Five-color fluorescence in situ hybridization system for karyotyping of Panax ginseng
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RESEARCH REPORT
Five-color fluorescence in situ hybridization system for karyotyping of Panax ginseng Nomar Espinosa Waminal1 · Tae‑Jin Yang2 · Jun‑Gyo In3 · Hyun Hee Kim1 Received: 25 February 2020 / Revised: 18 May 2020 / Accepted: 17 June 2020 © Korean Society for Horticultural Science 2020
Abstract Chromosomal mapping of repetitive elements is invaluable in understanding genome structure and evolution. Repetitive elements constitute ~ 80% of the allotetraploid ginseng (Panax ginseng) genome. Preparing sporophytic metaphase chromosomes of ginseng is laborious; therefore, it would be advantageous to maximize the information obtained from a single slide. Here, we investigated the suitability of simultaneous five-color fluorescence in situ hybridization using major ginseng repeats, namely PgDel1, PgDel2, PgTel, and Pg167TR. For Pg167TR, we generated two degenerate probe libraries (Pg167TRa and Pg167TRb) based on the chromosomal target coverage. We labeled the probes with dark-red, blue, red, orange, and green fluorochromes and used excitation/emission filter sets specific to each fluorochrome to detect fluorescence in situ hybridization signals. PgDel1 was distributed across all 24 chromosome pairs, except for the secondary constriction region of chromosome 16, whereas PgDel2 was distributed over 12 of the 24 pairs. PgTel was localized in the termini of chromosomes and in an intercalary region in chromosome 13. Pg167TRa and Pg167TRb were distributed among 22 chromosome pairs with loci polymorphisms. These results showed the utility of five-color fluorescence in situ hybridization for chromosomal mapping of five repeats to expedite karyotyping and facilitate genome evolution studies in ginseng and other plant species. Keywords Five-color FISH · Karyotyping · Multicolor FISH · Oligoprobe · Panax ginseng
1 Introduction Fluorescence in situ hybridization (FISH) is invaluable for chromosomal mapping of repetitive elements (REs). Since its first successful use in the early 1980s, it has been modified for use in a variety of different applications (Bauman Communicated by Sung-Chur Sim. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s13580-020-00267-1) contains supplementary material, which is available to authorized users. * Hyun Hee Kim [email protected] 1
Department of Chemistry and Life Science, Chromosome Research Institute, Sahmyook University, Seoul, Korea
2
Department of Plant Science, Plant Genomics and Breeding Institute, Research Institute of Agriculture and Life Sciences, College of Agriculture and Life Sciences, Seoul National University, Seoul, Korea
3
Laboratory of Analysis, R&D Headquarters, Korea Ginseng Corporation, Daejeon, Korea
et al. 1980; Pinkel et al. 1986; Lilly et al. 2001; Nakanishi et al. 2002; Levsky and Singer 2003; Baker 2012; Martis et al. 2012; Chamala et al. 2013; Hu et al. 2014). It is often performed using probes labeled with either haptens or fluorochromes (Nath and Johnson 1998; Bishop 2010). Two of the most-used hapt
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