A biomimetic platform for studying root-environment interaction

Kumari, Pallavi; Ginzburg, Neta; Sayas, Tali; Saphier, Sigal; Bucki, Patricia; Miyara, Sigal Brown; Caldwell, Denise; Iyer‐Pascuzzi, Anjali S.; Kleiman, Maya

doi:10.1007/s11104-019-04390-6

Cited by 8 publications

(9 citation statements)

References 53 publications

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“…cellulase secretion, as it occurs. This work is a continuation of our previous work 27 where we built a synthetic replication of the root surface microstructure to better understand how microtopography influences microorganism location on the root surface. This system was built as a simplified model of a root-microorganism interaction, focusing on the structural/physical aspect of this interaction.…”

Section: Resultsmentioning

confidence: 98%

“…Additionally, synthetic systems were proven efficient in depicting the structural preference in leaves 25,26 . We recently introduced a synthetic method to study structural preferences in soil borne pathogens 27 . Our method involves the replication of the root surface microstructure in a synthetic system in order to study the spatial preferences in root pathogens localization on the root surface.…”

Section: Introductionmentioning

confidence: 99%

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Developing a method for real-time visualization of cellulase activity

Kumari

Sayas

Bucki

et al. 2020

Preprint

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AbstractStudying the interactions between microorganisms and plant roots is crucial for understanding a variety of phenomena concerning crop yield and health. The role of root surface properties in these interactions, is rarely addressed. To this end, we previously built a synthetic system, from the inert polymer polydimethyl siloxane (PDMS), mimicking the root surface microstructure, using a replication technique. This replica enables the study of isolated effects of surface structure on microorganism-plant interactions. Since the root surface is composed mostly of cellulose, using cellulose-like materials as our replica, instead of PDMS, is the next logical step. This will enable following the hydrolysis of such surfaces as a result of microorganisms secreting Plant Cell Wall Degrading Enzymes (PCWDE), and in particular, cellulase. Visualization of such hydrolysis in a synthetic system can assist in studying the localization and activity of microorganisms and how they correlate with surface microtopography, separately from chemical plant signals.In this work, we modified the known carboxymethyl cellulase (CMC) hydrolysis visualization method to enable real-time tracking of cellulase activity of microorganisms on the surface. Surface was formed from pure CMC, rather than CMC incorporated in agar as is often done, and by that, eliminating diffusion issues. Acridine orange dye, which is compatible, at low concentrations, with microorganisms, as opposed to other routinely used dyes, was incorporated into the film. 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 the plant root, we showed it was possible to follow microorganism cellulase secretion on the surface in the form of CMC film hydrolysis. Furthermore, the addition of natural additives, in the form of root extract was also shown to be an option and resulted in an increased RKN response. We tested our newly developed method by changing temperature and pH conditions and by characterization of the hydrolyzed surface using both Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM).This method will be implemented in the future on a root surface microstructure replica. We believe the combination of this new method with our previously developed root surface microstructure replication technique can open a new avenue of research in the field of plant root-microorganism interactions.

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Section: Resultsmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Developing a method for real-time visualization of cellulase activity

Kumari

Sayas

Bucki

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…This work is a continuation of our previous work [35], where a synthetic replication of the root surface microstructure was built to better understand how microtopography influences microorganism location on the root surface. This system was built as a simplified model of a root-microorganism interaction, focusing on the structural/physical aspect of this interaction.…”

Section: Discussionmentioning

confidence: 97%

“…Additionally, synthetic systems were proven efficient in depicting the structural preference in leaves [33,34]. We recently introduced a synthetic method to study structural preferences in soil-borne pathogens [35]. This method involves the replication of the root surface microstructure in a synthetic system in order to study the spatial preferences in root pathogens localization on the root surface.…”

Section: Introductionmentioning

confidence: 99%

Real-Time Visualization of Cellulase Activity by Microorganisms on Surface

Kumari

Sayas

Bucki

et al. 2020

IJMS

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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.

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“…Biomimetic replicated surface is a powerful tool for understanding plant–environment interactions 14 , 15 . For example, replicating the hierarchical microstructures of lotus leaves have enabled the fabrication of superhydrophobic surfaces for practical applications, including anticorrosive, antifogging, antifouling, and oil/water separation purposes 16 – 18 .…”

Section: Introductionmentioning

confidence: 99%

Micro-replication platform for studying the structural effect of seed surfaces on wetting properties

Shin

Choi

Baasanmunkh

et al. 2022

Sci Rep

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Biological surfaces in plants are critical for controlling essential functions such as wettability, adhesion, and light management, which are linked to their adaptation, survival, and reproduction. Biomimetically patterned surfaces replicating the microstructures of plant surfaces have become an emerging tool for understanding plant–environment interactions. In this study, we developed a two-step micro-replication platform to mimic the microstructure of seed surfaces and demonstrated that this initial platform can be used to study seed surface–environment interactions. The two-step process involved the extraction of a simplified seed surface model from real seeds and micro-replication of the simplified seed surface model using nanoimprint lithography. Using Allium seeds collected from Mongolia and Central Asia as the model system, we studied the wettability of biological and synthetic seed surfaces. We could independently control the material properties of a synthetic seed surface while maintaining the microstructures and, thereby, provide clear evidence that Allium seed surfaces were highly wettable owing to the high surface energy in the epidermal material rather than a microstructural effect. We expect that this platform can facilitate study of the independent effect of microstructure on the interaction of seed surfaces with their surroundings and contribute to research on the evolution of plant–environment interactions.

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A biomimetic platform for studying root-environment interaction

Cited by 8 publications

References 53 publications

Developing a method for real-time visualization of cellulase activity

Developing a method for real-time visualization of cellulase activity

Real-Time Visualization of Cellulase Activity by Microorganisms on Surface

Micro-replication platform for studying the structural effect of seed surfaces on wetting properties

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