Abstract
This study investigated developmental changes in cold stressed microspores of Indica rice variety At 303. After 3 d at 10°C, approximately 76% of microspores were in the late uni-nucleate stage. Even after 5 d, 49% of viable cells remained in the late uni-nucleate stage without advancing to the bi-nucleate stage. In comparison, microspores undergoing normal gametogenesis in planta progressed rapidly from uni-nucleate to bi-nucleate stage eventually forming tri-nucleate pollen during this period. Thus, cold stress prevented normal microspore development and retained cells in the late uni-nucleate stage which is the most favorable stage for in vitro induction of sporophytic development in rice. Uni-nucleate microspores subjected to cold stress showed a characteristic pattern consisting of several minute vacuoles surrounding a centrally positioned nucleus, which can be interpreted as an early indicator of sporophytic determination in Indica rice microspores. During in vitro culture phase, freshly plated yellow anthers became brown. After 4 wk in culture, 51% of the anthers had discolored. Significantly, all yellow anthers contained only non-viable cells whereas 10% of the brown anthers had few viable cells. Some microspores in brown anthers underwent division on callus induction medium. The first division was symmetrical and occurred after 2 wk. The second division occurred after 4 wk and resulted in four-celled structures. Anther-derived callus was either compact or friable. Histo-differentiation occurred mostly from compact callus. Cell clusters, each delimited by a protoderm, were observed in histological sections of callus grown for 2 to 4 wk on regeneration medium. Within a cellular unit, two heterogeneous cell populations were arranged in concentric rings with larger cells in the center and smaller cells towards the periphery. However, an apical-basal polarity that is present in embryo-like structures was not observed. Therefore, it may be surmised that in Indica rice, regeneration from anther-derived callus takes place not by the formation of somatic embryos but by direct organogenesis.
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Acknowledgements
Contributions provided by Ms. N. P. S. De Silva, Regional Rice Research and Development Center, Bombuwela, Sri Lanka and Ms. R. K. Maddumage, the Department of Plant Sciences, Faculty of Science, University of Colombo, Sri Lanka, are acknowledged.
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This study is funded by the National Science Foundation (NSF) of Sri Lanka (research grant number: RG/2011/BT/10).
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Dr. D.M.R.G. Mayakaduwa reports a grant from the National Science Foundation (NSF) of Sri Lanka (grant number RG/ 2011/BT/10) during the conduct of the study. Prof. T.D. Silva reports a grant from the National Science Foundation (NSF) of Sri Lanka (grant number RG/2011/BT/10) during the conduct of the study.
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Mayakaduwa, D.M.R.G., Silva, T.D. In vitro response of Indica rice microspores subjected to cold stress: a cytological and histological perspective. In Vitro Cell.Dev.Biol.-Plant 57, 843–855 (2021). https://doi.org/10.1007/s11627-021-10177-1
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DOI: https://doi.org/10.1007/s11627-021-10177-1