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麻疯树是一种能适应多种恶劣环境条件的能源植物,目前关于其抵抗重金属胁迫的分子调控机理尚不清楚。从组学水平整体分析其基因表达模式对于筛选关键基因、解析镉胁迫响应调控网络和促进分子育种具有重要意义。利用Illumina测序技术对水培条件下培养的处理组(Cd100)和对照组(CK)麻疯树幼苗叶的转录组进行高通量测序,两个数据库的测序数据经de novo组装得到50448条高质量的Unigene。两个样品中发现了2551条差异表达基因,其中539条上调基因,2012条下调基因。根据不同数据库的注释信息,发现麻疯树镉胁迫引起叶片中多种代谢途径的变化,包括碳代谢,光合作用,植物激素信号转导以及植物病原响应途径。DAVID分析显示镉胁迫引起了麻疯树叶中与离子转运相关基因的变化,导致叶片中Na离子和铁离子稳态的变化。转录因子分析发现WRKY和ZIP在镉胁迫中发挥重要作用。用qRT-PCR技术对随机挑选的5个基因进行荧光定量验证,结果与测序数据一致,证实了差异表达基因数据的有效性。深入探讨了麻疯树镉胁迫的分子机理,为进一步应用于基因工程和植物修复提供基础。
Jatropha curcas is a kind of energy plant that can adapt to a variety of harsh environmental conditions. At present, the molecular regulation mechanism of its resistance to heavy metal stress is not yet clear. It is of great significance to analyze the gene expression patterns at the level of omics for screening key genes, analyzing the regulation network of cadmium stress response and promoting molecular breeding. Illumina sequencing was used to carry out high-throughput sequencing of the transcriptome of the leaves of the treatment group (Cd100) and the control group (CK) cultured in hydroponics. The sequencing data of the two databases were 50448 high Quality Unigene. In the two samples, 2551 differentially expressed genes were found, including 539 up-regulated genes and 2012 down-regulated genes. According to the annotation information of different databases, it was found that the cadmium stress in J. curcas caused changes in many metabolic pathways including carbon metabolism, photosynthesis, plant hormone signal transduction and plant pathogen response. DAVID analysis showed that cadmium stress caused the changes of ion transport related genes in Jatropha curcas leaves, resulting in the steady state changes of Na ions and Fe ions in leaves. Transcription factor analysis revealed that WRKY and ZIP play an important role in cadmium stress. The qRT-PCR technique was used to confirm the random selection of five genes by fluorescence quantitative analysis. The results were consistent with the sequencing data, which confirmed the validity of the differentially expressed gene data. The molecular mechanism of cadmium stress in Jatropha curcas was further discussed, which provided the basis for further application in genetic engineering and plant restoration.