| Are seeds suitable for flow cytometric estimation of plant genome size? | |
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MedLine Citation:
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PMID: 15739186 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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BACKGROUND: Nuclear DNA content in plants is commonly estimated using flow cytometry (FCM). Plant material suitable for FCM measurement should contain the majority of its cells arrested in the G0/G1 phase of the cell cycle. Usually young, rapidly growing leaves are used for analysis. However, in some cases seeds would be more convenient because they can be easily transported and analyzed without the delays and additional costs required to raise seedlings. Using seeds would be particularly suitable for species that contain leaf cytosol compounds affecting fluorochrome accessibility to the DNA. Therefore, the usefulness of seeds or their specific tissues for FCM genome size estimation was investigated, and the results are presented here. METHODS: The genome size of six plant species was determined by FCM using intercalating fluorochrome propidium iodide for staining isolated nuclei. Young leaves and different seed tissues were used as experimental material. Pisum sativum cv. Set (2C = 9.11 pg) was used as an internal standard. For isolation of nuclei from species containing compounds that interfere with propidium iodide intercalation and/or fluorescence, buffers were used supplemented with reductants. RESULTS: For Anethum graveolens, Beta vulgaris, and Zea mays, cytometrically estimated genome size was the same in seeds and leaves. For Helianthus annuus, different values for DNA amounts in seeds and in leaves were obtained when using all but one of four nuclei isolation buffers. For Brassica napus var. oleifera, none of the applied nuclei isolation buffers eliminated differences in genome size determined in the seeds and leaves. CONCLUSIONS: The genome size of species that do not contain compounds that influence fluorochrome accessibility appears to be the same when estimated using specific seed tissues and young leaves. Seeds can be more suitable than leaves, especially for species containing staining inhibitors in the leaf cytosol. Thus, use of seeds for FCM nuclear DNA content estimation is recommended, although for some species a specific seed tissue (usually the radicle) should be used. Protocols for preparation of samples from endospermic and endospermless seeds have been developed. |
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Authors:
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Elwira Sliwinska; Elzbieta Zielinska; Iwona Jedrzejczyk |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Cytometry. Part A : the journal of the International Society for Analytical Cytology Volume: 64 ISSN: 1552-4922 ISO Abbreviation: Cytometry A Publication Date: 2005 Apr |
Date Detail:
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Created Date: 2005-03-23 Completed Date: 2005-09-16 Revised Date: 2007-07-24 |
Medline Journal Info:
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Nlm Unique ID: 101235694 Medline TA: Cytometry A Country: United States |
Other Details:
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Languages: eng Pagination: 72-9 Citation Subset: IM |
Copyright Information:
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Copyright 2005 Wiley-Liss, Inc. |
Affiliation:
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Laboratory of Molecular Biology and Cytometry, Department of Genetics and Plant Breeding, University of Technology and Agriculture, Bydgoszcz, Poland. elwira@mail.atr.bydgoszcz.pl |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Anethum graveolens
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genetics Beta vulgaris / genetics Brassica napus / genetics Cell Nucleus / chemistry, genetics DNA, Plant / analysis*, chemistry Flow Cytometry / methods* Genome, Plant* Helianthus / genetics Peas / genetics Plant Leaves / genetics Plants / genetics* Ploidies Propidium / chemistry Seeds / genetics* Zea mays / genetics |
| Chemical | |
Reg. No./Substance:
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0/DNA, Plant; 36015-30-2/Propidium |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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