In vitro effect of biogenic silver nanoparticles on sterilisation of tobacco leaf explants and for higher yield of protoplasts.

Abstract

Isolation of protoplasts from leaves is useful in plant research. The standard reference methods for isolation of protoplasts are tedious, cause cell damage, are low-yield, time consuming and prone to microbial contamination. To overcome this problem, the authors used silver nanoparticles (AgNPs) for the control of microbial contamination and with low concentration of enzyme mixture for rapid release of protoplasts. The leaf explants were sterilised with 95% ethanol for 30 s followed by biologically synthesised AgNPs (1, 5, 10 and 15 mg/l) for 10 to 20 min. The authors found that 10 mg/l concentration of AgNPs treatment on explants showed remarkable inhibitory effect on microbial contamination with high level of tolerance. Moreover, during protoplasts isolation, the addition of 10 mg/l AgNPs in leaf incubation buffer yielded 34% viable protoplasts in 3 h. This is the first report of AgNPs synthesis from waste plant medium, which was applied for the sterilisation of explants and rapid isolation of protoplasts.