A modified staining technique for arbuscular mycorrhiza compatible with molecular probes.
Identifieur interne : 002B44 ( Main/Corpus ); précédent : 002B43; suivant : 002B45A modified staining technique for arbuscular mycorrhiza compatible with molecular probes.
Auteurs : M. Pitet ; A. Camprubí ; C. Calvet ; V. EstaúnSource :
- Mycorrhiza [ 0940-6360 ] ; 2009.
English descriptors
- KwdEn :
- DNA, Fungal (genetics), DNA, Fungal (isolation & purification), DNA, Ribosomal (genetics), DNA, Ribosomal (isolation & purification), Fungi (genetics), Molecular Probes (genetics), Mycorrhizae (genetics), Plant Roots (microbiology), Sensitivity and Specificity (MeSH), Staining and Labeling (methods).
- MESH :
- chemical , genetics : DNA, Fungal, DNA, Ribosomal, Molecular Probes.
- chemical , isolation & purification : DNA, Fungal, DNA, Ribosomal.
- genetics : Fungi, Mycorrhizae.
- methods : Staining and Labeling.
- microbiology : Plant Roots.
- Sensitivity and Specificity.
Abstract
The effects of the different steps of the root staining on the arbuscular mycorrhizal (AM) fungal rDNA extraction and amplification have been assessed. The results obtained using molecular techniques are compared with those obtained from fresh, non-stained leek roots. A modified staining procedure that eliminates heating, the use of hydrochloric acid and trypan blue, has been proved to be the most adequate to observe the AM colonisation in different plant species with/without lignified roots allowing at the same time the subsequent rDNA extraction and amplification from the stained roots. The staining technique decreased the sensitivity of the process and a higher number of roots had to be used to obtain enough material for a positive amplification. The extraction and amplification process was reliable up to 3 days after staining. A week after staining, the amplification was not dependable and after 2 weeks there was no amplification from stained material.
DOI: 10.1007/s00572-008-0206-1
PubMed: 18958505
Links to Exploration step
pubmed:18958505Le document en format XML
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Calvet</name><affiliation><nlm:affiliation>Plant Protection IRTA, Ctra. de Cabrils Km 2, E-08348 Cabrils, Barcelona, Spain.</nlm:affiliation></affiliation></author><author><name sortKey="Estaun, V" sort="Estaun, V" uniqKey="Estaun V" first="V" last="Estaún">V. Estaún</name><affiliation><nlm:affiliation>Plant Protection IRTA, Ctra. de Cabrils Km 2, E-08348 Cabrils, Barcelona, Spain. victoria.estaun@irta.cat.</nlm:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2009">2009</date><idno type="RBID">pubmed:18958505</idno><idno type="pmid">18958505</idno><idno type="doi">10.1007/s00572-008-0206-1</idno><idno type="wicri:Area/Main/Corpus">002B44</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">002B44</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">A modified staining technique for arbuscular mycorrhiza compatible with molecular probes.</title><author><name sortKey="Pitet, M" sort="Pitet, M" uniqKey="Pitet M" first="M" last="Pitet">M. Pitet</name><affiliation><nlm:affiliation>Plant Protection IRTA, Ctra. de Cabrils Km 2, E-08348 Cabrils, Barcelona, Spain.</nlm:affiliation></affiliation></author><author><name sortKey="Camprubi, A" sort="Camprubi, A" uniqKey="Camprubi A" first="A" last="Camprubí">A. Camprubí</name><affiliation><nlm:affiliation>Plant Protection IRTA, Ctra. de Cabrils Km 2, E-08348 Cabrils, Barcelona, Spain.</nlm:affiliation></affiliation></author><author><name sortKey="Calvet, C" sort="Calvet, C" uniqKey="Calvet C" first="C" last="Calvet">C. Calvet</name><affiliation><nlm:affiliation>Plant Protection IRTA, Ctra. de Cabrils Km 2, E-08348 Cabrils, Barcelona, Spain.</nlm:affiliation></affiliation></author><author><name sortKey="Estaun, V" sort="Estaun, V" uniqKey="Estaun V" first="V" last="Estaún">V. Estaún</name><affiliation><nlm:affiliation>Plant Protection IRTA, Ctra. de Cabrils Km 2, E-08348 Cabrils, Barcelona, Spain. victoria.estaun@irta.cat.</nlm:affiliation></affiliation></author></analytic><series><title level="j">Mycorrhiza</title><idno type="ISSN">0940-6360</idno><imprint><date when="2009" type="published">2009</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>DNA, Fungal (genetics)</term><term>DNA, Fungal (isolation & purification)</term><term>DNA, Ribosomal (genetics)</term><term>DNA, Ribosomal (isolation & purification)</term><term>Fungi (genetics)</term><term>Molecular Probes (genetics)</term><term>Mycorrhizae (genetics)</term><term>Plant Roots (microbiology)</term><term>Sensitivity and Specificity (MeSH)</term><term>Staining and Labeling (methods)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA, Fungal</term><term>DNA, Ribosomal</term><term>Molecular Probes</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>DNA, Fungal</term><term>DNA, Ribosomal</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Fungi</term><term>Mycorrhizae</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Staining and Labeling</term></keywords><keywords scheme="MESH" qualifier="microbiology" xml:lang="en"><term>Plant Roots</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Sensitivity and Specificity</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The effects of the different steps of the root staining on the arbuscular mycorrhizal (AM) fungal rDNA extraction and amplification have been assessed. The results obtained using molecular techniques are compared with those obtained from fresh, non-stained leek roots. A modified staining procedure that eliminates heating, the use of hydrochloric acid and trypan blue, has been proved to be the most adequate to observe the AM colonisation in different plant species with/without lignified roots allowing at the same time the subsequent rDNA extraction and amplification from the stained roots. The staining technique decreased the sensitivity of the process and a higher number of roots had to be used to obtain enough material for a positive amplification. The extraction and amplification process was reliable up to 3 days after staining. A week after staining, the amplification was not dependable and after 2 weeks there was no amplification from stained material.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">18958505</PMID><DateCompleted><Year>2009</Year><Month>03</Month><Day>24</Day></DateCompleted><DateRevised><Year>2019</Year><Month>12</Month><Day>10</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Print">0940-6360</ISSN><JournalIssue CitedMedium="Print"><Volume>19</Volume><Issue>2</Issue><PubDate><Year>2009</Year><Month>Feb</Month></PubDate></JournalIssue><Title>Mycorrhiza</Title><ISOAbbreviation>Mycorrhiza</ISOAbbreviation></Journal><ArticleTitle>A modified staining technique for arbuscular mycorrhiza compatible with molecular probes.</ArticleTitle><Pagination><MedlinePgn>125-131</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1007/s00572-008-0206-1</ELocationID><Abstract><AbstractText>The effects of the different steps of the root staining on the arbuscular mycorrhizal (AM) fungal rDNA extraction and amplification have been assessed. The results obtained using molecular techniques are compared with those obtained from fresh, non-stained leek roots. A modified staining procedure that eliminates heating, the use of hydrochloric acid and trypan blue, has been proved to be the most adequate to observe the AM colonisation in different plant species with/without lignified roots allowing at the same time the subsequent rDNA extraction and amplification from the stained roots. The staining technique decreased the sensitivity of the process and a higher number of roots had to be used to obtain enough material for a positive amplification. The extraction and amplification process was reliable up to 3 days after staining. A week after staining, the amplification was not dependable and after 2 weeks there was no amplification from stained material.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Pitet</LastName><ForeName>M</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Plant Protection IRTA, Ctra. de Cabrils Km 2, E-08348 Cabrils, Barcelona, Spain.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Camprubí</LastName><ForeName>A</ForeName><Initials>A</Initials><AffiliationInfo><Affiliation>Plant Protection IRTA, Ctra. de Cabrils Km 2, E-08348 Cabrils, Barcelona, Spain.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Calvet</LastName><ForeName>C</ForeName><Initials>C</Initials><AffiliationInfo><Affiliation>Plant Protection IRTA, Ctra. de Cabrils Km 2, E-08348 Cabrils, Barcelona, Spain.</Affiliation></AffiliationInfo></Author><Author 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