Ma Keshi, Shi Wenshuo, Xu Mengyue, Liu Jiaxi, Zhang Feixiong
College of Life Sciences, Capital Normal University, Beijing, China.
College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, China.
Front Plant Sci. 2018 Mar 6;9:244. doi: 10.3389/fpls.2018.00244. eCollection 2018.
It remains unclear whether plant lncRNAs are responsive to Ca-channel blocking. When using the Ca-channel blocker, LaCl, to treat germinated wheat seeds for 24 h, we found that both root length and mitosis were inhibited in the LaCl-treated groups. The effect of the Ca-channel blocker was verified in three ways: a [Ca] decrease detected using Fluo-3/AM staining, a decrease in the Ca content measured using inductively coupled plasma mass spectrometry, and an inhibition of Ca influx detected using Non-invasive Micro-test Technology. Genome-wide high throughput RNA-seq and bioinformatical methods were used to identify lncRNAs, and found 177 differentially expressed lncRNAs that might be in responsive to Ca-channel blocking. Among these, 108 were up-regulated and 69 were down-regulated. The validity of identified lncRNAs data from RNA-seq was verified using qPCR. GO and KEGG analysis indicated that a number of lncRNAs might be involved in diverse biological processes upon Ca-channel blocking. Further GO analysis showed that 23 lncRNAs might play roles as transcription factor (TF); Moreover, eight lncRNAs might participate in cell cycle regulation, and their relative expressions were detected using qPCR. This study also provides diverse data on wheat lncRNAs that can deepen our understanding of the function and regulatory mechanism of Ca-channel blocking in plants.
目前尚不清楚植物长链非编码RNA(lncRNAs)是否对钙通道阻断有反应。当使用钙通道阻滞剂LaCl处理萌发的小麦种子24小时时,我们发现LaCl处理组的根长和有丝分裂均受到抑制。通过三种方式验证了钙通道阻滞剂的作用:使用Fluo-3/AM染色检测到的[Ca]降低、使用电感耦合等离子体质谱法测量的钙含量降低以及使用非损伤微测技术检测到的钙内流抑制。利用全基因组高通量RNA测序和生物信息学方法鉴定lncRNAs,发现177个差异表达的lncRNAs可能对钙通道阻断有反应。其中,108个上调,69个下调。使用qPCR验证了RNA测序鉴定的lncRNAs数据的有效性。基因本体(GO)和京都基因与基因组百科全书(KEGG)分析表明,许多lncRNAs可能在钙通道阻断后参与多种生物学过程。进一步的GO分析表明,23个lncRNAs可能作为转录因子(TF)发挥作用;此外,8个lncRNAs可能参与细胞周期调控,并使用qPCR检测了它们的相对表达。这项研究还提供了关于小麦lncRNAs的多种数据,可加深我们对植物中钙通道阻断的功能和调控机制的理解。