Evidence map›Paper›PMID 27275186›Full record

ArticleMolecular cytogenetics2016

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

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

Open access · goldAbstract read
In one paragraph

Article in Molecular cytogenetics, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.

0numbers the graph read from it
0cells of the map it votes in
5citing papers in PubMed
4.1field-weighted citation impact, top 7% of its field
1 · What the graph read from it

What it found

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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

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3 · Its place in the literature

Who cites it

5 citing papers in PubMed, 11 citations in OpenAlex.

  1. Article
  2. Critical Steps in DAPI and FISH Imaging of Chromosome Spread Preparations.Methods in molecular biology (Clifton, N.J.) · 2023
    Article
  3. Fluorescence in situ hybridization in plants: recent developments and future applications.Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology · 2019
    Review
  4. Article
  5. Article
4 · The record

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5 · Who and what money

Authors and funding

4 authors at 2 institutions in 1 country.

Yi-Tzu KuoDepartment of Life Sciences, National Cheng Kung University, No.1, University Road, Tainan City, 701 Taiwan.
Hui-Lan HsuDepartment of Life Sciences, National Cheng Kung University, No.1, University Road, Tainan City, 701 Taiwan.
Chih-Hsin YehTaoyuan District Agricultural Research and Extension Station, Council of Agriculture, Executive Yuan, 139, Sec. 2, Dongfu Rd., Houzhuang, Sinwu Dist., Taoyuan City, 32745 Taiwan.
Song-Bin ChangDepartment of Life Sciences, National Cheng Kung University, No.1, University Road, Tainan City, 701 Taiwan.
National Cheng Kung University · TWCouncil of Agriculture · TW

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

backgroundPreparation 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.

resultsTwo 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.

conclusionThis 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.

Indexed as

Drop methodFISH mappingFluorescence in situ hybridization (FISH)OrchidPachytene chromosomePhalaenopsis

Identifiers

PMID27275186
PMCPMC4893830
OpenAlexW2419079336

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LicenceCC BY
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.