Abstract
Nicotiana alata anthers cultured on different modified media based on MS, MT and N were used to obtain haploid plants through direct and indirect ways. The haploid plants resulting on MS medium ranged from 52% - 80%, on MT medium ranged from 32% - 52% and on N medium ranged from 28% - 44%. Accordingly, the best medium used for haploid induction was MS supplemented with 0.2 mgl-l NAA + 0.5 mgl-l KIN. On the other hand, MS medium supplemented with 0.4 mgl-l NAA + 0.5 mgl-l KIN or 1.0 mgl-l BAP + 0.5 mgl-l NAA were the best mediums for callus induction and plant regeneration, respectively. Morphologically, the leaf size, stem highest and diameter, flower size and diameter, anther length and number were about 67% of the diploid plants growth. Irradiated anthers with doses of 0, 2.5, 5, 7.5, 10, 15, 20 and 25 Gy caused reducing the number of haploid plants with increasing gamma radiation dose. For the haploid plants irradiated with same doses, the mortality percentage of bud survival was increasing with increasing gamma radiation dose. The irradiated callus with doses of 0, 5, 10, 15 and 20 Gy was affected negatively on growth rate and morphology. Proline content in irradiated plantlets increased with increasing gamma radiation dose. As well, total soluble protein content was increased with gamma irradiation up to 10 Gy. However, the higher doses caused a severe decrease of total soluble proteins. The production of proline and total soluble proteins in haploid plants were 48.6% and 69.5%, respectively comparing with diploid plants.
Highlights
Nicotiana alata is an ornamental plant, member from Solanaceae family and the diploid cells contain 18 chromosomes
The results showed that higher haploid plant numbers were on MS medium, whereas the haploid plants produced a range from 13 to 20 anther cultures (52% - 80%), MT medium with a range of 8 to 13 anther cultures (32% - 52%) and the last performant was N medium with a of range 7-11 anther culture
It has been reported that flower buds which have developed after plant regeneration give rise to pollen grains of flower androgenetic ability (Maheshvari et al, 1980; Shtereva et al, 1998; Zhou, 1996)
Summary
Nicotiana alata is an ornamental plant, member from Solanaceae family and the diploid cells contain 18 chromosomes. Haploid plants can be obtained by isolation of anthers in vitro in two ways: directly, with formation of embryo from the pollen grains (microspore), and indirectly, with callus development and formation of haploid embryoids or adventive buds (Pierik, 1979). Gamma rays are often used on plants in developing varieties that are agriculturally an economically important and have high productivity potential (Jain et al., 1998). They are useful for mutations in breeding programs and in vitro mutagenesis in order to develop required features of plants and increase the genetic variability. The protocol of mutation induction at haploid level has not been established
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