Optimization of growth regulators and silver nitrate for micropropagation of <Emphasis Type="Italic">Dianthus caryophyllus</Emphasis> L. with the aid of a response surface experimental design |
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Authors: | Federico A Gutiérrez-Miceli Lourdes Arias Nicolás Juarez-Rodríguez Miguel Abud-Archila Aldo Amaro-Reyes Luc Dendooven |
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Institution: | 1.Instituto Tecnológico de Tuxtla Gutiérrez,Tuxtla Gutiérrez,México;2.Laboratorio de Cultivo de Tejidos Vegetales,Instituto para el Fomento de la Agricultura Tropical,Tuxtla-Gutiérrez,México;3.Laboratory of Soil Ecology, Dept. Biotechnology and Bioengineering,Cinvestav,México DF,México |
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Abstract: | This paper reports on the optimum concentrations of naphthalene acetic acid (NAA) and 6-benzyladenine (BA) to stimulate callus
growth and NAA; kinetin and silver nitrate (AgNO3) for callus redifferentiation in Dianthus caryophyllus L. Meristems were excised and placed in MS medium with 30 g l−1 sucrose and 9.0 μM 2,4-d. Callus clusters were transferred to MS medium containing NAA (0, 1.7, 3.3, and 5.0 μM) and BA (0, 1.7, 3.3, and 5.0 μM)
for proliferation and to MS medium with 30 g l−1 sucrose, 2.5 g l−1 phytagel, kinetin (0, 33, and 66 μM); NAA (0, 7.95, and 15.9 μM) and AgNO3 (0, 23.54 and 47.08 μM) for shoot and root induction. Treatments were applied according to a Box–Behnken design. After callus
growth and redifferentiation, plants were incubated in the greenhouse at 18 ± 2°C for 4 wk and at 20–26°C for 4 wk. Finally,
plants were changed to near-commercial greenhouse conditions with different day (30–35°C) and night (16–24°C) temperatures.
Results showed better callus growth at higher NAA concentrations. A maximum callus weight was found with 5.0 μM NAA but without
BA. A maximum of 78% calluses with shoots was obtained with 15.9 μM NAA, 47.08 μM AgNO3, and 0.74 μM kinetin and 58% with roots with 15.7 μM NAA and 47.08 μM AgNO3, but without kinetin. The shoots obtained showed little hyperhydricity. Vigorous plants were obtained after gradual acclimatization
with an 80% survival rate under nursery conditions. |
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