Hydrogen Peroxide as Chemical “sterilizer” for Lilium Micropropagation

NÉSTOR CURVETTO,, PABLO MARINANGELI, GABRIELA MOCKEL

Agricell Report

Año: 2007 vol. 48 p. 41 - 42

At Argentina´s Universidad Nacional del Sur and CERZOS, N. Curvetto, P. Marinangeli, and G. Mockel have compared tradicional micropropagation methods with a thecnique that uses inexpensive and easily available materials to disifest materials and culture media, and addition of hydrogen peroxide into the nutrient media to prevent growth of fungal and bacterial contaminants during micropropagation of Lilium longiflorum (Biocell 30 (3): 497-500, 2006). As a traditional method (control), the culture medium was sterilized in an autoclave. Scale segments were disinfested by treatment with ethanol 70% for one minute followed by immersion in sodium hypochlorite solution (1.6 grams active chloride per liter) containing two drops of Tween 20 per liter for 20 minutes, and washed three times with sterile distilled water, then sliced into explants. In the hydrogen peroxide technique, the culture medium was sterilized in a home pressure cooker. Scales were placed in sterile Petri dishes containing a 0.03% hydrogen peroxide solution then cut into approximately 2 mm transverse sections and placed on nutrient medium containing hydrogen peroxide at a concentrations of 0.005, 0.010, 0.015 or 0.020% to control contamination during in vitro establishment and subsequent culture. Curvetto et al. found that the level of culture contamination was within acceptable limits in all of the treatments, although it was higher in the hydrogen peroxide treatments (40%) as compared with the traditional protocols (20%). No significant differences in the number of bulblets per explant were observed and at the end of the multiplication phase, bulblets treated with 0.02% hydrogen peroxide showed a greater biomass as compared with other treatments. These bulblets also had a higher relative growth ratio with respect to the traditional method when cultured for an additional period of time and showed the highest average bulblet fresh weight. The authors conclude that their results “suggest that hydrogen peroxide can be used as a low cost alternative method to traditional micropropagation techniques by hobbysts or small lily producers to micropropagate L. longiflorum. Hydrogen peroxide up to 0.02% has no deleterious effects on bulblet differentiation, but it does have a stimulatory effect on bulblet growth. The higher bulblet mass obtained after 18 week cultivation period in the case of the 0.02% hydrogen peroxide treatment would eventually result in a better plantlet performance growth ex vitro.Lilium longiflorum (Biocell 30 (3): 497-500, 2006). As a traditional method (control), the culture medium was sterilized in an autoclave. Scale segments were disinfested by treatment with ethanol 70% for one minute followed by immersion in sodium hypochlorite solution (1.6 grams active chloride per liter) containing two drops of Tween 20 per liter for 20 minutes, and washed three times with sterile distilled water, then sliced into explants. In the hydrogen peroxide technique, the culture medium was sterilized in a home pressure cooker. Scales were placed in sterile Petri dishes containing a 0.03% hydrogen peroxide solution then cut into approximately 2 mm transverse sections and placed on nutrient medium containing hydrogen peroxide at a concentrations of 0.005, 0.010, 0.015 or 0.020% to control contamination during in vitro establishment and subsequent culture. Curvetto et al. found that the level of culture contamination was within acceptable limits in all of the treatments, although it was higher in the hydrogen peroxide treatments (40%) as compared with the traditional protocols (20%). No significant differences in the number of bulblets per explant were observed and at the end of the multiplication phase, bulblets treated with 0.02% hydrogen peroxide showed a greater biomass as compared with other treatments. These bulblets also had a higher relative growth ratio with respect to the traditional method when cultured for an additional period of time and showed the highest average bulblet fresh weight. The authors conclude that their results “suggest that hydrogen peroxide can be used as a low cost alternative method to traditional micropropagation techniques by hobbysts or small lily producers to micropropagate L. longiflorum. Hydrogen peroxide up to 0.02% has no deleterious effects on bulblet differentiation, but it does have a stimulatory effect on bulblet growth. The higher bulblet mass obtained after 18 week cultivation period in the case of the 0.02% hydrogen peroxide treatment would eventually result in a better plantlet performance growth ex vitro.(Biocell 30 (3): 497-500, 2006). As a traditional method (control), the culture medium was sterilized in an autoclave. Scale segments were disinfested by treatment with ethanol 70% for one minute followed by immersion in sodium hypochlorite solution (1.6 grams active chloride per liter) containing two drops of Tween 20 per liter for 20 minutes, and washed three times with sterile distilled water, then sliced into explants. In the hydrogen peroxide technique, the culture medium was sterilized in a home pressure cooker. Scales were placed in sterile Petri dishes containing a 0.03% hydrogen peroxide solution then cut into approximately 2 mm transverse sections and placed on nutrient medium containing hydrogen peroxide at a concentrations of 0.005, 0.010, 0.015 or 0.020% to control contamination during in vitro establishment and subsequent culture. Curvetto et al. found that the level of culture contamination was within acceptable limits in all of the treatments, although it was higher in the hydrogen peroxide treatments (40%) as compared with the traditional protocols (20%). No significant differences in the number of bulblets per explant were observed and at the end of the multiplication phase, bulblets treated with 0.02% hydrogen peroxide showed a greater biomass as compared with other treatments. These bulblets also had a higher relative growth ratio with respect to the traditional method when cultured for an additional period of time and showed the highest average bulblet fresh weight. The authors conclude that their results “suggest that hydrogen peroxide can be used as a low cost alternative method to traditional micropropagation techniques by hobbysts or small lily producers to micropropagate L. longiflorum. Hydrogen peroxide up to 0.02% has no deleterious effects on bulblet differentiation, but it does have a stimulatory effect on bulblet growth. The higher bulblet mass obtained after 18 week cultivation period in the case of the 0.02% hydrogen peroxide treatment would eventually result in a better plantlet performance growth ex vitro.et al. found that the level of culture contamination was within acceptable limits in all of the treatments, although it was higher in the hydrogen peroxide treatments (40%) as compared with the traditional protocols (20%). No significant differences in the number of bulblets per explant were observed and at the end of the multiplication phase, bulblets treated with 0.02% hydrogen peroxide showed a greater biomass as compared with other treatments. These bulblets also had a higher relative growth ratio with respect to the traditional method when cultured for an additional period of time and showed the highest average bulblet fresh weight. The authors conclude that their results “suggest that hydrogen peroxide can be used as a low cost alternative method to traditional micropropagation techniques by hobbysts or small lily producers to micropropagate L. longiflorum. Hydrogen peroxide up to 0.02% has no deleterious effects on bulblet differentiation, but it does have a stimulatory effect on bulblet growth. The higher bulblet mass obtained after 18 week cultivation period in the case of the 0.02% hydrogen peroxide treatment would eventually result in a better plantlet performance growth ex vitro.L. longiflorum. Hydrogen peroxide up to 0.02% has no deleterious effects on bulblet differentiation, but it does have a stimulatory effect on bulblet growth. The higher bulblet mass obtained after 18 week cultivation period in the case of the 0.02% hydrogen peroxide treatment would eventually result in a better plantlet performance growth ex vitro.ex vitro.