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Title: CONTROL OF IN VITRO CONTAMINATION OF EXPLANTS FROM GREENHOUSE AND FIELD GROWN TREES

Author
item Niedz, Randall
item Bausher, Michael

Submitted to: In Vitro Cellular and Developmental Biology - Plants
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/5/2002
Publication Date: 9/1/2002
Citation: Niedz,Randall P.,Bausher,Michael G.2002.Control of In vitro contamination of explants from greenhouse- and field-grown trees.In Vitro Cell Dev.Biol.Plant 38:468-471. Abstract

Interpretive Summary: Before plant cell, tissue, or organ cultures can be established for commercial or research purposes it is necessary to remove bacteria and fungi. Eliminating bacteria and fungi from plant tissue derived from greenhouse or field sources can be extremely difficult. Treatments that are often effective in removing contaminants are also toxic to the plant tissue. Or, contaminants may be effectively removed from the surface of the plant pieces, but internal contaminants (if present) will remain and will eventual grow and ruin the cultures. Plant Preservative Mixture (PPM) containing two isothiazolone biocides was added to tissue culture medium at high concentrations (up to 20 mls/L). PPM reduced contamination of greenhouse and field grown buds of citrus trees placed into tissue culture from 92% to as low as 37%. Sodium dichloroisocyanurate (NaDCC) a chlorine compound commonly used as a swimming pool disinfectant was also effective in removing surface contaminants from greenhouse and field grown buds. Only 4% of buds treated with 300 ppm NaDCC for 48 hours were contaminated after 45 days.

Technical Abstract: Controlling fungal and bacterial contamination of woody plant material can be extremely difficult. Isothiazolone biocides and sodium dichloroisocyanurate (NaDCC) have ben used singly and in combination to reduce microbial contamination to less than 5% in bud explants derived from field-grown citrus trees. Coating the explant with a slurry containing 20 mL.L-1 Plant Preservative Mixture (PPM), a mixture of tow isothiazolones, and culturing on medium containing 5 mL.L-1 PPM resulted in 63% clean explants compared to >90% contamination with standard disinfestation prodedures. Explants treated for 48 h with 100 or 300 ppm NaDCC resulted in 83% and 96% clean explants, respectively.