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. 2025 Jun 25:16:1621415.
doi: 10.3389/fpls.2025.1621415. eCollection 2025.

Establishment of a genome map-based karyotype of Artemisia argyi and identification of a new octoploid

Lina Li #  1 , Pei Du #  1 , Dahui Liu  2 , Xiangyang Li  1 , Guixiao La  1 , Guixia Shi  1 , Dandan Dai  1 , Tiegang Yang  1
Affiliations

Establishment of a genome map-based karyotype of Artemisia argyi and identification of a new octoploid

Lina Li et al. Front Plant Sci. .

Abstract

Artemisia argyi, an essential plant in traditional Chinese medicine, encounters significant challenges in the development of germplasm resources and cytological research. This study employed the A. argyi reference genome to develop 20 repetitive sequence oligonucleotide (oligo) probes, all of which produced clear signals on the chromosomes of the cultivar Qicun Xiang Ai (QCXA). These probes were configured into two probe cocktails (Multiplex #1 and Multiplex #2) that effectively generated chromosome signals under non-denaturing hybridization conditions through probe staining. By integrating probe staining with 45S rDNA fluorescence in situ hybridization (FISH) and electronic localization techniques, we established a genome map-based karyotype for QCXA that corresponded to its genomic sequence map. Utilizing this karyotype, we identified almost all chromosomes of the cultivars Wan Ai Ls-9 (WALs-9), Anguo Qi Ai (AGQA), and Anyang Bei Ai (AYBA) and investigated meiotic chromosome pairing behavior in WALs-9. These findings suggest that A. argyi may be a distinctive allotetraploid with a base chromosome number of 17. while AYBA (x = 8) appears to be a related species. Furthermore, a novel octoploid germplasm (APLs-9) is successfully generated and characterized through chromosomal doubling, demonstrating significantly enhanced moxa length and moxa content per leaf area - traits with substantial potential for improving both quality and yield. The developed octoploid and high-resolution karyotyping system are poised to significantly advance A. argyi breeding and production.

Keywords: Artemisia argyi; karyotype; moxa; octoploid; repetitive oligonucleotide.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Signals of each probe on the chromosomes of the cultivar Qichun Xiang Ai (QCXA) based on fluorescence in situ hybridization (FISH). (A) C1-1 (green); (B) C1-12 (green); (C) C5-2 (red); (D) Tel-1 (green). Scale bar: 10 μm.
Figure 2
Figure 2
Probe staining and 45S rDNA FISH and karyotype and chromosome plots of the cultivar QCXA. (A) Probe staining using Multiplex #1 (red) and Multiplex #2 (green); (B) FISH using 45S rDNA (green) as the probe; (C) karyotypes corresponding to chromosomes in the sequencing map of QCXA. The yellow dotted line shows the corresponding oligo signals, and the red arrows indicate the non-corresponding oligo signals. Scale bar: 10 μm.
Figure 3
Figure 3
Karyotypes of the distribution of all 20 oligos on QCXA chromosomes by oligo probe staining using Multiplex #1 (red) and Multiplex #2 (green) and sequential FISH using 45S rDNA (green) probes. Scale bar: 10 μm.
Figure 4
Figure 4
Sequential FISH karyotypes of WALs-9, AGQA and AYBA and their comparison with the karyotype idiogram of QCXA. (A–C) Probe staining of WALs-9, AGQA and AYBA using Multiplex #1 (red) and Multiplex #2 (green) and sequential FISH using the 45S rDNA probe (green); (D) karyotypes of WALs-9, AGQA and AYBA; (E) idiograms of the karyotypes of QCXA, WALs-9, AGQA and AYBA. Blue arrows indicated inconsistent signals compared to QCXA. Bar=10 μm.
Figure 5
Figure 5
Meiotic chromosome pairing analysis from two different cells (A, B) in WALs-9 and their probe staining using Multiplex #1 (red) and Multiplex #2 (green).
Figure 6
Figure 6
Karyotype identification of APLs-9 (A–C) and comparison of APLs-9 with Ls-9 in terms of pollen fertility (D), plant type (E), leaves (F), and moxa length (G). The probe and signal colors in (A–C) are the same as those in Figure 2 .
See this image and copyright information in PMC

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