Abstract
Two different androgenesis pathways, including shed-microspore culture (SMC) and anther culture (AC), were developed for haploid induction in four mutagenized M1 rice cultivars. The effect of gum arabic (GA), plant growth regulators, cold shock, culture medium, carbohydrate source, and genotype were evaluated in both induction and regeneration phases in both methods. The SMC method is reported for the first time in rice. In AC pathway, the highest number of regenerated embryos was obtained in MS medium supplemented with 2 mg/L Kinetin + 1 mg/L NAA + 10 mg/L GA. In SMC pathway, the highest number of regenerated embryos was obtained with 3% maltose and 10 mg/L GA in the upper medium and the activated charcoal in the lower medium. The average number of regenerated embryos in SMC was more than the AC pathway. Haploid plants were confirmed by chromosome counting and flow cytometry analysis. The highest percent of haploid plantlets was obtained in SMC of Hashemi cultivar. These protocols can be utilized to develop successful haploids induction in rice. Completely homozygous doubled haploid lines, subsequent to chromosome doubling, could be useful in mutation and selection breeding programs.
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Data Availability
The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- AC:
-
Anther culture
- AGP:
-
Arabinogalactan protein
- ANOVA:
-
Analysis of variance
- CRD:
-
Completely randomized design
- DH:
-
Doubled haploid
- GA:
-
Gum arabic
- HI:
-
Haploid induction
- IAA:
-
Indole acetic acid
- IMC:
-
Isolated microspore culture
- Kin:
-
Kinetin
- MS:
-
Murashige and Skoog medium
- NAA:
-
Naphthalene acetic acid
- PGR:
-
Plant growth regulator
- QTL:
-
Quantitative trait loci
- SMC:
-
Shed-microspore culture
- Zea:
-
Zeatin
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
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Acknowledgements
This work has been supported by the International Atomic Energy Agency (IAEA) CRP Code: D25005 and Contract Number: 20742. The authors are thankful to Eissa Zarifi for chromosome studies in Cytogenetic Lab. of National Gene Bank of Iran and Laura Morales for her kind assistance in the English Language editing of the manuscript.
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ST performed anther culture and shed-microspore culture experiments, MES (as the contract holder of IAEA) and AMAG (as the CRP project leader of IAEA) supervised the whole project and corrected the manuscript, MN drafted the whole body of manuscript and done all statistical analyses, BF supervised the PhD program of the first author and revised the manuscript for important intellectual content, NM and AK pour advised the PhD program of the first author and revised the manuscript for important intellectual content, and II contributed in anther culture experiments.
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Tajedini, S., Fakheri, B., Niazian, M. et al. Efficient Microspore Embryogenesis and Haploid Induction in Mutant Indica Rice (Oryza sativa L.) Cultivars. J Plant Growth Regul 42, 2345–2359 (2023). https://doi.org/10.1007/s00344-022-10709-y
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DOI: https://doi.org/10.1007/s00344-022-10709-y