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DTAF: an efficient probe to study cyanobacterial-plant interaction using confocal laser scanning microscopy (CLSM).

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Title: DTAF: an efficient probe to study cyanobacterial-plant interaction using confocal laser scanning microscopy (CLSM).
Authors: Ahmed, Mehboob, Stal, Lucas1, Hasnain, Shahida2 genetic@brain.net.pk
Source: Journal of Industrial Microbiology & Biotechnology. Jan2011, Vol. 38 Issue 1, p249-255. 7p. 3 Diagrams, 1 Chart.
Subjects: Cyanobacteria, Plant cell interaction, Plant roots, Microscopical technique, Fluorescence microscopy, Tissue engineering, Molecular biology, Polysaccharides
Abstract: variety of microscopic techniques have been utilized to study cyanobacterial associations with plant roots, but confocal laser scanning microscopy (CLSM) is the least used due to the unavailability of a suitable fluorescent dye. Commonly used lectins have problems with their binding ability with root cells and their visualization under CLSM. DTAF (5-(4,6-dichlorotriazinyl) aminofluorescein) is a fluorescent dye that has been widely used for staining various biological samples for fluorescent microscopy. It reacts with polysaccharides and peptides at ordinary conditions. The possible application and efficiency of DTAF for CLSM studies were examined in various aspects of cyanobacterial-plant interactions. Seedlings of Pisum sativum, Vigna rediata and Triticum aestivum were co-cultivated and stained with DTAF as a fluorochrome. Extracellular and intracellular interactions of cyanobacteria and the plant root surface were observed by CLSM. Results were compared with staining by other commonly used lectins. Advantages of the use of DTAF over other stains are its penetration into root tissues and binding with polysaccharides, mainly the cellulose. The staining was smooth, which clearly showed minute details on the cell of surface and root hairs with higher resolution. The emission wavelength for DTAF is 517 nm, which is highly advantageous as cyanobacteria have auto-fluorescence at 665 nm, and both can be simultaneously used in CLSM by visualizing in different channels. This worked efficiently with all three plants used and with filamentous and unicellular cyanobacterial strains. Cyanobacterial presence was not only clearly observed on the root surface, but also inside the root tissue and epidermal cells. The easy protocol and absence of tissue processing make DTAF a useful probe for studies of cyanobacterial associations with plant roots by CLSM. [ABSTRACT FROM AUTHOR]
Copyright of Journal of Industrial Microbiology & Biotechnology is the property of Oxford University Press / USA and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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  Data: DTAF: an efficient probe to study cyanobacterial-plant interaction using confocal laser scanning microscopy (CLSM).
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  Data: <searchLink fieldCode="AR" term="%22Ahmed%2C+Mehboob%22">Ahmed, Mehboob</searchLink><br /><searchLink fieldCode="AR" term="%22Stal%2C+Lucas%22">Stal, Lucas</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Hasnain%2C+Shahida%22">Hasnain, Shahida</searchLink><relatesTo>2</relatesTo><i> genetic@brain.net.pk</i>
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  Data: <searchLink fieldCode="JN" term="%22Journal+of+Industrial+Microbiology+%26+Biotechnology%22">Journal of Industrial Microbiology & Biotechnology</searchLink>. Jan2011, Vol. 38 Issue 1, p249-255. 7p. 3 Diagrams, 1 Chart.
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  Data: <searchLink fieldCode="DE" term="%22Cyanobacteria%22">Cyanobacteria</searchLink><br /><searchLink fieldCode="DE" term="%22Plant+cell+interaction%22">Plant cell interaction</searchLink><br /><searchLink fieldCode="DE" term="%22Plant+roots%22">Plant roots</searchLink><br /><searchLink fieldCode="DE" term="%22Microscopical+technique%22">Microscopical technique</searchLink><br /><searchLink fieldCode="DE" term="%22Fluorescence+microscopy%22">Fluorescence microscopy</searchLink><br /><searchLink fieldCode="DE" term="%22Tissue+engineering%22">Tissue engineering</searchLink><br /><searchLink fieldCode="DE" term="%22Molecular+biology%22">Molecular biology</searchLink><br /><searchLink fieldCode="DE" term="%22Polysaccharides%22">Polysaccharides</searchLink>
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  Label: Abstract
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  Data: variety of microscopic techniques have been utilized to study cyanobacterial associations with plant roots, but confocal laser scanning microscopy (CLSM) is the least used due to the unavailability of a suitable fluorescent dye. Commonly used lectins have problems with their binding ability with root cells and their visualization under CLSM. DTAF (5-(4,6-dichlorotriazinyl) aminofluorescein) is a fluorescent dye that has been widely used for staining various biological samples for fluorescent microscopy. It reacts with polysaccharides and peptides at ordinary conditions. The possible application and efficiency of DTAF for CLSM studies were examined in various aspects of cyanobacterial-plant interactions. Seedlings of Pisum sativum, Vigna rediata and Triticum aestivum were co-cultivated and stained with DTAF as a fluorochrome. Extracellular and intracellular interactions of cyanobacteria and the plant root surface were observed by CLSM. Results were compared with staining by other commonly used lectins. Advantages of the use of DTAF over other stains are its penetration into root tissues and binding with polysaccharides, mainly the cellulose. The staining was smooth, which clearly showed minute details on the cell of surface and root hairs with higher resolution. The emission wavelength for DTAF is 517 nm, which is highly advantageous as cyanobacteria have auto-fluorescence at 665 nm, and both can be simultaneously used in CLSM by visualizing in different channels. This worked efficiently with all three plants used and with filamentous and unicellular cyanobacterial strains. Cyanobacterial presence was not only clearly observed on the root surface, but also inside the root tissue and epidermal cells. The easy protocol and absence of tissue processing make DTAF a useful probe for studies of cyanobacterial associations with plant roots by CLSM. [ABSTRACT FROM AUTHOR]
– Name: AbstractSuppliedCopyright
  Label:
  Group: Ab
  Data: <i>Copyright of Journal of Industrial Microbiology & Biotechnology is the property of Oxford University Press / USA and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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        Value: 10.1007/s10295-010-0820-8
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        Text: English
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      – SubjectFull: Cyanobacteria
        Type: general
      – SubjectFull: Plant cell interaction
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      – SubjectFull: Plant roots
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      – SubjectFull: Microscopical technique
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      – SubjectFull: Fluorescence microscopy
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      – SubjectFull: Tissue engineering
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      – SubjectFull: Molecular biology
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      – SubjectFull: Polysaccharides
        Type: general
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      – TitleFull: DTAF: an efficient probe to study cyanobacterial-plant interaction using confocal laser scanning microscopy (CLSM).
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            NameFull: Ahmed, Mehboob
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              Text: Jan2011
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