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Cytogenetic assessment of Haworthia using flow cytometry and fluorescence in situ hybridization

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Abstract

The purpose of this study was to investigate cytogenetic variations between the nine Haworthia species necessary for subsequent breeding. Flow cytometric analysis of H. badia ‘Murasaki’, H. splendens, H. truncata ‘Seiko’, H. cooperi var. obtusa, H. cymbiformis, H. mirabilis var. mundula, and H. retusa showed a wide range of DNA content from 10.17 to 12.17 pg/1C, whereas two tetraploids, H. limifolia and H. angustifolia var. baylisii, showed higher DNA content of 16.81 and 12.64 pg/1C, respectively. Seven diploid Haworthia expressed three karyotype formulas, whereas two tetraploid Haworthia each expressed a different karyotype formula. Secondary constriction was observed in H. truncata ‘Seiko’ and H. cooperi var. obtusa. Results of fluorescence in situ hybridization (FISH) showed a major difference between the diploid and tetraploid Haworthia in the distribution of 5S and 18S ribosomal DNA (rDNA) loci. There were eight loci in two out of seven diploids and both tetraploid Haworthia, while H. cymbiformis and H. mirabilis var. mundula each showed seven 18S rDNA loci. H. badia ‘Murasaki’, H. splendens, and H. retusa had four, three, and five 18S rDNA loci, respectively. In addition, a single pair of 5S rDNA loci was observed in all diploid Haworthia except H. cooperi var. obtusa, which had four 5S rDNA loci. Each tetraploid Haworthia had four 5S rDNA loci, with two strong and two weak signals. All 5S and 18S rDNA loci were observed on the long arm, in the subtelomeric or telomeric region, while a significant variation in chromosome length was observed among the Haworthia. This cytological analysis provides the necessary genetic information to elucidate the genetic makeup of newly introduced Haworthia hybrids.

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Acknowledgements

This work was supported by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, and Forestry (iPET) through an Agri-Bio industry Technology Development Program, funded by Ministry of Agriculture, Food, and Rural Affairs (MAFRA) (Grant No. IPET318021-4).

The research being report in this publication was supported by Ministry of Agriculture, Food and Rural Affairs. I hereby disclose all of my conflicts of interest and other potentially conflicting interests, including specific financial interests and relationships and affiliations relevant to Horticulture, Environment, and Biotechnology (HEAB).

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RY and MI designed the experimental structure and ideas. DMD, H-YK, C-KK, and LK-B verified the analytical methods. K-BL encouraged and supervised the findings of this work. All authors discussed the results and contributed to the final manuscript.

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Correspondence to Ki Byung Lim.

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The authors have declared no conflict of interest.

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Communicated by Jongyun Kim, Ph.D.

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Yesmin, R., Islam, M.M., Deepo, D.M. et al. Cytogenetic assessment of Haworthia using flow cytometry and fluorescence in situ hybridization. Hortic. Environ. Biotechnol. 63, 115–123 (2022). https://doi.org/10.1007/s13580-021-00353-y

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  • DOI: https://doi.org/10.1007/s13580-021-00353-y

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