Two New Nuclear Isolation Buffers for Plant DNA Flow Cytometry: A Test with 37 Species
Identifieur interne : 002346 ( Main/Exploration ); précédent : 002345; suivant : 002347Two New Nuclear Isolation Buffers for Plant DNA Flow Cytometry: A Test with 37 Species
Auteurs : Joo Loureiro [Portugal] ; Eleazar Rodriguez [Portugal] ; Jaroslav Doleel [République tchèque] ; Conceio Santos [Portugal]Source :
- Annals of Botany [ 0305-7364 ] ; 2007-10.
Abstract
Background and Aims After the initial boom in the application of flow cytometry in plant sciences in the late 1980s and early 1990s, which was accompanied by development of many nuclear isolation buffers, only a few efforts were made to develop new buffer formulas. In this work, recent data on the performance of nuclear isolation buffers are utilized in order to develop new buffers, general purpose buffer (GPB) and woody plant buffer (WPB), for plant DNA flow cytometry. Methods GPB and WPB were used to prepare samples for flow cytometric analysis of nuclear DNA content in a set of 37 plant species that included herbaceous and woody taxa with leaf tissues differing in structure and chemical composition. The following parameters of isolated nuclei were assessed: forward and side light scatter, propidium iodide fluorescence, coefficient of variation of DNA peaks, quantity of debris background, and the number of particles released from sample tissue. The nuclear genome size of 30 selected species was also estimated using the buffer that performed better for a given species. Key Results In unproblematic species, the use of both buffers resulted in high quality samples. The analysis of samples obtained with GPB usually resulted in histograms of DNA content with higher or similar resolution than those prepared with the WPB. In more recalcitrant tissues, such as those from woody plants, WPB performed better and GPB failed to provide acceptable results in some cases. Improved resolution of DNA content histograms in comparison with previously published buffers was achieved in most of the species analysed. Conclusions WPB is a reliable buffer which is also suitable for the analysis of problematic tissues/species. Although GPB failed with some plant species, it provided high-quality DNA histograms in species from which nuclear suspensions are easy to prepare. The results indicate that even with a broad range of species, either GPB or WPB is suitable for preparation of high-quality suspensions of intact nuclei suitable for DNA flow cytometry.
Url:
- https://api.istex.fr/document/8D45F11E10F8D95E119EF4EF2786200B56E51A93/fulltext/pdf
- http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2749623
DOI: 10.1093/aob/mcm152
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: 001557
- to stream Istex, to step Curation: 001556
- to stream Istex, to step Checkpoint: 000624
- to stream Pmc, to step Corpus: 000C17
- to stream Pmc, to step Curation: 000C16
- to stream Pmc, to step Checkpoint: 001100
- to stream Ncbi, to step Merge: 000729
- to stream Ncbi, to step Curation: 000729
- to stream Ncbi, to step Checkpoint: 000729
- to stream Main, to step Merge: 002430
- to stream Main, to step Curation: 002346
Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title>Two New Nuclear Isolation Buffers for Plant DNA Flow Cytometry: A Test with 37 Species</title>
<author><name sortKey="Loureiro, Joo" sort="Loureiro, Joo" uniqKey="Loureiro J" first="Joo" last="Loureiro">Joo Loureiro</name>
</author>
<author><name sortKey="Rodriguez, Eleazar" sort="Rodriguez, Eleazar" uniqKey="Rodriguez E" first="Eleazar" last="Rodriguez">Eleazar Rodriguez</name>
</author>
<author><name sortKey="Doleel, Jaroslav" sort="Doleel, Jaroslav" uniqKey="Doleel J" first="Jaroslav" last="Doleel">Jaroslav Doleel</name>
</author>
<author><name sortKey="Santos, Conceio" sort="Santos, Conceio" uniqKey="Santos C" first="Conceio" last="Santos">Conceio Santos</name>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">ISTEX</idno>
<idno type="RBID">ISTEX:8D45F11E10F8D95E119EF4EF2786200B56E51A93</idno>
<date when="2007" year="2007">2007</date>
<idno type="doi">10.1093/aob/mcm152</idno>
<idno type="url">https://api.istex.fr/document/8D45F11E10F8D95E119EF4EF2786200B56E51A93/fulltext/pdf</idno>
<idno type="wicri:Area/Istex/Corpus">001557</idno>
<idno type="wicri:Area/Istex/Curation">001556</idno>
<idno type="wicri:Area/Istex/Checkpoint">000624</idno>
<idno type="wicri:doubleKey">0305-7364:2007:Loureiro J:two:new:nuclear</idno>
<idno type="wicri:source">PMC</idno>
<idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2749623</idno>
<idno type="RBID">PMC:2749623</idno>
<idno type="wicri:Area/Pmc/Corpus">000C17</idno>
<idno type="wicri:Area/Pmc/Curation">000C16</idno>
<idno type="wicri:Area/Pmc/Checkpoint">001100</idno>
<idno type="wicri:Area/Ncbi/Merge">000729</idno>
<idno type="wicri:Area/Ncbi/Curation">000729</idno>
<idno type="wicri:Area/Ncbi/Checkpoint">000729</idno>
<idno type="wicri:doubleKey">0305-7364:2007:Loureiro J:two:new:nuclear</idno>
<idno type="wicri:Area/Main/Merge">002430</idno>
<idno type="wicri:Area/Main/Curation">002346</idno>
<idno type="wicri:Area/Main/Exploration">002346</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title level="a">Two New Nuclear Isolation Buffers for Plant DNA Flow Cytometry: A Test with 37 Species</title>
<author><name sortKey="Loureiro, Joo" sort="Loureiro, Joo" uniqKey="Loureiro J" first="Joo" last="Loureiro">Joo Loureiro</name>
<affiliation wicri:level="1"><country xml:lang="fr">Portugal</country>
<wicri:regionArea>Laboratory of Biotechnology and Cytomics, CESAM and Department of Biology, University of Aveiro, Campus Universitrio de Santiago, 3810-193 Aveiro</wicri:regionArea>
<wicri:noRegion>3810-193 Aveiro</wicri:noRegion>
</affiliation>
<affiliation wicri:level="1"><country wicri:rule="url">Portugal</country>
</affiliation>
</author>
<author><name sortKey="Rodriguez, Eleazar" sort="Rodriguez, Eleazar" uniqKey="Rodriguez E" first="Eleazar" last="Rodriguez">Eleazar Rodriguez</name>
<affiliation wicri:level="1"><country xml:lang="fr">Portugal</country>
<wicri:regionArea>Laboratory of Biotechnology and Cytomics, CESAM and Department of Biology, University of Aveiro, Campus Universitrio de Santiago, 3810-193 Aveiro</wicri:regionArea>
<wicri:noRegion>3810-193 Aveiro</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Doleel, Jaroslav" sort="Doleel, Jaroslav" uniqKey="Doleel J" first="Jaroslav" last="Doleel">Jaroslav Doleel</name>
<affiliation wicri:level="1"><country xml:lang="fr">République tchèque</country>
<wicri:regionArea>Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovsk, Olomouc, CZ-77200</wicri:regionArea>
<wicri:noRegion>CZ-77200</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Santos, Conceio" sort="Santos, Conceio" uniqKey="Santos C" first="Conceio" last="Santos">Conceio Santos</name>
<affiliation wicri:level="1"><country xml:lang="fr">Portugal</country>
<wicri:regionArea>Laboratory of Biotechnology and Cytomics, CESAM and Department of Biology, University of Aveiro, Campus Universitrio de Santiago, 3810-193 Aveiro</wicri:regionArea>
<wicri:noRegion>3810-193 Aveiro</wicri:noRegion>
</affiliation>
</author>
</analytic>
<monogr></monogr>
<series><title level="j">Annals of Botany</title>
<idno type="ISSN">0305-7364</idno>
<idno type="eISSN">1095-8290</idno>
<imprint><publisher>Oxford University Press</publisher>
<date type="published" when="2007-10">2007-10</date>
<biblScope unit="volume">100</biblScope>
<biblScope unit="issue">4</biblScope>
<biblScope unit="page" from="875">875</biblScope>
<biblScope unit="page" to="888">888</biblScope>
</imprint>
<idno type="ISSN">0305-7364</idno>
</series>
<idno type="istex">8D45F11E10F8D95E119EF4EF2786200B56E51A93</idno>
<idno type="DOI">10.1093/aob/mcm152</idno>
<idno type="ArticleID">mcm152</idno>
</biblStruct>
</sourceDesc>
<seriesStmt><idno type="ISSN">0305-7364</idno>
</seriesStmt>
</fileDesc>
<profileDesc><textClass></textClass>
<langUsage><language ident="en">en</language>
</langUsage>
</profileDesc>
</teiHeader>
<front><div type="abstract">Background and Aims After the initial boom in the application of flow cytometry in plant sciences in the late 1980s and early 1990s, which was accompanied by development of many nuclear isolation buffers, only a few efforts were made to develop new buffer formulas. In this work, recent data on the performance of nuclear isolation buffers are utilized in order to develop new buffers, general purpose buffer (GPB) and woody plant buffer (WPB), for plant DNA flow cytometry. Methods GPB and WPB were used to prepare samples for flow cytometric analysis of nuclear DNA content in a set of 37 plant species that included herbaceous and woody taxa with leaf tissues differing in structure and chemical composition. The following parameters of isolated nuclei were assessed: forward and side light scatter, propidium iodide fluorescence, coefficient of variation of DNA peaks, quantity of debris background, and the number of particles released from sample tissue. The nuclear genome size of 30 selected species was also estimated using the buffer that performed better for a given species. Key Results In unproblematic species, the use of both buffers resulted in high quality samples. The analysis of samples obtained with GPB usually resulted in histograms of DNA content with higher or similar resolution than those prepared with the WPB. In more recalcitrant tissues, such as those from woody plants, WPB performed better and GPB failed to provide acceptable results in some cases. Improved resolution of DNA content histograms in comparison with previously published buffers was achieved in most of the species analysed. Conclusions WPB is a reliable buffer which is also suitable for the analysis of problematic tissues/species. Although GPB failed with some plant species, it provided high-quality DNA histograms in species from which nuclear suspensions are easy to prepare. The results indicate that even with a broad range of species, either GPB or WPB is suitable for preparation of high-quality suspensions of intact nuclei suitable for DNA flow cytometry.</div>
</front>
</TEI>
<affiliations><list><country><li>Portugal</li>
<li>République tchèque</li>
</country>
</list>
<tree><country name="Portugal"><noRegion><name sortKey="Loureiro, Joo" sort="Loureiro, Joo" uniqKey="Loureiro J" first="Joo" last="Loureiro">Joo Loureiro</name>
</noRegion>
<name sortKey="Loureiro, Joo" sort="Loureiro, Joo" uniqKey="Loureiro J" first="Joo" last="Loureiro">Joo Loureiro</name>
<name sortKey="Rodriguez, Eleazar" sort="Rodriguez, Eleazar" uniqKey="Rodriguez E" first="Eleazar" last="Rodriguez">Eleazar Rodriguez</name>
<name sortKey="Santos, Conceio" sort="Santos, Conceio" uniqKey="Santos C" first="Conceio" last="Santos">Conceio Santos</name>
</country>
<country name="République tchèque"><noRegion><name sortKey="Doleel, Jaroslav" sort="Doleel, Jaroslav" uniqKey="Doleel J" first="Jaroslav" last="Doleel">Jaroslav Doleel</name>
</noRegion>
</country>
</tree>
</affiliations>
</record>
Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Wicri/Bois/explor/OrangerV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 002346 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 002346 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien |wiki= Wicri/Bois |area= OrangerV1 |flux= Main |étape= Exploration |type= RBID |clé= ISTEX:8D45F11E10F8D95E119EF4EF2786200B56E51A93 |texte= Two New Nuclear Isolation Buffers for Plant DNA Flow Cytometry: A Test with 37 Species }}
![]() | This area was generated with Dilib version V0.6.25. | ![]() |