Elimination of in vitro bacterial contaminants in shoot cultures of ‘MRS 2/5’ plum hybrid by the use of Melia azedarach extracts

Abstract

The antimicrobial activity of leaf and callus extracts of Melia azedarach was tested on in vitro shoot cultures of the peach rootstoch ‘MRS 2/5’ (Prunus cerasifera × Prunus spinosa) that were heavily contaminated with Sphingomonas paucimobilis (Sp) and Bacillus circulans (Bc). The extracts were filter-sterilised and added at 0%, 1%, 5%, 10% and 20% to a modified Murashige and Skoog proliferation medium previously autoclave-sterilised. Up to about 17% shoots died with 10–20% extract, except for Sp-contaminated shoots, whose survival was reduced to 50% after treatment with 20% extract. No shoots died with 1% to 5% supplement. The undiluted leaf extract showed bactericidal activity on plated Sp and Bc isolates. The homogenates of shoots randomly collected from treated cultures were processed for bacterial colony counting. Thus the 10% supplement was the best treatment for ridding Bc-contaminated cultures of bacteria (although 5% had a similar bactericidal effect), and allowing shoot growth and proliferation comparable to controls at the fifth subculture on a standard medium, while 20% extract was needed to eliminate Sp, and could induce higher growth and proliferation rates in surviving shoots than in untreated cultures. Callus extract was ineffective. The bactericidal activity of the leaf extract seemed attributable to a synergistic effect of azadirachtin with other unidentified compounds present in the extract.