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- W2606340917 abstract "A practical scheme for developing Fiji disease virus (FDV) - free sugarcane plants and a reliable diagnostic method to test for the presence of the virus in transported planting material is of key importance to the Australian quarantine and sugar industry. This project aimed at; (1) Developing in vitro techniques for regeneration of genetically stable sugarcane plants free of bacteria, fungi and FDV. These techniques included axillary bud-culture and embryogenie callus-culture; (2) Developing viral diagnostic assays to detect the presence of FDV in the in vitro regenerants. These diagnostic tests included an ELISA and a nucleic acid hybridization dot-blot assay. Axillary bud-cultures of field-grown FDV-infected sugarcane plants cv. NCo310 were routinely established. The optimum culture conditions included; decapitation of the donor plants and stripping off all other leaves. The naked stems were sprayed with the fungicide Beniate (0.6 g l-1) and the insecticide Metasystox (1 ml l-1) 3 to 5 days prior to harvesting. Stem pieces, 3-5 cm long each containing a single bud, were thoroughly washed and given a hot water treatment at 50oC for 2 hours. Excised buds were then surface sterilized by washing in 70% ethanol for 1 min., rinsed in 4% chlorine (NaOCL) for 5 min. and washed 3 times in sterile distilled water. Buds were then separately heat treated at 61oC for 30 min. in a half strength -MS medium, pH 5.3, containing the anticontaminants Benlate (100 ppm benomyl), Nystatin (25 ppm), and Carbenicillin (100 ppm). The heat treated buds were cultured, until 8 to 15 cm long, on an agar-MS medium containing NAA (2 mg l-1), malt extract (500 mg l-1) and the antioxidant diethyldithiocarbamic acid (250 mg l-1) to control browning of the culture media. Roots were then developed on shoot-sprouted buds by transfer to a tube containing sterile dis tilled water. These procedures for bud-culture resulted in a decontamination rate of 90% from which 75% were regenerated into plants phenotypically identical to the mother cultivar. n" @default.
- W2606340917 created "2017-04-28" @default.
- W2606340917 creator A5078435427 @default.
- W2606340917 date "2016-12-05" @default.
- W2606340917 modified "2023-09-27" @default.
- W2606340917 title "The establishment and use of in vitro tissue culture techniques for the production of pathogen and Fiji disease virus-free clones of sugarcane" @default.
- W2606340917 doi "https://doi.org/10.14264/uql.2016.1104" @default.
- W2606340917 hasPublicationYear "2016" @default.
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