A Fumigation-Based Surface Sterilization Approach for Plant Tissue Culture

Abstract

Plant tissue culture has led to breakthroughs in understanding and applying the fundamental knowledge towards harnessing more from plants. Microbial contamination is one of the serious problems limiting the successful extrapolation of plant tissue culture practices. Sources of in vitro contamination include culture containers, media, explants, equipment, the environment of the culture room and transfer area, and operating personnel. The successful initiation of in vitro culture mostly depends on surface sterilization of explants because this is the primary source. Usually, surface sterilization is done using chemicals, or toxic nanomaterials, this is the first time such an approach has been demonstrated. Numerous surface microflora attached to plant surfaces grow faster than the cultured explants and release phytotoxic substances into the culture media, hindering positive outcomes. In the current work, for the first time, the applicability of turmeric and benzoin resin-based fumigation of explants is demonstrated. The results showed that fumigation methods for surface sterilization were promising and could lead to fifty and even 100% contamination-free plant tissue culture. Nanoparticulate carbon was identified in the turmeric and benzoin smoke and coined the key player in the surface sterilization effect. These studies open a whole new avenue for the use of fumigation-based methods for riddance of microbial contamination.

Keywords: antimicrobials; carbon nanodots; fumigation; plant tissue culture; surface sterilization.

Figure 1

Schematic of the experimental methodology. BR—Benzoin resin

Figure 2

Graph showing results of the surface sterilization effect following fumigation.

Figure 3

Photographs showing the results of the fumigation effect on explants versus the control. (A) Explant 1, (B) Explant 2, and (C) Explant 3; (a) Control (b) Tur (c) BR. Details are given in above text.

Figure 4

(A) UV–Vis curve of Tur and BR smoke. (B) FTIR of Tur and BR smoke.

Figure 5

TEM micrographs (A) Turmeric and (B) BR (Benzoin resin) smudges.

Figure 6

Cartoon speculating the mechanism of fumigation-based surface sterilization.