Abstract
In the present paper attempts were made to induce chromosome doubling of ‘Meiwa’ kumquat (Fortunella crassifolia) protoplasts and ‘Frost’ navel orange (Citrus sinensis Osbeck) embryogenic callus via colchicine treatment. Colchicine decreased protoplast viability, delayed protoplast division and inhibited callus growth, indicating presence of toxicity to cells. Cell lines established from ‘Meiwa’ protoplasts treated with 0.01 and 0.1% colchicine for 8, 16 and 24 h at each concentration showed different responses when they were cultured on embryoid-induction medium. Flow cytometry (FCM) demonstrated that tetraploids were detected in cell lines and embryoids from all of the treatments, with the highest frequency being 19.23%. As for ‘Frost’, tetraploid cells were only detected when the callus was treated with 0.1% colchicine for either 4 or 8 days, from which plantlets were regenerated. FCM and chromosome counting confirmed them as true tetraploids. The diploid cells were more active in mitotic division during a 12-day culture and smaller in size than their tetraploid counterpart. Potential applications of the novel tetraploid germplasms obtained through in vitro chromosome doubling to citrus cultivar improvement are discussed.
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Abbreviations
- DAPI:
-
4′, 6-Diamidino-2-phenylindole
- FCM:
-
Flow cytometry
- FDA:
-
Fluorescein diacetate
- MI:
-
Mitotic index
- MT:
-
Murashige and Tucker
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Acknowledgements
This research was financially supported by Natural Science Foundation of China, Program for Innovative Research Team in University (IRT0548) and partially by Hubei Provincial Natural Science Foundation (2005ABC001, 2006ABB026). The authors are grateful to Drs. JH Wu (Hortresearch, New Zealand) and XP Wen (Guizhou University, China) for their critical reading of the paper.
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Zeng, SH., Chen, CW., Hong, L. et al. In vitro induction, regeneration and analysis of autotetraploids derived from protoplasts and callus treated with colchicine in Citrus . Plant Cell Tiss Organ Cult 87, 85–93 (2006). https://doi.org/10.1007/s11240-006-9142-y
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DOI: https://doi.org/10.1007/s11240-006-9142-y