Five-color fl uorescence in situ hybridization system for karyotyping of Panax ginseng

다국어 초록 (Multilingual Abstract)

Chromosomal mapping of repetitive elements is invaluable in understanding genome structure and evolution. Repetitive elementsconstitute ~ 80% of the allotetraploid ginseng ( Panax ginseng ) genome. Preparing sporophytic metaphase chromosomesof ginseng is laborious; therefore, it would be advantageous to maximize the information obtained from a single slide. Here,we investigated the suitability of simultaneous fi ve-color fl uorescence in situ hybridization using major ginseng repeats,namely PgDel1 , PgDel2 , PgTel, and Pg167TR. For Pg167TR, we generated two degenerate probe libraries (Pg167TRa andPg167TRb) based on the chromosomal target coverage. We labeled the probes with dark-red, blue, red, orange, and greenfl uorochromes and used excitation/emission fi lter sets specifi c to each fl uorochrome to detect fl uorescence in situ hybridizationsignals. PgDel1 was distributed across all 24 chromosome pairs, except for the secondary constriction region of chromosome16, whereas PgDel2 was distributed over 12 of the 24 pairs. PgTel was localized in the termini of chromosomesand in an intercalary region in chromosome 13. Pg167TRa and Pg167TRb were distributed among 22 chromosome pairswith loci polymorphisms. These results showed the utility of fi ve-color fl uorescence in situ hybridization for chromosomalmapping of fi ve repeats to expedite karyotyping and facilitate genome evolution studies in ginseng and other plant species.

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