. and R. idaeus strigosus (Michx.). No commercial cultivars had chloroplast DNA patterns that were the same as an accession of the R. idaeus strigosus subspecies.
Lumariz Hernandez Rosario, Juan O. Rodríguez Padilla, Desiree Ramos Martínez, Alejandra Morales Grajales, Joel A. Mercado Reyes, Gabriel J. Veintidós Feliu, Benjamin Van Ee, and Dimuth Siritunga
and Solanum polygamum , are not well studied with no molecular data available in public databases such as GeneBank. Therefore, we apply molecular techniques, specifically a DNA barcoding approach, to better understand the species relationships and
R.N. Trigiano, B.H. Ownley, A.N. Trigiano, J. Coley, K.D. Gwinn, and J.K. Moulton
Biotechnology is a rapidly evolving field of science that combines cellular and molecular biology with applications in genetic engineering and recombinant DNA technology. The tools of biotechnology have come to play an increasingly important role in
Seth D. Wannemuehler, Chengyan Yue, Wendy K. Hoashi-Erhardt, R. Karina Gallardo, and Vicki McCracken
). This homology enables the development of DNA markers or specific pieces of DNA located near genes controlling traits of interest. Associations between DNA markers and variation of traits in a population for a trait of interest are analyzed using