Articles
BIOTECHNOLOGY OF ANNUAL FLOWER PLANTS: MICROPROPAGATION OF RUDBECKIA SP.
Article number
725_77
Pages
527 – 548
Language
English
Abstract
We conducted experiments for developing an in vitro micropropagation protocol starting from meristems of Rudbeckia hirta L. seedlings.
We pre-soaked the seeds in sterile ion-exchanged water for 17 hours, and then achieved surface disinfection in two separate steps.
First, we used concentrated household sodium-hypochloride solution for 20 minutes and, also for 20 minutes, we applied hydrogen peroxide of 10%, which was followed by washing with sterile ion-exchanged water three times.
For the propagation of seedling meristems, the combination of half-strength solid Murashige and Skoog (1962) culture medium containing 10 mg/L of kinetin and 2 mg/L of kinetin + 0.1 mg/L of 2iP proved to be the most suitable.
The average number of shoot-buds developed from the seedling axillary meristem in the best culture media varied between 5 and 17. Without separating them, we inoculated the shoot-bud clusters on MS culture medium containing 2 mg/L of IAA. After four weeks of incubation we obtained elongated shoots which we separated and inoculated into a new culture medium and we obtained elongated roots.
The rooted plants were gradually acclimatised in the cultivation room, potted and carried to a greenhouse, and then planted in open field for subsequent observation.
By adopting this method, our laboratory started the micropropagation of the superior and/or elite genotypes of the Rudbeckia hirta L. being of special value in respect of breeding.
We pre-soaked the seeds in sterile ion-exchanged water for 17 hours, and then achieved surface disinfection in two separate steps.
First, we used concentrated household sodium-hypochloride solution for 20 minutes and, also for 20 minutes, we applied hydrogen peroxide of 10%, which was followed by washing with sterile ion-exchanged water three times.
For the propagation of seedling meristems, the combination of half-strength solid Murashige and Skoog (1962) culture medium containing 10 mg/L of kinetin and 2 mg/L of kinetin + 0.1 mg/L of 2iP proved to be the most suitable.
The average number of shoot-buds developed from the seedling axillary meristem in the best culture media varied between 5 and 17. Without separating them, we inoculated the shoot-bud clusters on MS culture medium containing 2 mg/L of IAA. After four weeks of incubation we obtained elongated shoots which we separated and inoculated into a new culture medium and we obtained elongated roots.
The rooted plants were gradually acclimatised in the cultivation room, potted and carried to a greenhouse, and then planted in open field for subsequent observation.
By adopting this method, our laboratory started the micropropagation of the superior and/or elite genotypes of the Rudbeckia hirta L. being of special value in respect of breeding.
Authors
P. Szarvas, A. Zsila-André, M.G. Fari, Z. Kovats
Keywords
in vitro, organogenesis, seedling parts, Marygold, Hungary
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