An improved staining protocol for the assessment of arbuscular mycorrhizal in bryophytes
The most accepted method for staining arbuscular mycorrhiza (AM) in vascular plants has been one proposed by Phillips & Hayman in 1970. In particular, for the study of AM in bryophytes (s.l.) [Anthocerotophyta, Bryophyta (s.s.), Marchantiophyta] some authors have introduced modifications to...
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| Formato: | Artículo revista |
| Lenguaje: | Inglés |
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Sociedad Argentina de Botánica
2018
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| Acceso en línea: | https://revistas.unc.edu.ar/index.php/BSAB/article/view/20577 |
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I10-R325-article-205772019-12-14T13:46:33Z An improved staining protocol for the assessment of arbuscular mycorrhizal in bryophytes Un protocolo de tinción mejorado para la visualización de micorrizas arbusculares en briofitas. Cottet, Agustina C. Scervino, Jose M. Messuti, María I. Anthocerotophyta Bryophyta Marchantiophyta staining protocol. Anthocerotophyta Bryophyta Marchantiophyta protocolo de tinción. The most accepted method for staining arbuscular mycorrhiza (AM) in vascular plants has been one proposed by Phillips & Hayman in 1970. In particular, for the study of AM in bryophytes (s.l.) [Anthocerotophyta, Bryophyta (s.s.), Marchantiophyta] some authors have introduced modifications to this technique. Even though all these protocols stain AM, their main disadvantage is related to the result of material maceration (e.g. over-softening or completely destroying plant cells due to the high temperatures used, the high concentrations of reagents or the long-term exposure to aggressive chemicals). In order to optimise the results for the observation of AM in this group of plants, a modification is presented to the traditional staining technique. In the protocol here proposed, 70% ethanol is used as fixative and first clarifier, 1% potassium hydroxide (KOH) (80 °C, 20 min) as a second clarifier; 1% hydrochloric acid (HCl) (50 °C, 10 min) as an acidifier and 0.05% trypan blue (60 °C, 20 min) for dyeing. This improved protocol is not destructive, it is fast to perform and it is of wide application since it allows staining the AM in bryophytes. Un protocolo de tinción mejorado para la visualización de micorrizas arbusculares en briofitas. El método tradicional más aceptado para la tinción de micorrizas arbusculares (MA) en plantas vasculares es el propuesto por Phillips & Hayman en 1970. En particular, para el estudio de las MA en briofitas (s.l.) [Anthocerotophyta, Bryophyta (s.s.), Marchantiophyta] algunos autores introdujeron modificaciones a esta técnica. Aunque todos estos protocolos tiñen las MA, su principal desventaja está relacionada con el resultado de la maceración del material (e.g. reblandecimiento excesivo o destrucción de las células vegetales por altas temperaturas utilizadas, altas concentraciones de reactivos o exposición prolongada a sustancias químicas agresivas). Para optimizar la observación de las MA en briofitas (s.l.), se desarrolló una adaptación a la técnica de tinción tradicional y sus modificaciones. En el protocolo aquí propuesto, se usa etanol 70% para fijar y como primer clarificador, hidróxido de potasio (KOH) 1% (80 °C, 20 min) como segundo clarificador; ácido clorhídrico (HCl) 1% (50 °C, 10 min) como acidificante y azul de tripán 0,05% (60 °C, 20 min) para teñir. Este protocolo mejorado no es destructivo, es rápido de realizar y es de amplia aplicación ya que permite teñir las MA en briófitas (s.l.). Sociedad Argentina de Botánica 2018-07-06 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Artículo original application/pdf https://revistas.unc.edu.ar/index.php/BSAB/article/view/20577 10.31055/1851.2372.v53.n2.20577 Boletín de la Sociedad Argentina de Botánica (Journal of the Argentine Botanical Society; Vol. 53 No. 2 (2018): June; 201-206 Boletín de la Sociedad Argentina de Botánica; Vol. 53 Núm. 2 (2018): Junio; 201-206 Boletín de la Sociedad Argentina de Botánica; v. 53 n. 2 (2018): Junio; 201-206 1851-2372 0373-580X 10.31055/1851.2372.v53.n2 eng https://revistas.unc.edu.ar/index.php/BSAB/article/view/20577/20249 Derechos de autor 2018 Boletín de la Sociedad Argentina de Botánica |
| institution |
Universidad Nacional de Córdoba |
| institution_str |
I-10 |
| repository_str |
R-325 |
| container_title_str |
Boletín de la Sociedad Argentina de Botánica |
| language |
Inglés |
| format |
Artículo revista |
| topic |
Anthocerotophyta Bryophyta Marchantiophyta staining protocol. Anthocerotophyta Bryophyta Marchantiophyta protocolo de tinción. |
| spellingShingle |
Anthocerotophyta Bryophyta Marchantiophyta staining protocol. Anthocerotophyta Bryophyta Marchantiophyta protocolo de tinción. Cottet, Agustina C. Scervino, Jose M. Messuti, María I. An improved staining protocol for the assessment of arbuscular mycorrhizal in bryophytes |
| topic_facet |
Anthocerotophyta Bryophyta Marchantiophyta staining protocol. Anthocerotophyta Bryophyta Marchantiophyta protocolo de tinción. |
| author |
Cottet, Agustina C. Scervino, Jose M. Messuti, María I. |
| author_facet |
Cottet, Agustina C. Scervino, Jose M. Messuti, María I. |
| author_sort |
Cottet, Agustina C. |
| title |
An improved staining protocol for the assessment of arbuscular mycorrhizal in bryophytes |
| title_short |
An improved staining protocol for the assessment of arbuscular mycorrhizal in bryophytes |
| title_full |
An improved staining protocol for the assessment of arbuscular mycorrhizal in bryophytes |
| title_fullStr |
An improved staining protocol for the assessment of arbuscular mycorrhizal in bryophytes |
| title_full_unstemmed |
An improved staining protocol for the assessment of arbuscular mycorrhizal in bryophytes |
| title_sort |
improved staining protocol for the assessment of arbuscular mycorrhizal in bryophytes |
| description |
The most accepted method for staining arbuscular mycorrhiza (AM) in vascular plants has been one proposed by Phillips & Hayman in 1970. In particular, for the study of AM in bryophytes (s.l.) [Anthocerotophyta, Bryophyta (s.s.), Marchantiophyta] some authors have introduced modifications to this technique. Even though all these protocols stain AM, their main disadvantage is related to the result of material maceration (e.g. over-softening or completely destroying plant cells due to the high temperatures used, the high concentrations of reagents or the long-term exposure to aggressive chemicals). In order to optimise the results for the observation of AM in this group of plants, a modification is presented to the traditional staining technique. In the protocol here proposed, 70% ethanol is used as fixative and first clarifier, 1% potassium hydroxide (KOH) (80 °C, 20 min) as a second clarifier; 1% hydrochloric acid (HCl) (50 °C, 10 min) as an acidifier and 0.05% trypan blue (60 °C, 20 min) for dyeing. This improved protocol is not destructive, it is fast to perform and it is of wide application since it allows staining the AM in bryophytes. |
| publisher |
Sociedad Argentina de Botánica |
| publishDate |
2018 |
| url |
https://revistas.unc.edu.ar/index.php/BSAB/article/view/20577 |
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2024-09-03T22:49:05Z |
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