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Real-time visualization of cellulase activity by microorganisms on surface
Year:
2020
Authors :
Bucki, Patricia
;
.
Kleiman, Maya
;
.
Kumari, Palavi
;
.
Sayas, Tali
;
.
Volume :
21
Co-Authors:

Kumari, P. - Institute of Plant Sciences, Agricultural Research Organization (Volcani Center), Rishon Lezion, 7505101, Israel

Sayas, T - Institute of Plant Sciences, Agricultural Research Organization (Volcani Center), Rishon Lezion, 7505101, Israel

Bucki, P. - Institute of Plant Protection, Agricultural Research Organization (Volcani Center), Rishon Lezion, 7505101, Israel

Brown-Miyara, S. - Institute of Plant Protection, Agricultural Research Organization (Volcani Center), Rishon Lezion, 7505101, Israel

Kleiman, M. - aInstitute of Plant Sciences, Agricultural Research Organization (Volcani Center), Rishon Lezion, 7505101, Israel; cAgro-NanoTechnology and Advanced Materials Center, Agricultural Research Organization (Volcani Center), Rishon Lezion, 7505101, Israel

Facilitators :
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Total pages:
1
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Abstract:

A variety of methods to detect cellulase secretion by microorganisms has been developed over the years, none of which enables the real-time visualization of cellulase activity on a surface. This visualization is critical to study the interaction between soil-borne cellulase-secreting microorganisms and the surface of plant roots and specifically, the effect of surface features on this interaction. Here, we modified the known carboxymethyl cellulase (CMC) hydrolysis visualization method to enable the real-time tracking of cellulase activity of microorganisms on a surface. A surface was formed using pure CMC with acridine orange dye incorporated in it. The dye disassociated from the film when hydrolysis occurred, forming a halo surrounding the point of hydrolysis. This enabled real-time visualization, since the common need for post hydrolysis dyeing was negated. Using root-knot nematode (RKN) as a model organism that penetrates plant roots, we showed that it was possible to follow microorganism cellulase secretion on the surface. Furthermore, the addition of natural additives was also shown to be an option and resulted in an increased RKN response. This method will be implemented in the future, investigating different microorganisms on a root surface microstructure replica, which can open a new avenue of research in the field of plant root–microorganism interactions.

Note:
Related Files :
Hydrolysis
microorganisms
plant root
Real-time
Visualization
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More details
DOI :
10.3390/ijms21186593
Article number:
0
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
50314
Last updated date:
02/03/2022 17:27
Creation date:
21/09/2020 13:32
Scientific Publication
Real-time visualization of cellulase activity by microorganisms on surface
21

Kumari, P. - Institute of Plant Sciences, Agricultural Research Organization (Volcani Center), Rishon Lezion, 7505101, Israel

Sayas, T - Institute of Plant Sciences, Agricultural Research Organization (Volcani Center), Rishon Lezion, 7505101, Israel

Bucki, P. - Institute of Plant Protection, Agricultural Research Organization (Volcani Center), Rishon Lezion, 7505101, Israel

Brown-Miyara, S. - Institute of Plant Protection, Agricultural Research Organization (Volcani Center), Rishon Lezion, 7505101, Israel

Kleiman, M. - aInstitute of Plant Sciences, Agricultural Research Organization (Volcani Center), Rishon Lezion, 7505101, Israel; cAgro-NanoTechnology and Advanced Materials Center, Agricultural Research Organization (Volcani Center), Rishon Lezion, 7505101, Israel

Real-time visualization of cellulase activity by microorganisms on surface

A variety of methods to detect cellulase secretion by microorganisms has been developed over the years, none of which enables the real-time visualization of cellulase activity on a surface. This visualization is critical to study the interaction between soil-borne cellulase-secreting microorganisms and the surface of plant roots and specifically, the effect of surface features on this interaction. Here, we modified the known carboxymethyl cellulase (CMC) hydrolysis visualization method to enable the real-time tracking of cellulase activity of microorganisms on a surface. A surface was formed using pure CMC with acridine orange dye incorporated in it. The dye disassociated from the film when hydrolysis occurred, forming a halo surrounding the point of hydrolysis. This enabled real-time visualization, since the common need for post hydrolysis dyeing was negated. Using root-knot nematode (RKN) as a model organism that penetrates plant roots, we showed that it was possible to follow microorganism cellulase secretion on the surface. Furthermore, the addition of natural additives was also shown to be an option and resulted in an increased RKN response. This method will be implemented in the future, investigating different microorganisms on a root surface microstructure replica, which can open a new avenue of research in the field of plant root–microorganism interactions.

Scientific Publication
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