Peniton Eliazar Alumbro, Nguyen Hong Thi, Waminal Nomar Espinosa, Yang Tae-Jin, Kim Hyun Hee
Department of Chemistry & Life Science, Chromosome Research Institute, Sahmyook University, Seoul, 01795, Republic of Korea.
Molecular Biology and Biotechnology, Genetics, and Microbiology Division, Institute of Biological Sciences, Central Mindanao University, Musuan, Philippines.
Sci Rep. 2024 Dec 5;14(1):30289. doi: 10.1038/s41598-024-75004-0.
Aralia elata is closely related to Panax ginseng and contains high levels of saponins and other medicinal compounds. Successful A. elata micropropagation is commercially significant; however, the genomic stability of tissue culture-derived regenerants is unclear. In this study, callus-derived regenerated A. elata plants were obtained, and their cytogenomic constitutions were assessed. Using RepeatExplorer, pre-labeled oligonucleotide probes (PLOPs) were developed with newly mined tandem repeats from < 1× NGS whole-genome short reads, fluorescence in situ hybridization (FISH) was performed using six repeat probes, including three universal PLOPs, and genomic DNA content was estimated using flow cytometry. Regenerated A. elata plants (50) exhibited consistent ploidy, repeat distribution, and genome sizes compared with those exhibited by the mother plant. Six repeat probes were detected using FISH. Tandem repeat AeTR49 was identified as an excellent cytogenetic marker for homologous chromosomes, and AeTR161 and AeTR178 were localized in the centromeric and telomeric sections, respectively. Genomic DNA content (2C) was estimated at 2.46 ± 0.04 pg in the mother plant and 2.41 ± 0.05 pg in regenerated plants, with no significant variations in genome size or chromosome length. These results demonstrate that cytogenomics can be used to effectively evaluate chromosome-level genomic stability in regenerated A. elata plants.
辽东楤木与人参密切相关，含有高水平的皂苷和其他药用化合物。成功实现辽东楤木的微繁殖具有重要的商业意义；然而，组织培养再生植株的基因组稳定性尚不清楚。在本研究中，获得了愈伤组织来源的辽东楤木再生植株，并对其细胞基因组组成进行了评估。利用RepeatExplorer，从<1×NGS全基因组短读段中新挖掘串联重复序列，开发了预标记寡核苷酸探针（PLOPs），使用六种重复探针（包括三种通用PLOPs）进行荧光原位杂交（FISH），并使用流式细胞术估计基因组DNA含量。与母本植株相比，50株再生辽东楤木植株在倍性、重复序列分布和基因组大小方面表现一致。利用FISH检测到六种重复探针。串联重复序列AeTR49被鉴定为同源染色体的优良细胞遗传学标记，AeTR161和AeTR178分别定位于着丝粒和端粒区域。母本植株的基因组DNA含量（2C）估计为2.46±0.04 pg，再生植株为2.41±0.05 pg，基因组大小或染色体长度无显著差异。这些结果表明，细胞基因组学可用于有效评估辽东楤木再生植株染色体水平的基因组稳定性。
