Synchronization of cell division in root tips of seven major cereal species for high yields of metaphase chromosomes for flow-cytometric analysis and sorting

1997 ◽  
Vol 15 (2) ◽  
pp. 141-147 ◽  
Author(s):  
H. F. Kaeppler ◽  
S. M. Kaeppler ◽  
J. H. Lee ◽  
K. Arumuganathan
Keyword(s):  
Cell Division ◽  
Flow Cytometric Analysis ◽  
Root Tips ◽  
Metaphase Chromosomes ◽  
Flow Cytometric ◽  
Cereal Species ◽  
High Yields ◽  
Synchronization Of Cell Division

Cell synchronization and isolation of metaphase chromosomes from maize (Zea mays L.) root tips for flow cytometric analysis and sorting

Genome ◽  
1996 ◽  
Vol 39 (4) ◽  
pp. 697-703 ◽  
Author(s):  
Jai-Heon Lee ◽  
K. Arumuganathan ◽  
S. M. Kaeppler ◽  
C. M. Papa ◽  
H. F. Kaeppler
Keyword(s):  
Cell Cycle ◽  
Flow Cytometric Analysis ◽  
Maize Root ◽  
Root Tip ◽  
Root Tips ◽  
Chromosome Sorting ◽  
Synthesis Inhibitor ◽  
Metaphase Chromosomes ◽  
Flow Karyotype ◽  
Flow Cytometric

Accumulation of cells containing metaphase chromosomes is an important step in cytological analyses and chromosome sorting procedures. The goal of this research was to optimize treatment parameters to synchronize the cell cycle of maize root tip meristem cells. Levels of hydroxyurea, a DNA synthesis inhibitor, were assessed for their utility in accumulating cells at the G1 phase of the cell cycle. Trifluralin, amiprophos-methyl, and colchicine were used to accumulate cells containing metaphase chromosomes upon release from hydroxyurea inhibition. Optimal mitotic indices were achieved by treating seedlings with 5 mM hydroxyurea for 18 h, incubating for 1 h without chemical treatment to release the hydroxyurea block, and then treating emerging roots with 1 μM trifluralin for 4 h. The mitotic index of synchronized maize root tips was over 70%. Uniformity of synchronization depended upon selection of seeds with emerging radicles that were similar in length at the time of treatment. Suspensions of intact chromosomes were prepared by a simple slicing procedure. The chromosome preparations were found to be suitable for flow cytometric characterization and sorting. Chromosome peaks of the observed flow karyotype resembled the predicted flow karyotype calculated on the basis of maize chromosome size. Key words : flow karyotype, hydroxyurea, plant chromosome sorting, trifluralin.

Root tip cell cycle synchronization and metaphase-chromosome isolation suitable for flow sorting in common wheat (Triticum aestivum L.)

Genome ◽  
1997 ◽  
Vol 40 (5) ◽  
pp. 633-638 ◽  
Author(s):  
Jai-Heon Lee ◽  
K. Arumuganathan ◽  
Y. Yen ◽  
S. Kaeppler ◽  
H. Kaeppler ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Common Wheat ◽  
Univariate Analysis ◽  
Flow Cytometric Analysis ◽  
Root Tip ◽  
Bivariate Analysis ◽  
Root Tips ◽  
Metaphase Chromosomes ◽  
Cell Cycle Synchronization ◽  
Flow Cytometric

An efficient procedure for cell-cycle synchronization in meristematic root tips was achieved in common wheat. Treatment parameters for synchronizing the cell cycle of root tip meristem cells, such as time-course and applied concentrations of various chemicals, were systematically tested and optimized by flow cytometric analysis of isolated nuclei. High mitotic indices (69.5% in the root tip meristematic area) were routinely obtained by treating germinating seeds with 1.25 mM hydroxyurea for 16 h, followed by incubation in a hydroxyurea-free solution for 2 h, and treatment with 1 μM trifluralin for 4 h. Uniform seed germination prior to treatment is very important for achieving consistently high metaphase indices in the root tips. Large numbers of metaphase chromosomes, suitable for flow cytometric analysis and sorting, were isolated from synchronized root tip cells. Flow sorted wheat chromosomes, via univariate and bivariate analysis, showed four major chromosome peaks. Each discrete peak may represent wheat chromosome types with similar DNA content. Bivariate flow karyotyping based on AT and GC content did not improve the separation of wheat chromosomes.Key words: flow cytometry, trifluralin, hydroxyurea, univariate analysis, bivariate analysis.

Preparation and flow cytometric analysis of metaphase chromosomes of tomato

1991 ◽  
Vol 82 (1) ◽  
pp. 101-111 ◽  
Author(s):  
K. Arumuganathan ◽  
J. P. Slattery ◽  
S. D. Tanksley ◽  
E. D. Earle
Keyword(s):  
Flow Cytometric Analysis ◽  
Metaphase Chromosomes ◽  
Flow Cytometric

Cell synchronization and isolation of chromosomes from Chinese fir root tips for flow cytometric analysis

2015 ◽  
Vol 37 (6) ◽  
pp. 1309-1314 ◽  
Author(s):  
Jie Shen ◽  
Jin Xu ◽  
Jinhui Chen ◽  
Renhua Zheng ◽  
Jisen Shi
Keyword(s):  
Flow Cytometric Analysis ◽  
Chinese Fir ◽  
Root Tips ◽  
Cell Synchronization ◽  
Flow Cytometric

scholarly journals Isolated metaphase chromosomes. II. Proteins of Chinese hamster chromosomes.

1979 ◽  
Vol 27 (1) ◽  
pp. 454-457 ◽  
Author(s):  
W Wray ◽  
V P Wray
Keyword(s):  
Flow Cytometric Analysis ◽  
Chinese Hamster ◽  
High Capacity ◽  
Microscopic Analysis ◽  
Metaphase Chromosomes ◽  
Chromosomal Proteins ◽  
Quantitative Evidence ◽  
Flow Cytometric ◽  
Velocity Gradients

The proteins on metaphase chromosomes theoretically may be distributed ubiquitously throughout the karyotype, may be present uniquely on individual chromosomes or classes of chromosomes, or may exist in any combination of the above. Separation of chromosomes according to size using sucrose velocity gradients in high capacity zonal centrifuge rotors allows sufficient fractionation of the genome to indicate the distribution of proteins within the karyotype. Flow cytometric analysis and direct microscopic analysis were used to evaluate qualitatively the types of chromosomes present in the fractions obtained. This report is the first quantitative evidence that some of the chromosomal proteins are not distributed ubiquitously on all of the chromosomes of the karyotype.

Application of flow cytometric cell cycle analysis to the assessment of condition and growth in larvae of a freshwater teleost Galaxias olidus

2000 ◽  
Vol 57 (4) ◽  
pp. 732-741 ◽  
Author(s):  
Don Bromhead ◽  
John Kalish ◽  
Paul Waring
Keyword(s):  
Cell Cycle ◽  
Cell Division ◽  
Larval Fish ◽  
Fish Larvae ◽  
Flow Cytometric Analysis ◽  
Geographic Location ◽  
Time Of Day ◽  
Flow Cytometric ◽  
Effects Of Temperature ◽  
The Brain

Through its ability to measure cell DNA content, flow cytometric analysis (FCA) is a technique capable of accurately assessing the position of cells in the cell cycle. Using FCA to measure the proportion of dividing and nondividing cells, an index was created that allows the amount of cell division within larval fish tissues to be quantified. To assess the suitability of the cell division index (CDI) as an indicator of growth and condition in fish larvae, analyses were divided into four parts. These examined the effects of temperature, nutrition, time of day, and geographic location on the CDI of brain tissue from Galaxias olidus larvae. The index was sensitive to, firstly, differences in the brain CDI of larvae reared at 12 and 20°C and, secondly, to significant fluctuations in mean brain CDI from larvae sampled over 24 h. FCA also revealed significant differences in the CDI of starving and fed larvae. Overall, this study indicates that FCA may be suitable as an indicator of growth and condition in both laboratory-reared and wild fish larvae.

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