The long history of asexual reproduction of garlic using garlic cloves has resulted in virus accumulation and genetic depression. Propagation of garlic seedlings by tissue culture can both eliminate viruses and improve breeding efficiency. Aerial bulbs are the first-choice materials for breeding virus-free garlic seedlings under external conditions, but they show dormancy just like garlic bulbs. However, low temperatures can quickly break dormancy. In this research, we used a high-throughput sequencing method to sequence aerial bulbs during dormancy and after low-temperature-induced breaking of dormancy to screen out the key differentially expressed genes (DEGs) associated with low temperature and to provide a theoretical basis for exploring the molecular mechanism of low-temperature-induced breaking of aerial bulb dormancy. The high-throughput transcriptome sequencing results showed that 6,675 DEGs were upregulated and 36,596 DEGs were downregulated in the aerial bulbs given low-temperature treatment. Then, 19,507 DEGs were assigned KEGG annotations, among which most DEGs were annotated to the metabolism pathway (11,817 genes, accounting for 60.58%), followed by the genetic information processing pathway (4,521 genes, accounting for 23.18%). The DEGs were mostly concentrated in pathways such as protein processing in the endoplasmic reticulum, plant-pathogen interaction, plant hormone signal transduction, and ribosome biogenesis in eukaryotes, with significant enrichment. The key DEGs related to calcium signaling, hormonal signaling, and transcription factors were screened out, including CaM, CDPK, and CML in accessory pathways of calcium signaling; GA20ox, GAI1, and GA2ox in accessory pathways of hormonal signaling; and transcription factor genes such as MYB, AP2/ERF, bHLH, MADS, and bZIP. qRT-PCR verification results were consistent with the sequencing results, indicating that the transcriptome sequencing data were accurate and reliable. Our results provide a theoretical basis for breaking the dormancy of aerial bulbs with low-temperature treatment to produce virus-free seedlings and increase the output and quality of garlic.

中文翻译：

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低温诱导打破大蒜空中鳞茎休眠的转录组分析。

使用蒜瓣进行大蒜无性繁殖的悠久历史导致了病毒积累和遗传抑制。采用组织培养法繁殖大蒜苗，既可以消除病毒，又可以提高养殖效率。气生球茎是在外界条件下培育脱毒大蒜苗的首选材料，但其与大蒜球茎一样呈现休眠状态。然而，低温会很快打破休眠状态。本研究采用高通量测序方法对休眠期间和低温诱导休眠打破后的气生鳞茎进行测序，筛选出与低温相关的关键差异表达基因（DEG），为后续研究提供理论依据。探索低温诱导打破空中鳞茎休眠的分子机制。高通量转录组测序结果显示，经过低温处理的气生球茎中有6,675个DEG上调，36,596个DEG下调。然后，对19,507个DEG进行KEGG注释，其中最多的DEG被注释到代谢途径（11,817个基因，占60.58%），其次是遗传信息处理途径（4,521个基因，占23.18%）。DEGs主要集中在真核生物内质网蛋白质加工、植物与病原体相互作用、植物激素信号转导、核糖体生物发生等途径，且富集显着。筛选出与钙信号、激素信号、转录因子相关的关键DEG，包括钙信号旁路中的CaM、CDPK、CML ；激素信号转导旁路中的GA20ox、GAI1和GA2ox ；以及转录因子基因，例如MYB、AP2/ERF、bHLH、MADS和bZIP。qRT-PCR验证结果与测序结果一致，表明转录组测序数据准确可靠。本研究结果为低温处理打破气生球茎休眠、培育脱毒苗、提高大蒜产量和品质提供了理论依据。