Abstract
Lilium candidum L. is a species which grows in the South West Anatolia region of Turkey. It is a bulbous plant with beautifully scented flowers and is used in the floral industry. The bulbs are produced by using traditional propagation and in vitrotechniques. Micropropagation is a rapid propagation technique, but the greatest problem is contamination with fungi and bacteria. Antibiotic and fungicide treatments were done after sterilization for micropropagation of L. candidum. Fungal contaminants formed during the culture were determined. Bulb scales were used as explants (5 - 10 mm width) and were cultured in photoperiodic conditions (16 h light, 8 h dark) or complete darkness. Bulb scales rinsed in water were surface sterilized, then solutions containing chemotherapeutic substances (Benomyl, Nystatin, Streptomycin, Penicillin) in different combinations were applied for 30 min and subsequently were cultured in MS medium with supplement 0.1 mg dm-3 NAA + 0.01 mg dm-3 BA. During the experiment, fungal contaminants were observed in full treatments. Determined contaminants were identified according to their morphological and cultural characteristics by cultivation and were comprised of: Fusarium, Penicillium, Alternaria, Rhizopus, Cylindrocarpon and Aspergillusspecies. The most effective treatment against fungal contaminations was achieved by utilizing a Benomyl (100 mg dm-3) + Nystatin (100 mg dm-3) treatment combination. Key words: Contaminating microorganisms, Lilium candidum, bulb scale, in vitroculture, treatment, antibiotic, fungicide.
Highlights
Lilium are bulbous plants with many different species that have a special importance as cut flowers among ornamental plants (Uzun, 1984) and are economically important due to their attractive flowers (Nhut, 1998)
Bulb scales rinsed in water were surface sterilized, solutions containing chemotherapeutic substances (Benomyl, Nystatin, Streptomycin, Penicillin) in different combinations were applied for 30 min and subsequently were cultured in Murashige and Skoog (MS) medium with supplement 0.1 mg dm-3 naphthalene acetic acid (NAA) + 0.01 mg dm-3 BA
Medium was prepared and used in the following mixture: 30 g dm-3 sugar, 8 g dm-3 agar was added to MS medium (MurashigeSkoog, 1962) containing 0.1 mg dm-3 NAA + 0.01 mg dm-3 BA plant growth regulator were prepared according to Franklin and Dixon (1994) and pH adjusted to 5.7 before autoclaving for 15 min
Summary
Lilium are bulbous plants with many different species that have a special importance as cut flowers among ornamental plants (Uzun, 1984) and are economically important due to their attractive flowers (Nhut, 1998). Even though it is possible to produce a large number of plants by micropropagation, the greatest problem in this technique is contamination. Shields et al (1984) analysed the effects of a number of fungicides against in vitro fungal contaminants and their toxicity in tobacco cultures. They recommend 2 fungicides, carbendazim and fenbendazole (30 μg cm-3). It has been determined that 10 μg cm-3 Rifampin + 1 g dm-3 Benomyl has been an important effective combination in the control of fungal and bacterial contaminants in Camellia cultures (Haldeman et al, 1987). The identification of fungal contamination emerging during micropropagation of Lilium candidum and the effects of antibiotic and fungicide treatments after sterilization was determined
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