Department of Ornamental Horticulture, Sichuan Agricultural University, Chengdu, People's Republic of China.
DNA Cell Biol. 2018 Dec;37(12):1016-1030. doi: 10.1089/dna.2018.4352. Epub 2018 Oct 16.
Salt response has long been considered a polygenic-controlled character in plants. Under salt stress conditions, plants respond by activating a great amount of proteins and enzymes. To develop a better understanding of the molecular mechanism and screen salt responsive genes in chrysanthemum under salt stress, we performed the RNA sequencing (RNA-seq) on both salt-processed chrysanthemum seedling roots and the control group, and gathered six cDNA databases eventually. Moreover, to overcome the Illumina HiSeq technology's limitation on sufficient length of reads and improve the quality and accuracy of the result, we combined Illumina HiSeq with single-molecule real-time sequencing (SMRT-seq) to decode the full-length transcripts. As a result, we successfully collected 550,823 unigenes, and from which we selected 48,396 differentially expressed genes (DEGs). Many of these DEGs were associated with the signal transduction, biofilm system, antioxidant system, and osmotic regulation system, such as mitogen-activated protein kinase (MAPK), Acyl-CoA thioesterase (ACOT), superoxide (SOD), catalase (CAT), peroxisomal membrane protein (PMP), and pyrroline-5-carboxylate reductase (P5CR). The quantitative real-time polymerase chain reaction (qRT-PCR) analysis of 15 unigenes was performed to test the data validity. The results were highly consistent with the RNA-seq results. In all, these findings could facilitate further detection of the responsive molecular mechanism under salt stress. They also provided more accurate candidate genes for genetic engineering on salt-tolerant chrysanthemums.
盐响应长期以来被认为是植物的多基因控制性状。在盐胁迫条件下,植物通过激活大量蛋白质和酶来做出响应。为了更好地理解分子机制并筛选盐胁迫下菊花中的盐响应基因,我们对盐处理的菊花幼苗根系和对照组进行了 RNA 测序(RNA-seq),最终收集了六个 cDNA 数据库。此外,为了克服 Illumina HiSeq 技术在读取长度上的局限性,并提高结果的质量和准确性,我们将 Illumina HiSeq 与单分子实时测序(SMRT-seq)相结合,以解码全长转录本。结果,我们成功地收集了 550823 个 unigene,并从中选择了 48396 个差异表达基因(DEGs)。这些 DEGs 中的许多与信号转导、生物膜系统、抗氧化系统和渗透调节系统有关,如丝裂原活化蛋白激酶(MAPK)、酰基辅酶 A 硫酯酶(ACOT)、超氧化物(SOD)、过氧化氢酶(CAT)、过氧化物酶体膜蛋白(PMP)和吡咯啉-5-羧酸还原酶(P5CR)。对 15 个 unigene 进行了定量实时聚合酶链反应(qRT-PCR)分析,以测试数据的有效性。结果与 RNA-seq 结果高度一致。总之,这些发现有助于进一步检测盐胁迫下的响应分子机制,为菊花的耐盐遗传工程提供了更准确的候选基因。