College of Life Science, Capital Normal University, Beijing 100048, China.
BMC Plant Biol. 2014 Jan 13;14:20. doi: 10.1186/1471-2229-14-20.
Wheat seed germination directly affects wheat yield and quality. Although transcriptome and proteome analyses during seed germination have been reported in some crop plant species, dynamic transcriptome characterization during wheat seed germination has not been conducted. We performed the first comprehensive dynamic transcriptome analysis during different seed germination stages of elite Chinese bread wheat cultivar Jimai 20 using the Affymetrix Wheat Genome Array.
A total of 61,703 probe sets representing 51,411 transcripts were identified during the five seed germination stages of Jimai 20, of which 2,825 differential expression probe sets corresponding to 2,646 transcripts with different functions were declared by ANOVA and a randomized variance model. The seed germination process included a rapid initial uptake phase (0-12 hours after imbibition [HAI]), a plateau phase (12-24 HAI), and a further water uptake phase (24-48 HAI), corresponding to switches from the degradation of small-molecule sucrose to the metabolism of three major nutrients and to photosynthesis. Hierarchical cluster and MapMan analyses revealed changes in several significant metabolism pathways during seed germination as well as related functional groups. The signal pathway networks constructed with KEGG showed three important genes encoding the phosphofructokinase family protein, with fructose-1, 6-bisphosphatase, and UTP-glucose-1-phosphate uridylyltransferase located at the center, indicating their pivotal roles in the glycolytic pathway, gluconeogenesis, and glycogenesis, respectively. Several significant pathways were selected to establish a metabolic pathway network according to their degree value, which allowed us to find the pathways vital to seed germination. Furthermore, 51 genes involved in transport, signaling pathway, development, lipid metabolism, defense response, nitrogen metabolism, and transcription regulation were analyzed by gene co-expression network with a k-core algorithm to determine which play pivotal roles in germination. Twenty-three meaningful genes were found, and quantitative RT-PCR analysis validated the expression patterns of 12 significant genes.
Wheat seed germination comprises three distinct phases and includes complicated regulation networks involving a large number of genes. These genes belong to many functional groups, and their co-regulations guarantee regular germination. Our results provide new insight into metabolic changes during seed germination and interactions between some significant genes.
小麦种子的萌发直接影响小麦的产量和品质。尽管在一些作物物种中已经报道了种子萌发过程中的转录组和蛋白质组分析,但尚未对小麦种子萌发过程中的动态转录组进行描述。我们使用 Affymetrix 小麦基因组芯片对中国优质面包小麦品种金麦 20 的五个种子萌发阶段进行了首次全面的动态转录组分析。
在金麦 20 的五个种子萌发阶段共鉴定出 61703 个探针组,代表 51411 个转录本,其中 2825 个差异表达探针组通过 ANOVA 和随机方差模型鉴定为 2646 个具有不同功能的转录本。种子萌发过程包括快速初始吸水阶段(吸胀后 0-12 小时)、平台阶段(12-24 小时)和进一步的吸水阶段(24-48 小时),对应于从小分子蔗糖的降解到三大营养物质代谢和光合作用的转变。层次聚类和 MapMan 分析显示,种子萌发过程中几个重要代谢途径以及相关功能群发生了变化。KEGG 构建的信号通路网络显示,三个重要基因编码磷酸果糖激酶家族蛋白,果糖-1,6-二磷酸酶和 UTP-葡萄糖-1-磷酸尿苷酰转移酶位于中心,分别表明它们在糖酵解途径、糖异生和糖生成中的关键作用。根据其度值选择了几个重要的途径来建立一个代谢途径网络,这使我们能够找到对种子萌发至关重要的途径。此外,通过 k 核算法的基因共表达网络分析了 51 个参与运输、信号通路、发育、脂质代谢、防御反应、氮代谢和转录调控的基因,以确定哪些基因在萌发过程中发挥关键作用。发现了 23 个有意义的基因,并通过定量 RT-PCR 分析验证了 12 个显著基因的表达模式。
小麦种子萌发包括三个不同的阶段,涉及大量基因的复杂调控网络。这些基因属于许多功能群,它们的共同调控保证了正常的萌发。我们的研究结果为种子萌发过程中的代谢变化以及一些重要基因之间的相互作用提供了新的认识。