Breeding using the double haploid approach has been used in accelerating the rice breeding process. However, the main obstacle in the application of this technique in rice is the low level of callus formation and their regeneration into haploid plants. Based on these issues, this research is aimed at developing the best method for obtaining double haploid plants through anther culture using three kinds of aromatic rice of Indonesian local varieties. Anthers of Pendok, Genjah Arum, and Merah Wangi varieties were cultured in callus induction media which were either supplemented with different types of carbon sources, or auxins and ethylene inhibitors. However, the application of different sugar types did not significantly affect callus formation of Pendok and Genjah Arum. However, auxin treatment showed NAA to be more efficient for the callus induction of Pendok, while 2,4-D had no effect on callus induction in all tested varieties. The addition of ethylene inhibitor successfully induced callus formation of Merah Wangi varieties, while the rates of Pendok and Genjah Arum were not significantly affected. Callus of three plant lines were then regenerated into plantlet in regeneration media. Of the three callus varieties used, only Pendok could regenerate into plantlets and grow into adult haploid plants.
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This research was supported by Penelitian Dasar Perguruan Tinggi (PDUPT) Grant 0551/UN25.3.1/LT/2017 from the Ministry of Research, Technology and Higher Education of the Republic of Indonesia.
Conflict of interest
The authors declare that they have no competing interest.
Pendok, Genjah Arum, and Merah are aromatic rice varieties originated from East Java, Indonesia. We subjected these three rice varieties to anther culture optimization using culture media supplemented with either of ethylene inhibitor, auxin, and sugar. Among the three varieties, we found that Pendok showed the highest rate in callus formation and plant regeneration, whereas Merah Wangi showed the lack of response to those two parameters. Furthermore, maltose is the best sugar for callus induction in Pendok, and 2,4-D is the suitable auxin for callus induction. Finally, we found that ethylene inhibitor was only effective for callus induction rate of recalcitrant-type Merah Wangi.
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Arisandi, D.P., Paradisa, F.V., Sugiharto, B. et al. Effect of ethylene inhibitor, type of auxin, and type of sugar on anther culture of local East Java aromatic rice varieties. J. Crop Sci. Biotechnol. 23, 367–373 (2020). https://doi.org/10.1007/s12892-020-00045-6
- Aromatic rice
- Anther culture optimization
- Double haploid