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Application of a modified drop method for high-resolution pachytene chromosome spreads in two Phalaenopsis species

Overview of attention for article published in Molecular Cytogenetics, June 2016
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Title
Application of a modified drop method for high-resolution pachytene chromosome spreads in two Phalaenopsis species
Published in
Molecular Cytogenetics, June 2016
DOI 10.1186/s13039-016-0254-8
Pubmed ID
Authors

Yi-Tzu Kuo, Hui-Lan Hsu, Chih-Hsin Yeh, Song-Bin Chang

Abstract

Preparation of good chromosome spreads without cytoplasmic contamination is the crucial step in cytogenetic mapping. To date, cytogenetic research in the Orchidaceae family has been carried out solely on mitotic metaphase chromosomes. Well-spread meiotic pachytene chromosomes can provide higher resolution and fine detail for analysis of chromosomal structure and are also beneficial for chromosomal FISH (fluorescence in situ hybridization) mapping. However, an adequate method for the preparation of meiotic pachytene chromosomes in orchid species has not yet been reported. Two Taiwanese native Phalaenopsis species were selected to test the modified drop method for preparation of meiotic pachytene chromosomes from pollinia. In this modified method, pollinia were ground and treated with an enzyme mixture to completely remove cell walls. Protoplasts were resuspended in ethanol/glacial acetic acid and dropped onto a wet inclined slide of 30° from a height of 0.5 m. The sample was then flowed down the inclined plane to spread the chromosomes. Hundreds of pachytene chromosomes with little to no cytoplasmic contamination were well spread on each slide. We also showed that the resolution of 45S rDNA-containing chromosomes at the pachytene stage was up to 20 times higher than that at metaphase. Slides prepared following this modified drop method were amenable to FISH mapping of both 45S and 5S rDNA on pachytene chromosomes and, after FISH, the chromosomal structure remained intact for further analysis. This modified drop method is suitable for pachytene spreads from pollinia of Phalaenopsis orchids. The large number and high-resolution pachytene spreads, with little or no cytoplasmic contamination, prepared by the modified drop method could be used for FISH mapping of DNA fragments to accelerate the integration of cytogenetic and molecular research in Phalaenopsis orchids.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 19 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 19 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 4 21%
Student > Doctoral Student 3 16%
Student > Bachelor 3 16%
Student > Ph. D. Student 2 11%
Lecturer 1 5%
Other 1 5%
Unknown 5 26%
Readers by discipline Count As %
Agricultural and Biological Sciences 9 47%
Biochemistry, Genetics and Molecular Biology 3 16%
Neuroscience 1 5%
Unknown 6 32%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 09 June 2016.
All research outputs
#20,332,117
of 22,876,619 outputs
Outputs from Molecular Cytogenetics
#298
of 402 outputs
Outputs of similar age
#293,313
of 340,764 outputs
Outputs of similar age from Molecular Cytogenetics
#8
of 9 outputs
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