Fluorescent labeling of the root cap cells with the bioactive NBD-S chemical probe based on the cellulose biosynthesis inhibition herbicides

Shigeru Tamogami; Ganesh K Agrawal; Randeep Rakwal

doi:10.1016/j.bbrep.2021.101063

Fluorescent labeling of the root cap cells with the bioactive NBD-S chemical probe based on the cellulose biosynthesis inhibition herbicides

Biochem Biophys Rep. 2021 Jun 29:27:101063. doi: 10.1016/j.bbrep.2021.101063. eCollection 2021 Sep.

Authors

Shigeru Tamogami¹, Ganesh K Agrawal^{2

3}, Randeep Rakwal^{2

3

4}

Affiliations

¹ Laboratory of Biologically Active Compounds, Department of Biological Production, Akita Prefectural University, Akita 010-0195, Japan.
² Research Laboratory for Biotechnology and Biochemistry (RLABB), Kathmandu, Nepal.
³ GRADE Academy Private Limited, Adarsh Nagar-13, Birgunj, Nepal.
⁴ Faculty of Health and Sport Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, Japan.

Abstract

Development of the methods to examine the molecular targets of biologically active compounds is one of the most important subjects in experimental biology/biochemistry. To evaluate the usability of the (7-nitro-2,1,3-benzoxadiazole)-thioether (NBD-S) probe for this purpose, bioactive chemical probe (1) as the cellulose biosynthesis (CB) inhibitor was synthesized and tested. As a result, a variety of fluorescently-labeled particles and organelles were found in the columella root cap cells of radish plants. Of note, well-defined cellular organelles were clearly recognized in the detaching root cap cells (border-like cells). These results imply that the bioactive NBD-S chemical probe could be a valuable direct-labeling reagent. Analysis of these fluorescent substances would be helpful in providing new information on defined molecular targets and events.

Keywords: CB, cellulose biosynthesis; CW, cell wall; Chemical probe; Cys, cysteine; DIEA, N,N-diisopropylethylamine; DMSO, N,N-dimethylsulfoxide; Fluorescence; HEPES, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; Indaziflam; Lys, lysine; NBD; NBD, nitrobenzoxadaizole; NBD-Cl, 4-chloro-7-nitro-2,1,3-benzoxadiazole; NBD-N, (7-nitro-2,1,3-benzoxadiazole)-amine; NBD-O, (7-nitro-2,1,3-benzoxadiazole)-ether; NBD-S, (7-nitro-2,1,3-benzoxadiazole)-thioether; Root cap; Triaziflam.