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Lihong Hao, Hui Ma, Jaime A. Teixeira da Silva, and XiaoNan Yu

26 herbaceous peony cultivars with different ploidy levels were examined to provide a useful palynological basis for studying kinship and taxonomy among peonies and to advance hybrid peony breeding. Materials and Methods Plant materials. Fresh pollen

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Jason D. Lattier and Ryan N. Contreras

used to reduce or eliminate seed production, ploidy manipulation remains one of the more reliable tools for creating seedless or near-seedless clones, as illustrated by ornamental taxa such as Hypericum ( Trueblood et al., 2010 ), Buddleja ( Smith

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Juan Chen, Nianhe Xia, Jietang Zhao, Jianjun Chen, and Richard J. Henny

n = 84 for C. kwangsiensis . Chromosome numbers of other Chinese Curcumas have not been documented such as C. flaviflora S. Q. Tong and C. sichuanensis . Chromosome numbers and ploidy levels are important information for plant taxonomy

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Chao Yu, Le Luo, Hui-tang Pan, Yun-ji Sui, Run-hua Guo, Jin-yao Wang, and Qi-xiang Zhang

of the long generation time and the frequent cross-incompatibility and hybrid sterility between parents of different ploidy levels. Therefore, there is interest in the variation of ploidy within and among wild Rosa species. Despite the development

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Mark K. Ehlenfeldt and James L. Luteyn

had the potential to act as a bridge between taxonomic sections and ploidies in Vaccinium ( Ehlenfeldt et al., 2018 ). In this study, we sought to determine the feasibility of crossing 4 x V. meridionale with 4 x V. corymbosum to facilitate

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Peter H. Velguth and Harold Pellett

We evaluated flow cytometric measurement of nuclear DNA content to determine ploidy level in azalea. If ploidy level correlates with DNA content, ploidy level could be determined more readily than by direct chromosome counts and assist in planning crosses and evaluating progeny. Tested plants included azalea cultivars, materials from the azalea breeding project at the Univ. of Minnesota, and species from the Rhododendron Species Botanic Garden and the North Carolina Arboretum. Data compiled from DNA assays of practically all material analyzed fell into distinct groups consistent with their being either diploid, triploid, or tetraploid. Additionally, a known diploid plant of each of four diploid species, together with a natural or derived tetraploid plant of each of these species was obtained. Results showed that the four diploids had a similar DNA content compared to one another. DNA content of the tetraploids was also similar, and the tetraploid's DNA content was approximately twice that of the diploids, as expected. Unfortunately, success with direct chromosome counts in other material has proven elusive, currently precluding direct correlation of DNA amount with ploidy level across other species and cultivars. Although many cases exist in the literature where DNA content has a direct relationship to ploidy level, this does not always hold. Although the majority of plants tested fell into a diploid, triploid, or tetraploid grouping based on DNA content, further study is required to determine the exact relationship between ploidy level and DNA content in azalea.

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Mary Ann Start, James Luby, Robert Guthrie, and Debby Filler

The hardy Actinidia species represent a source of genetic diversity for improving A. deliciosa (kiwifruit) as well as for creating new economically important cultivars through intra- and interspecific crosses. Attempts at breeding in Actinidia have been complicated by the existence of intraspecific as well as interspecific variation in ploidy. The haploid chromosome number in Actinidia is 29 and diploid (2n=2x=58), tetraploid (2n=4x=116), and hexaploid (2n=6x=174) levels have been identified. Because of the problems encountered when crossing parents differing in ploidy level, it is desirable to know the ploidy levels of plants to be used in breeding. We determined the ploidy levels of 61 Actinidia accessions currently available in the U.S., including primarily accessions of relatively winter-hardy species. The 61 accessions, representing eight species and three interspecific hybrids, were screened for ploidy using flow cytometry. Mitotic root tip cells from one plant from each putative ploidy level were examined microscopically to confirm the ploidy level derived from flow cytometry. There were 17 diploids, 40 tetraploids, and 4 hexaploids. Intraspecific variation was not found among accessions of the species arguta, callosa, deliciosa, kolomikta, melanandra, polygama, or purpurea. All kolomikta and polygama accessions were diploid. All arguta, callosa, melanandra, and purpurea accessions were tetraploid. Actinidia deliciosa was hexaploid. One chinensis accession was tetraploid. Two accessions (NGPR 0021.14 and 0021.3), acquired as chinensis, were hexaploid and may, in fact, be A. deliciosa based on their morphology. `Issai' (arguta × polygama) was hexaploid and `Ken's Red' and `Red Princess' (both melanandra × arguta) were tetraploid.

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Michael E. Compton, J.W. Harris, and D.J. Gray

Ploidy of in vitro watermelon plantlets was estimated by painting the lower epidermis of leaves with fluorescein diacetate (FDA) and observing fluorescence of guard cell chloroplasts with a microscope and UV light. Leaves from shoot-tip cultures of known diploid and tetraploid cultivars were used to establish the mean number of chloroplasts per guard cell pair for in vitro plantlets. Leaves from diploid and tetraploid plantlets had 9.7 and 17.8 chloroplasts per guard cell pair, respectively. This method was used to estimate ploidy of shoots regenerated from cotyledon explants of the diploid cultivar Minilee. Approximately 10.6% of regenerated shoots were classified as tetraploid while still in vitro. Putative tetraploids were transplanted to the field and self-pollinated. A majority of polyploids identified in vitro were true breeding, nonchimeric tetraploids. This study demonstrate that FDA can be used to estimate ploidy of in vitro shoots of watermelon prior to acclimatization and transfer of plants to the greenhouse or field.

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Rengong Meng and Chad Finn

Nuclear DNA flow cytometry was used to differentiate ploidy level and determine nuclear DNA content in Rubus. Nuclei suspensions were prepared from leaf discs of young leaves following published protocols with modifications. DNA was stained with propidium iodide. Measurement of fluorescence of 40 genotypes, whose published ploidy ranged from diploid to dodecaploid, indicated that fluorescence increased with an increase in chromosome number. Ploidy level accounted for 99% of the variation in fluorescence intensity (r 2 = 0.99) and variation among ploidy levels was much higher than within ploidy levels. This protocol was used successfully for genotypes representing eight different Rubus subgenera. Rubus ursinus Cham. and Schldl., a native blackberry species in the Pacific Northwest, which has been reported to have 6x, 8x, 9x, 10x, 11x, and 12x forms, was extensively tested. Genotypes of R. ursinus were predominantly 12x, but 6x, 7x, 8x, 9x, 11x, and 13x forms were found as well. Attempts to confirm the 13x estimates with manual counts were unsuccessful. Ploidy level of 103 genotypes in the USDA-ARS breeding program was determined by flow cytometry. Flow cytometry confirmed that genotypes from crosses among 7x and 4x parents had chromosome numbers that must be the result of nonreduced gametes. This technique was effective in differentiating chromosome numbers differing by 1x, but was not able to differentiate aneuploids. Nuclear DNA contents of 21 diploid Rubus species from five subgenera were determined by flow cytometry. Idaeobatus, Chamaebatus, and Anaplobatus were significantly lower in DNA content than those of Rubus and Cylactis. In the Rubus subgenus, R. hispidus and R. canadensis had the lowest DNA content and R. sanctus had the highest DNA content, 0.59 and 0.75 pg, respectively. Idaeobatus had greater variation in DNA content among diploid species than the Rubus subgenus, with the highest being from R. ellipticus (0.69 pg) and lowest from R. illecebrosus (0.47 pg).

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P. Salas and L. Mansur

Gene flow between species of different ploidy levels is important in plant evolution and breeding. A cytological study was conducted on a natural population with individuals belonging to the diploid L. purpurea Gay (2n = 10) and to the tetraploid L. coquimbensis F. Phil (2n = 18) species, as well as intermediate phenotypes of apparent hybrid origin. The genus Leucocoryne is endemic to Chile and it exhibits heterogeneity, presumably genetic, for shape, size, and color of its flowers. The objective of the study was to determine if there is gene flow between species having different ploidy levels. From the karyotypic analyses of the seeds, only parental types having 2n = 10 and 2n = 18 individuals were observed. However, from the bulb analyses, 2n = 10, 2n = 18, 2n = 14, and 2n = 22 individuals were encountered. The karyotypes of the 2n = 14 and 2n = 22 individuals suggest the occurrence of natural interespecific hybridization between species with different ploidy levels in nature. Models which may account for the origin of these genotypes are proposed.