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Ploidy doubling by in vitro culture of excised protocorms or protocorm-like bodies in Phalaenopsis species

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Abstract

Endopolyploidy was observed in the protocorms of diploid Phalaenopsis aphrodite subsp. formosana with ploidy doubling achieved by in vitro regeneration of excised protocorms, or protocorm-like bodies (PLBs). Thirty-four per cent of the PLBs regenerated from the first cycle of sectioned protocorms were found to be polyploids with ploidy doubled once or twice as determined by flow-cytometry. The frequency of ploidy doubling increased as the sectioning cycles increased and was highest in diploid followed by the triploid and tetraploid. Regeneration of the endopolyploid cells in the tissue of the protocorms or PLBs is proposed as the source of the development of ploidy doubled plantlets. The frequency of ploidy doubling was similar in seven other Phalaenopsis species, although the rate of increase within cycles was genotype specific. In two species, a comparison of five parameters between 5-month-old diploid and tetraploid potted plants showed only the stomata density differed significantly. The flowers of the tetraploid plant were larger and heavier than those of the diploids. This ploidy doubling method is a simple and effective means to produce large number of polyploid Phalaenopsis species plants as well as their hybrids. The method will be beneficial to orchid breeding programs especially for the interspecific hybridization between varieties having different chromosome sizes and ploidy levels.

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Abbreviations

DAPI:

4,6-Diamidino-2-phenylindol

FCM:

Flow cytometry

MS:

Murashige and Skoog (1962) medium

PLBs:

Protocorm-like bodies

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Acknowledgments

This work was supported by the National Science Council, Taiwan (NSC-93-2317-B-390-002 and NSC-96-2317-B-390-004). The authors would like to thank Dr M.C. Palada (Senior Scientist, The World Vegetable Center, Hsin Hua, Taiwan) for his critical review of this manuscript, Mr G.T. Jean for his contribution of the photograph in Fig. 7 and Miss Y.C. Chu for her assistance in tissue culture.

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  1. Department of Life Sciences, National University of Kaohsiung, No. 700, Kaohsiung University Road, Nan-Tze District, Kaohsiung, 811, Taiwan, ROC

    Wen Huei Chen, Ching Yan Tang & Yu Lin Kao

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  1. Wen Huei Chen
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Correspondence to Yu Lin Kao.

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Chen, W.H., Tang, C.Y. & Kao, Y.L. Ploidy doubling by in vitro culture of excised protocorms or protocorm-like bodies in Phalaenopsis species. Plant Cell Tiss Organ Cult 98, 229–238 (2009). https://doi.org/10.1007/s11240-009-9557-3

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