Zeng Fansuo, Sun Fengkun, Li Leilei, Liu Kun, Zhan Yaguang
State Key Laboratory of Tree Genetics and Breeding (Northeast Forestry University), Harbin 150040, China; College of Life Science, Northeast Forestry University, Harbin 150040, China.
College of Life Science, Northeast Forestry University, Harbin 150040, China.
PLoS One. 2014 Dec 31;9(12):e116157. doi: 10.1371/journal.pone.0116157. eCollection 2014.
Evidence supporting nitric oxide (NO) as a mediator of plant biochemistry continues to grow, but its functions at the molecular level remains poorly understood and, in some cases, controversial. To study the role of NO at the transcriptional level in Betula platyphylla cells, we conducted a genome-scale transcriptome analysis of these cells. The transcriptome of untreated birch cells and those treated by sodium nitroprusside (SNP) were analyzed using the Solexa sequencing. Data were collected by sequencing cDNA libraries of birch cells, which had a long period to adapt to the suspension culture conditions before SNP-treated cells and untreated cells were sampled. Among the 34,100 UniGenes detected, BLASTX search revealed that 20,631 genes showed significant (E-values≤10-5) sequence similarity with proteins from the NR-database. Numerous expressed sequence tags (i.e., 1374) were identified as differentially expressed between the 12 h SNP-treated cells and control cells samples: 403 up-regulated and 971 down-regulated. From this, we specifically examined a core set of NO-related transcripts. The altered expression levels of several transcripts, as determined by transcriptome analysis, was confirmed by qRT-PCR. The results of transcriptome analysis, gene expression quantification, the content of triterpenoid and activities of defensive enzymes elucidated NO has a significant effect on many processes including triterpenoid production, carbohydrate metabolism and cell wall biosynthesis.
支持一氧化氮(NO)作为植物生物化学介质的证据不断增加,但其在分子水平上的功能仍知之甚少,在某些情况下还存在争议。为了研究NO在白桦细胞转录水平上的作用,我们对这些细胞进行了全基因组规模的转录组分析。使用Solexa测序分析了未处理的桦树细胞以及用硝普钠(SNP)处理的细胞的转录组。通过对桦树细胞的cDNA文库进行测序来收集数据,在对SNP处理的细胞和未处理的细胞进行采样之前,桦树细胞有很长一段时间适应悬浮培养条件。在检测到的34,100个单基因中,BLASTX搜索显示,有20,631个基因与NR数据库中的蛋白质具有显著(E值≤10-5)的序列相似性。在12小时SNP处理的细胞和对照细胞样本之间,鉴定出许多差异表达的表达序列标签(即1374个):403个上调,971个下调。据此,我们专门研究了一组核心的NO相关转录本。转录组分析确定的几种转录本表达水平的变化,通过qRT-PCR得到了证实。转录组分析、基因表达定量、三萜含量和防御酶活性的结果表明,NO对包括三萜产生、碳水化合物代谢和细胞壁生物合成在内的许多过程都有显著影响。