scholarly journals Determination of ploidy level and nuclear DNA content in blueberry by flow cytometry

1993 ◽  
Vol 86 (8) ◽  
pp. 1001-1006 ◽  
Author(s):  
D. E. Costich ◽  
R. Ortiz ◽  
T. R. Meagher ◽  
L. P. Bruederle ◽  
N. Vorsa
Keyword(s):  
Flow Cytometry ◽  
Ploidy Level ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content

scholarly journals Determination of Ploidy Level and Nuclear DNA Content in the Droseraceae by Flow Cytometry

CYTOLOGIA ◽  
2017 ◽  
Vol 82 (3) ◽  
pp. 321-327 ◽  
Author(s):  
Yoshikazu Hoshi ◽  
Masako Azumatani ◽  
Chika Suyama ◽  
Lubomίr Adamec
Keyword(s):  
Flow Cytometry ◽  
Ploidy Level ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content

Nuclear DNA amount in pure species and hybrid willows (Salix): a flow cytometric investigation

1998 ◽  
Vol 76 (1) ◽  
pp. 157-165 ◽  
Author(s):  
Jérôme Thibault
Keyword(s):  
Flow Cytometry ◽  
Ploidy Level ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Natural Hybridization ◽  
Ploidy Levels ◽  
Key Words Flow Cytometry ◽  
Dna Values ◽  
Dna Amounts

Flow cytometry (FCM) has been used to estimate the nuclear DNA content of 11 Salix species and 5 hybrids. One hundred and sixty nine individuals were studied including 159 individuals from a sequence of 32 communities along a stretch of river in France and 10 individuals from French and English collections for comparison. Isolated nuclei were stained with propidium iodide. FCM was a significantly more practical and rapid technique than that of establishing the karyotype to survey many samples of Salix for variation in ploidy. The 2C DNA amounts for diploid species ranged from 0.76 to 0.98 pg, and tetraploid values ranged from 1.62 to 1.80 pg. The DNA values were consistent with the known ploidy levels. With the exception of a doubtful Salix xquercifolia, ploidy levels and DNA amounts of hybrids were intermediate compared with those of their parents. Intraspecific variation of nuclear DNA values including instrumental variation was low (i.e., 6-11% at the same ploidy level). FCM appeared to be an accurate tool for determination of Salix triploid hybrids. However, it remains limited concerning hybrids from crosses between species of the same ploidy level. Results suggest that natural hybridization might not be frequent in the communities studied, although they have been subject to disturbance. Previous overestimates of hybridization frequency in willows were probably due to misinterpretation of the effects of the environment on Salix spp. morphology; however, the extent and mechanisms of introgression in the genus remain to be further investigated. Key words: flow cytometry, Salix, hybridization, nuclear DNA content, riparian vegetation, disturbance.

scholarly journals Chromosome Number, Ploidy Level, and Nuclear DNA Content in 23 Species of Echeveria (Crassulaceae)

Genes ◽  
2021 ◽  
Vol 12 (12) ◽  
pp. 1950
Author(s):  
Guadalupe Palomino ◽  
Javier Martínez-Ramón ◽  
Verónica Cepeda-Cornejo ◽  
Miriam Ladd-Otero ◽  
Patricia Romero ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Chromosome Number ◽  
Ploidy Level ◽  
Dna Content ◽  
Chromosome Numbers ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Root Tips ◽  
Ploidy Levels ◽  
Dna Amounts

Echeveria is a polyploid genus with a wide diversity of species and morphologies. The number of species registered for Echeveria is approximately 170; many of them are native to Mexico. This genus is of special interest in cytogenetic research because it has a variety of chromosome numbers and ploidy levels. Additionally, there are no studies concerning nuclear DNA content and the extent of endopolyploidy. This work aims to investigate the cytogenetic characteristics of 23 species of Echeveria collected in 9 states of Mexico, analyzing 2n chromosome numbers, ploidy level, nuclear DNA content, and endopolyploidy levels. Chromosome numbers were obtained from root tips. DNA content was obtained from the leaf parenchyma, which was processed according to the two-step protocol with Otto solutions and propidium iodide as fluorochrome, and then analyzed by flow cytometry. From the 23 species of Echeveria analyzed, 16 species lacked previous reports of 2n chromosome numbers. The 2n chromosome numbers found and analyzed in this research for Echeveria species ranged from 24 to 270. The range of 2C nuclear DNA amounts ranged from 1.26 pg in E. catorce to 7.70 pg in E. roseiflora, while the 1C values were 616 Mbp and 753 Mbp, respectively, for the same species. However, differences in the level of endopolyploidy nuclei were found, corresponding to 4 endocycles (8C, 16C, 32C and 64C) in E. olivacea, E. catorce, E. juarezensis and E. perezcalixii. In contrast, E. longiflora presented 3 endocycles (8C, 16C and 32C) and E. roseiflora presented 2 endocycles (8C and 16C). It has been suggested that polyploidization and diploidization processes, together with the presence of endopolyploidy, allowed Echeveria species to adapt and colonize new adverse environments.

scholarly journals NUCLEAR DNA CONTENT OF DENDROBIUM ORCHID SPECIES AS DETERMINED BY LASER FLOW CYTOMETRY

HortScience ◽  
1996 ◽  
Vol 31 (3) ◽  
pp. 322c-322
Author(s):  
W.E. Jones ◽  
A.R. Kuehnle ◽  
K. Arumuganathan
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Nuclear Dna ◽  
Evolutionary Relationship ◽  
Internal Standard ◽  
Nuclear Dna Content ◽  
Orchid Species ◽  
Group Assignment ◽  
Taxonomic Groups

Flow cytometry (FC) has proven to be an efficient and reliable method to estimate nuclear DNA content (genome size) in quantifiable units useful for genetic and molecular biology studies. This method also makes possible determination of the variation in nuclear DNA content between related taxa, which gives insights into the process of speciation. In this study, DNA content was determined in nuclei isolated from leaves of 21 Dendrobium species representing each of the major taxonomic groups used in the Univ. of Hawaii breeding program. Nuclei were mechanically isolated, stained with the nucleic acid-specific fluorochrom propidium iodide, and DNA content determined using a Coulter Epics 753 laser flow cytometer. Chicken erythrocyte nuclei (2C = 2.33 pg DNA) were used as an internal standard for direct comparative measurement. The mean diploid genome (2C) values for Dendrobium species ranged from 3.36 to 5.06 pg. Genome sizes were evaluated for possible use as discrete characters for taxonomic group assignment and compared to previous data on breeding compatibility and evolutionary relationship between species.

scholarly journals Determining Ploidy Level and Nuclear DNA Content in Rubus by Flow Cytometry

2002 ◽  
Vol 127 (5) ◽  
pp. 767-775 ◽  
Author(s):  
Rengong Meng ◽  
Chad Finn
Keyword(s):  
Flow Cytometry ◽  
Pacific Northwest ◽  
Ploidy Level ◽  
Dna Content ◽  
Chromosome Numbers ◽  
Nuclear Dna ◽  
Diploid Species ◽  
Nuclear Dna Content ◽  
Dna Flow Cytometry ◽  
Ploidy Levels

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 (r2 = 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).

Determination of Ploidy Level and Nuclear DNA Content in Tunisian Populations of Atriplex halimus L.

2005 ◽  
Vol 53 (1) ◽  
pp. 1-5 ◽  
Author(s):  
Kheiria Hcini ◽  
David J. Walker ◽  
Sadok Bouzid ◽  
Elena González ◽  
Nora Frayssinet ◽  
...  
Keyword(s):  
Ploidy Level ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Atriplex Halimus

scholarly journals Nuclear DNA content determination of paraffin-embedded liver tissue using flow cytometry.

Kanzo ◽  
1988 ◽  
Vol 29 (5) ◽  
pp. 637-642
Author(s):  
Shuichi KINO ◽  
Mitsuo KUSANO ◽  
Michio MITO
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Liver Tissue ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Content Determination

scholarly journals Variation in Ploidy Levels and Reproductive Pathways Among Flowering Crabapples

HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 773B-773
Author(s):  
Thomas Ranney* ◽  
Thomas Eaker
Keyword(s):  
Ploidy Level ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Unreduced Gametes ◽  
Breeding Programs ◽  
Ploidy Levels ◽  
Level Measurement ◽  
Dna Contents

Information on ploidy levels is extremely valuable for use in plant breeding programs. Fertility, crossability, and heritability of traits are all influenced by ploidy levels. Knowledge of reproductive pathways, including occurrence of apomixes, pseudogamy, and formation of unreduced gametes can also be important information for developing breeding strategies. Although ploidy level can be determined by counting chromosomes, flow cytometry provides a reliable and much faster means for determination of nuclear DNA content and associated ploidy level. Measurement of ploidy levels of seeds (embryo and endosperm) can also provide useful insights into reproductive pathways. The objective of this study was to determine the approximate genome size, estimated ploidy level, and range of reproductive pathways of a diverse collection of flowering crapbapples (Malus spp.). Genome sizes were calculated as nuclear DNA content for unreduced tissue (2C). Results from the taxa included in our survey showed DNA contents ranging from 1.52 to 1.82 for diploids, 2.40 to 2.62 for triploids, and 3.36 to 3.74 pg/2C for tetraploids. Based on these ranges, we identified 43 diploid, 10 triploid, and 4 tetraploid crabapple taxa in this collection. Results from open pollinated seeds and seedlings demonstrated a variety of reproductive pathways including apomixes and unreduced gametes. This research provides information on ploidy levels and reproductive pathways of flowering crabapples and will allow for more systematic and efficient progress in the development of improved cultivars.

scholarly journals Determination of Nuclear DNA Content and Ploidy Level in Rubus by Flow Cytometry

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 514D-514
Author(s):  
Rengong Meng ◽  
Chad E. Finn ◽  
Robert P. Doss
Keyword(s):  
Flow Cytometry ◽  
Chromosome Number ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Dna Flow Cytometry ◽  
Chromosome Counting ◽  
Mature Leaves ◽  
Microscopic Evaluation

Knowledge of the chromosome number in Rubus would be valuable when planning crosses and identifying plants, etc., however, preparation of tissue for microscopic evaluation and chromosome counting is difficult and time-consuming. Flow cytometry offers a more-efficient approach to this task. DNA flow cytometry was used to determine the nuclear DNA content in 22 Rubus genotypes. The genotypes represented a range of reported chromosome numbers from 2x to 12x. Six of the genotypes were representatives of Rubus ursinus, which is reported to have both 8x and 12x forms. Samples of nuclei were prepared from leaf discs of newly emerged and mature leaves following published protocols with some modifications. The DNA content was estimated by comparison of the fluorescence of Rubus nuclei with an internal DNA standard. There was an increase in nuclear DNA content concurrent with the increase in chromosome number. In these studies DNA flow cytometry could differentiate genotypes that differed by 2x, such as 6x and 8x, but could not reliably distinguish genotypes that differed by 1x, such as 7x vs. 8x or 6x. Aneuploids cannot be differentiated at this time.

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