PDF(Pubmed)
Abstract：
Cold stress can impact plant biology at both the molecular and morphological levels. We cultivated two different types of tobacco seedlings using distinct seeding methods, observing significant differences in their cold tolerance at 4 °C. After 12 h cold stress, shallow water seeding cultivation treatment demonstrates a relatively good growth state with slight wilting of the leaves. Tobacco grown using the float system exhibited short, thick roots, while those cultivated through shallow water seeding had elongated roots with more tips and forks. After cold stress, the shallow water seeding cultivation treatment demonstrated higher antioxidant enzyme activity, and lower malondialdehyde (MDA) content.Transcriptome analysis was performed on the leaves of these tobacco seedlings at three stages of cold treatment (before cold stress, after cold stress, and after 3 days of recovery). Upon analyzing the raw data, we found that the shallow water seeding cultivation treatment was associated with significant functional enrichment of nicotinamide adenine dinucleotide (NAD) biosynthesis and NAD metabolism before cold stress, enrichment of functions related to the maintenance of cellular structure after cold stress, and substantial functional enrichment related to photosynthesis during the recovery period. Weighted gene co-expression network analysis (WGCNA) was conducted, identifying several hub genes that may contribute to the differences in cold tolerance between the two tobacco seedlings. Hub genes related to energy conversion were predominantly identified in shallow water seeding cultivation treatment during our analysis, surpassing findings in other areas. These include the AS gene, which controls the synthesis of NAD precursors, the PED1 gene, closely associated with fatty acid β-oxidation, and the RROP1 gene, related to ATP production.Overall, our study provides a valuable theoretical basis for exploring improved methods of cultivating tobacco seedlings. Through transcriptome sequencing technology, we have elucidated the differences in gene expression in different tobacco seedlings at three time points, identifying key genes affecting cold tolerance in tobacco and providing possibilities for future gene editing.
摘要：
冷胁迫可以在分子和形态水平上影响植物生物学。我们使用不同的播种方法种植了两种不同类型的烟草幼苗，在4°C下观察到它们的耐寒性的显著差异。冷应激12小时后，浅水播种栽培处理显示出相对较好的生长状态，叶片略有枯萎。使用浮法系统种植的烟草表现得很短，粗根,而那些通过浅水播种种植的人的根细长，有更多的尖端和叉子。在寒冷的压力之后，浅水播种栽培处理表现出更高的抗氧化酶活性，丙二醛(MDA)含量较低。在冷处理的三个阶段（冷胁迫前，在寒冷的压力之后，和恢复3天后)。在分析原始数据后，我们发现，浅水播种栽培处理与低温胁迫前烟酰胺腺嘌呤二核苷酸（NAD）生物合成和NAD代谢的显著功能富集有关，冷应激后与维持细胞结构相关的功能的富集，以及恢复期与光合作用相关的大量功能富集。进行加权基因共表达网络分析(WGCNA),鉴定几个可能导致两种烟草幼苗之间耐寒性差异的hub基因。在我们的分析过程中，与能量转换相关的集线器基因主要在浅水播种栽培处理中得到鉴定，超越其他领域的发现。这些包括AS基因，控制NAD前体的合成，PED1基因,与脂肪酸β-氧化密切相关，和RROP1基因，与ATP生产有关。总的来说,本研究为探索烟苗培育的改良方法提供了有价值的理论依据。通过转录组测序技术,我们已经阐明了在三个时间点不同烟草幼苗中基因表达的差异，确定影响烟草耐寒性的关键基因，为未来的基因编辑提供可能。
